WO2025251009A1 - Méthodes de traitement du cancer de la vessie à l'aide d'un virus oncolytique et d'un agent améliorant la transduction - Google Patents

Méthodes de traitement du cancer de la vessie à l'aide d'un virus oncolytique et d'un agent améliorant la transduction

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Publication number
WO2025251009A1
WO2025251009A1 PCT/US2025/031754 US2025031754W WO2025251009A1 WO 2025251009 A1 WO2025251009 A1 WO 2025251009A1 US 2025031754 W US2025031754 W US 2025031754W WO 2025251009 A1 WO2025251009 A1 WO 2025251009A1
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individual
bcg
administration
months
bladder
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Vijay K. KASTURI
III Kirk A. KEEGAN
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CG Oncology Inc
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CG Oncology Inc
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • C12N15/86—Viral vectors
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
    • A61K31/7032—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a polyol, i.e. compounds having two or more free or esterified hydroxy groups, including the hydroxy group involved in the glycosidic linkage, e.g. monoglucosyldiacylglycerides, lactobionic acid, gangliosides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/76—Viruses; Subviral particles; Bacteriophages
    • A61K35/761—Adenovirus
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0031—Rectum, anus
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0034—Urogenital system, e.g. vagina, uterus, cervix, penis, scrotum, urethra, bladder; Personal lubricants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
    • C12N2710/00011—Details
    • C12N2710/10011—Adenoviridae
    • C12N2710/10311—Mastadenovirus, e.g. human or simian adenoviruses
    • C12N2710/10332—Use of virus as therapeutic agent, other than vaccine, e.g. as cytolytic agent
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
    • C12N2710/00011—Details
    • C12N2710/10011—Adenoviridae
    • C12N2710/10311—Mastadenovirus, e.g. human or simian adenoviruses
    • C12N2710/10341—Use of virus, viral particle or viral elements as a vector
    • C12N2710/10343—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector

Definitions

  • the present invention relates to methods of treating bladder cancer using an oncolytic virus, such as cretostimogene grenadenorepvec.
  • Non-muscle invasive bladder cancer (stages Ta, Tl, or carcinoma in situ) accounts for 70-80% of bladder cancer cases, while muscle invasive disease (stage T2 and above) and metastatic disease make up the remaining 20-30%.
  • Bladder cancers are divided into 2 subtypes based on their distinct cellular growth patterns: papillary tumors and flat tumors.
  • Carcinoma in situ (CIS) is a flat tumor confined to the surface layer of the bladder lumen. Compared to papillary tumors of the Ta and Tl stage, bladder CIS is more difficult to diagnose, and has a high risk of progression to muscle invasive bladder cancer.
  • Intravesical Bacillus Calmette-Guerin is the standard-of-care treatment for non-muscle invasive bladder cancer (NMIBC). Patients typically receive an induction course consisting of weekly intravesical installations of BCG for 6 weeks, followed by monthly maintenance treatments for 6 to 12 months, with each maintenance course typically consisting of weekly intravesical instillations for 3 weeks. However, about 30-40% of patients classify do not respond to the induction course of BCG treatment and are classified as having BCG-resistant NMIBC (Kodera, et al. (2023) "The management of Bacillus Calmette -Guerin (BCG) failure in high-risk non-muscle invasive bladder cancer: A review article.” Cureus 15.6).
  • NMIBC non-muscle invasive bladder cancer
  • cystectomy including partial and radical cystectomy, remains the treatment of choice.
  • cystectomy is associated with severe side effects and adversely affects patients’ quality of life (Maibom et al. BMI Open.
  • a transduction enhancing agent such as n-Dodecyl [3-D-maltoside (DDM)
  • DDM n-Dodecyl [3-D-maltoside
  • the present disclosure is based, at least in part, on the discovery that a modified two-step instillation procedure, wherein a single instillation of a transduction enhancing agent prior to each instillation of the oncolytic virus, with no intervening saline wash, is safe and effective for treating bladder cancer.
  • this novel instillation procedure reduces the total administration time of oncolytic viral therapy and increases patient comfort.
  • each two-step instillation regimen comprises: intravesically administering a pretreatment composition comprising a transductionenhancing agent to the individual; and subsequently intravesically administering an oncolytic virus to the individual directly after administering the pretreatment composition, wherein there is no intervening bladder wash between the administration of the pretreatment composition and the administration of the oncolytic virus, wherein the oncolytic virus comprises a viral vector comprising a tumor cell-specific promoter operably linked to a viral gene essential for replication of the oncolytic virus, and a heterologous gene encoding an immune-related molecule.
  • each two-step instillation regimen consists of: intravesically administering the pretreatment composition to the individual; and subsequently intravesically administering the oncolytic virus to the individual directly after administering the pretreatment composition.
  • the method further comprises one or more bladder washes with saline prior to the two-step instillation regimen. In some embodiments, the method further comprises one or more bladder washes with the pretreatment composition prior to the two-step instillation regimen.
  • the transduction enhancing agent is N-Dodecyl-[3-D- maltoside (DDM).
  • the pretreatment composition comprises DDM at a concentration of about 0.01% to about 0.5%. In certain embodiments, the pretreatment composition comprises DDM at a concentration of about 0.05% to about 0.2%. In certain embodiments, the pretreatment composition comprises DDM at a concentration of about 0.1%.
  • the pretreatment composition is administered at a volume of between about 50 mb and about 100 mb. In some embodiments, the pretreatment composition is administered at a volume of between about 85 mb and about 100 mb.
  • the administration of the pretreatment composition comprises allowing the pretreatment composition to dwell in the bladder for more than 5 minutes after completion of the administration of the full volume of the pretreatment composition into the bladder. In some embodiments, the administration of the pretreatment composition comprises allowing the pretreatment composition to dwell in the bladder for between about 10 minutes and about 20 minutes after completion of the administration of the full volume of the pretreatment composition into the bladder. In some embodiments, the administration of the pretreatment composition comprises allowing the pretreatment composition to dwell in the bladder for about 15 minutes after completion of the administration of the full volume of the pretreatment composition into the bladder.
  • the administration of the oncolytic virus is administered at a dose of about 1 x 10 8 to about 1 x 10 14 viral particles. In certain embodiments, the administration of the oncolytic virus is administered at a dose of about 1 x 10 12 viral particles. In some embodiments, the oncolytic virus is administered in a volume of between about 50 mb and about 100 mb. In some embodiments, the oncolytic virus is administered in a volume of between about 85 mb and about 100 mb. In some embodiments, the administration of the oncolytic virus comprises allowing the oncolytic virus to dwell in the bladder for between about 55 minutes and about 65 minutes after completion of the administration of the full volume of the oncolytic virus into the bladder. In some embodiments, the administration of the oncolytic virus comprises allowing the oncolytic virus to dwell in the bladder for about 60 minutes after completion of the administration of the full volume of the oncolytic virus into the bladder.
  • the administration of the full volume of the oncolytic virus into the bladder is completed within about 35 minutes after the administration of the full volume of the pretreatment composition into the bladder is completed. In certain embodiments, the administration of the full volume of the oncolytic virus into the bladder is completed within about 30 minutes after the administration of the full volume of the pretreatment composition into the bladder is completed. In some embodiments, the full volume of the pretreatment composition and the full volume of the oncolytic virus are both administered into the bladder within about 35 minutes. In certain embodiments, the full volume of the pretreatment composition and the full volume of the oncolytic virus are both administered into the bladder within about 30 minutes. In some embodiments, the administration of the oncolytic virus is initiated within about 30 minutes of the initiation of the administration of the pretreatment composition.
  • the administration of the oncolytic virus is initiated within about 20 minutes of the initiation of the administration of the pretreatment composition. In certain embodiments, the administration of the oncolytic virus is initiated within about 15 minutes of the initiation of the administration of the pretreatment composition.
  • the individual has non-muscle invasive bladder cancer (NMIBC). In some embodiments, the individual has carcinoma in situ. In certain embodiments, the individual has carcinoma in situ and has a concurrent papillary carcinoma of Ta or T1 stage (e.g, high-grade Ta or Tl). In certain other embodiments, the individual has carcinoma in situ and does not have a concurrent papillary carcinoma of Ta or Tl stage (e.g, high-grade Ta or Tl).
  • the individual has a papillary carcinoma of Ta or Tl stage (e.g, high-grade Ta or Tl). In certain embodiments, the individual has a papillary carcinoma of Ta or Tl stage (e.g, high-grade Ta or Tl) and does not have concurrent carcinoma in situ.
  • a papillary carcinoma of Ta or Tl stage e.g, high-grade Ta or Tl
  • the individual has a papillary carcinoma of Ta or Tl stage (e.g, high-grade Ta or Tl) and does not have concurrent carcinoma in situ.
  • the individual a) has not received prior intravesical Bacillus Calmette-Guerin (BCG) therapy; b) has not received intravesical BCG therapy within 24 months prior to the most recent diagnosis of high-risk NMIBC; or c) has received a maximum of 1 or 2 doses of BCG within 24 months prior to the most recent diagnosis of high-risk NMIBC.
  • BCG Bacillus Calmette-Guerin
  • the individual has CIS with or without concomitant Ta or Tl disease.
  • the individual has high-grade Ta or Tl disease without CIS.
  • the individual has been previously treated intravesically with Bacillus Calmette-Guerin (BCG) therapy.
  • BCG Bacillus Calmette-Guerin
  • the individual was responsive to BCG therapy.
  • the individual experienced reduced bladder cancer symptoms, reduced size or number of bladder tumors, or an absence of bladder tumors after intravesical BCG therapy.
  • the individual was BCG-unresponsive.
  • the individual has: a) received at least 5 of 6 doses of a BCG induction course and at least 2 doses of 1 course of either reinduction or maintenance BCG and experienced persistent or recurrent carcinoma in situ (CIS), with or without concomitant Ta or Tl disease, within 12 months following the last dose of BCG; b) received at least 5 of 6 doses of a BCG induction course and at least 2 doses of 1 course of either reinduction or maintenance BCG and experienced recurrence of high-grade papillary-only Ta or Tl stage disease within 6 months following the last dose of BCG; or c) received at least 5 of 6 doses of a BCG induction course and experienced high-grade Tl disease at the first evaluation following the BCG induction course.
  • CIS persistent or recurrent carcinoma in situ
  • the individual has high-risk NMIBC.
  • the individual has high-grade Ta or Tl disease without CIS, wherein the individual has: a) received at least 5 of 6 doses of a BCG induction course and at least 2 doses of one course of either reinduction or maintenance BCG and experienced recurrence of high-grade papillary- only Ta or T1 stage disease within 6 months following the last dose of BCG; or b) received at least 5 of 6 doses of a BCG induction course and experienced high-grade T1 disease at the first evaluation following the BCG induction course.
  • the individual has: a) received at least 5 of 6 doses of a BCG induction course and experienced persistent or recurrent high-grade papillary carcinoma of Ta stage or carcinoma in situ (CIS) at a first evaluation following the BCG induction course; b) received at least 5 of 6 doses of a BCG induction course and at least 2 doses of 1 course of either reinduction or maintenance BCG, achieved an initial complete response, and subsequently experienced recurrence of a highgrade Ta or T1 disease more than 6 months after and within 24 months after the last dose of BCG; c) received at least 5 of 6 doses of a BCG induction course and at least 2 doses of 1 course of either reinduction or maintenance BCG, achieved an initial complete response, and subsequently experienced recurrence of CIS more than 12 months after and within 24 months after the last dose of BCG; or d) received 3 to 6 doses of a BCG induction course and no BCG reinduction or maintenance course, achieved an initial complete response, and subsequently experienced
  • the individual has intermediate-risk NMIBC.
  • the intermediate-risk NMIBC comprises: a) a low-grade tumor of Ta stage, wherein the tumor of Ta stage is recurrent within about 12 months of resection of a prior low- grade tumor of Ta stage or solitary high-grade tumor of Ta stage having a diameter of 3 cm or smaller; b) a solitary low-grade tumor of Ta stage having a diameter of greater than 3 cm; c) two or more low-grade tumors of Ta stage; d) a primary and solitary high-grade tumor of Ta stage having a diameter of less than 3 cm; e) a low-grade tumor of Tl stage; or f) any combination of (a), (c), and (e).
  • the method comprises one or more treatment courses each comprising intravesically administering the two-step instillation regimen weekly.
  • each treatment courses comprises administering the two-step instillation regimen weekly for about 1 week to about 6 weeks.
  • the method comprises at least a first induction phase comprising administering the two-step instillation regimen to the individual once per week for six weeks.
  • the method comprises a maintenance phase subsequent to the induction phase, wherein the maintenance phase comprises administering the two-step instillation regimen to the individual every three or six months.
  • the maintenance phase comprises administering the two-step instillation regimen weekly for one to three weeks every three or six months.
  • the start of the first induction phase and the start of the maintenance phase are separated by about three months or about six months.
  • the two-step instillation regimen is administered to the individual once per week for six weeks on month zero during the first induction phase and the individual begins the maintenance phase at month three or around week 13 following the start of the first induction phase.
  • the two-step instillation regimen is administered to the individual once per week for six weeks on month zero during the first induction phase and the individual is reevaluated at month three or around week 11 following the start of the first induction phase.
  • the individual begins the maintenance phase at month three or around week 13 following the start of the first induction phase.
  • the maintenance phase comprises administering the two-step instillation regimen once per week for three weeks every three months for nine months and subsequently administering the two-step instillation regimen once per week for three weeks every six months.
  • the maintenance phase comprises administering the two- step instillation regimen once per week for three weeks every three months for six months and subsequently administering the two-step instillation regimen once every three months for six months.
  • the individual receives a second induction phase comprising administration of the two-step instillation regimen once per week for six weeks at month three or around week 13 following the start of the first induction phase.
  • the maintenance phase comprises administering the two-step instillation regimen once per week for three weeks every three months for six months and subsequently administering the two-step instillation regimen once per week for three weeks every six months.
  • the maintenance phase comprises: (i) administering the two- step instillation regimen once per week for three weeks every three months for six to nine months, and subsequently (ii) administering the two-step instillation regimen once per week for three weeks every six months.
  • the method further comprises administering to the individual an additional therapeutic agent.
  • the additional therapeutic agent is an immune checkpoint modulator.
  • the immune checkpoint modulator comprises an immune checkpoint inhibitor.
  • the immune checkpoint modulator comprises an inhibitor of an immune checkpoint molecule selected from the group consisting of CTLA-4, PD-1, PD-L1, PD-L2, TIM3, B7-H3, B7-H4, LAG-3, KIR, CD137, and ligands thereof.
  • the immune checkpoint inhibitor is an antibody that binds to an immune checkpoint molecule selected from the group consisting of CTLA-4, PD-1, PD-L1, PD-L2, TIM3, B7-H3, B7-H4, LAG-3, KIR, and ligands thereof
  • the immune checkpoint modulator is an anti-PD-1 antibody.
  • the anti-PD-1 antibody is nivolumab, pidilizumab, pembrolizumab, BMS- 936559, atezolizumab, lambrolizumab, MK- 3475, AMP -224, AMP-514, STI-Al l 10, TSR- 042, or any combination thereof.
  • the additional therapeutic agent is a chemotherapeutic agent.
  • the chemotherapeutic agent comprises gemcitabine, cisplatin, carboplatin, paclitaxel, docetaxel, ifosfamide, doxorubicin, methotrexate, vinblastine, mitomycin, 5 -fluorouracil (5-FU), or any combination thereof.
  • the chemotherapeutic agent comprises: a) gemcitabine and cisplatin; b) dose-dense methotrexate, vinblastine, doxorubicin (Adriamycin), and cisplatin (DDMVAC); or c) gemcitabine and paclitaxel.
  • the chemotherapeutic agent is gemcitabine.
  • the individual is human.
  • kits for treating bladder cancer in an individual comprising: a) an oncolytic virus; b) a pretreatment composition comprising a transduction enhancing agent; and c) a device for intravesically administering the oncolytic adenovirus, the pretreatment composition, or both to the individual, wherein the oncolytic adenovirus comprises a viral vector comprising a tumor cell-specific promoter operably linked to a viral gene essential for replication of the oncolytic adenovirus, and a heterologous gene encoding an immune-related molecule.
  • the kit comprises a dosage unit of the oncolytic adenovirus in an amount of between about 1 x 10 8 and about 1 x 10 14 viral particles, optionally around 1 x 10 12 viral particles.
  • the transduction enhancing agent is N-Dodecyl-[3-D-maltoside (DDM).
  • the pretreatment composition comprises DDM at a concentration of about 0.1%.
  • the oncolytic virus is selected from the group consisting of adenovirus, herpes simplex virus, vaccinia virus, mumps virus, Newcastle disease virus, polio virus, measles virus, Seneca valley virus, coxsackie virus, reovirus, vesicular stomatitis virus, maraba and rhabdovirus, and parvovirus.
  • the oncolytic virus comprises a viral vector comprising a tumor cell-specific promoter operably linked to a viral gene essential for replication of the oncolytic virus, and a heterologous gene encoding an immune-related molecule.
  • the oncolytic virus preferentially replicates in a cancer cell.
  • the cancer cell is defective in the Rb pathway.
  • the tumor cell-specific promoter is an E2F-1 promoter.
  • the E2F-1 promoter comprises the nucleotide sequence set forth in SEQ ID NO: 1.
  • the immune-related molecule is selected from the group consisting of GM-CSF, IL-2, IL-12, interferon, CCL4, CCL19, CCL21, CXCL13, TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, TLR10, RIG-I, MDA5, LGP2, and LTap.
  • the immune-related molecule is GM-CSF.
  • the heterologous gene is operably linked to a viral promoter.
  • the oncolytic virus is an oncolytic adenovirus.
  • the oncolytic virus is an oncolytic adenovirus and the viral gene essential for replication of the oncolytic virus is selected from the group consisting of E1A, E1B, and E4.
  • the oncolytic virus is an oncolytic adenovirus and the heterologous gene is operably linked to an El promoter or an E3 promoter.
  • the oncolytic virus is an adenovirus serotype 5, wherein the endogenous Ela promoter of a native adenovirus serotype 5 is replaced by the human E2F-1 promoter, and the endogenous E3 19kD coding region of the native adenovirus serotype 5 is replaced by a nucleic acid encoding human GM-CSF.
  • the oncolytic virus is cretostimogene.
  • FIG. 1 is a schematic diagram of cretostimogene grenadenorepvec (also referred to as cretostimogene or CG0070) and wild type adenovirus type 5.
  • Cretostimogene is based on adenovirus serotype 5, but the endogenous Ela promoter and E3 19kD coding region have been replaced by the human E2F-1 promoter and a cDNA coding region of human GM-CSF, respectively.
  • FIG. 2A depicts a flow chart showing administration of a clinical trial evaluating cretostimogene monotherapy (Mono) and combination therapy with one or more additional therapeutic agents (IO, such as an immune checkpoint modulator or chemotherapeutic agent) based on a determination of urinary genomic disease burden (GDB) and an associated risk status of low, intermediate (int), or high for the individual being treated.
  • IO cretostimogene monotherapy
  • GDB urinary genomic disease burden
  • 2B depicts a flow chart showing administration of a clinical trial evaluating cretostimogene monotherapy (Mono creto) and combination therapy with one or more additional therapeutic agents (X, such as an immune checkpoint modulator or chemotherapeutic agent) based on a determination of urinary genomic disease burden (GDB) and an associated minimum residual disease (MRD) status of positive (+) or negative (-) for the individual being treated.
  • additional therapeutic agents such as an immune checkpoint modulator or chemotherapeutic agent
  • FIG. 3 depicts a chart showing the criteria for the patient groups of the Phase 2 clinical trial described in Example 3, including BCG-naive, BCG-unresponsive, and BCG- exposed patient groups.
  • FIG. 4 depicts a chart summarizing the cohorts evaluated in the Phase 2 clinical trial described in Example 3. The number of patients depicted in each cohort is an estimate of patients to be recruited for the trial.
  • FIG. 5A depicts the treatment schedule to be used for Cohort A (all arms), Cohort B (all arms), and Arm 1 of Cohort CX of the Phase 2 clinical trial described in Example 3.
  • Each instillation is represented by a line topped with a circle.
  • each instillation corresponds to an instillation of DDM followed by an instillation of cretostimogene.
  • Cohort CX Arm 1, each instillation correspond to an instillation of DDM, followed by an instillation of cretostimogene, followed by an instillation of gemcitabine.
  • FIG. 5B depicts the treatment schedule to be used for Arm 2 of Cohort CX of the Phase 2 clinical trial described in Example 3.
  • Instillations of cretostimogene are represented by a line topped with a circle and correspond to an instillation of DDM followed by an instillation of cretostimogene.
  • Instillations of gemcitabine are represented by a line topped with a diamond and correspond to an instillation of gemcitabine only.
  • FIGS. 6A-6B depict results from genomic disease burden (GDB) analysis of patients in the BOND-003 trial.
  • FIG. 6A depicts results from patients prior to the first administration of cretostimogene.
  • FIG. 6B depicts results from patients 6-months after the first administration of cretostimogene.
  • Each column in the bar chart corresponds to a single patient.
  • SNV single-nucleotide variants
  • InDei insertions/deletions
  • CNV copy number variants
  • aneuploidy is shown in red.
  • Genomic disease burden (GDB) for each patient is shown below the bar chart, with low to high GDB shown in a gradient from black, to purple, to red, to yellow.
  • Minimum residual disease (MRD) status for each patient is shown below GDB, with white indicating MRD-negative and red indicating MRD-positive patients.
  • FIGS. 7A-7B depict results from GDB analysis of patients in the CORE-001 trial.
  • FIG. 7A depicts results from patients prior to the first administration of cretostimogene.
  • FIG. 7B depicts results from patients 6-months after the first administration of cretostimogene.
  • Each column in the bar chart corresponds to a single patient.
  • GDB Genomic disease burden
  • MRD Minimum residual disease
  • FIG. 8 depicts the fraction of patients with bladder cancer evaluated using UroAmp® in the BOND-003 and CORE-001 trials that were found to have variants in bladder-cancer-associated genes at baseline prior to the initiation of treatment.
  • Singlenucleotide variants (SNV) and insertions/deletions (InDei) are shown as blue dots; copy number variants (CNV) are shown as orange dots; and aneuploidy is shown as a red dot.
  • SNV Singlenucleotide variants
  • CNV copy number variants
  • aneuploidy is shown as a red dot.
  • FIG. 9 depicts a graph showing the probability of high-grade recurrence -free survival (RFS) over time in MRD-negative and MRD-positive patients in the BOND-003 trial who have been treated with cretostimogene and achieved a complete response at or around month 3.
  • the graph represents a 12-month RFS Kaplan-Meier model, predicted based on the Uro Amp® molecular signature (MRD profile) obtained at the Week 13 assessment.
  • MRD- negative patients are shown in blue, and MRD-positive patients are shown in red.
  • FIG. 10 depicts the maximum variant allele frequency (VAF) analyzed over about 12 months (49 weeks) in patients in the BOND-003 trial that have been stratified according to their response to cretostimogene therapy.
  • VAF maximum variant allele frequency
  • All CR refers to patients that received a weekly x6 induction course of cretostimogene administration and achieved a complete response (CR) at three months (week 13) and did not receive a second induction course. All patients in the “All CR” group maintained complete response (CR) throughout the 12-month period shown.
  • NR to CR refers to patients that initially received a weekly x6 induction course of cretostimogene administration, were not responsive, and subsequently received a second weekly x6 induction course at three months (week 13), and then achieved CR by the six- month (week 25) evaluation.
  • the “NR to CR” group then maintained CR throughout to the 12-month timepoint depicted in the figure.
  • CRto NR refers to patients that received a single weekly x6 induction course of cretostimogene administration, achieved CR at three months (week 13), and then experienced disease recurrence within 12 months (by week 49) from the first dose of cretostimogene
  • All NR refers to patients that that received at least one weekly x6 induction course of cretostimogene, did not achieve CR at any point in the study, and discontinued the study after three months (week 13).
  • FIG. 11 depicts a Kaplan-Meier estimate for high-grade recurrence-free survival based on the first 24 patients evaluated in Cohort P of the BOND-003 trial.
  • Non-muscle invasive bladder cancer refers to bladder cancer where the cancer cells/tumors are confined to the bladder wall and are not present muscle surrounding the bladder.
  • NMIBC includes papillary carcinomas of Ta and T1 stage, carcinoma in situ (CIS), and combinations thereof.
  • High-risk NMIBC refers to bladder cancer comprising high-grade papillary carcinomas of Ta and T1 stage, carcinoma in situ (CIS), or a combination thereof.
  • Papillary carcinoma refers to bladder tumors that grow in slender, finger-like projections from the inner surface of the bladder toward the hollow center. Papillary tumors often grow toward the center of the bladder without growing into the deeper bladder layers. These tumors are called non-invasive papillary cancers. Very low-grade (slow growing), non- invasive papillary cancer is sometimes called papillary urothelial neoplasm of low-malignant potential (PUNLMP).
  • PUNLMP papillary urothelial neoplasm of low-malignant potential
  • Ta stage papillary carcinoma also referred to as “Ta disease,” refers to papillary carcinoma that is found on the surface of the inner lining of the bladder. Cancer cells are grouped together and can often be easily removed. The cancer has not invaded the muscle or connective tissue of the bladder wall. Ta papillary carcinoma is also referred to as noninvasive papillary urothelial carcinoma, or Stage Oa bladder cancer.
  • ‘Tl” stage papillary carcinoma also referred to as “T1 disease,” refers to papillary carcinoma that has grown through the inner lining of the bladder into the lamina basement. It has not spread to the thick layer of muscle in the bladder wall or to lymph nodes or other organs. In some embodiments, the carcinoma has invaded subepithelial connective tissue.
  • “bladder carcinoma in situ,” “bladder CIS,” and “carcinoma in situ of the bladder” are used interchangeably to refer to the clinical stage of bladder cancer characterized by a flat (i.e., non-papillary) lesion comprising of cytologically malignant cells which may involve either full or partial thickness of the urothelium.
  • low-grade tumor refers to a well-differentiated tumor
  • high-grade tumor refers to a poorly-differentiated tumor, as determined by a pathologist.
  • a high-grade tumor is often faster-growing, more likely to spread, more likely to recur after treatment, and cells look more abnormal compared to a low-grade tumor.
  • tumor grading is as described in Humphrey PA, et al. The 2016 WHO Classification of Tumours of the Urinary System and Male Genital Organs-Part B: Prostate and Bladder Tumours. Eur Urol. 2016 Jul;70(l): 106-119.
  • treatment is an approach for obtaining beneficial or desired results including clinical results.
  • beneficial or desired clinical results include, but are not limited to, one or more of the following: alleviating one or more symptoms resulting from the bladder cancer, diminishing the extent of the bladder cancer, stabilizing the bladder cancer (e.g., preventing or delaying the worsening of the bladder cancer), preventing or delaying the spread (e.g., metastasis) of the bladder cancer, preventing or delaying the recurrence of the bladder cancer, reducing recurrence rate of the bladder cancer, delay or slowing the progression of the bladder cancer, ameliorating the bladder cancer state, providing a remission (partial or total) of the bladder cancer, decreasing the dose of one or more other medications required to treat the bladder cancer, delaying the progression of the bladder cancer, increasing the quality of life, and/or prolonging survival.
  • treatment is a reduction of pathological consequence of bladder cancer.
  • BCG-naive refers to an individual who has not received prior intravesical therapy with Bacillus Calmette -Guerin (BCG) for the treatment of bladder cancer (also known as “completely BCG-naive”), or who has not received prior intravesical BCG therapy within the past 24 months (e.g., within the last 24 months prior to their current pathological diagnosis), or has received a maximum of 1 or 2 doses of BCG therapy within the past 24 months (e.g., within the last 24 months prior to their current pathological diagnosis).
  • BCG Bacillus Calmette -Guerin
  • Adjuvant setting refers to a clinical setting in which an individual has had a history of bladder cancer, and generally (but not necessarily) been responsive to therapy, which includes, but is not limited to, surgery (e.g., transurethral resection of bladder tumor (“TURBT”), partial cystectomy, or radical cystectomy), radiotherapy, and chemotherapy.
  • Treatment or administration in the “adjuvant setting” refers to a subsequent mode of treatment.
  • Neoadjuvant setting refers to a clinical setting in which the method is carried out before the primary/defmitive therapy.
  • the term “individual,” “subject,” and “patient” are used interchangeably herein to describe a mammal, including humans.
  • An individual includes, but is not limited to, human, bovine, horse, feline, canine, rodent, or primate.
  • the individual is human.
  • an individual suffers from bladder cancer.
  • the individual is in need of treatment.
  • “delaying” the development of bladder cancer means to defer, hinder, slow, retard, stabilize, and/or postpone development of the bladder cancer. This delay can be of varying lengths of time, depending on the history of the disease and/or individual being treated. As is evident to one skilled in the art, a sufficient or significant delay can, in effect, encompass prevention, in that the individual does not develop the disease.
  • a method that “delays” development of bladder cancer is a method that reduces probability of disease development in a given time frame and/or reduces the extent of the bladder cancer in a given time frame, when compared to not using the method. Such comparisons are typically based on clinical studies, using a statistically significant number of subjects.
  • Bladder cancer development can be detectable using standard methods, including, but not limited to, urinary cytology, urethra-cystoscopy (UCS), computed tomography (CT Scan, e.g., helical spiral CT scan), endoscopic ultrasound (EUS), endoscopic retrograde cholangiopancreatography (ERCP), laparoscopy, or biopsy (e.g., percutaneous needle biopsy or fine needle aspiration). Development may also refer to bladder cancer progression that may be initially undetectable and includes recurrence.
  • UCS urethra-cystoscopy
  • CT Scan computed tomography
  • EUS endoscopic ultrasound
  • ERCP endoscopic retrograde cholangiopancreatography
  • laparoscopy e.g., percutaneous needle biopsy or fine needle aspiration.
  • biopsy e.g., percutaneous needle biopsy or fine needle aspiration.
  • combination therapy is meant that a first agent be administered in conjunction with another agent.
  • “In conjunction with” refers to administration of one treatment modality in addition to another treatment modality.
  • “in conjunction with” refers to administration of one treatment modality before, during, or after delivery of the other treatment modality to the individual.
  • monotherapy refers to administration of a single therapeutic agent, such as the oncolytic virus, which is not in conjunction with another treatment modality, such as an immune checkpoint modulator.
  • a monotherapy with the oncolytic virus can be administered with a pretreatment composition, such as a transduction enhancing agent (e.g., N-Dodecyl-P-D-maltoside [DDM]), which is not considered as a different treatment modality for the purpose of this invention.
  • a transduction enhancing agent e.g., N-Dodecyl-P-D-maltoside [DDM]
  • telomere binding means that the compound preferably interacts with (e.g., binds to, modulates, and inhibits) a particular target (e.g., a protein and an enzyme) than a non-target.
  • a particular target e.g., a protein and an enzyme
  • an effective amount refers to an amount of an agent (such as the oncolytic virus described herein) or composition sufficient to treat a specified disorder, condition or disease such as ameliorate, palliate, lessen, and/or delay one or more of its symptoms.
  • an effective amount comprises an amount sufficient to cause a tumor to shrink and/or to decrease the growth rate of the tumor (such as to suppress tumor growth) or to prevent or delay other unwanted cell proliferation in cancer.
  • an effective amount is an amount sufficient to delay development of bladder cancer.
  • an effective amount is an amount sufficient to prevent or delay recurrence.
  • an effective amount is an amount sufficient to reduce recurrence rate in the individual.
  • the effective amount is an amount sufficient to inhibit tumor metastasis in the individual.
  • An effective amount can be administered in one or more administrations.
  • the effective amount of the agent or composition may: (i) reduce the number of cancer cells; (ii) reduce tumor size; (iii) inhibit, retard, slow to some extent and preferably stop cancer cell infiltration into peripheral organs; (iv) inhibit (i.e., slow to some extent and preferably stop) tumor metastasis; (v) inhibit tumor growth; (vi) prevent occurrence and/or recurrence of tumor; (vii) delay occurrence and/or recurrence of tumor; (viii) reduce recurrence rate of tumor, and/or (ix) relieve to some extent one or more of the symptoms associated with the cancer.
  • an “effective amount” may be in one or more doses, i.e., a single dose or multiple doses may be required to achieve the desired treatment endpoint.
  • the term “simultaneous administration,” as used herein, means that a first therapy and second therapy in a combination therapy are administered at the same time.
  • the first and second therapies may be contained in the same composition (e.g., a composition comprising both a first and second therapy) or in separate compositions (e.g., a first therapy is contained in one composition and a second therapy is contained in another composition).
  • the term “sequential administration” or “in sequence” means that the first therapy and second therapy in a combination therapy are administered with a time separation, for example, of more than about 1 minute, such as more than about any of 5, 10, 15, 20, 30, 40, 50, 60, or more minutes.
  • the term “sequential administration” means that the first therapy and second therapy in a combination therapy are administered with a time separation of more than about 1 day, such as more than about any of 1 day to 1 week, 2 weeks, 3 weeks, 4 weeks, 8 weeks, 12 weeks, or more weeks. Either the first therapy or the second therapy may be administered first.
  • the first and second therapies are contained in separate compositions, which may be contained in the same or different packages or kits.
  • administered immediately prior to means that the first therapy is administered no more than about 15 minutes, such as no more than about any of 10, 5 or 1 minutes before administration of the second therapy.
  • administered immediately after means that the first therapy is administered no more than about 15 minutes, such as no more than about any of 15, 10 or 1 minutes after administration of the second therapy.
  • pharmaceutically acceptable or “pharmacologically compatible” is meant a material that is not biologically or otherwise undesirable, e.g., the material may be incorporated into a pharmaceutical composition administered to a patient without causing any significant undesirable biological effects or interacting in a deleterious manner with any of the other components of the composition in which it is contained.
  • Pharmaceutically acceptable carriers or excipients have preferably met the required standards of toxicological and manufacturing testing and/or are included on the Inactive Ingredient Guide prepared by the U.S. Food and Drug administration.
  • An “adverse event” or “AE” as used herein refers to any untoward medical occurrence in an individual receiving a marketed pharmaceutical product or in an individual who is participating on a clinical trial who is receiving an investigational or non- investigational pharmaceutical agent.
  • the AE does not necessarily have a causal relationship with the individual’s treatment. Therefore, an AE can be any unfavorable and unintended sign, symptom, or disease temporally associated with the use of a medicinal product, whether or not considered to be related to the medicinal product.
  • An AE includes but is not limited to: an exacerbation of a pre-existing illness; an increase in frequency or intensity of a preexisting episodic event or condition; a condition detected or diagnosed after study drug administration even though it may have been present prior to the start of the study; and continuously persistent disease or symptoms that were present at baseline and worsen following the start of the study.
  • An AE generally does not include: medical or surgical procedures (e.g., surgery, endoscopy, tooth extraction, or transfusion); however, the condition that leads to the procedure is an adverse event; pre-existing diseases, conditions, or laboratory abnormalities present or detected at the start of the study that do not worsen; hospitalizations or procedures that are done for elective purposes not related to an untoward medical occurrence (e.g., hospitalizations for cosmetic or elective surgery or social/convenience admissions); the disease being studied or signs/symptoms associated with the disease unless more severe than expected for the individual's condition; and overdose of study drug without any clinical signs or symptoms.
  • medical or surgical procedures e.g., surgery, endoscopy, tooth extraction, or transfusion
  • pre-existing diseases, conditions, or laboratory abnormalities present or detected at the start of the study that do not worsen e.g., hospitalizations for cosmetic or elective surgery or social/convenience admissions
  • the disease being studied or signs/symptoms associated with the disease unless more severe than
  • a “serious adverse event” or (SAE) as used herein refers to any untoward medical occurrence at any dose including, but not limited to, that: a) is fatal; b) is life-threatening (defined as an immediate risk of death from the event as it occurred); c) results in persistent or significant disability or incapacity; d) requires in-patient hospitalization or prolongs an existing hospitalization (exception: Hospitalization for elective treatment of a pre-existing condition that did not worsen during the study is not considered an adverse event.
  • AEs Complications that occur during hospitalization are AEs and if a complication prolongs hospitalization, then the event is serious); e) is a congenital anomaly/birth defect in the offspring of an individual who received medication; or f) conditions not included in the above definitions that may jeopardize the individual or may require intervention to prevent one of the outcomes listed above unless clearly related to the individual’s underlying disease.
  • “Lack of efficacy” progressive disease
  • the signs and symptoms or clinical sequelae resulting from lack of efficacy should be reported if they fulfill the AE or SAE definitions.
  • response assessments may be used to evaluate a nontarget lesion: “complete response” or “CR” refers to disappearance of all non-target lesions; “stable disease” or “SD” refers to the persistence of one or more non-target lesions not qualifying for CR or PD; and “progressive disease” or “PD” refers to the “unequivocal progression” of existing non-target lesion(s) or appearance of one or more new lesion(s) is considered progressive disease (if PD for the individual is to be assessed for a time point based solely on the progression of non-target lesion(s), then additional criteria are required to be fulfilled.
  • Progression free survival indicates the length of time during and after treatment that the cancer does not grow. Progression-free survival includes the amount of time individuals have experienced a complete response or a partial response, as well as the amount of time individuals have experienced stable disease.
  • Cystectomy free survival indicates the length of time during and after treatment that a cystectomy is not required for the patient as determined by the physician.
  • “High-grade event-free-survival (HG EFS)” refers to the duration from the first treatment on study during which a patient does not experience high-grade recurrence or persistence of NMIBC, disease progression, radical cystectomy, or death from any cause.
  • “Bladder cancer-specific survival” indicates the length of time from the time of first treatment to the time of death due to bladder cancer.
  • Predicting or “prediction” is used herein to refer to the likelihood that an individual is likely to respond either favorably or unfavorably to a treatment regimen.
  • “at the time of starting treatment” or “baseline” refers to the time period at or prior to the first exposure to the treatment.
  • sample refers to a composition which contains a molecule which is to be characterized and/or identified, for example, based on physical, biochemical, chemical, physiological, and/or genetic characteristics.
  • reference to "not" a value or parameter generally means and describes "other than” a value or parameter.
  • the method is not used to treat cancer of type X means the method is used to treat cancer of types other than X.
  • Described herein is a method of treating bladder cancer in an individual (e.g., a human individual), comprising intravesically administering to the individual a two-step instillation regimen one or more times, wherein each two-step instillation regimen comprises: intravesically administering a pretreatment composition comprising a transductionenhancing agent (e.g., DDM) to the individual; and subsequently intravesically administering an oncolytic virus to the individual directly after administering the pretreatment composition, wherein there is no intervening bladder wash between the administration of the pretreatment composition and the administration of the oncolytic virus.
  • a transductionenhancing agent e.g., DDM
  • the oncolytic virus comprises a viral vector comprising a tumor cell-specific promoter operably linked to a viral gene essential for replication of the oncolytic virus, and a heterologous gene encoding an immune-related molecule.
  • each two-step instillation regimen consists of: intravesically administering the pretreatment composition to the individual; and subsequently intravesically administering the oncolytic virus to the individual directly after administering the pretreatment composition.
  • the method further comprises one or more bladder washes with saline prior to the two-step instillation regimen.
  • the method further comprises one or more bladder washes with the pretreatment composition prior to the two-step instillation regimen.
  • the method comprises one or more treatment courses each comprising intravesically administering the two-step instillation regimen weekly. In certain embodiments, each treatment courses comprises administering the two-step instillation regimen weekly for about 1 week to about 6 weeks. [0084] In some embodiments, the method comprises at least a first induction phase comprising administering the two-step instillation regimen to the individual once per week for six weeks. In some embodiments, the method comprises a maintenance phase subsequent to the induction phase, wherein the maintenance phase comprises administering the two-step instillation regimen to the individual every three or six months. In certain embodiments, the maintenance phase comprises administering the two-step instillation regimen weekly for one to three weeks every three or six months.
  • the start of the first induction phase and the start of the maintenance phase are separated by about three months or about six months.
  • the two-step instillation regimen is administered to the individual once per week for six weeks on month zero during the first induction phase and the individual begins the maintenance phase at month three or around week 13 following the start of the first induction phase.
  • the two-step instillation regimen is administered to the individual once per week for six weeks on month zero during the first induction phase and the individual is reevaluated at month three or around week 11 following the start of the first induction phase.
  • the individual begins the maintenance phase at month three or around week 13 following the start of the first induction phase.
  • the maintenance phase comprises administering the two-step instillation regimen once per week for three weeks every three months for nine months and subsequently administering the two-step instillation regimen once per week for three weeks every six months.
  • the maintenance phase comprises administering the two- step instillation regimen once per week for three weeks every three months for six months and subsequently administering the two-step instillation regimen once every three months for six months.
  • the individual receives a second induction phase comprising administration of the two-step instillation regimen once per week for six weeks at month three or around week 13 following the start of the first induction phase.
  • the maintenance phase comprises administering the two-step instillation regimen once per week for three weeks every three months for six months and subsequently administering the two-step instillation regimen once per week for three weeks every six months.
  • the maintenance phase comprises: (i) administering the two- step instillation regimen once per week for three weeks every three months for six to nine months, and subsequently (ii) administering the two-step instillation regimen once per week for three weeks every six months.
  • Any of the methods described herein may be useful for inhibiting growth of a bladder tumor, inhibiting metastasis of a bladder tumor, prolonging survival (such as disease- free survival, progression-free survival, recurrence-free survival, cystectomy-free survival, or high-grade event-free survival) of an individual having bladder cancer, causing disease remission in an individual having bladder cancer, preventing disease progression of an individual having bladder cancer, and/or improving quality of life of an individual having bladder cancer.
  • the method is bladder-preserving or bladder-sparing.
  • the bladder-preserving or bladder-sparing method is useful for improving the quality of life of the individual.
  • the methods described herein comprises intravesically administering to the individual a two-step instillation regimen one or more times, wherein each two-step instillation regimen comprises (e.g., consisting of or consisting essentially of): intravesically administering a pretreatment composition comprising a transduction-enhancing agent to the individual, and subsequently intravesically administering an oncolytic virus to the individual directly after administering the pretreatment composition, wherein there is no intervening bladder wash between the administration of the pretreatment composition and the administration of the oncolytic virus.
  • the method further comprises one or more bladder washes with saline prior to the two-step instillation regimen.
  • the method further comprises one or more bladder washes with the pretreatment composition prior to the two-step instillation regimen.
  • the transduction enhancing agent is N-Dodecyl-[3-D- maltoside (DDM).
  • DDM is a nonionic surfactant comprised of a maltose derivatized with a single twelve-carbon chain, and acts as a mild detergent and solubilizing agent. It has been used as a food additive and is known to enhance mucosal surface permeation in rodents, probably due to its effect on membrane associated GAG and tight junctions.
  • the pretreatment composition comprises DDM at a concentration of about 0.01% to about 0.5%, including for example any of about 0.01% to about 0.05%, about 0.05% to about 0.1%, about 0.1% to about 0.2%, about 0.2% to about 0.3%, about 0.3% to about 0.4%, and about 0.4% to about 0.5%.
  • the pretreatment composition comprises DDM at a concentration of about 0.05% to about 0.2%.
  • the pretreatment composition comprises DDM at a concentration of about 0.1%.
  • the method comprises one or more intravesical instillations of DDM.
  • the intravesical instillation of DDM comprises administering the DDM into the bladder and allowing the DDM to dwell in the bladder for a dwell time of at least about 5 minutes before draining the bladder.
  • the intravesical instillation of DDM comprises allowing the DDM to dwell in the bladder for a dwell time of about 5, 10, 15, 20, 25, or 30 minutes.
  • the intravesical instillation of DDM comprises allowing the DDM to dwell in the bladder for a dwell time of about 15 minutes, or between about 10 minutes and about 20 minutes.
  • the intravesical instillation comprises allowing the DDM to dwell in the bladder for a dwell time of between 5 and 10 minutes, between 5 and 15 minutes, between 5 and 20 minutes, between 5 and 25 minutes, between 5 and 30 minutes, between 10 and 15 minutes, between 10 and 20 minutes, between 10 and 25 minutes, between 10 and 30 minutes, between 15 and 20 minutes, between 15 and 25 minutes, between 15 and 30 minutes, between 20 and 25 minutes, or between 25 and 30 minutes.
  • each dose of DDM is administered as an intravesical instillation of DDM comprising allowing the DDM to dwell in the bladder for a dwell time of at least about 5 minutes (e.g., about 15 minutes, or between about 10 minutes and about 20 minutes).
  • the method comprises one or more bladder washes with DDM or saline.
  • the bladder wash comprises administering the DDM or saline into the bladder and then draining the bladder shortly thereafter, e.g., immediately after or within about 5, 4, 3, 2 or 1 minute or less after administration.
  • the methods described herein comprise administering one or more doses (e.g. , instillations and/or washes) of DDM to the individual prior to administering the oncolytic virus.
  • the methods described herein comprises administering a single dose (e.g., a single instillation) of DDM prior to administering the oncolytic virus.
  • the methods described herein comprise a single bladder wash with DDM and a single instillation of DDM prior to administering the oncolytic virus.
  • the method comprises one or more saline washes before or after instillation of DDM and prior to administering the oncolytic virus.
  • the method does not comprise any saline washes in between instillation of DDM and the administration of the oncolytic vims.
  • a method of treating bladder cancer comprising administering DDM and an oncolytic vims (e.g, cretostimogene), wherein the method comprises a five-step instillation regimen.
  • a method of treating bladder cancer comprising administering DDM and an oncolytic vims (e.g., cretostimogene), wherein the method comprises administering: 1) a first bladder wash with normal saline; 2) a second bladder wash with DDM; 3) instillation of DDM; 4) a third bladder was with normal saline; and 5) instillation of cretostimogene.
  • the first and third bladder washes are with between 50 mb and 100 mL normal saline (e.g., about 100 mL normal saline).
  • the second bladder wash is with between 50 mL and 100 mL (e.g., about 75 mL) of a solution of DDM.
  • the instillation of DDM is instillation is with between 50 mL and 100 mL (e.g., about 85 mL to 100 mL, or about 100 mL) of a solution of DDM.
  • the instillation of cretostimogene is with between 50 mL and 100 mL (e.g., about 85 mL to 100 mL, or about 100 mL) of a solution of cretostimogene.
  • the solution of DDM is 0.1% w/w DDM in normal saline.
  • the solution of cretostimogene comprises between 1 x 10 8 and 1 x 10 14 viral particles (e.g., around 1 x 10 12 viral particles).
  • the instillation of cretostimogene is administered no later than 60 minutes (e.g., no later than 60 minutes, 50 minutes, 40 minutes, 30 minutes, or 20 minutes) after the first bladder wash. In certain embodiments, the instillation of cretostimogene is administered no later than about 30 minutes (e.g., 25 minutes to 35 minutes) after the first bladder wash.
  • a method of treating bladder cancer comprising administering DDM and an oncolytic virus (e.g, cretostimogene), wherein the method comprises a two-step instillation regimen.
  • the two-step instillation regimen does not comprise bladder washes with saline.
  • a method of treating bladder cancer comprising administering DDM and an oncolytic virus (e.g., cretostimogene), wherein the method comprises administering: 1) a single instillation of DDM; and 2) instillation of cretostimogene, wherein the method does not comprise a bladder wash with saline.
  • the method does not comprise a bladder wash with DDM.
  • the instillation of DDM is instillation is with between 50 mL and 100 mL (e.g., about 85 mL to 100 mL, or about 100 mL) of a solution of DDM.
  • the instillation of cretostimogene is with between 50 mL and 100 mL (e.g., about 85 mb to 100 mb, or about 100 mb) of a solution of cretostimogene.
  • the solution of DDM is 0.1% w/w DDM in normal saline.
  • the solution of cretostimogene comprises between 1 x 10 8 and 1 x 10 14 viral particles (e.g., around 1 x 10 12 viral particles).
  • the instillation of cretostimogene is administered no later than 60 minutes (e.g, no later than 60 minutes, 50 minutes, 40 minutes, 30 minutes, or 20 minutes) after the instillation of DDM.
  • the instillation of cretostimogene is administered no later than about 30 minutes (e.g., 25 minutes to 35 minutes) after the instillation of DDM.
  • the method comprises intravesically administering at least one dose of DDM to the individual prior to administering the oncolytic virus.
  • the oncolytic virus is administered directly after the at least one dose of DDM, without intravesical administration of a saline wash.
  • the method comprises administering a first and second dose of DDM to the individual prior to administering the oncolytic virus.
  • the method comprises intravesically administering a saline wash after only the first DDM wash.
  • the oncolytic virus is administered directly after the second dose of DDM, without intravesical administration of a saline wash after the second dose of DDM and before administration of the oncolytic virus.
  • the DDM is administered at a concentration of about any one of 0.01%, 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 1%, 2%, 3%, 4%, or 5%. In certain embodiments, the DDM is administered at a volume of between about 50 mL and about 100 mL.
  • the methods described herein comprise administering a first and second dose of DDM to the individual prior to administering an oncolytic virus, wherein the oncolytic virus comprises a viral vector comprising a tumor cell-specific promoter operably linked to a viral gene essential for replication of the oncolytic virus, and a heterologous gene encoding an immune-related molecule.
  • the method comprises intravesically administering a saline wash after only the first DDM wash.
  • the oncolytic virus is administered directly after the second dose of DDM, without intravesical administration of a saline wash after the second dose of DDM and administration of the oncolytic virus.
  • the oncolytic virus is administered directly after the at least one dose of DDM, without intravesical administration of a saline wash.
  • the DDM is administered at a concentration of about 0.1%.
  • the DDM is administered at a volume of between about 50 mL and about 100 mL.
  • each two-step instillation regimen comprises or consists of: a) intravesically administering a pretreatment composition comprising a transduction-enhancing agent to the individual; and subsequently b) intravesically administering an oncolytic virus to the individual directly after administering the pretreatment composition, wherein there is no intervening bladder wash between the administration of the pretreatment composition and the administration of the oncolytic virus.
  • the oncolytic virus comprises a viral vector comprising a tumor cellspecific promoter operably linked to a viral gene essential for replication of the oncolytic virus, and a heterologous gene encoding an immune -related molecule.
  • the oncolytic virus is cretostimogene.
  • the transduction- enhanding agent is N-Dodecyl-[3-D-maltoside (DDM).
  • the method further comprises one or more bladder washes with saline prior to the two-step instillation regimen (e.g., prior to each two-step instillation regimen). In some embodiments, the method further comprises one or more bladder washes with the pretreatment composition prior to the two-step instillation regimen (e.g., prior to each two-step instillation regimen).
  • the pretreatment composition comprises DDM at a concentration of about 0.01% to about 0.5%. In certain embodiments, the pretreatment composition comprises DDM at a concentration of about 0.05% to about 0.2%. IN certain embodiments, the pretreatment composition comprises DDM at a concentration of about 0.1%.
  • the pretreatment composition comprises DDM at a concentration of about 0.01% to about 0.05%, about 0.01% to about 0.1%, about 0.01% to about 0.15%, about 0.01% to about 0.2%, about 0.01% to about 0.3%, about 0.01% to about 0.4%, about 0.01% to about 0.5%, about 0.05% to about 0.1%, about 0.05% to about 0.15%, about 0.05% to about 0.2%, about 0.05% to about 0.3%, about 0.05% to about 0.4%, about 0.05% to about 0.5%, about 0. 1% to about 0. 15%, about 0. 1% to about 0.2%, about 0. 1% to about 0.3%, about 0.1% to about 0.4%, about 0. 1% to about 0.5%, about 0. 0.1% to about 0.5%, about 0. 0.
  • the pretreatment composition is administered at a volume of between about 50 mb and about 100 mb. In certain embodiments, the pretreatment composition is administered at a volume of between about 85 mb and about 100 mb. In certain embodiments, the pretreatment composition is administered at a volume of between about 50 mb and about 60 mb, between about 50 mb and about 70 mb, between about 50 mb and about 80 mb, between about 50 mb and about 90 mb, between about 50 mb and about 100 mb, between about 60 mb and about 70 mb, between about 60 mb and about 80 mb, between about 60 mb and about 90 mb, between about 60 mb and about 100 mb, between about 70 mb and about 80 mb, between about 70 mb and about 90 mb, between about 70 mb and about 100 mb, between about 80 mb and about 90 mb, between about 80 mb and about 90 mb, between about 80
  • the administration of the pretreatment composition comprises allowing the pretreatment composition to dwell in the bladder for more than 5 minutes after completion of the administration of the full volume of the pretreatment composition into the bladder. In certain embodiments, the administration of the pretreatment composition comprises allowing the pretreatment composition to dwell in the bladder for more than 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 minutes after completion of the administration of the full volume of the pretreatment composition into the bladder. In some embodiments, the administration of the pretreatment composition comprises allowing the pretreatment composition to dwell in the bladder for between about 10 minutes and about 20 minutes after completion of the administration of the full volume of the pretreatment composition into the bladder.
  • the administration of the pretreatment composition comprises allowing the pretreatment composition to dwell in the bladder for about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 minutes after completion of the administration of the full volume of the pretreatment composition into the bladder. In certain embodiments, the administration of the pretreatment composition comprises allowing the pretreatment composition to dwell in the bladder for about 15 minutes after completion of the administration of the full volume of the pretreatment composition into the bladder.
  • the administration of the oncolytic virus comprises allowing the oncolytic virus to dwell in the bladder for between about 55 minutes and about 65 minutes after completion of the administration of the full volume of the oncolytic virus into the bladder. In certain embodiments, the administration of the oncolytic virus comprises allowing the oncolytic virus to dwell in the bladder for about 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, or 65 minutes after completion of the administration of the full volume of the oncolytic virus into the bladder. In certain embodiments, the administration of the oncolytic virus comprises allowing the oncolytic virus to dwell in the bladder for about 60 minutes after completion of the administration of the full volume of the oncolytic virus into the bladder.
  • the administration of the full volume of the oncolytic virus into the bladder is completed within about 60 minutes after the administration of the full volume of the pretreatment composition into the bladder is completed. In certain embodiments, the administration of the full volume of the oncolytic virus into the bladder is completed within about 60, 55, 50, 45, 40, 35, 30, 25, 20, 15, 10, or 5 minutes after the administration of the full volume of the pretreatment composition into the bladder is completed. In certain embodiments, the administration of the full volume of the oncolytic virus into the bladder is completed within about 30 minutes after the administration of the full volume of the pretreatment composition into the bladder is completed. In certain embodiments, the administration of the full volume of the oncolytic virus into the bladder is completed within about 35 minutes after the administration of the full volume of the pretreatment composition into the bladder is completed.
  • the full volume of the pretreatment composition and the full volume of the oncolytic virus are both administered into the bladder within about 60 minutes. In certain embodiments, he full volume of the pretreatment composition and the full volume of the oncolytic virus are both administered into the bladder within about 60, 55, 50, 45, 40, 35, 30, 25, 20, 15, 10, or 5 minutes. In certain embodiments, the full volume of the pretreatment composition and the full volume of the oncolytic virus are both administered into the bladder within about 35 minutes. In certain embodiments, the full volume of the pretreatment composition and the full volume of the oncolytic virus are both administered into the bladder within about 30 minutes.
  • the administration of the oncolytic virus is initiated within about 60 minutes after the initiation of the administration of the pretreatment composition. In certain embodiments, the administration of the oncolytic virus is initiated within about 60, 55, 50, 45, 40, 35, 30, 25, 20, 15, 10, or 5 minutes after the initiation of the administration of the pretreatment composition. In some embodiments, the administration of the oncolytic virus is initiated within about 30 minutes after the initiation of the administration of the pretreatment composition. In some embodiments, the administration of the oncolytic virus is initiated within about 20 minutes after the initiation of the administration of the pretreatment composition. In some embodiments, the administration of the oncolytic virus is initiated within about 15 minutes after the initiation of the administration of the pretreatment composition.
  • the pretreatment composition is administered at a volume of between about 50 mb and about 100 mb, such as any of about 55 mb, 60 mb, 65 mb, 70 mb 75 mb, 80 mb, 85 mb, 90 mb, 95 mb, or 100 mb. In some embodiments, the pretreatment composition is administered at a volume of between about 85 mb and about 100 mb.
  • the administration of the pretreatment composition comprises allowing the pretreatment composition to dwell in the bladder for more than 5 minutes (e.g., more than about any of 6, 7, 8, 9, 10, 15, 20, or 30 minutes) after completion of the administration of the full volume of the pretreatment composition into the bladder. In some embodiments, the administration of the pretreatment composition comprises allowing the pretreatment composition to dwell in the bladder for between about 10 minutes and about 20 minutes after completion of the administration of the full volume of the pretreatment composition into the bladder. In some embodiments, the administration of the pretreatment composition comprises allowing the pretreatment composition to dwell in the bladder for about 15 minutes after completion of the administration of the full volume of the pretreatment composition into the bladder.
  • the administration of the oncolytic virus is administered at a dose of about 1 x 10 8 to about 1 x 10 14 viral particles. In certain embodiments, the administration of the oncolytic virus is administered at a dose of about 1 x 10 12 viral particles. In some embodiments, the oncolytic virus is administered in a volume of between about 50 mb and about 100 mb. In some embodiments, the oncolytic virus is administered in a volume of between about 85 mb and about 100 mb. In some embodiments, the administration of the oncolytic virus comprises allowing the oncolytic virus to dwell in the bladder for between about 55 minutes and about 65 minutes after completion of the administration of the full volume of the oncolytic virus into the bladder. In some embodiments, the administration of the oncolytic virus comprises allowing the oncolytic virus to dwell in the bladder for about 60 minutes after completion of the administration of the full volume of the oncolytic virus into the bladder.
  • the administration of the full volume of the oncolytic virus into the bladder is completed within about 35 minutes after the administration of the full volume of the pretreatment composition into the bladder is completed. In certain embodiments, the administration of the full volume of the oncolytic virus into the bladder is completed within about 30 minutes after the administration of the full volume of the pretreatment composition into the bladder is completed. In some embodiments, the full volume of the pretreatment composition and the full volume of the oncolytic virus are both administered into the bladder within about 35 minutes. In certain embodiments, the full volume of the pretreatment composition and the full volume of the oncolytic virus are both administered into the bladder within about 30 minutes. In some embodiments, the administration of the oncolytic virus is initiated within about 30 minutes of the initiation of the administration of the pretreatment composition.
  • the administration of the oncolytic virus is initiated within about 20 minutes of the initiation of the administration of the pretreatment composition. In certain embodiments, the administration of the oncolytic virus is initiated within about 15 minutes of the initiation of the administration of the pretreatment composition.
  • Suitable concentration of the pretreatment composition include, but are not limited to, about any one of 0.01%, 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 1%, 2%, 3%, 4%, or 5% of the transducing enhancing agent (such as DDM).
  • the pretreatment composition comprises any of about 0.01% to about 0.05%, about 0.05% to about 0. 1%, about 0.
  • the transduction enhancing agent such as DDM
  • the pretreatment (such as DDM) is administered immediately (such as no more than 5 minutes) prior to the administration of the oncolytic virus.
  • the pretreatment (such as DDM) is administered no more than about any of 5 minutes, 10 minutes, 15 minutes, 20 minutes, 30 minutes, 45 minutes, 1 hour, 90 minutes, 2 hours, 3 hours or 4 hours before the administration of the oncolytic virus.
  • the pretreatment (such as DDM) is administered no more than about 2 hours before the administration of the oncolytic virus.
  • Suitable dosages for the pretreatment composition include, but are not limited to, about any of 0.1 mg/kg, 0.5 mg/kg, 1 mg/kg, 1.5mg/kg, 2 mg/kg, 2.5 mg/kg, 5mg/kg, 10 mg/kg, 25 mg/kg, 50 mg/kg, 100 mg/kg, 150 mg/kg, 200 mg/kg, 250 mg/kg, 300 mg/kg, 400 mg/kg, 500 mg/kg, 0.
  • 1 mg/kg to 0.5 mg/kg 0.5 mg/kg to 1 mg/kg, 0.5 mg/kg to 1 mg/kg, 1 mg/kg to 2 mg/kg, 2 mg/kg to 5mg/kg, 5mg/kg to 10 mg/kg, 10 mg/kg to 25 mg/kg, 25 mg/kg to 50 mg/kg, 50 mg/kg to 100 mg/kg, 100 mg/kg to 150 mg/kg, 150 mg/kg to 200 mg/kg, 200 mg/kg to 250 mg/kg, 250 mg/kg to 500 mg/kg, or 0.5 mg/kg to about 5 mg/kg.
  • a suitable dosage for the pretreatment composition is about any one of 0.1 g, 0.2 g, 0.5g, 0.75 g, 1 g, 1.5 g, 2 g, 2.5 g, 5 g, or 10 g of the transduction enhancing agent (such as DDM).
  • the transduction enhancing agent such as DDM
  • kits for treating bladder cancer in an individual comprising intravesical administration of an effective amount of oncolytic virus in combination with administration of an effective amount of additional therapeutic agent.
  • the additional therapeutic agent comprises a chemotherapeutic agent.
  • the additional therapeutic agent comprises an immune checkpoint modulator.
  • the oncolytic virus and the additional therapeutic agent are administered sequentially, z.e., the oncolytic virus is administered before or after the administration of the additional therapeutic agent. In some embodiments, the oncolytic virus is administered prior to the administration of the additional therapeutic agent. In some embodiments, the oncolytic virus is administered no more than about any of 15 minutes, 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 12 hours, or 24 hours prior to the administration of the additional therapeutic agent.
  • the oncolytic virus is administered about days or weeks (such as about any of 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 4 weeks, or more) prior to the administration of the additional therapeutic agent. In some embodiments, the oncolytic virus is administered after the administration of the additional therapeutic agent. In some embodiments, the oncolytic virus is administered no more than about any of 15 minutes, 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 12 hours, or 24 hours after the administration of the additional therapeutic agent.
  • the oncolytic virus is administered about days or weeks (such as about any of 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 4 weeks, or more) after the administration of the additional therapeutic agent.
  • the oncolytic virus and the additional therapeutic agent are administered with one immediately after another (e.g., within 5 minutes or less between the two administrations).
  • the oncolytic virus is administered immediately before the administration of the additional therapeutic agent.
  • the oncolytic virus is administered immediately after the administration of the additional therapeutic agent.
  • the oncolytic vims and the additional therapeutic agent are administered simultaneously.
  • the oncolytic vims and the additional therapeutic agent are administered simultaneously via separate compositions. In some embodiments, the oncolytic vims and the additional therapeutic agent are administered simultaneously via the same composition. In some embodiments, the oncolytic vims and the additional therapeutic agent are both administered intravesically via the same composition.
  • exemplary routes of administration of additional therapeutic agents include, but are not limited to, intratumoral, intravesical, intramuscular, intraperitoneal, intravenous, intra-arterial, intracranial, intrapleural, subcutaneous, and epidermal routes, or be delivered into lymph glands, body spaces, organs or tissues known to contain such live cancer cells.
  • the additional therapeutic agent e.g., immune checkpoint modulator or chemotherapeutic agent
  • the additional therapeutic agent is administered locally to the site of the tumor or to the tissue having the tumor.
  • the additional therapeutic agent e.g, immune checkpoint modulator or chemotherapeutic agent
  • the additional therapeutic agent is administered by direct injection of the agent(s) into the tumor.
  • the additional therapeutic agent e.g, immune checkpoint modulator or chemotherapeutic agent
  • the additional therapeutic agent e.g., immune checkpoint modulator or chemotherapeutic agent
  • the additional therapeutic agent e.g., immune checkpoint modulator or chemotherapeutic agent
  • the additional therapeutic agent is administered systemically.
  • the additional therapeutic agent e.g., immune checkpoint modulator or chemotherapeutic agent
  • the method comprises administration of a single additional therapeutic agent (e.g., immune checkpoint modulator or chemotherapeutic agent).
  • the additional therapeutic agent is a chemotherapeutic agent.
  • the additional therapeutic agent is an immune checkpoint modulator (e.g. an immune checkpoint inhibitor).
  • the immune checkpoint inhibitor is selected from the immune checkpoint inhibitors listed in Table 1, wherein the immune checkpoint inhibitor is administered with the same route of administration, and/or dose, and/or dosing frequency, and/or duration, and/or maintenance schedule as listed in Table 1.
  • the immune checkpoint inhibitor is selected from the immune checkpoint inhibitors listed in Table 1, wherein the immune checkpoint inhibitor is administered with the different route of administration, and/or dose, and/or dosing frequency, and/or duration, and/or maintenance schedule as listed in Table 1. In some embodiments, the immune checkpoint inhibitor is not a molecule selected from Table 1.
  • the method comprises administration of at least two (such as any of 2, 3, 4, 5, 6, or more) additional therapeutic agents (e.g., immune checkpoint modulators and/or chemotherapeutic agents). In some embodiments, all or part of the at least two additional therapeutic agents are administered simultaneously, such as in a single composition. In some embodiments, all or part of the at least two additional therapeutic agents are administered sequentially. In some embodiments, the method comprises administration of a combination of additional therapeutic agents comprising an immune checkpoint inhibitor, a chemotherapeutic agent, or both. In some embodiments, the method comprises administration of a combination of additional therapeutic agents comprising two or more (such as any of 2, 3, 4, 5, 6, or more) immune checkpoint inhibitors.
  • additional therapeutic agents e.g., immune checkpoint modulators and/or chemotherapeutic agents.
  • the method comprises administration of a combination of additional therapeutic agents comprising two or more (such as any of 2, 3, 4, 5, 6, or more) chemotherapeutic agents. In some embodiments, the method comprises administration of a combination of additional therapeutic agents comprising any number (such as any of 1, 2, 3, 4, 5, 6, or more) of immune checkpoint inhibitors and any number (such as any of 2, 3, 4, 5, 6, or more) of chemotherapeutic agents. In some embodiments, the at least two additional therapeutic agents comprise one or more immune checkpoint inhibitors selected from Table 1 and/or one or more chemotherapeutic agents described herein. In some embodiments, the at least two additional therapeutic agents are each independently administered systemically (e.g., intravenously) or locally (e.g., intravesically).
  • the method comprises systemic (e.g., intravenous) administration of an immune checkpoint modulator, local (e.g., intravesical) administration of an immune checkpoint modulator, systemic (e.g., intravenous) administration of a chemotherapeutic agent, local (e.g., intravesical) administration of a chemotherapeutic agent, or any combination thereof.
  • the method comprises intravenous administration of an immune checkpoint modulator and intravesical administration of a chemotherapeutic agent.
  • the at least two additional therapeutic agents are administered sequentially.
  • the at least two additional therapeutic agents are administered simultaneously.
  • the at least two additional therapeutic agents are administered simultaneously via separate compositions.
  • the at least two different therapeutic agents are administered simultaneously via the same composition.
  • the administration of the additional therapeutic agents can be of any sequence, including simultaneous systemic administration of the first additional therapeutic agent and local administration of the second additional therapeutic agent, and sequential administration of the additional therapeutic agents, among which at least one additional therapeutic agent is administered systemically, for example, first administering the second additional therapeutic agent locally (such as intravesically) to the site of the tumor followed by systemic (such as intravenous) administration of the first additional therapeutic agent, or first administering the first additional therapeutic agent systemically (such as intravenously) followed by local (such as intratumoral) administration of the second additional therapeutic agent.
  • Additional therapeutic agents administered simultaneously via the same administration route may be administered as a single composition.
  • the additional therapeutic agents can be admixed prior to (such as immediately prior to, e.g., within less than about 10, 5, or 1 minutes before) the administration of the single composition.
  • a method of treating bladder cancer comprising intravesically administering an oncolytic virus described herein and intravesically administering one or more additional therapeutic agents described herein.
  • the additional therapeutic agent is administered concurrently with the oncolytic virus.
  • the additional therapeutic agent and the oncolytic virus are administered sequentially.
  • the method comprises intravesical administration of the oncolytic virus followed by intravesical administration of the additional therapeutic agent on the same day of treatment. In some embodiments, the method comprises intravesical administration of the oncolytic virus followed immediately by intravesical administration of the additional therapeutic agent. In some embodiments, wherein the method comprises at least a first 6-week induction phase comprising administering the oncolytic virus and additional therapeutic agent to the individual on weeks 1, 2, 3, 4, 5, and 6. In some embodiments, the method comprises a maintenance phase subsequent to the induction phase, wherein the maintenance phase comprises a 3 -week treatment comprising administering the oncolytic virus and additional therapeutic agent to the individual every three or six months.
  • the 3 -week treatment comprises administering the oncolytic virus and additional therapeutic agent weekly on weeks 1, 2, and 3.
  • the start of the first 6-week induction phase and the start of the maintenance phase are separated by about three or six months.
  • the individual is reevaluated at month three or around week 11 following the start of the first 6-week induction phase.
  • the individual begins the maintenance phase at month three or around week 13 following the start of the first 6-week induction phase.
  • the maintenance phase comprises administering the 3-week treatment every three months for nine months, followed by administering the 3-week treatment every six months.
  • the individual receives a second 6-week induction phase at month three or around week 13 following the start of the first 6-week induction phase.
  • the maintenance phase comprises administering the 3-week treatment every three months for six months, followed by administering the 3-week treatment every six months.
  • the additional therapeutic agent and the oncolytic virus are administered on different days or different weeks of a treatment course.
  • the method comprises at least a first 6-week induction phase comprising: i) administering the oncolytic virus to the individual on weeks 1, 2, 4 and 5; and ii) administering additional therapeutic agent to the individual on weeks 3 and 6.
  • the method comprises a maintenance phase subsequent to the induction phase, wherein the maintenance phase comprises a 3-week treatment comprising administering the oncolytic virus and additional therapeutic agent to the individual every three or six months.
  • the 3-week treatment comprises administering the oncolytic virus weekly on weeks 1 and 2 followed by administering additional therapeutic agent in week 3.
  • the start of the first 6-week induction phase and the start of the maintenance phase are separated by about three or six months.
  • the individual is reevaluated at month three or around week 11 following the start of the first 6- week induction phase.
  • the individual begins the maintenance phase at month three or around week 13 following the start of the first 6-week induction phase.
  • the maintenance phase comprises administering the 3-week treatment every three months for nine months, followed by administering the 3-week treatment every six months.
  • the individual receives a second 6-week induction phase at month three or around week 13 following the start of the first 6-week induction phase.
  • the maintenance phase comprises administering the 3-week treatment every three months for six months, followed by administering the 3-week treatment every six months.
  • the methods and compositions described herein are related to oncolytic viruses, for example, oncolytic adenovirus.
  • the oncolytic virus may be a naturally occurring virus, or a genetically modified virus, for example an attenuated virus, and/or a virus with additional favorable features (e.g., preferential replication in cancer cells, or encoding an immune- related molecule).
  • the oncolytic virus is a wild type oncolytic virus.
  • the oncolytic virus is genetically modified.
  • the oncolytic virus is attenuated (for example through multiple passages, inactivation or genetic modification).
  • the oncolytic virus is replication competent.
  • the oncolytic virus preferentially replicates in a cancer cell, such as a cancer cell defective in the Rb pathway.
  • the oncolytic virus (such as oncolytic adenovirus) comprises a viral vector comprising a tumor cell-specific promoter operably linked to a viral gene essential for replication of the oncolytic virus.
  • the tumor-specific promoter is an E2F-1 promoter, such as a human E2F-1 promoter or an E2F-1 promoter comprising the nucleotide sequence set forth in SEQ ID NO: 1 as shown below.
  • the viral gene essential for replication of the oncolytic virus is selected from the group consisting of E1A, E1B, and E4.
  • the oncolytic virus (such as oncolytic adenovirus) comprises a viral vector comprising a tumor-selective promoter operably linked to a viral gene essential for replication of the oncolytic virus.
  • the tumor-selective promoter is an E2F-1 promoter, such as a human E2F-1 promoter or an E2F-1 promoter comprising the nucleotide sequence set forth in SEQ ID NO: 1 as shown below.
  • the viral gene essential for replication of the oncolytic virus is selected from the group consisting ofElA, E1B, and E4.
  • the oncolytic virus a viral vector comprising a tumor cellspecific promoter operably linked to a viral gene essential for replication of the oncolytic virus.
  • the oncolytic virus comprises a tumor-selective promoter.
  • the oncolytic virus is an adenovirus serotype 5, wherein the endogenous Ela promoter of a native adenovirus serotype 5 is replaced by the human E2F-1 promoter, and the endogenous E3 19kD coding region of the native adenovirus serotype 5 is replaced by a nucleic acid encoding human GM-CSF.
  • the oncolytic virus is cretostimogene.
  • the tumor selective promoter allows preferential replication of the oncolytic virus in tumor cells.
  • the oncolytic virus preferentially replicates in a cancer cell, such as a cancer cell defective in the Rb pathway.
  • the oncolytic virus is selected from the group consisting of adenovirus, herpes simplex virus, vaccinia virus, mumps virus, Newcastle disease virus, polio virus, measles virus, Seneca valley virus, coxsackie virus, reovirus, vesicular stomatitis virus, maraba and rhabdovirus, and parvovirus.
  • the tumor-specific promoter is an E2F-1 promoter, such as a human E2F-1 promoter or an E2F-1 promoter comprising the nucleotide sequence set forth in SEQ ID NO: 1.
  • the viral gene essential for replication of the oncolytic virus is selected from the group consisting of E1A, E1B, and E4.
  • the oncolytic virus further comprises an immune-related molecule (such as cytokine, chemokine, or PRRago (i.e., pathogen recognition receptor agonist)).
  • an immune-related molecule such as cytokine, chemokine, or PRRago (i.e., pathogen recognition receptor agonist)
  • the immune-related molecule is not an immune checkpoint modulator.
  • the immune-related molecule is selected from the group consisting of GM-CSF, IL-2, IL-12, interferon (such as Type 1, Type 2 or Type 3 interferon, e g., interferon y), CCL4, CCL19, CCL21, CXCL13, TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, TLR10, RIG-I, MDA5, LGP2, and LTap.
  • interferon such as Type 1, Type 2 or Type 3 interferon, e g., interferon y
  • the immune-related molecule is selected from the group consisting of STING (i.e., stimulator of interferon genes) activators (such as CDN, i.e., cyclic dinucleotides), PRRago (such as CpG, Imiquimod, or Poly I:C), TLR stimulators (such as GS-9620, AED- 1419, CYT-003-QbG10, AVE-0675, or PF-7909), and RLR stimulators (such as RIG-I, Mda5, or LGP2 stimulators).
  • STING i.e., stimulator of interferon genes
  • activators such as CDN, i.e., cyclic dinucleotides
  • PRRago such as CpG, Imiquimod, or Poly I:C
  • TLR stimulators such as GS-9620, AED- 1419, CYT-003-QbG10, AVE-0675, or PF-7909
  • the immune -related molecule is expressed by the oncolytic virus.
  • the oncolytic virus may comprise a nucleic acid encoding the immune- related molecule, and the nucleic acid can be in the viral vector or on a separate vector.
  • the oncolytic virus is a virus comprising a viral vector, and wherein the viral vector comprises the nucleic acid encoding the immune-related molecule.
  • the nucleic acid encoding the immune-related molecule is operably linked to a viral promoter, such as an El promoter, or an E3 promoter.
  • the immune -related molecule enhances an immune response in the individual.
  • Immune-related molecules may include, but are not limited to, a cytokine, a chemokine, a stem cell growth factor, a lymphotoxin, an hematopoietic factor, a colony stimulating factor (CSF), erythropoietin, thrombopoietin, tumor necrosis factor-alpha (TNF), TNF-beta , granulocyte-colony stimulating factor (G-CSF), granulocyte macrophage-colony stimulating factor (GM-CSF), interferon-alpha, interferon-beta, interferon-gamma, interferon- lambda, stem cell growth factor designated "SI factor", human growth hormone, N- methionyl human growth hormone, bovine growth hormone, parathyroid hormone, thyroxine, insulin, proinsulin, relaxin, prorelaxin, follicle stimulating
  • the oncolytic virus is an adenovirus serotype 5.
  • the endogenous Ela promoter and E3 19kD coding region of a native adenovirus is replaced by the human E2F-1 promoter and a nucleic acid encoding human GM-CSF.
  • a polyadenylation signal (PA) is inserted 5’ of the E2F-1 promoter.
  • the nucleic acid encoding human GM-CSF is operably linked to the E3 promoter.
  • the vector backbone of the adenovirus serotype 5 further comprises E2, E4, late protein regions or inverted terminal repeats (ITRs) identical to the wildtype adenovirus serotype 5 genome.
  • the oncolytic virus has the genomic structure as shown in Figure 1.
  • the oncolytic virus is conditionally replicating.
  • the oncolytic virus preferentially replicates in cancer cells.
  • the cancer cells are Rb pathway-defective cancer cells.
  • the oncolytic virus is cretostimogene.
  • the oncolytic virus is an oncolytic adenovirus comprising a tumor cell-specific promoter operably linked to a viral gene essential for replication of the oncolytic adenovirus, and a heterologous gene encoding an immune-related molecule.
  • the immune-related molecule is GM-CSF and the heterologous gene encoding the GM-CSF is operably linked to an El promoter or an E3 promoter.
  • the tumor cell-specific promoter is an E2F-1 promoter (e.g., an E2F-1 promoter comprising the nucleotide sequence set forth in SEQ ID NO: 1) and the viral gene essential for replication of the oncolytic adenovirus is selected from the group consisting of E1A, E1B, and E4.
  • E2F-1 promoter e.g., an E2F-1 promoter comprising the nucleotide sequence set forth in SEQ ID NO: 1
  • the viral gene essential for replication of the oncolytic adenovirus is selected from the group consisting of E1A, E1B, and E4.
  • the immune-related molecule is GM-CSF and the heterologous gene encoding the GM-CSF is operably linked to an El promoter or an E3 promoter
  • the tumor cell-specific promoter is an E2F-1 promoter (e.g., an E2F-1 promoter comprising the nucleotide sequence set forth in SEQ ID NO: 1)
  • the viral gene essential for replication of the oncolytic adenovirus is selected from the group consisting of E1A, E1B, and E4.
  • the oncolytic adenovirus is an adenovirus serotype 5.
  • the endogenous E la promoter of the native adenovirus serotype 5 is replaced by the human E2F-1 promoter.
  • the endogenous E3 19kD coding region of the native adenovirus serotype 5 is replaced by a nucleic acid encoding human GM-CSF.
  • the endogenous Ela promoter of the native adenovirus serotype 5 is replaced by the human E2F-1 promoter and the endogenous E3 19kD coding region of the native adenovirus serotype 5 is replaced by a nucleic acid encoding human GM-CSF.
  • a method of treating bladder cancer in an individual comprising intravesically administering to the individual an effective amount of an adenovirus serotype 5, wherein the endogenous Ela promoter and E3 19kD coding region of a native adenovirus is replaced by the human E2F-1 promoter and a nucleic acid encoding an immune-related molecule (such as cytokine or chemokine, for example, GM-CSF).
  • the tumor-specific promoter is a human E2F-1 promoter or an E2F-1 promoter comprising the nucleotide sequence set forth in SEQ ID NO: 1.
  • the method further comprises administering to the individual a transduction enhancing agent (such as DDM) prior to the administration of the adenovirus.
  • the method comprises intravesically administering at least one dose of DDM to the individual prior to administering the oncolytic virus.
  • the oncolytic virus is administered directly after the at least one dose of DDM, without intravesical administration of a saline wash.
  • the method comprises intravesically administering a saline wash after only the first DDM wash.
  • the oncolytic virus is administered directly after the second dose of DDM, without intravesical administration of a saline wash after the second dose of DDM and administration of the oncolytic virus.
  • the adenovirus is administered at a dose of about 1 x 10 8 to about 1 x 10 14 viral particles (such as about IxlO 12 viral particles). In some embodiments, the adenovirus is administered weekly.
  • the IR- NMIBC comprises a tumor of low-grade Ta stage, wherein the tumor of Ta stage is recurrent within about 12 months of resection of a prior low-grade tumor of Ta stage or high-grade tumor of Ta stage having a diameter of 3 cm or smaller.
  • the IR- NMIBC comprises a single tumor of low grade Ta stage having a diameter of greater than 3 cm.
  • the IR-NMIBC comprises two or more low-grade tumors of Ta stage.
  • the IR-NMIBC comprises a primary and solitary high-grade tumor of Ta stage having a diameter of less than 3 cm.
  • the IR-NMIBC comprises a low-grade tumor of T1 stage.
  • the oncolytic vims (such as oncolytic adenovirus) comprises a viral vector comprising a tumor cell-specific promoter operably linked to a viral gene essential for replication of the oncolytic vims and a nucleic acid encoding an immune-related molecule (such as cytokine or chemokine) operably linked to a viral promoter.
  • the tumor-specific promoter is an E2F-1 promoter, such as a human E2F-1 promoter or an E2F-1 promoter comprising the nucleotide sequence set forth in SEQ ID NO: 1.
  • the viral gene essential for replication of the oncolytic vims is selected from the group consisting of E1A, E1B, and E4.
  • the viral promoter operably linked to the nucleic acid encoding the immune -related molecule is the E3 promoter.
  • the immune-related molecule is GM-CSF.
  • the oncolytic vims (such as oncolytic adenovims) comprises a viral vector comprising a tumor cell-selective promoter operably linked to a viral gene essential for replication of the oncolytic vims and a nucleic acid encoding an immune-related molecule (such as cytokine or chemokine) operably linked to a viral promoter.
  • the tumor-selective promoter is an E2F-1 promoter, such as a human E2F-1 promoter or an E2F-1 promoter comprising the nucleotide sequence set forth in SEQ ID NO: 1.
  • the viral gene essential for replication of the oncolytic vims is selected from the group consisting of E1A, E1B, and E4.
  • the viral promoter operably linked to the nucleic acid encoding the immune -related molecule is the E3 promoter.
  • the immune-related molecule is GM-CSF.
  • the oncolytic vims is an adenovims serotype 5, wherein the endogenous Ela promoter and E3 19kD coding region of a native adenovims is replaced by the human E2F-1 promoter and a nucleic acid encoding an immune-related molecule (such as cytokine or chemokine, for example, GM-CSF).
  • the tumor-specific promoter is a human E2F-1 promoter or an E2F-1 promoter comprising the nucleotide sequence set forth in SEQ ID NO: 1.
  • the oncolytic vims is cretostimogene, an exemplary adenovims serotype 5 which has an E2F promoter at the Ela gene and a GM-CSF expression at the E3 gene.
  • Cretostimogene also known as CG0070 or cretostimogene grenadenorepvec
  • Cretostimogene is a conditionally replicating oncolytic adenovims (serotype 5) designed to preferentially replicate in and kill Rb pathway-defective cancer cells.
  • This vector is transcriptionally regulated by a promoter (e.g., E2F-1 promoter) that is up-regulated in Rb-pathway-detective tumor cells.
  • a promoter e.g., E2F-1 promoter
  • one or more genes of the Rb pathway such as the tumor suppressor Rb gene, are mutated.
  • cretostimogene also encodes the human cytokine GM-CSF, which is expressed selectively in the infected tumor cells to stimulate immune responses against uninfected distant (such as metastases) and local tumor foci.
  • Cretostimogene The genomic structure of the oncolytic adenoviral vector cretostimogene is shown schematically in FIG. 1.
  • Products of the adenoviral early El A gene are essential for efficient expression of other regions of the adenoviral genome.
  • Cretostimogene has been engineered to express the E1A gene under control of the human E2F-1 promoter, which provides tumor specificity to the El A gene product.
  • a polyadenylation signal (PA) was inserted 5' of the E2F-1 promoter.
  • Cretostimogene includes the entire wild type E3 region except for the 19kD-coding region.
  • cretostimogene carries the cDNA for human GM-CSF under the control of the endogenous E3 promoter (E3P). Since the E3 promoter is in turn activated by El A, both viral replication and GM-CSF expression are ultimately under the control of the E2F-1 promoter.
  • E3P endogenous E3 promoter
  • Cretostimogene is manufactured in a human cancer cell line and released from infected cells by detergent lysis. Cretostimogene is purified from the lysate by chromatography, and then formulated in 5% sucrose, 10 mM Tris, 0.05% polysorbate-80, 1 % glycine, 1 mM magnesium chloride, pH 7.8.
  • Cretostimogene is supplied as a sterile, slightly opalescent, frozen liquid in stoppered glass vials.
  • the particle concentration per mb (vp/mL) is stated on the Certificate of Analysis for each lot of cretostimogene.
  • Cretostimogene has additional potential anti-tumor activity in that it carries the cDNA for human GM-CSF, a key cytokine for generating long-lasting anti-tumor immunity.
  • cretostimogene is a selectively replicating oncolytic vector with the potential for attacking the tumor by two mechanisms: direct cytotoxicity as a replicating vector and induction of a host immune response.
  • direct cytotoxicity as a replicating vector
  • induction of a host immune response In vitro and in vivo studies have been conducted to characterize the tumor selectivity and anti-tumor activity and safety of cretostimogene. See, for example, U.S. Patent No. 11,596,660, which incorporated herein by reference in its entirety.
  • cretostimogene grenadenorepvec As used herein, the terms “cretostimogene grenadenorepvec,” “cretostimogene,” and “CG0070” are used interchangeably.
  • the methods described herein comprise administration of an immune checkpoint modulator in combination with intravesical administration of an oncolytic virus.
  • Immune checkpoint modulators of particular interest in the present invention include immune -stimulating agents and immune checkpoint inhibitors.
  • immune checkpoint inhibitors As used herein, the term “immune checkpoint inhibitors,” “checkpoint inhibitors,” and the like refers to compounds that inhibit the activity of control mechanisms of the immune system. Immune system checkpoints, or immune checkpoints, are inhibitory pathways in the immune system that generally act to maintain self-tolerance or modulate the duration and amplitude of physiological immune responses to minimize collateral tissue damage.
  • Checkpoint inhibitors can inhibit an immune system checkpoint by stimulating the activity of a stimulatory checkpoint molecule, or inhibiting the activity of an inhibitory checkpoint molecule in the pathway.
  • Stimulatory checkpoint molecules are molecules, such as proteins, that stimulate or positively regulate the immune system.
  • Inhibitory checkpoint molecules are molecules, such as proteins, that inhibit or negatively regulate the immune system.
  • Immune system checkpoint molecules include, but are not limited to, cytotoxic T-lymphocyte antigen 4 (CTLA-4), programmed cell death 1 protein (PD-1), programmed cell death 1 ligand 1 (PD- Ll), programmed cell death 1 ligand 2 (PD-L2), lymphocyte activation gene 3 (LAG3), B7-1, B7-H3, B7-H4, T cell membrane protein 3 (TIM3), B- and T-lymphocyte attenuator (BTLA), V-domain immunoglobulin (Ig) -containing suppressor of T-cell activation (VISTA), Killercell immunoglobulin-like receptor (KIR), and A2A adenosine receptor (A2aR).
  • CTLA-4 cytotoxic T-lymphocyte antigen 4
  • PD-1 programmed cell death 1 protein
  • PD- Ll programmed cell death 1 ligand 1
  • PD-L2 programmed cell death 1 ligand 2
  • LAG3 lymphocyte activation gene 3
  • TIM3 B7
  • checkpoint inhibitors include antagonists of CTLA-4, PD-1, PD-L1, PD-L2, LAG3, B7-1, B7-H3, B7-H4, BTLA, VISTA, KIR, A2aR, or TIM3.
  • CTLA-4, PD-1, PD-L1, PD-L2, LAG3, B7-1, B7-H3, B7-H4, BTLA, VISTA, KIR, A2aR, or TIM3 are checkpoint inhibitors.
  • any molecule e.g., peptide, nucleic acid, small molecule, etc.
  • the immune checkpoint modulator is an immune checkpoint inhibitor.
  • the immune checkpoint inhibitor is a natural or engineered ligand of an inhibitory immune checkpoint molecule, including, for example, ligands of CTLA-4 (e.g., B7.1, B7.2), ligands of TIM3 (e.g., Galectin-9), ligands of A2a Receptor (e.g., adenosine, Regadenoson), ligands of LAG3 (e.g., MHC class I or MHC class II molecules), ligands of BTLA (e g., HVEM, B7-H4), ligands of KIR (e.g., MHC class I or MHC class II molecules), ligands of PD-1 (e.g., PD-L1, PD-L2), ligands of IDO (e.g., NKTR-218, Indoximod, NLG919), ligand
  • the immune checkpoint inhibitor is an antibody that targets an inhibitory immune checkpoint protein.
  • the immune checkpoint inhibitor is an antibody selected from the group consisting of anti-CTLA-4 (e.g., Ipilimumab, Tremelimumab, KAHR-102), anti-TIM3 (e.g., F38-2E2, ENUM005), anti- LAG3 (e.g., BMS-986016, IMP701, IMP321, C9B7W), anti-KIR (e.g., Lirilumab, IPH2101, IPH4102), anti-PD-1 (e.g., Nivolumab, Pidilizumab, Pembrolizumab, BMS-936559, atezolizumab, Lambrolizumab, MK-3475, AMP-224, AMP-514, STI-Al l 10, TSR-042), anti- PD-L1 (e.g., KY-1003 (EP20120194977
  • the antibody is an antagonistic antibody. In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody is an antigen-binding fragment selected from the group consisting of Fab, Fab’, F(ab’)2, Fv, scFv, VHH (nanobody), and other antigen-binding subsequences of the full length antibody. In some embodiments, the antibody is a human, humanized, or chimeric antibody. In some embodiments, the antibody is a bispecific antibody, a multispecific antibody, a single domain antibody, a fusion protein comprising an antibody portion, or any other functional variants or derivatives thereof.
  • the immune checkpoint inhibitor can be of any one of the molecular modalities known in the art, including, but not limited to, aptamer, mRNA, siRNA, microRNA, shRNA, peptide, antibody, anticalin, spherical nucleic acid, TALEN, Zinc Finger Nuclease, CRISPR/Cas9, and small molecule.
  • the immune checkpoint inhibitors can be used singly or in combination. For example, any number (such as any of 1, 2, 3, 4, 5, 6, or more) of immune checkpoint inhibitors can be used simultaneously or sequentially. Sequential administration of immune checkpoint inhibitors can be separated by hours, days or weeks. The administration route(s) for two or more immune checkpoint inhibitors can be the same or different.
  • one immune checkpoint inhibitors can be administered locally (e.g., intravesically or intratumorally), and a second immune checkpoint inhibitors can be administered systemically (e.g., intravenously); two immune checkpoint inhibitors can both be administered locally (e.g., intravesically or intratumorally); or two immune checkpoint inhibitors can both be administered systemically (e.g, intravenously).
  • two or more immune checkpoint inhibitors are administered simultaneously in the same composition.
  • two or more immune checkpoint inhibitors are administered simultaneously in separate compositions.
  • immune checkpoint molecules and inhibitors thereof are discussed below. It is understood that other suitable immune checkpoint molecules and immune checkpoint inhibitors known in the art are also within the scope of the present application.
  • the immune checkpoint inhibitor is an inhibitor of PD-1.
  • the inhibitor of PD-1 is an anti-PD-1 antibody.
  • Any of the anti-PD-1 antibodies known in the art may be used in the present invention, including, but not limited to, nivolumab, pembrolizumab, pidilizumab, BMS-936559, and atezolizumab, lambrolizumab, MK-3475, AMP-224, AMP-514, STI-A1110, and TSR-042.
  • the anti-PD-1 antibody is a monoclonal antibody or a polyclonal antibody.
  • the anti-PD- 1 antibody is an antigen-binding fragment selected from the group consisting of Fab, Fab’, F(ab’)2, Fv, scFv, and other antigen-binding subsequences of the full-length anti-PD-1 antibody.
  • the anti-PD-1 antibody is a human, humanized, or chimeric antibody.
  • the anti-PD-1 antibody is a bispecific antibody, a multispecific antibody, a single domain antibody, a fusion protein comprising an antibody portion, or any other variants or derivatives thereof.
  • the inhibitor of PD-1 is a natural or engineered ligand of PD-1, such as PD-L1 or PD-L2.
  • the inhibitor of PD-1 is an inhibitor of the interaction between PD-1 and its ligand, for example, an inhibitor of PD-1/PD-L1 interaction or an inhibitor of PD-1/PD-L2 interaction.
  • the inhibitor of PD-1 is an inhibitor of a PD-1 ligand, such as an inhibitor of PD-L1 (e.g., anti-PD-Ll antibody) or an inhibitor of PD-L2 (e.g., anti-PD-L2 antibody). Any of the inhibitors of interaction between PD-1 and its ligand may be used in the present invention, see, for example, U.S. Patent No.
  • the inhibitor of PD-1 is an Fc fusion protein comprising a PD-1 ligand, such as an Fc-fusion of PD-L2 (e.g., AMP -224).
  • the inhibitor of PD-1 is pembrolizumab or nivolumab.
  • PD-1 is a part of the B7/CD28 family of co-stimulatory molecules that regulate T- cell activation and tolerance, and thus antagonistic anti-PD-1 antibodies can be useful for overcoming tolerance.
  • PD-1 has been defined as a receptor for B7-4.
  • B7-4 can inhibit immune cell activation upon binding to an inhibitory receptor on an immune cell.
  • Anti-PD- 1 antibodies are provided in Table 1. Any of the anti-PD- 1 antibodies known in the art may be used in the present invention, for example, see US Patent Nos.
  • nivolumab is a human monoclonal antibody to PD-1 that is FDA approved for the treatment of unresectable or metastatic melanoma, squamous non-small cell lung cancer, and urothelial carcinoma.
  • Another exemplary anti-PD-1 antibody, pembrolizumab is a humanized (from mouse) monoclonal antibody to PD- 1 that is FDA approved for the treatment of a variety of cancer types, including high-risk, non-muscle invasive bladder cancer and advanced bladder cancer.
  • PD-L1/PD-L2 is a human monoclonal antibody to PD-1 that is FDA approved for the treatment of a variety of cancer types, including high-risk, non-muscle invasive bladder cancer and advanced bladder cancer.
  • the immune checkpoint inhibitor is an inhibitor of PD-1 ligand (e.g., PD-L1 and/or PD-L2).
  • the inhibitor of PD-1 ligand is an anti-PD-Ll antibody.
  • the inhibitor of PD-1 ligand is an anti-PD-L2 antibody.
  • Exemplary anti-PD-Ll antibodies include, but are not limited to, KY-1003, MCLA-145, RG7446 (also known as atezolizumab), BMS935559 (also known as MDX- 1105), MPDL3280A, MEDI4736, Avelumab (also known as MSB0010718C), and STI- A1010.
  • the anti-PD-Ll or anti-PD-L2 is a monoclonal antibody or a polyclonal antibody. In some embodiments, the anti-PD-Ll or anti-PD-L2 is an antigenbinding fragment selected from the group consisting of Lab, Lab’, P(ab’)2, Fv, scFv, VHH (nanobody) and other antigen-binding subsequences of the full-length anti-PD-Ll or anti-PD- L2 antibody. In some embodiments, the anti-PD-Ll or anti-PD-L2 antibody is a human, humanized, or chimeric antibody.
  • the anti-PD-Ll or anti-PD-L2 antibody is a bispecific antibody, a multispecific antibody, a single domain antibody, a fusion protein comprising an antibody portion, or any other variants or derivatives thereof.
  • the inhibitor of PD-1 ligand is an inhibitor (e.g., peptide, protein or small molecule) of both PD-L1 and PD-L2.
  • Exemplary inhibitors of both PD-L1 and PD-L2 include, but are not limited to, AUR-012, and AMP -224.
  • the inhibitor of PD-L 1 and the inhibitor of PD-L2 can be used interchangeably in any of the methods of treatment described herein.
  • PD-L1 (Programmed cell death-ligand 1) is also known as cluster of differentiation 274 (CD274) or B7 homolog 1 (B7-H1).
  • PD-L1 serves as a ligand for PD-1 to play a major role in suppressing the immune system during particular events such as pregnancy, tissue allographs, autoimmune disease and other disease states such as hepatitis and cancer.
  • the formation of PD-1 receptor/PD-Ll ligand complex transmits an inhibitory signal which reduces the proliferation of CD8+ T cells at the lymph nodes.
  • any of the known anti-PD-Ll antibodies may be used in the present invention, see, for example, U.S. Patent Nos. US7943743, US7722868, US8217149, US8383796, US8552154, and US9102725; and U.S. Patent Application Publication Nos. US20140341917, and US20150203580; and International Patent Application No. PCT/US2001/020964.
  • anti-PD-Ll antibodies that are in clinical development include BMS935559 (also known as MDX-1105), MPDL3280A, MEDI4736, Avelumab (also known as MSB0010718C), KY-1003, MCLA-145, RG7446 (also known as atezolizumab), and STI-A1010.
  • PD-L2 (Programmed cell death 1 ligand 2) is also known as B7-DC. PD-L2 serves as a ligand for PD- 1. Under certain circumstances, PD-L2 and its inhibitor can be used as a substitute for PD-L1 and its inhibitor respectively.
  • the immune checkpoint inhibitor is an inhibitor of CTLA-4.
  • the inhibitor of CTLA-4 is an anti-CTLA-4 antibody. Any of the anti- CTLA-4 antibodies that are known in the art may be used in the present invention, including, but not limited to, Ipilimumab, Tremelimumab, and KAHR-102.
  • the anti-CTLA-4 antibody is YERVOY® (Ipilimumab).
  • the anti-CTLA-4 antibody is a monoclonal antibody or a polyclonal antibody.
  • the anti- CTLA-4 antibody is an antigen-binding fragment selected from the group consisting of Fab, Fab’, F(ab’)2, Fv, scFv, VHH (nanobody) and other antigen-binding subsequences of the full length anti-CTLA-4 antibody.
  • the anti-CTLA-4 antibody is a human, humanized, or chimeric antibody.
  • the anti-CTLA-4 antibody is a bispecific antibody, a multispecific antibody, a single domain antibody, a fusion protein comprising an antibody portion, or any other functional variants or derivatives thereof.
  • the inhibitor of CTLA-4 is an engineered lipocalin protein specifically recognizing CTLA-4 (such as an anticalin molecule that specifically binds to CTLA-4).
  • the inhibitor of CTLA-4 is a natural or engineered ligand of CTLA-4, such as B7.1 or B7.2.
  • CTLA-4 is an immune checkpoint molecule, which is up-regulated on activated T- cells.
  • An anti-CTLA-4 mAb can block the interaction of CTLA-4 with CD80/86 and switch off the mechanism of immune suppression and enable continuous stimulation of T-cells by DCs.
  • Examples of anti-CTLA-4 antibodies are Ipilimumab (see U.S. Patent Nos. 6,984,720, 7,452,535, 7,605,238, 8,017,114 and 8,142,778), Tremilimumab (see U.S. Patent No.
  • Ipilimumab Two IgG mAb directed against CTLA-4, Ipilimumab and Tremelimumab, have been tested in clinical trials for a number of indications.
  • Ipilimumab is approved by the FDA for the treatment of melanoma, e.g., for late stage melanoma patients.
  • the complete prescribing information is fully described in the packaging insert of YERVOY® (Bristol Meyers).
  • YERVOY® (Ipilimumab) comes in 50mg single use vials.
  • Anticalins are engineered proteins that are able to recognize and bind specific targets with high affinity. They are antibody mimetics, but they are not structurally related to antibodies. Instead, they are derived from human lipocalins, which are a family of naturally binding proteins. Anticalins are being used in lieu of monoclonal antibodies, but are about eight times smaller than monoclonal antibodies with a size of about 180 amino acids and a mass of about 20 kDa. Anticalins have been described in U.S. Patent No. 7,250, 297.
  • CTLA-4-binding anticalins Any of the CTLA-4-binding anticalins may be used in the present application.
  • the CTLA-4 binding anticalin is PRS-010 (Piers AG).
  • Table 1 below summarizes examples of commercially available immune checkpoint inhibitors that have been approved by the FDA or are involved in clinical trial studies. Any one or any combination of the immune checkpoint inhibitors in Table 1 may be used in the methods described herein, using the same or different administration routes, and/or dosages, and/or dosing frequency, and/or duration, and/or maintenance schedule as listed in Table 1.
  • the methods described herein comprise administration of a chemotherapeutic agent in combination with intravesical administration of an oncolytic virus.
  • Chemotherapeutic agents that may be administered in the methods describe herein include, for example, gemcitabine, cisplatin, carboplatin, paclitaxel, docetaxel, ifosfamide, doxorubicin, methotrexate, vinblastine, mitomycin, 5 -fluorouracil (5-FU), and the like.
  • the chemotherapeutic agent comprises gemcitabine, cisplatin, carboplatin, paclitaxel, docetaxel, ifosfamide, doxorubicin, methotrexate, vinblastine, mitomycin, 5 -fluorouracil (5-FU), or any combination thereof.
  • the chemotherapeutic agent comprises gemcitabine.
  • the chemotherapeutic agent comprises cisplatin.
  • the chemotherapeutic agent comprises carboplatin.
  • the chemotherapeutic agent comprises paclitaxel.
  • the chemotherapeutic agent comprises docetaxel.
  • the chemotherapeutic agent comprises ifosfamide. In certain embodiments, the chemotherapeutic agent comprises doxorubicin. In certain embodiments, the chemotherapeutic agent comprises methotrexate. In certain embodiments, the chemotherapeutic agent comprises vinblastine. In certain embodiments, the chemotherapeutic agent comprises mitomycin (e.g., mitomycin C). In certain embodiments, the chemotherapeutic agent comprises 5 -fluorouracil (5-FU). In certain embodiments, the chemotherapeutic comprises a combination of gemcitabine and cisplatin.
  • the chemotherapeutic agent comprises a combination of dose-dense methotrexate, vinblastine, doxorubicin (Adriamycin), and cisplatin (DDMVAC). In certain embodiments, the chemotherapeutic agent comprises a combination of gemcitabine and paclitaxel. In some embodiments, the chemotherapeutic agent comprises interferon (such as interferon-a). In some embodiments, the chemotherapeutic agent comprises platinum-based agents, for example, cisplatin, carboplatin, oxaliplatin, or any combination thereof. In some embodiments, the chemotherapeutic agent comprises mitomycin (e.g., mitomycin C) and thiotepa. In some embodiments, the chemotherapeutic agent comprises cisplatin, doxorubicin, gemcitabine, and valrubicin.
  • interferon such as interferon-a
  • the chemotherapeutic agent comprises platinum-based agents, for example, cisp
  • chemotherapeutic agents described herein can be used singly or in combination. For example, any number (such as any of 1, 2, 3, 4, 5, 6, or more) of chemotherapeutic agents can be administered simultaneously or sequentially. Sequential administration chemotherapeutic agents can be separated by hours, days or weeks. The administration route(s) for two or more chemotherapeutic agents can be the same or different.
  • one chemotherapeutic agents can be administered locally (e.g., intravesically or intratumorally), and a chemotherapeutic agents can be administered systemically (e.g., intravenously); two chemotherapeutic agents can both be administered locally (e.g., intravesically or intratumorally); or two chemotherapeutic agents can both be administered systemically (e.g., intravenously).
  • two or more chemotherapeutic agents are administered simultaneously in the same composition.
  • two or more chemotherapeutic agents are administered simultaneously in separate compositions.
  • the chemotherapeutic agent is gemcitabine. Accordingly, in some embodiments, provided herein is a method of treating bladder cancer comprising intravesically administering an oncolytic virus and intravesically administering cretostimogene.
  • the bladder cancer is NMIBC. In certain embodiments, the bladder cancer is high-risk NMIBC.
  • a method of treating high-risk NMIBC in an individual comprising intravesically administering an oncolytic adenovirus to the individual and intravesically administering gemcitabine to the individual.
  • the oncolytic adenovirus comprises a tumor cell-specific promoter operably linked to a viral gene essential for replication of the oncolytic adenovirus, and a heterologous gene encoding an immune-related molecule.
  • the oncolytic adenovirus preferentially replicates in a cancer cell.
  • the cancer cell is defective in the Rb pathway.
  • the tumor cell-specific promoter is an E2F-1 promoter (e.g., an E2F-1 promoter comprising the nucleotide sequence set forth in SEQ ID NO: 1).
  • the immune-related molecule is GM-CSF.
  • the heterologous gene is operably linked to a viral promoter.
  • the viral gene essential for replication of the oncolytic adenovirus is selected from the group consisting of E1A, E1B, and E4.
  • the heterologous gene is operably linked to an El promoter or an E3 promoter.
  • the endogenous E3 19kD coding region of the native adenovirus serotype 5 is replaced by a nucleic acid encoding human GM-CSF.
  • the endogenous Ela promoter of a native adenovirus serotype 5 is replaced by the human E2F-1 promoter, and the endogenous E3 19kD coding region of the native adenovirus serotype 5 is replaced by a nucleic acid encoding human GM-CSF.
  • the oncolytic adenovirus is cretostimogene.
  • a method of treating bladder cancer in an individual comprising intravesically administering cretostimogene to the individual and intravesically administering gemcitabine to the individual.
  • the bladder cancer is NMIBC.
  • the bladder cancer is high-risk NMIBC.
  • the individual has had at least one diagnosis of high-risk NMIBC.
  • the individual has high-risk high-grade BCG- unresponsive NMIBC or high-risk NMIBC after prior therapy with BCG.
  • the gemcitabine is administered concurrently with the cretostimogene.
  • the gemcitabine and the cretostimogene are administered sequentially.
  • a method of treating high-risk NMIBC in an individual comprising intravesically administering cretostimogene to the individual and intravesically administering gemcitabine to the individual.
  • the individual has had at least one diagnosis of high-risk NMIBC and, prior to the administration of the oncolytic adenovirus and gemcitabine, the individual has: a) received at least 5 of 6 doses of a BCG induction course and at least 2 doses of 1 course of either reinduction or maintenance BCG and experienced persistent or recurrent carcinoma in situ (CIS), with or without concomitant Ta or T1 disease, within 12 months following the last dose of BCG; b) received at least 5 of 6 doses of a BCG induction course and at least 2 doses of 1 course of either reinduction or maintenance BCG and experienced recurrence of highgrade papillary-only Ta or T1 stage disease within 6 months following the last dose of BCG; c) received at least 5 of 6 dose
  • a method of treating bladder cancer e.g, high-risk NMIBC in an individual comprising intravesically administering an oncolytic virus and gemcitabine to the individual, wherein the individual: a) received at least 5 of 6 doses of a BCG induction course and at least 2 doses of 1 course of either reinduction or maintenance BCG and experienced persistent or recurrent carcinoma in situ (CIS), with or without concomitant Ta or Tl disease, within 12 months following the last dose of BCG; b) received at least 5 of 6 doses of a BCG induction course and at least 2 doses of 1 course of either reinduction or maintenance BCG and experienced recurrence of high-grade papillary-only Ta or Tl stage disease within 6 months following the last dose of BCG; c) received at least 5 of 6 doses of a BCG induction course and experienced high-grade Tl disease at a first evaluation (e.g., a 3-month evaluation) following the BCG induction course;
  • a first evaluation e.g., a
  • the BCG maintenance course comprises 2 or 3 weekly intravesical instillations of BCG.
  • the oncolytic virus comprises a viral vector comprising a tumor cell-specific promoter (e.g., a human E2F-1 promoter) operably linked to a viral gene essential for replication of the oncolytic virus, and a heterologous gene encoding an immune-related molecule (e.g, GM-CSF).
  • the oncolytic virus is an adenovirus serotype 5, wherein the endogenous Ela promoter of a native adenovirus serotype 5 is replaced by the human E2F-1 promoter, and the endogenous E3 19kD coding region of the native adenovirus serotype 5 is replaced by a nucleic acid encoding human GM-CSF.
  • the oncolytic virus is cretostimogene.
  • the method comprises intravesically administering at least one dose ofN-Dodecyl-P-D-maltoside (DDM) to the individual prior to administering the oncolytic virus.
  • DDM N-Dodecyl-P-D-maltoside
  • the method comprises intravesically administering a single dose of DDM to the individual prior to each administration of the oncolytic virus. In certain embodiments, the method does not comprise administering a saline bladder wash before or after the administration of the DDM and before the administration of the oncolytic virus. In some embodiments, the individual has non-muscle invasive bladder cancer (NMIBC). In certain embodiments, the individual has high-risk NMIBC. In some embodiments, the method comprises administration of one or more additional therapeutic agents (e.g., an immune checkpoint modulator or chemotherapeutic agent described herein, e.g., gemcitabine). In some embodiments, the method comprises an induction phase comprising administering the oncolytic virus to the individual once per week for six weeks.
  • NMIBC non-muscle invasive bladder cancer
  • the method comprises administration of one or more additional therapeutic agents (e.g., an immune checkpoint modulator or chemotherapeutic agent described herein, e.g., gemcitabine).
  • the method comprises an induction phase compris
  • the method comprises a maintenance phase subsequent to the induction phase, wherein the maintenance phase comprises administering the oncolytic virus to the individual weekly for three weeks every three or six months.
  • the start of the induction phase and the start of the maintenance phase are separated by about three months or about six months.
  • the oncolytic virus is administered at a dose of about 1 x 10 8 to about 1 x 10 14 viral particles (e.g., about 1 x 10 12 viral particles).
  • the individual a) received at least 5 of 6 doses of a BCG induction course and at least 2 doses of 1 course of either reinduction or maintenance BCG and experienced persistent or recurrent carcinoma in situ (CIS), with or without concomitant Ta or T1 disease, within 12 months following the last dose of BCG; b) received at least 5 of 6 doses of a BCG induction course and at least 2 doses of 1 course of either reinduction or maintenance BCG and experienced recurrence of high-grade papillary-only Ta or T1 stage disease within 6 months following the last dose of BCG; c) received at least 5 of 6 doses of a BCG induction course and experienced high-grade T1 disease at a first evaluation (e.g., a 3- month evaluation) following the BCG induction course; d) received at least 5 of 6 doses of a BCG induction course and experienced persistent or recurrent high-grade papillary carcinoma of Ta stage or CIS at a first evaluation (e.g., a 3
  • the BCG induction course comprises 5 or 6 weekly intravesical instillations of BCG.
  • the BCG maintenance course comprises 2 or 3 weekly intravesical instillations of BCG.
  • the method comprises intravesically administering at least one dose of N-Dodecyl-[3-D-maltoside (DDM) to the individual prior to administering cretostimogene.
  • the method comprises intravesically administering a single dose of DDM to the individual prior to each administration of cretostimogene.
  • the method does not comprise administering a saline bladder wash before or after the administration of the DDM and before the administration of cretostimogene.
  • cretostimogene is administered at a dose of about 1 x 10 8 to about 1 x 10 14 viral particles (e.g., about 1 x 10 12 viral particles).
  • the method is according to Example 3, Cohort CX.
  • the method comprises intravesical administration of cretostimogene followed by intravesical administration of gemcitabine on the same day of treatment. In some embodiments, the method comprises intravesical administration of cretostimogene followed immediately by intravesical administration of gemcitabine. In some embodiments, wherein the method comprises at least a first 6-week induction phase comprising administering cretostimogene and additional therapeutic agent to the individual on weeks 1, 2, 3, 4, 5, and 6. In some embodiments, the method comprises a maintenance phase subsequent to the induction phase, wherein the maintenance phase comprises a 3-week treatment comprising administering cretostimogene and additional therapeutic agent to the individual every three or six months.
  • the 3- week treatment comprises administering cretostimogene and additional therapeutic agent weekly on weeks 1, 2, and 3.
  • the start of the first 6-week induction phase and the start of the maintenance phase are separated by about three or six months.
  • the individual is reevaluated at month three or around week 11 following the start of the first 6-week induction phase.
  • the individual begins the maintenance phase at month three or around week 13 following the start of the first 6-week induction phase.
  • the maintenance phase comprises administering the 3-week treatment every three months for nine months, followed by administering the 3-week treatment every six months.
  • the individual receives a second 6-week induction phase at month three or around week 13 following the start of the first 6-week induction phase.
  • the maintenance phase comprises administering the 3-week treatment every three months for six months, followed by administering the 3-week treatment every six months.
  • cretostimogene is administered at a dose of between 1 x 10 8 and 1 x 10 14 viral particles (e.g., about 1 x 10 12 viral particles).
  • gemcitabine is administered at a dose of between 0.1 g and 5 g (e.g., between 0.1 and 2 g or between 0.5 g and 1.5 g). In certain embodiments, gemcitabine is administered at a dose of around 1 g.
  • the method is according to Example 3, Cohort CX, Arm 1.
  • the method comprises at least a first 6-week induction phase comprising: i) administering cretostimogene to the individual on weeks 1, 2, 4 and 5; and ii) administering additional therapeutic agent to the individual on weeks 3 and 6.
  • the method comprises a maintenance phase subsequent to the induction phase, wherein the maintenance phase comprises a 3 -week treatment comprising administering cretostimogene and additional therapeutic agent to the individual every three or six months.
  • the 3 -week treatment comprises administering cretostimogene weekly on weeks 1 and 2 followed by administering additional therapeutic agent in week 3.
  • the start of the first 6-week induction phase and the start of the maintenance phase are separated by about three or six months.
  • the individual is reevaluated at month three or around week 11 following the start of the first 6-week induction phase.
  • the individual begins the maintenance phase at month three or around week 13 following the start of the first 6-week induction phase.
  • the maintenance phase comprises administering the 3-week treatment every three months for nine months, followed by administering the 3-week treatment every six months.
  • the individual receives a second 6-week induction phase at month three or around week 13 following the start of the first 6-week induction phase.
  • the maintenance phase comprises administering the 3-week treatment every three months for six months, followed by administering the 3-week treatment every six months.
  • cretostimogene is administered at a dose of between 1 x 10 8 and 1 x 10 14 viral particles (e.g., about 1 x 10 12 viral particles).
  • gemcitabine is administered at a dose of between 0. 1 g and 5 g (e.g., between 1 g and 3 g or between 1.5 g and 2.5 g). In certain embodiments, gemcitabine is administered at a dose of around 2 g.
  • the method is according to Example 3, Cohort CX, Arm 2.
  • gemcitabine is administered at a dose between 0.1 g and 5 g. In some embodiments, gemcitabine is administered at a dose of about 0.1 g, about 0.5 g, about 1 g, about 1.5 g, about 2 g, about 2.5 g, about 3 g, about 3.5 g, about 4 g, about 4.5 g, or about 5 g. In certain embodiments, gemcitabine is administered at a dose of about 1 g. In certain embodiments, gemcitabine is administered at a dose of about 2 g.
  • gemcitabine is administered at a dose of between 0.1 g and 0.5 g, between 0.1 g and 1 g, between 0.1 g and 1.5 g, between 0.1 g and 2 g, between 0.1 g and 2.5 g, between 0.1 g and 3 g, between 0.1 g and 3.5 g, between 0.1 g and 4 g, between 0.1 g and 4.5 g, between 0.1 g and 5 g, between 0.5 g and 1 g, between 0.5 g and 1.5 g, between 0.5 g and 2 g, between 0.5 g and 2.5 g, between 0.5 g and 3 g, between 0.5 g and 3.5 g, between 0.5 g and 4 g, between 0.5 g and 4.5 g, between 0.5 g and 5 g, between 1 g and 1.5 g, between 1 g and 2 g, between 1 g and 2.5 g, between 1 g and 3 g, between 1 g and 3 g, between 1 g
  • gemcitabine is administered at a dose of between 0.5 g and 2.5 g. In certain embodiments, gemcitabine is administered at a dose of between 0.5 g and 1.5 g. In certain embodiments, gemcitabine is administered at a dose of between 1 g and 3 g. In certain embodiments, gemcitabine is administered at a dose of between 1.5 g and 2.5 g.
  • each dose of gemcitabine is administered by intravesical instillation of between 0.1 g and 5 g (e.g., about 1 g or about 2 g) in between 50 and 100 mL normal saline (e.g., in about 50 mL, about 60 mL, about 70 mL, about 80 mL, about 90 mL, or about 100 mL normal saline).
  • each instillation of gemcitabine is maintained in the bladder for a dwell time of around 60 minutes (e.g., between 50 and 70 minutes or between 55 and 65 minutes) before draining.
  • the methods described herein comprise administering an immunomodulatory agent.
  • the immunomodulatory agent is an immune -stimulating agent.
  • the immune -stimulating agent is a natural or engineered ligand of an immune stimulatory molecule, including, for example, ligands of 0X40 (e.g., OX40L), ligands of CD-28 (e.g., CD80, CD86), ligands of ICOS (e.g., B7RP1), ligands of 4-1BB (e.g., 4-1BBL, Ultra4-1BBL), ligands of CD27 (e.g., CD70), ligands of CD40 (e.g., CD40L), and ligands of TCR (e.g., MHC class I or class II molecules, IMCgplOO).
  • 0X40 e.g., OX40L
  • CD-28 e.g., CD80, CD86
  • ICOS e.g., B7RP1
  • 4-1BB e.g., 4
  • the immune -stimulating agent is an antibody selected from the group consisting of anti-CD28 (e.g., TGN-1412), anti-OX40 (e.g., MEDI6469, MEDI-0562), anti-ICOS (e.g., MEDI-570), anti-GITR (e.g., TRX518, INBRX-110, NOV-120301), anti-41-BB (e.g., BMS-663513, PF- 05082566), anti-CD27 (e.g., BION-1402, Varlilumab and hCD27.15), anti-CD40 (e.g., CP870,893, BI-655064, BMS-986090, APX005, APX005M), anti-CD3 (e.g., blinatumomab, muromonab), and anti-HVEM.
  • anti-CD28 e.g., TGN-1412
  • anti-OX40 e
  • the antibody is an agonistic antibody. In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody is an antigen-binding fragment selected from the group consisting of Fab, Fab’, F(ab’)2, Fv, scFv, and other antigen-binding subsequences of the full length antibody. In some embodiments, the antibody is a human, humanized, or chimeric antibody. In some embodiments, the antibody is a bispecific antibody, a multispecific antibody, a single domain antibody, a fusion protein comprising an antibody portion, or any other functional variants or derivatives thereof.
  • the immune-stimulating agent is an activator of CD40.
  • the activator of CD40 is an agonistic anti-CD40 antibody.
  • Any of the known anti-CD40 antibodies may be used in the present invention, including, but not limited to, CP-870,893, Dacetuzumab (also known as SGN-40), ChiLob 7/4, APX005, and APX005M, BI-655064, and BMS-986090.
  • the agonistic anti-CD40 antibody is a monoclonal antibody or a polyclonal antibody.
  • the agonistic anti-CD40 antibody is an antigen-binding fragment selected from the group consisting of Fab, Fab’, F(ab’)2, Fv, scFv, and other antigen-binding subsequences of the full- length anti-CD40 antibody.
  • the agonistic anti-CD40 antibody is a human, humanized, or chimeric antibody.
  • the agonistic anti-CD40 antibody is a bispecific antibody, a multispecific antibody, a single domain antibody, a fusion protein comprising an antibody portion, or any other variants or derivatives thereof.
  • the activator of CD40 is a natural or engineered CD40 ligand, such as CD40L.
  • the activator of CD40 is an inhibitor of the interaction between CD40 and CD40L. In some embodiments, the activator of CD40 increases the signaling of CD40.
  • the immune-stimulating agent is an activator of 0X40. In some embodiments, the activator of 0X40 is an agonistic anti-OX40 antibody. Any of the known anti- 0X40 antibodies may be used in the present invention, including, but not limited to, MEDI6469, MEDI0562, MEDI6383, GSK3174998, KHK4083 and InVivoMM) clone OX-86.
  • the agonistic anti-OX40 antibody is a monoclonal antibody or a polyclonal antibody. In some embodiments, the agonistic anti-OX40 antibody is an antigenbinding fragment selected from the group consisting of Fab, Fab’, F(ab’)2, Fv, scFv, and other antigen-binding subsequences of the full-length anti-OX40 antibody. In some embodiments, the agonistic anti-OX40 antibody is a human, humanized, or chimeric antibody.
  • the agonistic anti-OX40 antibody is a bispecific antibody, a multispecific antibody, a single domain antibody, a fusion protein comprising an antibody portion, or any other variants or derivatives thereof.
  • the activator of 0X40 is a natural or engineered 0X40 ligand, such as OX40L.
  • the activator of 0X40 is an inhibitor of the interaction between 0X40 and OX40L. Any of the inhibitors of interaction between 0X40 and OX40L may be used in the present invention, see, for example, U.S Patent No. US8283450, US11867621, US7547438, US7063845,% US7537763 and US5801227.
  • the activator of 0X40 increases the signaling of 0X40.
  • the methods described herein comprise administering a targeted therapy agent.
  • Targeted therapy agents are a class of drugs specifically designed to interfere with molecular targets that drive cancer progression. Unlike traditional chemotherapy, which broadly attacks rapidly dividing cells, targeted therapies aim to disrupt cancer-specific pathways, offering improved precision, reduced toxicity, and often enhanced efficacy. These agents work by modulating distinct cancer hallmarks — biological capabilities acquired during tumor development — through inhibition or modulation of key signaling pathways, receptors, or genetic mutations (Wilkes. Targeted Therapy: Attacking Cancer with Molecular and Immunological Targeted Agents. Asia Pac J Oncol Nurs. 2018 Apr- Jun;5(2): 137-155).
  • the methods described herein can comprise administering any targeted therapy agent known in the art or described herein.
  • Table 2 summarizes examples of targeted therapy agents based on the cancer mechanism that they target. Any one or any combination of the targeted therapy agents in Table 2 may be used in the methods described herein, using any administration routes, and/or dosage, and/or dosing frequency, and/or duration, and/or maintenance schedule described herein.
  • the targeted therapy agent is a tyrosine kinase inhibitor.
  • the tyrosine kinase inhibitor targets a BCR-ABL fusion protein.
  • Tyrosine kinase inhibitors that target a BCR-ABL fusion protein include, for example, Imatinib (Gleevec®), also known as imatinib mesylate.
  • the tyrosine kinase inhibitor targets EGFR.
  • Tyrosine kinase inhibitors that target EGFR include, for example, Erlotinib (Tarceva®), Gefitinib (Iressa®), Afatinib (Gilotrif®), Osimertinib (Tagrisso®), and Lapatinib (Tykerb®).
  • the tyrosine kinase inhibitor targets HER2.
  • Tyrosine kinase inhibitors that target HER2 include, for example, Lapatinib (Tykerb®).
  • the tyrosine kinase inhibitor targets VEGFR.
  • Tyrosine kinase inhibitors that target VEGFR include, for example, Sunitinib (Sutent®), Sorafenib (Nexavar®), Pazopanib (Votrient®), and Axitinib (Inlyta®).
  • the tyrosine kinase inhibitor targets ALK and/or ROS 1.
  • Tyrosine kinase inhibitors that target ALK and/or ROS 1 include, for example, Crizotinib (Xalkori®), Ceritinib (Zykadia®), and Alectinib (Alecensa®).
  • the tyrosine kinase inhibitor targets TRK and/or ROS 1.
  • Tyrosine kinase inhibitors that target TRK and/or ROS 1 include, for example, Larotrectinib (Vitrakvi®) and Entrectinib (Rozlytrek®).
  • the targeted therapy agent is a monoclonal antibody.
  • the monoclonal antibody is an anti-HER2 antibody, such as Trastuzumab (Herceptin®) or Pertuzumab (Peg eta®).
  • the monoclonal antibody is an anti-EGFR antibody, such as Cetuximab (Erbitux®) or Panitumumab (Vectibix®).
  • the monoclonal antibody is an anti-VEGF antibody, such as Bevacizumab (A vastin®).
  • the monoclonal antibody is an anti-CD38 antibody, such as Daratumumab (Darzalex®).
  • the monoclonal antibody is an anti- CD20 antibody, such as Rituximab (Rituxan®) or Obinutuzumab (Gazyva®).
  • the targeted therapy agent is an mTOR and PI3K/AKT pathway inhibitor.
  • the mTOR and PI3K/AKT pathway inhibitor targets mTOR.
  • mTOR and PI3K/AKT pathway inhibitors that target mTOR include, for example, Everolimus (Afmitor®) and Temsirolimus (Torisel®).
  • the mTOR and PI3K/AKT pathway inhibitor targets PI3Ka.
  • mTOR and PI3K/AKT pathway inhibitors that target PI3Ka include, for example, Alpelisib (Piqray®).
  • the targeted therapy agent is a PARP inhibitor.
  • PARP inhibitors include, for example, Olaparib (Lynparza®), Rucaparib (Rubraca®), Niraparib (Zejula®), and Talazoparib (Talzenna®).
  • the targeted therapy agent comprises a BRAF inhibitor and/or a MEK inhibitor.
  • the targeted therapy agent comprises a BRAF inhibitor, such as an agent that targets a BRAF V600 mutant (e.g., V600E).
  • BRAF inhibitors include, for example, Vemurafenib (Zelboraf®), Dabrafenib (Tafmlar®), and Encorafenib (Braftovi®).
  • the targeted therapy agent comprises a MEK inhibitor.
  • MEK inhibitors include, for example, Trametinib (Mekinist®), Cobimetinib (Cotellic®), and Binimetinib (Mektovi®).
  • the targeted therapy agent comprises a BRAF inhibitor and MEK inhibitor.
  • the targeted therapy agent comprises Encorafenib (Braftovi®) and Binimetinib (Mektovi®).
  • the targeted therapy agent targets a hormonal or endocrine target.
  • the targeted therapy agent is an estrogen receptor modulator, such as tamoxifen.
  • the targeted therapy agent is an aromatase inhibitor, such as letrozole, anastrozole, or exemestane.
  • the targeted therapy agent is a CYP17 inhibitor, such as Abiraterone (Zytiga®).
  • the targeted therapy agent is an androgen receptor antagonist, such as Enzalutamide (Xtandi®).
  • the targeted therapy agent is an agent that targets mutated isocitrate dehydrogenase- 1 (IDH1), such as Ivosidenib (Tibsovo®).
  • the targeted therapy agent is an agent that targets mutated isocitrate dehydrogenase- 1 (IDH1), such as Enasidenib (Idhifa®).
  • the targeted therapy agent is an agent that targets FGFR (e.g, FGFR-mutated urothelial carcinoma), such as Erdafitinib (Balversa®).
  • the targeted therapy agent is an agent that targets HERZ, such as Tucatinib (Tukysa®).
  • the targeted therapy agent is an agent that targets RET, such as Selpercatinib (Retevmo®).
  • the individual has been subject to tumor site preparation prior to administration of the oncolytic virus, using one or more (such as 1, 2, 3, 4, 5, or more) treatment modalities, including, but are not limited to radiation therapy, administration of one or more immune-related molecules, administration of other therapeutic agents, and combinations thereof. It is believed that adding other pre-treatment preparations can increase the chance of success for the methods described above. Without being bound by any theory or hypothesis, for example, local radiation, with or without lymphodepletion effects, or chemotherapy, may increase the chance of the transfection of the oncolytic virus, and may deplete the more sensitive Treg cells at the tumor sites, thereby reviving the exhausted or telorized T memory cells.
  • one or more treatment modalities including, but are not limited to radiation therapy, administration of one or more immune-related molecules, administration of other therapeutic agents, and combinations thereof.
  • tumor site preparations prior to or in concomitant with the administration of the invention combination at the tumor site can involve cytokines, chemokines, small molecules and other well-known beneficial immunomodulators, such as IL2, IL12, 0X40, CD40 and 4-1BB agonist.
  • cytokines such as IL2, IL12, 0X40, CD40 and 4-1BB agonist.
  • a method of treating bladder cancer in an individual comprising intravesically administering to the individual a two-step instillation regimen one or more times, wherein each administration of the two-step instillation regimen comprises: a) intravesically administering a pretreatment composition comprising a transduction-enhancing agent to the individual; and subsequently b) intravesically administering an oncolytic virus to the individual directly after administering the pretreatment composition, wherein there is no intervening bladder wash between the administration of the pretreatment composition and the administration of the oncolytic virus, wherein the oncolytic virus comprises a viral vector comprising a tumor cell-specific promoter operably linked to a viral gene essential for replication of the oncolytic virus, and a heterologous gene encoding an immune-related molecule.
  • each administration of the two-step instillation regiment consists of a) intravesically administering a pretreatment composition comprising a transduction-enhancing agent to the individual; and subsequently b) intravesically administering an oncolytic virus to the individual directly after administering the pretreatment composition, wherein there is no intervening bladder wash between the administration of the pretreatment composition and the administration of the oncolytic virus.
  • the oncolytic virus is an oncolytic adenovirus.
  • the oncolytic virus is an adenovirus serotype 5, wherein the endogenous Ela promoter of a native adenovirus serotype 5 is replaced by the human E2F-1 promoter, and the endogenous E3 19kD coding region of the native adenovirus serotype 5 is replaced by a nucleic acid encoding human GM-CSF.
  • the oncolytic virus is cretostimogene.
  • the oncolytic virus is administered at a dose of about 1 x 10 8 to about 1 x 10 14 viral particles (e.g., at a dose of about 1 x 10 12 viral particles).
  • the oncolytic virus is administered in a volume of between about 50 mb and about 100 mb (e.g., in a volume of between about 85 mb and about 100 mb). In some embodiments, the administration of the oncolytic virus comprises allowing the oncolytic virus to dwell in the bladder for between about 55 minutes and about 65 minutes (e.g., for about 60 minutes) after completion of the administration of the full volume of the oncolytic virus into the bladder.
  • the transduction-enhancing agent is N-Dodecyl-P-D-maltoside (DDM).
  • the pretreatment composition comprises DDM at a concentration of about 0.01% to about 0.5% (e.g., about 0.05% to about 0.2%, or about 0.1%). In some embodiments, the pretreatment composition is administered at a volume of between about 50 mb and about 100 mb (e.g., between about 85 mb and about 100 mb). In some embodiments, the administration of the pretreatment composition comprises allowing the pretreatment composition to dwell in the bladder for more than 5 minutes (e.g., for between about 10 minutes and about 20 minutes, or for about 15 minutes) after completion of the administration of the full volume of the pretreatment composition into the bladder. In some embodiments, the bladder cancer is non-muscle invasive bladder cancer. In certain embodiments, the individual has a Ta or T1 papillary carcinoma without concurrent carcinoma in situ. In certain embodiments, the individual has carcinoma in situ, with or without concurrent Ta or T1 papillary carcinoma. In some embodiments, the individual is BCG-unresponsive.
  • a method of treating bladder cancer in an individual comprising intravesically administering to the individual a two-step instillation regimen one or more times, wherein each administration of the two-step instillation regimen comprises: a) intravesically administering a pretreatment composition comprising N- Dodecyl-P-D-maltoside (DDM) to the individual; and subsequently b) intravesically administering an oncolytic adenovirus to the individual directly after administering the pretreatment composition, wherein there is no intervening bladder wash between the administration of the pretreatment composition and the administration of the oncolytic adenovirus.
  • DDM N- Dodecyl-P-D-maltoside
  • the oncolytic adenovirus is an adenovirus serotype 5, wherein the endogenous Ela promoter of a native adenovirus serotype 5 is replaced by the human E2F-1 promoter, and the endogenous E3 19kD coding region of the native adenovirus serotype 5 is replaced by a nucleic acid encoding human GM-CSF.
  • the oncolytic adenovirus is cretostimogene.
  • each administration of the two-step instillation regiment consists of a) intravesically administering the pretreatment composition comprising DDM to the individual; and subsequently b) intravesically administering the oncolytic adenovirus to the individual directly after administering the pretreatment composition, wherein there is no intervening bladder wash between the administration of the pretreatment composition and the administration of the oncolytic adenovirus.
  • the oncolytic adenovirus is administered at a dose of about 1 x 10 8 to about 1 x 10 14 viral particles (e.g., at a dose of about 1 x 10 12 viral particles).
  • the oncolytic adenovirus is administered in a volume of between about 50 mb and about 100 mb (e.g., in a volume of between about 85 mb and about 100 mb).
  • the administration of the oncolytic adenovirus comprises allowing the oncolytic adenovirus to dwell in the bladder for between about 55 minutes and about 65 minutes (e.g., for about 60 minutes) after completion of the administration of the full volume of the oncolytic adenovirus into the bladder.
  • the pretreatment composition comprises DDM at a concentration of about 0.01% to about 0.5% (e.g., about 0.05% to about 0.2%, or about 0.1%).
  • the pretreatment composition is administered at a volume of between about 50 mb and about 100 mb (e.g., between about 85 mb and about 100 mb). In some embodiments, the administration of the pretreatment composition comprises allowing the pretreatment composition to dwell in the bladder for more than 5 minutes (e.g., for between about 10 minutes and about 20 minutes, or for about 15 minutes) after completion of the administration of the full volume of the pretreatment composition into the bladder.
  • the bladder cancer is non-muscle invasive bladder cancer.
  • the individual has a Ta or T1 papillary carcinoma without concurrent carcinoma in situ. In certain embodiments, the individual has carcinoma in situ, with or without concurrent Ta or T1 papillary carcinoma. In some embodiments, the individual is BCG-unresponsive.
  • a method of treating bladder cancer in an individual comprising intravesically administering to the individual a two-step instillation regimen one or more times, wherein each administration of the two-step instillation regimen comprises: a) intravesically administering a pretreatment composition comprising N- Dodecyl-P-D-maltoside (DDM) to the individual; and subsequently b) intravesically administering an oncolytic adenovirus to the individual directly after administering the pretreatment composition, wherein there is no intervening bladder wash between the administration of the pretreatment composition and the administration of the oncolytic adenovirus, and wherein the oncolytic adenovirus is an adenovirus serotype 5, wherein the endogenous Ela promoter of a native adenovirus serotype 5 is replaced by the human E2F-1 promoter, and the endogenous E3 19kD coding region of the native adenovirus serotype 5 is replaced by a nucleic
  • the oncolytic adenovirus is cretostimogene.
  • each administration of the two-step instillation regiment consists of a) intravesically administering the pretreatment composition comprising DDM to the individual; and subsequently b) intravesically administering the oncolytic adenovirus to the individual directly after administering the pretreatment composition, wherein there is no intervening bladder wash between the administration of the pretreatment composition and the administration of the oncolytic adenovirus.
  • the oncolytic adenovirus is administered at a dose of about 1 x 10 8 to about 1 x 10 14 viral particles (e.g., at a dose of about 1 x 10 12 viral particles).
  • the oncolytic adenovirus is administered in a volume of between about 50 mb and about 100 mb (e.g., in a volume of between about 85 mb and about 100 mb).
  • the administration of the oncolytic adenovirus comprises allowing the oncolytic adenovirus to dwell in the bladder for between about 55 minutes and about 65 minutes (e.g., for about 60 minutes) after completion of the administration of the full volume of the oncolytic adenovirus into the bladder.
  • the pretreatment composition comprises DDM at a concentration of about 0.01% to about 0.5% (e.g., about 0.05% to about 0.2%, or about 0.1%).
  • the pretreatment composition is administered at a volume of between about 50 mb and about 100 mb (e.g., between about 85 mb and about 100 mb). In some embodiments, the administration of the pretreatment composition comprises allowing the pretreatment composition to dwell in the bladder for more than 5 minutes (e.g., for between about 10 minutes and about 20 minutes, or for about 15 minutes) after completion of the administration of the full volume of the pretreatment composition into the bladder.
  • the bladder cancer is non-muscle invasive bladder cancer.
  • the individual has a Ta or T1 papillary carcinoma without concurrent carcinoma in situ. In certain embodiments, the individual has carcinoma in situ, with or without concurrent Ta or T1 papillary carcinoma. In some embodiments, the individual is BCG-unresponsive.
  • a method of treating bladder cancer in an individual comprising intravesically administering to the individual a two-step instillation regimen one or more times, wherein each administration of the two-step instillation regimen comprises: a) intravesically administering a pretreatment composition comprising N- Dodecyl-P-D-maltoside (DDM) to the individual; and subsequently b) intravesically administering cretostimogene to the individual directly after administering the pretreatment composition, wherein there is no intervening bladder wash between the administration of the pretreatment composition and the administration of the cretostimogene.
  • DDM N- Dodecyl-P-D-maltoside
  • each administration of the two-step instillation regiment consists of a) intravesically administering the pretreatment composition comprising DDM to the individual; and subsequently b) intravesically administering cretostimogene to the individual directly after administering the pretreatment composition, wherein there is no intervening bladder wash between the administration of the pretreatment composition and the administration of the cretostimogene.
  • the cretostimogene is administered at a dose of about 1 x 10 8 to about 1 x 10 14 viral particles (e.g., at a dose of about 1 x 10 12 viral particles).
  • the cretostimogene is administered in a volume of between about 50 mL and about 100 mL (e.g., in a volume of between about 85 mb and about 100 mb).
  • the administration of the cretostimogene comprises allowing the cretostimogene to dwell in the bladder for between about 55 minutes and about 65 minutes (e.g., for about 60 minutes) after completion of the administration of the full volume of the cretostimogene into the bladder.
  • the pretreatment composition comprises DDM at a concentration of about 0.01% to about 0.5% (e.g., about 0.05% to about 0.2%, or about 0.1%).
  • the pretreatment composition is administered at a volume of between about 50 mL and about 100 mL (e.g., between about 85 mL and about 100 mL). In some embodiments, the administration of the pretreatment composition comprises allowing the pretreatment composition to dwell in the bladder for more than 5 minutes (e.g., for between about 10 minutes and about 20 minutes, or for about 15 minutes) after completion of the administration of the full volume of the pretreatment composition into the bladder.
  • the bladder cancer is non-muscle invasive bladder cancer.
  • the individual has a Ta or T1 papillary carcinoma without concurrent carcinoma in situ. In certain embodiments, the individual has carcinoma in situ, with or without concurrent Ta or T1 papillary carcinoma. In some embodiments, the individual is BCG-unresponsive.
  • a method of treating non-muscle invasive bladder cancer (NMIBC) in an individual comprising intravesically administering to the individual a two-step instillation regimen one or more times, wherein each administration of the two-step instillation regimen comprises: a) intravesically administering a pretreatment composition comprising N-Dodecyl-[3-D-maltoside (DDM) to the individual; and subsequently b) intravesically administering cretostimogene to the individual directly after administering the pretreatment composition, wherein there is no intervening bladder wash between the administration of the pretreatment composition and the administration of the cretostimogene.
  • NMIBC non-muscle invasive bladder cancer
  • each administration of the two-step instillation regiment consists of a) intravesically administering the pretreatment composition comprising DDM to the individual; and subsequently b) intravesically administering cretostimogene to the individual directly after administering the pretreatment composition, wherein there is no intervening bladder wash between the administration of the pretreatment composition and the administration of the cretostimogene.
  • the cretostimogene is administered at a dose of about 1 x 10 8 to about 1 x 10 14 viral particles (e.g., at a dose of about 1 x 10 12 viral particles).
  • the cretostimogene is administered in a volume of between about 50 mL and about 100 mL (e.g., in a volume of between about 85 mL and about 100 mL).
  • the administration of the cretostimogene comprises allowing the cretostimogene to dwell in the bladder for between about 55 minutes and about 65 minutes (e.g, for about 60 minutes) after completion of the administration of the full volume of the cretostimogene into the bladder.
  • the pretreatment composition comprises DDM at a concentration of about 0.01% to about 0.5% (e.g., about 0.05% to about 0.2%, or about 0. 1%).
  • the pretreatment composition is administered at a volume of between about 50 mL and about 100 mL (e.g., between about 85 mL and about 100 mL). In some embodiments, the administration of the pretreatment composition comprises allowing the pretreatment composition to dwell in the bladder for more than 5 minutes (e.g., for between about 10 minutes and about 20 minutes, or for about 15 minutes) after completion of the administration of the full volume of the pretreatment composition into the bladder.
  • the individual has high-risk NMIBC. In some embodiments, the individual has a Ta or T1 papillary carcinoma without concurrent carcinoma in situ.
  • the individual has high-grade Ta or T1 disease without CIS, wherein the individual has: a) received at least 5 of 6 doses of a BCG induction course and at least 2 doses of one course of either reinduction or maintenance BCG and experienced recurrence of high-grade papillary-only Ta or T1 stage disease within 6 months following the last dose of BCG; or b) received at least 5 of 6 doses of a BCG induction course and experienced high-grade T1 disease at the first evaluation following the BCG induction course.
  • the individual a) has not received prior intravesical Bacillus Calmette -Guerin (BCG) therapy; b) has not received intravesical BCG therapy within 24 months prior to the most recent diagnosis of high-risk NMIBC; or c) has received a maximum of 1 or 2 doses of BCG within 24 months prior to the most recent diagnosis of high-risk NMIBC.
  • BCG Bacillus Calmette -Guerin
  • the individual has: a) received at least 5 of 6 doses of a BCG induction course and at least 2 doses of 1 course of either reinduction or maintenance BCG and experienced persistent or recurrent carcinoma in situ (CIS), with or without concomitant Ta or T1 disease, within 12 months following the last dose of BCG; b) received at least 5 of 6 doses of a BCG induction course and at least 2 doses of 1 course of either reinduction or maintenance BCG and experienced recurrence of highgrade papillary-only Ta or T1 stage disease within 6 months following the last dose of BCG; or c) received at least 5 of 6 doses of a BCG induction course and experienced high-grade T1 disease at the first evaluation following the BCG induction course.
  • CIS persistent or recurrent carcinoma in situ
  • the individual has: a) received at least 5 of 6 doses of a BCG induction course and experienced persistent or recurrent high-grade papillary carcinoma of Ta stage or carcinoma in situ (CIS) at a first evaluation following the BCG induction course; b) received at least 5 of 6 doses of a BCG induction course and at least 2 doses of 1 course of either reinduction or maintenance BCG, achieved an initial complete response, and subsequently experienced recurrence of a high-grade Ta or T1 disease more than 6 months after and within 24 months after the last dose of BCG; c) received at least 5 of 6 doses of a BCG induction course and at least 2 doses of 1 course of either reinduction or maintenance BCG, achieved an initial complete response, and subsequently experienced recurrence of CIS more than 12 months after and within 24 months after the last dose of BCG; or d) received 3 to 6 doses of a BCG induction course and no BCG reinduction or maintenance course, achieved an initial complete response, and subsequently experienced
  • the individual has intermediate-risk NMIBC.
  • the intermediate-risk NMIBC comprises: a) a low-grade tumor of Ta stage, wherein the tumor of Ta stage is recurrent within about 12 months of resection of a prior low-grade tumor of Ta stage or solitary high-grade tumor of Ta stage having a diameter of 3 cm or smaller; b) a solitary low-grade tumor of Ta stage having a diameter of greater than 3 cm; c) two or more low-grade tumors of Ta stage; d) a primary and solitary high-grade tumor of Ta stage having a diameter of less than 3 cm; e) a low-grade tumor of Tl stage; or f) any combination of (a), (c), and (e).
  • a method of treating non-muscle invasive bladder cancer (NMIBC) in an individual having a Ta or Tl papillary carcinoma without concurrent carcinoma in situ comprising intravesically administering to the individual a two-step instillation regimen one or more times, wherein each administration of the two-step instillation regimen comprises: a) intravesically administering a pretreatment composition comprising N-Dodecyl-[3-D-maltoside (DDM) to the individual; and subsequently b) intravesically administering cretostimogene to the individual directly after administering the pretreatment composition, wherein there is no intervening bladder wash between the administration of the pretreatment composition and the administration of the cretostimogene.
  • NMIBC non-muscle invasive bladder cancer
  • each administration of the two-step instillation regiment consists of a) intravesically administering the pretreatment composition comprising DDM to the individual; and subsequently b) intravesically administering cretostimogene to the individual directly after administering the pretreatment composition, wherein there is no intervening bladder wash between the administration of the pretreatment composition and the administration of the cretostimogene.
  • the cretostimogene is administered at a dose of about 1 x 10 8 to about 1 x 10 14 viral particles (e.g., at a dose of about 1 x 10 12 viral particles).
  • the cretostimogene is administered in a volume of between about 50 mb and about 100 mb (e.g., in a volume of between about 85 mb and about 100 mb).
  • the administration of the cretostimogene comprises allowing the cretostimogene to dwell in the bladder for between about 55 minutes and about 65 minutes (e.g, for about 60 minutes) after completion of the administration of the full volume of the cretostimogene into the bladder.
  • the pretreatment composition comprises DDM at a concentration of about 0.01% to about 0.5% (e.g., about 0.05% to about 0.2%, or about 0. 1%).
  • the pretreatment composition is administered at a volume of between about 50 mb and about 100 mb (e.g., between about 85 mb and about 100 mb). In some embodiments, the administration of the pretreatment composition comprises allowing the pretreatment composition to dwell in the bladder for more than 5 minutes (e.g., for between about 10 minutes and about 20 minutes, or for about 15 minutes) after completion of the administration of the full volume of the pretreatment composition into the bladder.
  • the individual has highgrade Ta or T1 disease without CIS, wherein the individual has: a) received at least 5 of 6 doses of a BCG induction course and at least 2 doses of one course of either reinduction or maintenance BCG and experienced recurrence of high-grade papillary-only Ta or T1 stage disease within 6 months following the last dose of BCG; or b) received at least 5 of 6 doses of a BCG induction course and experienced high-grade T1 disease at the first evaluation following the BCG induction course.
  • a method of treating non-muscle invasive bladder cancer (NMIBC) in an individual having had at least one diagnosis of high-risk NMIBC comprising intravesically administering to the individual a two-step instillation regimen one or more times, wherein each administration of the two-step instillation regimen comprises: a) intravesically administering a pretreatment composition comprising N- Dodecyl-P-D-maltoside (DDM) to the individual; and subsequently b) intravesically administering cretostimogene to the individual directly after administering the pretreatment composition, wherein there is no intervening bladder wash between the administration of the pretreatment composition and the administration of the cretostimogene, wherein the individual i) has not received prior intravesical Bacillus Calmette-Guerin (BCG) therapy; ii) has not received intravesical BCG therapy within 24 months prior to the most recent diagnosis of high-risk NMIBC; or iii) has received a maximum of 1
  • BCG Bacillus Calmette-Guerin
  • each administration of the two-step instillation regiment consists of a) intravesically administering the pretreatment composition comprising DDM to the individual; and subsequently b) intravesically administering cretostimogene to the individual directly after administering the pretreatment composition, wherein there is no intervening bladder wash between the administration of the pretreatment composition and the administration of the cretostimogene.
  • the cretostimogene is administered at a dose of about 1 x 10 8 to about 1 x 10 14 viral particles (e.g., at a dose of about 1 x 10 12 viral particles).
  • the cretostimogene is administered in a volume of between about 50 mL and about 100 mb (e.g., in a volume of between about 85 mL and about 100 mL).
  • the administration of the cretostimogene comprises allowing the cretostimogene to dwell in the bladder for between about 55 minutes and about 65 minutes (e.g., for about 60 minutes) after completion of the administration of the full volume of the cretostimogene into the bladder.
  • the pretreatment composition comprises DDM at a concentration of about 0.01% to about 0.5% (e.g., about 0.05% to about 0.2%, or about 0.1%).
  • the pretreatment composition is administered at a volume of between about 50 mL and about 100 mL (e.g., between about 85 mL and about 100 mL). In some embodiments, the administration of the pretreatment composition comprises allowing the pretreatment composition to dwell in the bladder for more than 5 minutes (e.g., for between about 10 minutes and about 20 minutes, or for about 15 minutes) after completion of the administration of the full volume of the pretreatment composition into the bladder.
  • a method of treating non-muscle invasive bladder cancer (NMIBC) in an individual comprising intravesically administering to the individual a two-step instillation regimen one or more times, wherein each administration of the two-step instillation regimen comprises: a) intravesically administering a pretreatment composition comprising N-Dodecyl-[3-D-maltoside (DDM) to the individual; and subsequently b) intravesically administering cretostimogene to the individual directly after administering the pretreatment composition, wherein there is no intervening bladder wash between the administration of the pretreatment composition and the administration of the cretostimogene, wherein the individual has: i) received at least 5 of 6 doses of a BCG induction course and at least 2 doses of 1 course of either reinduction or maintenance BCG and experienced persistent or recurrent carcinoma in situ (CIS), with or without concomitant Ta or T1 disease, within 12 months following the last dose of BCG; ii) received
  • each administration of the two- step instillation regiment consists of a) intravesically administering the pretreatment composition comprising DDM to the individual; and subsequently b) intravesically administering cretostimogene to the individual directly after administering the pretreatment composition, wherein there is no intervening bladder wash between the administration of the pretreatment composition and the administration of the cretostimogene.
  • the cretostimogene is administered at a dose of about 1 x 10 8 to about 1 x 10 14 viral particles (e.g., at a dose of about 1 x 10 12 viral particles).
  • the cretostimogene is administered in a volume of between about 50 mb and about 100 mb (e.g., in a volume of between about 85 mb and about 100 mb).
  • the administration of the cretostimogene comprises allowing the cretostimogene to dwell in the bladder for between about 55 minutes and about 65 minutes (e.g., for about 60 minutes) after completion of the administration of the full volume of the cretostimogene into the bladder.
  • the pretreatment composition comprises DDM at a concentration of about 0.01% to about 0.5% (e.g., about 0.05% to about 0.2%, or about 0.1%).
  • the pretreatment composition is administered at a volume of between about 50 mb and about 100 mb (e.g., between about 85 mb and about 100 mb). In some embodiments, the administration of the pretreatment composition comprises allowing the pretreatment composition to dwell in the bladder for more than 5 minutes (e.g., for between about 10 minutes and about 20 minutes, or for about 15 minutes) after completion of the administration of the full volume of the pretreatment composition into the bladder.
  • a method of treating non-muscle invasive bladder cancer (NMIBC) in an individual comprising intravesically administering to the individual a two-step instillation regimen one or more times, wherein each administration of the two-step instillation regimen comprises: a) intravesically administering a pretreatment composition comprising N-Dodecyl-[3-D-maltoside (DDM) to the individual; and subsequently b) intravesically administering cretostimogene to the individual directly after administering the pretreatment composition, wherein there is no intervening bladder wash between the administration of the pretreatment composition and the administration of the cretostimogene, wherein the individual the individual has: i) received at least 5 of 6 doses of a BCG induction course and experienced persistent or recurrent high-grade papillary carcinoma of Ta stage or carcinoma in situ (CIS) at a first evaluation following the BCG induction course; ii) received at least 5 of 6 doses of a BCG induction course
  • CIS persistent or recurrent high-grade pa
  • each administration of the two-step instillation regiment consists of a) intravesically administering the pretreatment composition comprising DDM to the individual; and subsequently b) intravesically administering cretostimogene to the individual directly after administering the pretreatment composition, wherein there is no intervening bladder wash between the administration of the pretreatment composition and the administration of the cretostimogene.
  • the cretostimogene is administered at a dose of about 1 x 10 8 to about 1 x 10 14 viral particles (e.g., at a dose of about 1 x 10 12 viral particles).
  • the cretostimogene is administered in a volume of between about 50 mb and about 100 mb (e.g., in a volume of between about 85 mb and about 100 mb).
  • the administration of the cretostimogene comprises allowing the cretostimogene to dwell in the bladder for between about 55 minutes and about 65 minutes (e.g., for about 60 minutes) after completion of the administration of the full volume of the cretostimogene into the bladder.
  • the pretreatment composition comprises DDM at a concentration of about 0.01% to about 0.5% (e.g., about 0.05% to about 0.2%, or about 0.1%).
  • the pretreatment composition is administered at a volume of between about 50 mb and about 100 mb (e.g., between about 85 mb and about 100 mb). In some embodiments, the administration of the pretreatment composition comprises allowing the pretreatment composition to dwell in the bladder for more than 5 minutes (e.g., for between about 10 minutes and about 20 minutes, or for about 15 minutes) after completion of the administration of the full volume of the pretreatment composition into the bladder.
  • a method of treating intermediate-risk non-muscle invasive bladder cancer (NMIBC) in an individual comprising intravesically administering to the individual a two-step instillation regimen one or more times, wherein each administration of the two-step instillation regimen comprises: a) intravesically administering a pretreatment composition comprising N-Dodecyl-P-D-maltoside (DDM) to the individual; and subsequently b) intravesically administering cretostimogene to the individual directly after administering the pretreatment composition, wherein there is no intervening bladder wash between the administration of the pretreatment composition and the administration of the cretostimogene.
  • NMIBC intermediate-risk non-muscle invasive bladder cancer
  • each administration of the two- step instillation regiment consists of a) intravesically administering the pretreatment composition comprising DDM to the individual; and subsequently b) intravesically administering cretostimogene to the individual directly after administering the pretreatment composition, wherein there is no intervening bladder wash between the administration of the pretreatment composition and the administration of the cretostimogene.
  • the cretostimogene is administered at a dose of about 1 x 10 8 to about 1 x 10 14 viral particles (e.g., at a dose of about 1 x 10 12 viral particles).
  • the cretostimogene is administered in a volume of between about 50 mb and about 100 mb (e.g., in a volume of between about 85 mb and about 100 mb).
  • the administration of the cretostimogene comprises allowing the cretostimogene to dwell in the bladder for between about 55 minutes and about 65 minutes (e.g., for about 60 minutes) after completion of the administration of the full volume of the cretostimogene into the bladder.
  • the pretreatment composition comprises DDM at a concentration of about 0.01% to about 0.5% (e.g., about 0.05% to about 0.2%, or about 0.1%).
  • the pretreatment composition is administered at a volume of between about 50 mb and about 100 mb (e.g., between about 85 mb and about 100 mb). In some embodiments, the administration of the pretreatment composition comprises allowing the pretreatment composition to dwell in the bladder for more than 5 minutes (e.g., for between about 10 minutes and about 20 minutes, or for about 15 minutes) after completion of the administration of the full volume of the pretreatment composition into the bladder.
  • the intermediate-risk NMIBC comprises: a) a low-grade tumor of Ta stage, wherein the tumor of Ta stage is recurrent within about 12 months of resection of a prior low- grade tumor of Ta stage or solitary high-grade tumor of Ta stage having a diameter of 3 cm or smaller; b) a solitary low-grade tumor of Ta stage having a diameter of greater than 3 cm; c) two or more low-grade tumors of Ta stage; d) a primary and solitary high-grade tumor of Ta stage having a diameter of less than 3 cm; e) a low-grade tumor of T1 stage; or f) any combination of (a), (c), and (e).
  • the individual has particular stage of bladder cancer.
  • the methods described herein can be used to treat a variety of bladder cancer conditions.
  • the methods described herein can be used to treat a variety of bladder cancer stages.
  • the bladder cancer is muscle invasive (e.g., stage T2, T3 or T4 tumors).
  • the bladder cancer is non-muscle invasive (e.g., stage Ta, Tl, Cis, Cis with Ta and/or T1 tumors, in the absence of stage T2, T3 or T4 tumors).
  • the methods described herein can be used to treat a variety of bladder cancer grades.
  • the grade of a bladder cancer tumor describes the microscopic appearance of cells from the tumor relative to normal bladder cells, particularly with respect to cell division.
  • the bladder cancer is graded using a numbered grading system.
  • the bladder cancer comprises a bladder tumor of grade 1, in which the tumor cells appear mostly well-differentiated, are slow-growing and unlikely to spread.
  • the bladder cancer comprises a bladder tumor of grade 2, in which the tumor cells appear somewhat differentiated and are growing faster than grade 1 cells.
  • the bladder cancer comprises a bladder tumor of grade 3, in which the tumor cells appear mostly undifferentiated and are growing faster than grade 2 cells.
  • the bladder cancer is graded using a binary high or low grading system.
  • the bladder cancer is a low-grade bladder cancer, in which the cancer cells are slow-growing and are less likely to spread. In some embodiments, the bladder cancer is a high-grade bladder cancer, the cancer cells grow more quickly and are more likely to spread. Bladder cancer comprising CIS is classified as high-grade.
  • the individual has carcinoma in situ. In some embodiments, the individual has carcinoma in situ without a concurrent Ta or Tl stage tumor. In some embodiments, the individual has carcinoma in situ and at least one concurrent Ta or Tl stage tumor. In some embodiments, the individual has a Ta or Tl stage tumor. In some embodiments, the individual has a Ta or Tl stage tumor without concurrent carcinoma in situ. In some embodiments, the individual has a Ta or Tl stage tumor and concurrent carcinoma in situ.
  • the individual has nonmuscle invasive bladder cancer (NMIBC).
  • NMIBC nonmuscle invasive bladder cancer
  • the individual has high-risk NMIBC.
  • the individual has intermediate-risk NMIBC.
  • the individual has low-risk NMIBC.
  • the individual has high-risk, intermediate-risk, or low-risk NMIBC as determined according to the TNM staging system by the American Joint Committee on Cancer (AJCC) guidelines.
  • the individual has high-risk NMIBC comprising a) CIS; b) two or more grade 2 stage T1 tumors; c) one or more grade 3 stage Ta and/or T1 tumors; or d) any combination of (a), (b), and (c).
  • the individual has high-risk, intermediate-risk, or low-risk NMIBC as determined according to American Urological Association (AU A) guidelines for NMIBC (https://www.auanet.org/guidelines-and-quality/guidelines/bladder- cancer-non-muscle-invasive-guideline).
  • AU A American Urological Association
  • the individual has high-risk NMIBC comprising a) one or more high-grade T1 tumors; b) any recurrent high-grade Ta tumors; c) a high-grade Ta tumor greater than 3 cm or multi-focal high-grade Ta tumors; d) any carcinoma in situ; e) one or more high-grade NMIBC tumors wherein the individual has failed BCG treatment; f) any variant histology; g) any lymphovascular invasion; and/or h) any high-grade prostatic urethral involvement.
  • the bladder cancer is transitional cell carcinoma or urothelial carcinoma (such as metastatic urothelial carcinoma), including, but not limited to, papillary tumors and flat carcinomas.
  • the bladder cancer is metastatic urothelial carcinoma.
  • the bladder cancer is urothelial carcinoma of the bladder.
  • the bladder cancer is urothelial carcinoma of the ureter.
  • the bladder cancer is urothelial carcinoma of the urethra.
  • the bladder cancer is urothelial carcinoma of the renal pelvis.
  • the bladder cancer is squamous cell carcinoma. In some embodiments, the bladder cancer is non-squamous cell carcinoma. In some embodiments, the bladder cancer is adenocarcinoma. In some embodiments, the bladder cancer is small cell carcinoma.
  • the bladder cancer is early stage bladder cancer, non- metastatic bladder cancer, non-invasive bladder cancer, non-muscle-invasive bladder cancer, primary bladder cancer, advanced bladder cancer, locally advanced bladder cancer (such as unresectable locally advanced bladder cancer), metastatic bladder cancer, or bladder cancer in remission.
  • the bladder cancer is localized resectable, localized unresectable, or unresectable.
  • the bladder cancer is a high grade, non- muscle-invasive cancer that has been refractory to standard intra-bladder infusion (intravesical) therapy.
  • the methods provided herein can be used to treat an individual (e.g., human) who has been diagnosed with or is suspected of having bladder cancer.
  • the individual has undergone a tumor resection.
  • the individual has refused surgery.
  • the individual is medically inoperable.
  • the individual is at a clinical stage of Ta, Tis, Tl, T2, T3a, T3b, or T4 bladder cancer.
  • the individual is at a clinical stage of carcinoma in situ (CIS, also known as Tis), Ta, or Tl.
  • CIS carcinoma in situ
  • the individual has been previously treated for bladder cancer (also referred to as the “prior therapy”).
  • individual has been previously treated with a standard therapy for bladder cancer.
  • the prior standard therapy is treatment with intravesical Bacillus Calmette-Guerin (BCG).
  • BCG Bacillus Calmette-Guerin
  • the prior standard therapy is treatment with mitomycin C.
  • the prior standard therapy is treatment with a chemotherapeutic agent, such as a chemotherapeutic agent described herein.
  • the prior standard therapy is treatment with mitomycin C, gemcitabine, a platinum-based agent (e.g., cisplatin), or another chemotherapeutic agent.
  • the individual has bladder cancer in remission, progressive bladder cancer, or recurrent bladder cancer. In some embodiments, the individual is resistant to treatment of bladder cancer with the prior therapy. In some embodiments, the individual is initially responsive to treatment of bladder cancer with the prior therapy but has progressed after treatment. In some embodiments, the individual has received a prior therapy prior to the administration of the oncolytic virus.
  • the individual has recurrent bladder cancer (such as a bladder cancer at the clinical stage of Ta, Tis, Tl, T2, T3a, T3b, or T4) after a prior therapy (such as prior standard therapy, for example treatment with BCG).
  • a prior therapy such as prior standard therapy, for example treatment with BCG.
  • the individual may be initially responsive to the treatment with the prior therapy, but develops bladder cancer after about any of about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 24, 36, 48, or 60 months upon the cessation of the prior therapy.
  • the prior therapy is radiation therapy (e.g., with or without chemotherapy). In some embodiments, the radiation therapy is in combination with chemotherapy. In some embodiments, the prior therapy is radiation therapy to the whole body. In some embodiments, the prior therapy is radiation therapy to only tumor sites. In some embodiments, the prior therapy is radiation therapy to tissues having the tumor. In some embodiments, the prior therapy is radiation therapy to only the site of the tumor selected for local administration of the oncolytic virus. In some embodiments, the prior therapy is radiation therapy to only a tissue having the tumor selected for local administration of the oncolytic virus. In some embodiments, the dose of the radiation therapy is insufficient to treat the tumor cells.
  • a suitable dosage of the radiation therapy is about any one of 1 Gy, 5 Gy, 10 Gy, 15 Gy, 20 Gy, 25 Gy, 30 Gy, 35 Gy, 40 Gy, 45 Gy, 50 Gy, 55 Gy, 60 Gy, 65 Gy, 70 Gy, 75 Gy, 80 Gy, 90 Gy or 100 Gy.
  • the dose of the radiation therapy is no more than about any one of 1 Gy, 5 Gy, 10 Gy, 15 Gy, 20 Gy, 25 Gy, 30 Gy, 35 Gy, 40 Gy, 45 Gy, 50 Gy, 55 Gy, 60 Gy, 65 Gy, 70 Gy, 75 Gy, 80 Gy, 90 Gy or 100 Gy.
  • the dose of the radiation therapy is any one of about 1 Gy to about 5 Gy, about 5 Gy to about 10 Gy, about 10 Gy to about 15 Gy, about 15 Gy to about 20 Gy, about 20 Gy to about 25 Gy, about 25 Gy to about 30 Gy, about 30 Gy to about 35 Gy, about 5 Gy to about 15 Gy, about 10 Gy to about 20 Gy, about 20 Gy to about 30 Gy, about 30 Gy to about 40 Gy, about 40 Gy to about 50 Gy, about 50 Gy to about 60 Gy, about 60 Gy to about 70 Gy, about 70 Gy to about 80 Gy, about 80 Gy to about 100 Gy, about 10 Gy to about 30 Gy, about 20 Gy to about 40 Gy, about IGy to about 25 Gy, about 25 Gy to about 50 Gy, about 30 Gy to about 60 Gy, about 60 Gy to about 80 Gy, or about 10 Gy to about 60 Gy.
  • the suitable dosage of the radiation therapy may also depend on the type, stage and location of the tumor.
  • the radiation therapy is administered in more than one fraction, such as about any one of 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 15, 16, 18, 20 or more fractions.
  • the radiation therapy fractions are administered over the course of about any one of 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks or more.
  • the radiation therapy fractions are administered over the course of any one of about 1 day to about 5 days, about 1 week to about 2 weeks, about 2 weeks to about 3 weeks, about 3 weeks to about 4 weeks, about 4 weeks to about 5 weeks, about 5 weeks to about 6 weeks, about 6 weeks to about 7 weeks, about 2 weeks to about 4 weeks, about 4 weeks to about 6 weeks, or about 1 week to about 6 weeks.
  • the radiation therapy is administered about two fractions per day.
  • each fraction of the radiation therapy is about 1.8 Gy to about 2 Gy per day, five days a week, for an adult, or about 1.5 Gy to about 1.8 Gy per day, five days a week for a child.
  • each fraction of the radiation therapy is about any one of 1 Gy, 1.5 Gy, 2 Gy, 2.5 Gy, 5 Gy, 10 Gy, 15 Gy, 20 Gy, 30 Gy, 40 Gy, 50 Gy or more. In some embodiments, each fraction of the radiation therapy is any one of about 1 Gy to about 1.5 Gy, about 1.5 Gy to about 2 Gy, about 1 Gy to about 2.5 Gy, about 2.5 Gy to about 5 Gy, about 5 Gy to about 10 Gy, about 10 Gy to about 15 Gy, about 15 Gy to about 20 Gy, about 20 Gy to about 30 Gy, about 25 Gy to about 50 Gy, about 1 Gy to about 10 Gy, or about 2 Gy to about 20 Gy. In some embodiments, the radiation therapy is administered in a single fraction.
  • any of the known methods of radiation therapy may be used in the present invention, including, but not limited to external beam radiation therapy (EBRT or XRT), tele therapy, brachytherapy, sealed source radiation therapy, systemic radioisotope therapy (RIT), unsealed source radiation therapy, intraoperative radiation therapy (IORT), targeted intraoperative radiation therapy (TARGIT), intensity-modulated radiation therapy (IMRT), volumetric modulated arc therapy (VMAT), particle therapy, and auger therapy.
  • EBRT or XRT external beam radiation therapy
  • tele therapy tele therapy
  • brachytherapy sealed source radiation therapy
  • RIT systemic radioisotope therapy
  • unsealed source radiation therapy unsealed source radiation therapy
  • intraoperative radiation therapy IORT
  • targeted intraoperative radiation therapy targeted intraoperative radiation therapy
  • IMRT intensity-modulated radiation therapy
  • VMAT volumetric modulated arc therapy
  • particle therapy and auger therapy.
  • the prior therapy comprises administration of a therapeutic agent.
  • the dosage of the therapeutic agent is sufficient to treat the tumor.
  • the dosage of the therapeutic agent is insufficient to treat the tumor.
  • the therapeutic agent is any one or combination of chemotherapeutic agents known in the art, for example, cyclophosphamide.
  • the therapeutic agent is any one or combination of agents targeting or blocking a cellular signaling pathway known in the art, for example, a BRAF inhibitor.
  • the therapeutic agent is any one or combination of cell therapies known in the art, for example, TIL cells, CAR/T cells, and/or TCR/T cells.
  • the therapeutic agent is an agent that increases the level of cytokines involved an immunogenic pathway.
  • cytokines such as IL6, IL8 and IL 18 (these cytokines can either have pro and/or anti-inflammatory actions, or some may promote new blood vessels formation and tumor growth), chemokines (such as CCL21 that can promote tumor spread by increase of lymphatic structures), growth factors (such as FLT3L), heat shock proteins, small molecule kinase inhibitors (such as JAK2 inhibitor), IAP inhibitors, STING activators (such as CDN), PRRago (such as CpG ODN (oligodeoxynucleotides), Imiquimod, or Poly I:C), TLR stimulators (such as GS-9620, AED- 1419, CYT-003-QbG10, AVE-0675, or PF-7909), and RLR stimulators (such as RIG
  • the therapeutic agent is an agent that causes dysfunction or damage to a structural component of a tumor.
  • agents include, but are not limited to, anti-VEGF antibody, a hyaluronidase, and n-dodecyl-P- maltoside.
  • the therapeutic agent induces immune cells, such as dendritic cells, B cells, and T cells (such as follicular T helper cells).
  • immune cells such as dendritic cells, B cells, and T cells (such as follicular T helper cells).
  • Any of the therapeutic agent/s described herein, e.g. chemotherapeutic agents, agents targeting or blocking cell signaling pathways, cytokines, chemokines, cell therapies, etc. can be administered locally (e.g., intravesically) or systemically (e.g. through intravenous administration) to the tumor site, either singly or in combination.
  • the prior standard therapy is treatment with intravesical Bacillus Calmette-Guerin (BCG).
  • BCG Bacillus Calmette-Guerin
  • the individual has been previously treated intravesically with BCG therapy.
  • the individual was responsive to BCG therapy (e.g, experienced complete response (CR) after BCG therapy).
  • the individual experienced reduced bladder cancer symptoms, reduced size or number of bladder tumors, or an absence of bladder tumors after intravesical BCG therapy.
  • the individual was unresponsive to BCG therapy (e.g., had persistent bladder cancer after BCG therapy).
  • the individual was previously treated with intravesical BCG therapy and was unresponsive to BCG therapy.
  • the individual has previously received a BCG induction course and had persistent or recurrent bladder cancer subsequent to the BCG induction course.
  • the individual has previously received a BCG induction course and at least one BCG maintenance course and had persistent or recurrent bladder cancer subsequent to the BCG maintenance course.
  • the BCG induction course comprises weekly intravesical instillation of BCG once per week for six weeks.
  • the individual received at least 5 of the 6 doses of the BCG induction course.
  • the individual has previously received a BCG induction course beginning at month 0 of a treatment regimen and had persistent bladder cancer at month 3 of the treatment regimen.
  • the BCG maintenance course comprises weekly intravesical instillation of BCG once per week for three weeks.
  • the individual received at least two of the three weekly intravesical instillations in a standard BCG maintenance course.
  • the individual had a prior diagnosis of high-grade NMIBC (e.g., CIS, high-grade Ta, and/or high-grade Tl) prior to receiving BCG therapy.
  • the individual had persistent or recurrent high-grade NMIBC after receiving BCG therapy.
  • the patient had recurrent CIS with or without concurrent Ta or Tl bladder cancer within 12 months of the final dose of adequate BCG therapy (e.g., at least 5 of 6 doses of a BCG induction course and at least 2 of 3 doses of a BCG maintenance course).
  • the patient had recurrent papillary-only high-grade Ta or Tl bladder cancer within six months of the final dose of adequate BCG therapy (e.g. , at least 5 of 6 doses of a BCG induction course and at least 2 of 3 doses of a BCG maintenance course).
  • the individual has previously received a BCG induction course beginning at month 0 of a treatment regimen and had persistent highgrade T1 bladder cancer at month 3 of the treatment regimen.
  • a standard BCG induction course is 6 weekly intravesical instillations of BCG
  • standard BCG maintenance course is 3 weekly intravesical instillations of BCG.
  • an individual may be treated with a second induction course (z.e., a reinduction course) of BCG.
  • the individual was previously treated with adequate intravesical BCG therapy, responded to BCG therapy, and subsequently experienced a delayed relapse of bladder cancer.
  • the individual has previously received a BCG induction course, experienced an absence of bladder tumors after the BCG induction course, subsequently received at least one BCG maintenance course, and further subsequently experienced a delayed relapse of bladder cancer.
  • the individual has previously received a BCG induction course, experienced an absence of bladder tumors after the BCG induction course, subsequently received at least one BCG maintenance course, and further subsequently experienced recurrence of a papillary carcinoma of Ta or T1 (e.g., one or more high grade Ta or T1 stage tumors) stage at least 6 months (e.g., at least 6, 9, 12, 18, 24, 30, or 36 months) after the BCG induction course (e.g., after the first administration of the BCG induction course) or after diagnosis.
  • a papillary carcinoma of Ta or T1 e.g., one or more high grade Ta or T1 stage tumors
  • the recurrence of the papillary carcinoma of Ta or T1 stage occurred between 6 and 9 months, between 6 and 12 months, between 6 and 18 months, between 6 and 24 months, between 6 and 30 months, between 6 and 36 months, between 9 and 12 months, between 9 and 18 months, between 9 and 24 months, between 9 and 30 months, between 9 and 36 months, between 12 and 18 months, between 12 and 24 months, between 12 and 30 months, between 12 and 36 months, between 18 and 24 months, between 18 and 30 months, between 18 and 36 months, between 24 and 30 months, between 24 and 36 months, or between 30 and 36 months, after the BCG induction course (e.g, after the first administration of the BCG induction course) or after diagnosis.
  • the BCG induction course e.g. after the first administration of the BCG induction course
  • diagnosis e.g., after diagnosis.
  • the recurrence of the papillary carcinoma of Ta or T1 stage occurred between 6 and 24 months after the BCG induction course (e.g., after the first administration of the BCG induction course) or after diagnosis.
  • the individual has previously received a BCG induction course, experienced an absence of bladder tumors after the BCG induction course, subsequently received at least one BCG maintenance course, and further subsequently experienced recurrence of carcinoma in situ at least 12 months (e.g., at least 12, 18, 24, 30, or 36 months) after the BCG induction course (e.g. , after the first administration of the BCG induction course) or after diagnosis.
  • the recurrence of the carcinoma in situ occurs between 12 and 18 months, between 12 and 24 months, between 12 and 30 months, between 12 and 36 months, between 18 and 24 months, between 18 and 30 months, between 18 and 36 months, between 24 and 30 months, between 24 and 36 months, or between 30 and 36 months, after the BCG induction course (e.g. , after the first administration of the BCG induction course) or after diagnosis.
  • the recurrence of the carcinoma in situ occurs between 12 and 24 after the BCG induction course (e.g. , after the first administration of the BCG induction course) or after diagnosis.
  • the BCG maintenance course comprises 3 weekly intravesical instillations of BCG.
  • the individual has received at least 2 of the 3 weekly intravesical instillations of a BCG maintenance course.
  • the BCG induction course comprises weekly intravesical instillation of BCG once per week for five or six weeks.
  • the individual received at least 5 of the 6 doses of the standard BCG induction course.
  • the individual has previously received a BCG induction course beginning at month 0 of a treatment regimen and had persistent bladder cancer at month 3 of the treatment regimen.
  • the individual was previously treated with inadequate intravesical BCG therapy, responded to the BCG therapy, and subsequently experienced a delayed relapse of bladder cancer.
  • the individual has previously received a BCG induction course, experienced an absence of bladder tumors after the BCG induction course, did not receive any BCG maintenance courses, and subsequently experienced recurrence of a papillary carcinoma of Ta or T1 stage, carcinoma in situ, or a combination thereof (e.g., at least 6, 9, 12, 18, 24, 30, or 36 months) after the BCG induction course (e.g. , after the first administration of the BCG induction course) or after diagnosis.
  • the recurrence of the papillary carcinoma of Ta or T1 stage occurred between 6 and 9 months, between 6 and 12 months, between 6 and 18 months, between 6 and 24 months, between 6 and 30 months, between 6 and 36 months, between 9 and 12 months, between 9 and 18 months, between 9 and 24 months, between 9 and 30 months, between 9 and 36 months, between 12 and 18 months, between 12 and 24 months, between 12 and 30 months, between 12 and 36 months, between 18 and 24 months, between 18 and 30 months, between 18 and 36 months, between 24 and 30 months, between 24 and 36 months, or between 30 and 36 months, after the BCG induction course (e.g.
  • the recurrence of the papillary carcinoma of Ta or T1 stage e.g., one or more high grade Ta or T1 stage tumors), carcinoma in situ, or combination thereof occurred between about 6 months and about 24 months after the BCG induction course (e.g., after the first administration of the BCG induction course) or after diagnosis.
  • the BCG induction course comprises weekly intravesical instillation of BCG once per week for five or six weeks.
  • the individual received at least 1, 2, or 3 but not all 6 of the doses of the standard BCG induction course. In certain embodiments, the individual received at least 5 of the 6 doses of the standard BCG induction course.
  • the individual received at least 3 of the 6 doses of the standard BCG induction course. In some variations, the individual received 3, 4, 5, or 6 of the 6 doses of the standard BCG induction course. In some embodiments, the individual has previously received a BCG induction course beginning at month 0 of a treatment regimen and had persistent bladder cancer at month 3 of the treatment regimen. In some embodiments, the individual did not receive a BCG maintenance course due to a shortage of BCG (e.g., a localized or global shortage of BCG).
  • a shortage of BCG e.g., a localized or global shortage of BCG.
  • a method of treating bladder cancer in an individual comprising intravesically administering an oncolytic virus to the individual, wherein the individual: a) has not received prior intravesical Bacillus Calmette- Guerin (BCG) therapy; b) has previously received a BCG induction course, experienced an absence of bladder tumors after the BCG induction course, did not receive any BCG maintenance courses, and subsequently experienced recurrence of a papillary carcinoma of Ta or T1 stage, carcinoma in situ, or a combination thereof at least 6 months after the BCG induction course; or c) has previously received a BCG induction course, experienced an absence of bladder tumors after the BCG induction course, subsequently received at least one BCG maintenance course, and further subsequently experienced: i) recurrence of a papillary carcinoma of Ta or T1 stage at least 6 months after the BCG induction course; or ii) recurrence of carcinoma in situ at least 12 months after the BCG
  • the BCG induction course comprises 5 or 6 weekly intravesical instillations of BCG.
  • the BCG maintenance course comprises 3 weekly intravesical instillations of BCG.
  • the oncolytic vims comprises a viral vector comprising a tumor cell-specific promoter (e.g. , a human E2F-1 promoter) operably linked to a viral gene essential for replication of the oncolytic vims, and a heterologous gene encoding an immune -related molecule (e.g., GM-CSF).
  • the oncolytic vims is an adenovims serotype 5, wherein the endogenous Ela promoter of a native adenovims serotype 5 is replaced by the human E2F-1 promoter, and the endogenous E3 19kD coding region of the native adenovims serotype 5 is replaced by a nucleic acid encoding human GM-CSF.
  • the oncolytic vims is cretostimogene.
  • the method comprises intravesically administering at least one dose of N-Dodecyl-P-D-maltoside (DDM) to the individual prior to administering the oncolytic vims (e.g., prior to each intravesical instillation of the oncolytic vims).
  • DDM N-Dodecyl-P-D-maltoside
  • the method comprises intravesically administering a first and second dose of DDM to the individual, and intravesically administering a saline wash after administering the first and second dose of DDM to the individual and prior to administering the oncolytic vims.
  • the method comprises intravesically administering a single dose of DDM to the individual directly prior to each instillation of the oncolytic vims, without administration of a saline wash between the single dose of DDM and the administration of the oncolytic vims.
  • the method comprises administering a first and second dose of DDM to the individual prior to administering the oncolytic vims, wherein the oncolytic vims is administered directly after the second dose of DDM (e.g., without intravesical administration of a saline wash after the second dose of DDM and before administration of the oncolytic vims).
  • the individual has non-muscle invasive bladder cancer (NMIBC).
  • the individual has high-risk NMIBC.
  • the method comprises administration of one or more additional therapeutic agents (e.g., an immune checkpoint modulator or chemotherapeutic agent described herein).
  • the additional therapeutic agent is administered based on a determination of GDB (e.g., a GDB value associated with intermediate or high risk) in a urine sample from the subject as described herein.
  • the method comprises an induction phase comprising administering the oncolytic virus to the individual once per week for six weeks.
  • the method comprises a maintenance phase subsequent to the induction phase, wherein the maintenance phase comprises administering the oncolytic virus to the individual weekly for three weeks every three or six months.
  • the start of the induction phase and the start of the maintenance phase are separated by about three months or about six months.
  • the oncolytic virus is administered at a dose of about 1 x 10 8 to about 1 x 10 14 viral particles (e.g, about 1 x 10 12 viral particles).
  • a method of treating bladder cancer e.g, high-risk NMIBC in an individual comprising intravesically administering an oncolytic virus to the individual, wherein the individual: a) has not received prior intravesical Bacillus Calmette-Guerin (BCG) therapy; b) has received at least one diagnosis of high-risk NMIBC and has not received intravesical BCG therapy within 24 months prior to the most recent diagnosis of high-risk NMIBC; c) has received at least one diagnosis of high-risk NMIBC and has received a maximum of 1 or 2 doses of BCG within 24 months prior to the most recent diagnosis of high-risk NMIBC; d) received at least 5 of 6 doses of a BCG induction course and at least 2 doses of 1 course of either reinduction or maintenance BCG and experienced persistent or recurrent carcinoma in situ (CIS), with or without concomitant Ta or T1 disease, within 12 months following the last dose of BCG; e) received at least 5 of 6 dose
  • BCG Bacillus Calmette-Guerin
  • the BCG induction course comprises 5 or 6 weekly intravesical instillations of BCG.
  • the BCG maintenance course comprises 2 or 3 weekly intravesical instillations of BCG.
  • the oncolytic virus comprises a viral vector comprising a tumor cell-specific promoter (e.g., a human E2F-1 promoter) operably linked to a viral gene essential for replication of the oncolytic virus, and a heterologous gene encoding an immune-related molecule (e.g, GM-CSF).
  • the oncolytic virus is an adenovirus serotype 5, wherein the endogenous Ela promoter of a native adenovirus serotype 5 is replaced by the human E2F-1 promoter, and the endogenous E3 19kD coding region of the native adenovirus serotype 5 is replaced by a nucleic acid encoding human GM-CSF.
  • the oncolytic virus is cretostimogene.
  • the method comprises intravesically administering at least one dose ofN-Dodecyl-P-D-maltoside (DDM) to the individual prior to administering the oncolytic virus.
  • DDM N-Dodecyl-P-D-maltoside
  • the method comprises intravesically administering: 1) a first bladder wash with normal saline; 2) a second bladder wash with DDM; 3) instillation of DDM; 4) a third bladder was with normal saline; and 5) instillation of cretostimogene.
  • the method comprises intravesically administering a single dose of DDM to the individual prior to each administration of the oncolytic virus.
  • the method does not comprise administering a saline bladder wash before or after the administration of the DDM and before the administration of the oncolytic virus.
  • the individual has non-muscle invasive bladder cancer (NMIBC). In certain embodiments, the individual has high-risk NMIBC.
  • the method comprises administration of one or more additional therapeutic agents (e.g., an immune checkpoint modulator or chemotherapeutic agent described herein, e.g., gemcitabine).
  • the method comprises an induction phase comprising administering the oncolytic virus to the individual once per week for six weeks.
  • the method comprises a maintenance phase subsequent to the induction phase, wherein the maintenance phase comprises administering the oncolytic virus to the individual weekly for three weeks every three or six months.
  • the start of the induction phase and the start of the maintenance phase are separated by about three months or about six months.
  • the oncolytic virus is administered at a dose of about 1 x 10 8 to about 1 x 10 14 viral particles (e.g., about 1 x 10 12 viral particles).
  • NMIBC high-risk non-muscle invasive bladder cancer
  • the individual a) has not received prior intravesical Bacillus Calmette-Guerin (BCG) therapy; b) has previously received a BCG induction course, experienced an absence of bladder tumors after the BCG induction course, did not receive any BCG maintenance courses, and subsequently experienced recurrence of a papillary carcinoma of Ta or T1 stage, carcinoma in situ, or a combination thereof at least 6 months after the BCG induction course; or c) has previously received a BCG induction course, experienced an absence of bladder tumors after the BCG induction course, subsequently received at least one BCG maintenance course, and further
  • the BCG induction course comprises 5 or 6 weekly intravesical instillations of BCG.
  • the BCG maintenance course comprises 3 weekly intravesical instillations of BCG.
  • the oncolytic virus comprises a viral vector comprising a tumor cell-specific promoter (e.g. , a human E2F- 1 promoter) operably linked to a viral gene essential for replication of the oncolytic virus, and a heterologous gene encoding an immune-related molecule (e.g., GM-CSF).
  • the oncolytic virus is an adenovirus serotype 5, wherein the endogenous Ela promoter of a native adenovirus serotype 5 is replaced by the human E2F-1 promoter, and the endogenous E3 19kD coding region of the native adenovirus serotype 5 is replaced by a nucleic acid encoding human GM-CSF.
  • the oncolytic virus is cretostimogene.
  • the method comprises intravesically administering at least one dose ofN-Dodecyl-P-D-maltoside (DDM) to the individual prior to administering the oncolytic virus (e.g., prior to each intravesical instillation of the oncolytic virus).
  • DDM N-Dodecyl-P-D-maltoside
  • the method comprises intravesically administering a first and second dose of DDM to the individual, and intravesically administering a saline wash after administering the first and second dose of DDM to the individual and prior to administering the oncolytic virus.
  • the method comprises intravesically administering a single dose of DDM to the individual directly prior to each instillation of the oncolytic virus, without administration of a saline wash between the single dose of DDM and the administration of the oncolytic virus.
  • the method comprises administering a first and second dose of DDM to the individual prior to administering the oncolytic virus, wherein the oncolytic virus is administered directly after the second dose of DDM (e.g., without intravesical administration of a saline wash after the second dose of DDM and before administration of the oncolytic virus).
  • the method comprises administration of one or more additional therapeutic agents (e.g., an immune checkpoint modulator or chemotherapeutic agent described herein).
  • the additional therapeutic agent is administered based on a determination of GDB (e.g., a GDB value associated with intermediate or high risk) in a urine sample from the subject as described herein.
  • the method comprises an induction phase comprising administering the oncolytic virus to the individual once per week for six weeks.
  • the method comprises a maintenance phase subsequent to the induction phase, wherein the maintenance phase comprises administering the oncolytic virus to the individual weekly for three weeks every three or six months.
  • the start of the induction phase and the start of the maintenance phase are separated by about three months or about six months.
  • the oncolytic virus is administered at a dose of about 1 x 10 8 to about 1 x 10 14 viral particles (e.g., about 1 x 10 12 viral particles).
  • a method of treating high-risk NMIBC in an individual comprising intravesically administering an oncolytic virus to the individual, wherein the individual: a) has not received prior intravesical Bacillus Calmette -Guerin (BCG) therapy; b) has received at least one diagnosis of high-risk NMIBC and has not received intravesical BCG therapy within 24 months prior to the most recent diagnosis of high-risk NMIBC; c) has received at least one diagnosis of high-risk NMIBC and has received a maximum of 1 or 2 doses of BCG within 24 months prior to the most recent diagnosis of high-risk NMIBC; d) received at least 5 of 6 doses of a BCG induction course and at least 2 doses of 1 course of either reinduction or maintenance BCG and experienced persistent or recurrent carcinoma in situ (CIS), with or without concomitant Ta or T1 disease, within 12 months following the last dose of BCG; e) received at least 5 of 6 doses of a BCG in
  • the BCG induction course comprises 5 or 6 weekly intravesical instillations of BCG.
  • the BCG maintenance course comprises 2 or 3 weekly intravesical instillations of BCG.
  • the oncolytic virus comprises a viral vector comprising a tumor cell-specific promoter (e.g. , a human E2F-1 promoter) operably linked to a viral gene essential for replication of the oncolytic virus, and a heterologous gene encoding an immune- related molecule (e.g, GM-CSF).
  • the oncolytic virus is an adenovirus serotype 5, wherein the endogenous Ela promoter of a native adenovirus serotype 5 is replaced by the human E2F-1 promoter, and the endogenous E3 19kD coding region of the native adenovirus serotype 5 is replaced by a nucleic acid encoding human GM-CSF.
  • the oncolytic virus is cretostimogene.
  • the method comprises intravesically administering at least one dose of N-Dodecyl-P-D-maltoside (DDM) to the individual prior to administering the oncolytic virus.
  • DDM N-Dodecyl-P-D-maltoside
  • the method comprises intravesically administering: 1) a first bladder wash with normal saline; 2) a second bladder wash with DDM; 3) instillation of DDM; 4) a third bladder was with normal saline; and 5) instillation of cretostimogene.
  • the method comprises intravesically administering a single dose of DDM to the individual prior to each administration of the oncolytic virus.
  • the method does not comprise administering a saline bladder wash before or after the administration of the DDM and before the administration of the oncolytic virus.
  • the method comprises administration of one or more additional therapeutic agents (e.g., an immune checkpoint modulator or chemotherapeutic agent described herein, e.g., gemcitabine).
  • the method comprises an induction phase comprising administering the oncolytic virus to the individual once per week for six weeks.
  • the method comprises a maintenance phase subsequent to the induction phase, wherein the maintenance phase comprises administering the oncolytic virus to the individual weekly for three weeks every three or six months.
  • the start of the induction phase and the start of the maintenance phase are separated by about three months or about six months.
  • the oncolytic virus is administered at a dose of about 1 x 10 8 to about 1 x 10 14 viral particles (e.g., about 1 x 10 12 viral particles).
  • a method of treating high-risk NMIBC in an individual comprising intravesically administering cretostimogene to the individual, wherein the individual: a) has not received prior intravesical Bacillus Calmette -Guerin (BCG) therapy; b) has received at least one diagnosis of high-risk NMIBC and has not received intravesical BCG therapy within 24 months prior to the most recent diagnosis of high-risk NMIBC; c) has received at least one diagnosis of high-risk NMIBC and has received a maximum of 1 or 2 doses of BCG within 24 months prior to the most recent diagnosis of high-risk NMIBC; d) received at least 5 of 6 doses of a BCG induction course and at least 2 doses of 1 course of either reinduction or maintenance BCG and experienced persistent or recurrent carcinoma in situ (CIS), with or without concomitant Ta or T1 disease, within 12 months following the last dose of BCG; e) received at least 5 of 6 doses of a BCG
  • the BCG induction course comprises 5 or 6 weekly intravesical instillations of BCG.
  • the BCG maintenance course comprises 2 or 3 weekly intravesical instillations of BCG.
  • the method comprises intravesically administering at least one dose ofN-Dodecyl-P-D-maltoside (DDM) to the individual prior to administering cretostimogene.
  • the method comprises intravesically administering: 1) a first bladder wash with normal saline; 2) a second bladder wash with DDM; 3) instillation of DDM; 4) a third bladder was with normal saline; and 5) instillation of cretostimogene.
  • the method comprises intravesically administering a single dose of DDM to the individual prior to each administration of cretostimogene. In certain embodiments, the method does not comprise administering a saline bladder wash before or after the administration of the DDM and before the administration of cretostimogene. In some embodiments, the method comprises administration of one or more additional therapeutic agents (e.g., an immune checkpoint modulator or chemotherapeutic agent described herein, e.g., gemcitabine). In some embodiments, the method comprises an induction phase comprising administering cretostimogene to the individual once per week for six weeks.
  • additional therapeutic agents e.g., an immune checkpoint modulator or chemotherapeutic agent described herein, e.g., gemcitabine.
  • the method comprises an induction phase comprising administering cretostimogene to the individual once per week for six weeks.
  • the method comprises a maintenance phase subsequent to the induction phase, wherein the maintenance phase comprises administering cretostimogene to the individual weekly for three weeks every three or six months.
  • the start of the induction phase and the start of the maintenance phase are separated by about three months or about six months.
  • cretostimogene is administered at a dose of about 1 x 10 8 to about 1 x 10 14 viral particles (e.g., about 1 x 10 12 viral particles).
  • a method of treating high-risk non-muscle invasive bladder cancer (NMIBC) in an individual comprising intravesically administering an oncolytic virus to the individual, wherein the individual a) has not received prior intravesical Bacillus Calmette-Guerin (BCG) therapy, b) has not received intravesical BCG therapy within 24 months prior to the most recent diagnosis of high-risk NMIBC, or c) has received a maximum of 1 or 2 doses of BCG within 24 months prior to the most recent diagnosis of high-risk NMIBC.
  • the oncolytic virus comprises a viral vector comprising a tumor cell-specific promoter (e.g.
  • the oncolytic virus is an adenovirus serotype 5, wherein the endogenous Ela promoter of a native adenovirus serotype 5 is replaced by the human E2F-1 promoter, and the endogenous E3 19kD coding region of the native adenovirus serotype 5 is replaced by a nucleic acid encoding human GM-CSF.
  • the oncolytic virus is cretostimogene.
  • the method comprises intravesically administering at least one dose of N-Dodecyl-[3-D-maltoside (DDM) to the individual prior to administering the oncolytic virus (e.g., prior to each intravesical instillation of the oncolytic virus).
  • the method comprises intravesically administering a first and second dose of DDM to the individual, and intravesically administering a saline wash after administering the first and second dose of DDM to the individual and prior to administering the oncolytic virus.
  • the method comprises intravesically administering a single dose of DDM to the individual directly prior to each instillation of the oncolytic virus, without administration of a saline wash between the single dose of DDM and the administration of the oncolytic virus.
  • the method comprises administering a first and second dose of DDM to the individual prior to administering the oncolytic virus, wherein the oncolytic virus is administered directly after the second dose of DDM (e.g., without intravesical administration of a saline wash after the second dose of DDM and before administration of the oncolytic virus).
  • the method comprises intravesically administering: 1) a first bladder wash with normal saline; 2) a second bladder wash with DDM; 3) instillation of DDM; 4) a third bladder was with normal saline; and 5) instillation of cretostimogene.
  • the method comprises intravesically administering a single dose of DDM to the individual prior to each administration of the oncolytic virus.
  • the method does not comprise administering a saline bladder wash before or after the administration of the DDM and before the administration of the oncolytic virus.
  • the method comprises administration of one or more additional therapeutic agents (e.g., an immune checkpoint modulator or chemotherapeutic agent described herein, e.g., gemcitabine).
  • the additional therapeutic agent is administered based on a determination of GDB (e.g., a GDB value associated with intermediate or high risk) in a urine sample from the subject as described herein.
  • the method comprises an induction phase comprising administering the oncolytic virus to the individual once per week for six weeks.
  • the method comprises a maintenance phase subsequent to the induction phase, wherein the maintenance phase comprises administering the oncolytic virus to the individual weekly for three weeks every three or six months.
  • the start of the induction phase and the start of the maintenance phase are separated by about three months or about six months.
  • the oncolytic virus is administered at a dose of about 1 x 10 8 to about 1 x 10 14 viral particles (e.g., about 1 x 10 12 viral particles).
  • the method is as described in Example 3, Cohort A.
  • a method of treating high-risk non-muscle invasive bladder cancer (NMIBC) in a BCG-naive individual comprising intravesically administering to the individual an oncolytic virus.
  • the BCG-naive individual has not received prior intravesical Bacillus Calmette-Guerin (BCG) therapy, also known as complete BCG-naive.
  • BCG Bacillus Calmette-Guerin
  • the BCG-naive individual has not received intravesical BCG therapy within 24 months prior to the most recent diagnosis of high-risk NMIBC.
  • the BCG-naive individual has received a maximum of 1 or 2 doses of BCG within 24 months prior to the most recent diagnosis of high-risk NMIBC.
  • the oncolytic virus comprises a viral vector comprising a tumor cell-specific promoter (e.g., a human E2F-1 promoter) operably linked to a viral gene essential for replication of the oncolytic virus, and a heterologous gene encoding an immune-related molecule (e.g., GM-CSF).
  • a tumor cell-specific promoter e.g., a human E2F-1 promoter
  • an immune-related molecule e.g., GM-CSF
  • the oncolytic virus is an adenovirus serotype 5, wherein the endogenous Ela promoter of a native adenovirus serotype 5 is replaced by the human E2F-1 promoter, and the endogenous E3 19kD coding region of the native adenovirus serotype 5 is replaced by a nucleic acid encoding human GM-CSF.
  • the oncolytic virus is cretostimogene.
  • the method comprises intravesically administering at least one dose of N-Dodecyl-P-D-maltoside (DDM) to the individual prior to administering the oncolytic virus (e.g., prior to each intravesical instillation of the oncolytic virus).
  • DDM N-Dodecyl-P-D-maltoside
  • the method comprises intravesically administering a first and second dose of DDM to the individual, and intravesically administering a saline wash after administering the first and second dose of DDM to the individual and prior to administering the oncolytic virus.
  • the method comprises intravesically administering a single dose of DDM to the individual directly prior to each instillation of the oncolytic virus, without administration of a saline wash between the single dose of DDM and the administration of the oncolytic virus.
  • the method comprises administering a first and second dose of DDM to the individual prior to administering the oncolytic virus, wherein the oncolytic virus is administered directly after the second dose of DDM (e.g., without intravesical administration of a saline wash after the second dose of DDM and before administration of the oncolytic virus).
  • the method comprises intravesically administering: 1) a first bladder wash with normal saline; 2) a second bladder wash with DDM; 3) instillation of DDM; 4) a third bladder was with normal saline; and 5) instillation of cretostimogene.
  • the method comprises intravesically administering a single dose of DDM to the individual prior to each administration of the oncolytic virus. In certain embodiments, the method does not comprise administering a saline bladder wash before or after the administration of the DDM and before the administration of the oncolytic virus. In some embodiments, the method comprises administration of one or more additional therapeutic agents (e.g., an immune checkpoint modulator or chemotherapeutic agent described herein, e.g, gemcitabine). In certain embodiments, the additional therapeutic agent is administered based on a determination of GDB (e.g., a GDB value associated with intermediate or high risk) in a urine sample from the subject as described herein.
  • GDB e.g., a GDB value associated with intermediate or high risk
  • the method comprises an induction phase comprising administering the oncolytic virus to the individual once per week for six weeks.
  • the method comprises a maintenance phase subsequent to the induction phase, wherein the maintenance phase comprises administering the oncolytic virus to the individual weekly for three weeks every three or six months.
  • the start of the induction phase and the start of the maintenance phase are separated by about three months or about six months.
  • the oncolytic virus is administered at a dose of about 1 x 10 8 to about 1 x 10 14 viral particles (e.g., about 1 x 10 12 viral particles).
  • the method is as described in Example 3, Cohort A.
  • BCG Bacillus Calmette -Guerin
  • the individual has concurrent high-trade Ta or T1 bladder cancer. In other embodiments, the individual does not have a concurrent high-trade Ta or T1 bladder cancer.
  • the method comprises intravesically administering at least one dose of N-Dodecyl-[3-D-maltoside (DDM) to the individual prior to administering the cretostimogene (e.g., prior to each intravesical instillation of the oncolytic virus).
  • the method comprises intravesically administering a first and second dose of DDM to the individual, and intravesically administering a saline wash after administering the first and second dose of DDM to the individual and prior to administering the oncolytic virus.
  • the method comprises intravesically administering a single dose of DDM to the individual directly prior to each instillation of the oncolytic virus, without administration of a saline wash between the single dose of DDM and the administration of the oncolytic virus.
  • the method comprises administering a first and second dose of DDM to the individual prior to administering cretostimogene, wherein the cretostimogene is administered directly after the second dose of DDM (e.g., without intravesical administration of a saline wash after the second dose of DDM and before administration of the cretostimogene).
  • the method comprises intravesically administering: 1) a first bladder wash with normal saline; 2) a second bladder wash with DDM; 3) instillation of DDM; 4) a third bladder was with normal saline; and 5) instillation of cretostimogene.
  • the method comprises intravesically administering a single dose of DDM to the individual prior to each administration of the cretostimogene.
  • the method does not comprise administering a saline bladder wash before or after the administration of the DDM and before the administration of the cretostimogene.
  • the method comprises an induction phase comprising administering the cretostimogene to the individual once per week for six weeks.
  • the method comprises a maintenance phase subsequent to the induction phase, wherein the maintenance phase comprises administering the cretostimogene to the individual weekly for three weeks every three or six months.
  • the start of the induction phase and the start of the maintenance phase are separated by about three months or about six months.
  • the cretostimogene is administered at a dose of about 1 x 10 8 to about 1 x 10 14 viral particles (e.g., about 1 x 10 12 viral particles).
  • the method is as described in Example 3, Cohort A, Arm 1 or Arm 2.
  • the individual has concurrent high-trade Ta or T1 bladder cancer. In other embodiments, the individual does not have a concurrent high-trade Ta or T1 bladder cancer.
  • the method comprises an induction phase comprising administering the cretostimogene to the individual once per week for six weeks. In certain embodiments, the method comprises a maintenance phase subsequent to the induction phase, wherein the maintenance phase comprises administering the cretostimogene to the individual weekly for three weeks every three or six months. In some embodiments, the start of the induction phase and the start of the maintenance phase are separated by about three months or about six months.
  • the cretostimogene is administered at a dose of about 1 x 10 8 to about 1 x 10 14 viral particles (e.g., about 1 x 10 12 viral particles).
  • the method is as described in Example 3, Cohort A, Arm 1.
  • BCG Bacillus Calmette -Guerin
  • the method does not comprise administering a saline bladder wash before or after the administration of the DDM and before the administration of the cretostimogene.
  • the individual has concurrent high-trade Ta or T1 bladder cancer.
  • the individual does not have a concurrent high-trade Ta or T1 bladder cancer.
  • the method comprises an induction phase comprising administering the cretostimogene to the individual once per week for six weeks.
  • the method comprises a maintenance phase subsequent to the induction phase, wherein the maintenance phase comprises administering the cretostimogene to the individual weekly for three weeks every three or six months.
  • the start of the induction phase and the start of the maintenance phase are separated by about three months or about six months.
  • the cretostimogene is administered at a dose of about 1 x 10 8 to about 1 x 10 14 viral particles (e.g., about 1 x 10 12 viral particles).
  • the method is as described in Example 3, Cohort A, Arm 2.
  • BCG Bacillus Calmette-Guerin
  • the method comprises intravesically administering a single dose of DDM to the individual prior to each administration of the cretostimogene. In certain embodiments, the method does not comprise administering a saline bladder wash before or after the administration of the DDM and before the administration of the cretostimogene. In some embodiments, the method comprises an induction phase comprising administering the cretostimogene to the individual once per week for six weeks. In certain embodiments, the method comprises a maintenance phase subsequent to the induction phase, wherein the maintenance phase comprises administering the cretostimogene to the individual weekly for three weeks every three or six months. In some embodiments, the start of the induction phase and the start of the maintenance phase are separated by about three months or about six months.
  • the cretostimogene is administered at a dose of about 1 x 10 8 to about 1 x 10 14 viral particles (e.g., about 1 x 10 12 viral particles).
  • the method is as described in Example 3, Cohort A, Arm 3.
  • a method of treating high-risk non-muscle invasive bladder cancer (NMIBC) in an individual that has received prior BCG therapy comprising intravesically administering to the individual an oncolytic virus.
  • the individual a) is BCG-resistant (e.g., experienced persistent or recurrent high-grade Ta or CIS following an BCG induction course); b) is experiencing delayed relapse after inadequate BCG (e.g., has previously received a BCG induction course, experienced an absence of bladder tumors after the BCG induction course, did not receive any BCG maintenance courses, and subsequently experienced recurrence of a papillary carcinoma of Ta or T1 stage, carcinoma in situ, or a combination thereof at least 6 months after the BCG induction course); or c) is experiencing delayed relapse after adequate BCG (e.g, has previously received a BCG induction course, experienced an absence of bladder tumors after the BCG induction course, subsequently received at least one B
  • BCG-resistant e.g., experienced persistent or recurrent
  • the individual a) received at least 5 of 6 doses of a BCG induction course and at least 2 doses of 1 course of either reinduction or maintenance BCG and experienced persistent or recurrent carcinoma in situ (CIS), with or without concomitant Ta or T1 disease, within 12 months following the last dose of BCG; b) received at least 5 of 6 doses of a BCG induction course and at least 2 doses of 1 course of either reinduction or maintenance BCG and experienced recurrence of highgrade papillary-only Ta or T1 stage disease within 6 months following the last dose of BCG; f) received at least 5 of 6 doses of a BCG induction course and experienced high-grade T1 disease at a first evaluation (e.g, a 3-month evaluation) following the BCG induction course; g) received at least 5 of 6 doses of a BCG induction course and experienced persistent or recurrent high-grade papillary carcinoma of Ta stage or CIS at a first evaluation (e.g, a 3- month evaluation) following
  • the BCG induction course comprises 5 or 6 weekly intravesical instillations of BCG.
  • the BCG maintenance course comprises 3 weekly intravesical instillations of BCG.
  • the oncolytic virus comprises a viral vector comprising a tumor cell-specific promoter (e.g., a human E2F-1 promoter) operably linked to a viral gene essential for replication of the oncolytic virus, and a heterologous gene encoding an immune-related molecule (e.g., GM-CSF).
  • the oncolytic virus is an adenovirus serotype 5, wherein the endogenous Ela promoter of a native adenovirus serotype 5 is replaced by the human E2F-1 promoter, and the endogenous E3 19kD coding region of the native adenovirus serotype 5 is replaced by a nucleic acid encoding human GM-CSF.
  • the oncolytic virus is cretostimogene.
  • the method comprises intravesically administering at least one dose of N-Dodecyl-P-D-maltoside (DDM) to the individual prior to administering the oncolytic virus (e.g., prior to each intravesical instillation of the oncolytic virus).
  • DDM N-Dodecyl-P-D-maltoside
  • the method comprises intravesically administering a first and second dose of DDM to the individual, and intravesically administering a saline wash after administering the first and second dose of DDM to the individual and prior to administering the oncolytic virus.
  • the method comprises intravesically administering a single dose of DDM to the individual directly prior to each instillation of the oncolytic virus, without administration of a saline wash between the single dose of DDM and the administration of the oncolytic virus.
  • the method comprises administering a first and second dose of DDM to the individual prior to administering the oncolytic virus, wherein the oncolytic virus is administered directly after the second dose of DDM (e.g., without intravesical administration of a saline wash after the second dose of DDM and before administration of the oncolytic virus).
  • the individual has non-muscle invasive bladder cancer (NMIBC).
  • NMIBC non-muscle invasive bladder cancer
  • the method comprises administration of one or more additional therapeutic agents (e.g., an immune checkpoint modulator or chemotherapeutic agent described herein).
  • the additional therapeutic agent is administered based on a determination of GDB (e.g., a GDB value associated with intermediate or high risk) in a urine sample from the subject as described herein.
  • the method comprises an induction phase comprising administering the oncolytic virus to the individual once per week for six weeks.
  • the method comprises a maintenance phase subsequent to the induction phase, wherein the maintenance phase comprises administering the oncolytic virus to the individual weekly for three weeks every three or six months.
  • the start of the induction phase and the start of the maintenance phase are separated by about three months or about six months.
  • the oncolytic virus is administered at a dose of about 1 x 10 8 to about 1 x 10 14 viral particles (e.g, about 1 x 10 12 viral particles).
  • a method of treating high-risk non-muscle invasive bladder cancer (NMIBC) in an individual that has received prior BCG therapy comprising intravesically administering to the individual an oncolytic virus.
  • the individual was treated with a BCG induction course and experienced persistent or recurrent bladder cancer after the induction course.
  • the individual was identified as having persistent or recurrent high-grade Ta or CIS at an evaluation at month three, where month 0 begins at the start of the BCG induction course.
  • the BCG induction course comprises 5 or 6 weekly intravesical instillations of BCG.
  • the oncolytic virus comprises a viral vector comprising a tumor cell-specific promoter (e.g. , a human E2F-1 promoter) operably linked to a viral gene essential for replication of the oncolytic virus, and a heterologous gene encoding an immune -related molecule (e.g., GM-CSF).
  • a tumor cell-specific promoter e.g. , a human E2F-1 promoter
  • a heterologous gene encoding an immune -related molecule e.g., GM-CSF
  • the oncolytic virus is an adenovirus serotype 5, wherein the endogenous Ela promoter of a native adenovirus serotype 5 is replaced by the human E2F-1 promoter, and the endogenous E3 19kD coding region of the native adenovirus serotype 5 is replaced by a nucleic acid encoding human GM-CSF.
  • the oncolytic virus is cretostimogene.
  • the method comprises intravesically administering at least one dose of N-Dodecyl-[3-D-maltoside (DDM) to the individual prior to administering the oncolytic virus (e.g., prior to each intravesical instillation of the oncolytic virus).
  • the method comprises intravesically administering a first and second dose of DDM to the individual, and intravesically administering a saline wash after administering the first and second dose of DDM to the individual and prior to administering the oncolytic virus.
  • the method comprises intravesically administering a single dose of DDM to the individual directly prior to each instillation of the oncolytic virus, without administration of a saline wash between the single dose of DDM and the administration of the oncolytic virus.
  • the method comprises administering a first and second dose of DDM to the individual prior to administering the oncolytic virus, wherein the oncolytic virus is administered directly after the second dose of DDM (e.g., without intravesical administration of a saline wash after the second dose of DDM and before administration of the oncolytic virus).
  • the individual has non-muscle invasive bladder cancer (NMIBC).
  • NMIBC non-muscle invasive bladder cancer
  • the individual has high-risk NMIBC.
  • the method comprises administration of one or more additional therapeutic agents (e.g., an immune checkpoint modulator or chemotherapeutic agent described herein).
  • the additional therapeutic agent is administered based on a determination of GDB (e.g., a GDB value associated with intermediate or high risk) in a urine sample from the subject as described herein.
  • the method comprises an induction phase comprising administering the oncolytic virus to the individual once per week for six weeks.
  • the method comprises a maintenance phase subsequent to the induction phase, wherein the maintenance phase comprises administering the oncolytic virus to the individual weekly for three weeks every three or six months.
  • the start of the induction phase and the start of the maintenance phase are separated by about three months or about six months.
  • the oncolytic virus is administered at a dose of about 1 x 10 8 to about 1 x 10 14 viral particles (e.g, about 1 x 10 12 viral particles).
  • a method of treating high-risk NMIBC in an individual having carcinoma in situ (CIS) and/or high-grade Ta or T1 stage bladder cancer that has received prior BCG therapy comprising intravesically administering cretostimogene to the individual, wherein the individual: a) received at least 5 of 6 doses of a BCG induction course and at least 2 doses of 1 course of either reinduction or maintenance BCG and experienced persistent or recurrent carcinoma in situ (CIS), with or without concomitant Ta or T1 disease, within 12 months following the last dose of BCG; b) received at least 5 of 6 doses of a BCG induction course and at least 2 doses of 1 course of either reinduction or maintenance BCG and experienced recurrence of high-grade papillary-only Ta or T1 stage disease within 6 months following the last dose of BCG; f) received at least 5 of 6 doses of a BCG induction course and experienced high-grade T1 disease at a first evaluation (e
  • the BCG induction course comprises 5 or 6 weekly intravesical instillations of BCG.
  • the BCG maintenance course comprises 2 or 3 weekly intravesical instillations of BCG.
  • the method comprises intravesically administering at least one dose of N-Dodecyl-P-D-maltoside (DDM) to the individual prior to administering the oncolytic virus.
  • the method comprises intravesically administering a single dose of DDM to the individual prior to each administration of the oncolytic virus.
  • the method does not comprise administering a saline bladder wash before or after the administration of the DDM and before the administration of the oncolytic virus.
  • the method comprises administration of one or more additional therapeutic agents (e.g., an immune checkpoint modulator or chemotherapeutic agent described herein, e.g., gemcitabine).
  • the method comprises an induction phase comprising administering the oncolytic virus to the individual once per week for six weeks.
  • the method comprises a maintenance phase subsequent to the induction phase, wherein the maintenance phase comprises administering the oncolytic virus to the individual weekly for three weeks every three or six months.
  • the start of the induction phase and the start of the maintenance phase are separated by about three months or about six months.
  • the oncolytic virus is administered at a dose of about 1 x 10 8 to about 1 x 10 14 viral particles (e.g., about 1 x 10 12 viral particles).
  • the method is according to Example 3, Cohort B.
  • a method of treating high-risk NMIBC in individual having carcinoma in situ (CIS) that has received prior BCG therapy comprising intravesically administering cretostimogene to the individual, wherein the individual: a) received at least 5 of 6 doses of a BCG induction course and at least 2 doses of 1 course of either reinduction or maintenance BCG and experienced persistent or recurrent carcinoma in situ (CIS), with or without concomitant Ta or T1 disease, within 12 months following the last dose of BCG; b) received at least 5 of 6 doses of a BCG induction course and at least 2 doses of 1 course of either reinduction or maintenance BCG and experienced recurrence of high-grade papillary-only Ta or T1 stage disease within 6 months following the last dose of BCG; f) received at least 5 of 6 doses of a BCG induction course and experienced high-grade T1 disease at a first evaluation (e.g., a 3-month evaluation) following the BCG
  • a first evaluation e.g., a 3-
  • the BCG induction course comprises 5 or 6 weekly intravesical instillations of BCG.
  • the BCG maintenance course comprises 2 or 3 weekly intravesical instillations of BCG.
  • the individual has concurrent high-grade Ta or Tl stage bladder cancer. In some embodiments, the individual does not have concurrent high-grade Ta or Tl stage bladder cancer.
  • the method comprises intravesically administering at least one dose ofN-Dodecyl-P-D-maltoside (DDM) to the individual prior to administering the oncolytic virus. In some embodiments, the method comprises intravesically administering a single dose of DDM to the individual prior to each administration of the oncolytic virus.
  • DDM N-Dodecyl-P-D-maltoside
  • the method does not comprise administering a saline bladder wash before or after the administration of the DDM and before the administration of the oncolytic virus.
  • the method comprises administration of one or more additional therapeutic agents (e.g., an immune checkpoint modulator or chemotherapeutic agent described herein, e.g., gemcitabine).
  • the method comprises an induction phase comprising administering the oncolytic virus to the individual once per week for six weeks.
  • the method comprises a maintenance phase subsequent to the induction phase, wherein the maintenance phase comprises administering the oncolytic virus to the individual weekly for three weeks every three or six months.
  • the start of the induction phase and the start of the maintenance phase are separated by about three months or about six months.
  • the oncolytic virus is administered at a dose of about 1 x 10 8 to about 1 x 10 14 viral particles (e.g, about 1 x 10 12 viral particles).
  • the method is according to Example 3, Cohort B, Arm 1.
  • a method of treating high-risk NMIBC in a individual having high-grade Ta or T1 stage bladder cancer without carcinoma in situ (CIS) that has received prior BCG therapy comprising intravesically administering cretostimogene to the individual, wherein the individual: a) received at least 5 of 6 doses of a BCG induction course and at least 2 doses of 1 course of either reinduction or maintenance BCG and experienced persistent or recurrent carcinoma in situ (CIS), with or without concomitant Ta or T1 disease, within 12 months following the last dose of BCG; b) received at least 5 of 6 doses of a BCG induction course and at least 2 doses of 1 course of either reinduction or maintenance BCG and experienced recurrence of high-grade papillary-only Ta or T1 stage disease within 6 months following the last dose of BCG; f) received at least 5 of 6 doses of a BCG induction course and experienced high-grade T1 disease at a first evaluation (e.
  • the BCG induction course comprises 5 or 6 weekly intravesical instillations of BCG.
  • the BCG maintenance course comprises 2 or 3 weekly intravesical instillations of BCG.
  • the method comprises intravesically administering at least one dose of N-Dodecyl-P-D-maltoside (DDM) to the individual prior to administering the oncolytic virus.
  • the method comprises intravesically administering a single dose of DDM to the individual prior to each administration of the oncolytic virus.
  • the method does not comprise administering a saline bladder wash before or after the administration of the DDM and before the administration of the oncolytic virus.
  • the method comprises administration of one or more additional therapeutic agents (e.g., an immune checkpoint modulator or chemotherapeutic agent described herein, e.g., gemcitabine).
  • the method comprises an induction phase comprising administering the oncolytic virus to the individual once per week for six weeks.
  • the method comprises a maintenance phase subsequent to the induction phase, wherein the maintenance phase comprises administering the oncolytic virus to the individual weekly for three weeks every three or six months.
  • the start of the induction phase and the start of the maintenance phase are separated by about three months or about six months.
  • the oncolytic virus is administered at a dose of about 1 x 10 8 to about 1 x 10 14 viral particles (e.g., about 1 x 10 12 viral particles).
  • the method is according to Example 3, Cohort B, Arm 2.
  • a method of treating high-risk non-muscle invasive bladder cancer (NMIBC) in an individual comprising intravesically administering an oncolytic virus to the individual, wherein the individual has previously received a BCG induction course, experienced an absence of bladder tumors after the BCG induction course, did not receive any BCG maintenance courses, and subsequently experienced recurrence of a papillary carcinoma of Ta or Tl stage, carcinoma in situ, or a combination thereof at least 6 months after the BCG induction course.
  • the BCG induction course comprises 5 or 6 weekly intravesical instillations of BCG.
  • the oncolytic virus comprises a viral vector comprising a tumor cell-specific promoter (e.g.
  • the oncolytic virus is an adenovirus serotype 5, wherein the endogenous Ela promoter of a native adenovirus serotype 5 is replaced by the human E2F-1 promoter, and the endogenous E3 19kD coding region of the native adenovirus serotype 5 is replaced by a nucleic acid encoding human GM-CSF.
  • the oncolytic virus is cretostimogene.
  • the method comprises intravesically administering at least one dose of N-Dodecyl-P-D-maltoside (DDM) to the individual prior to administering the oncolytic virus (e.g., prior to each intravesical instillation of the oncolytic virus).
  • DDM N-Dodecyl-P-D-maltoside
  • the method comprises intravesically administering a first and second dose of DDM to the individual, and intravesically administering a saline wash after administering the first and second dose of DDM to the individual and prior to administering the oncolytic virus.
  • the method comprises intravesically administering a single dose of DDM to the individual directly prior to each instillation of the oncolytic virus, without administration of a saline wash between the single dose of DDM and the administration of the oncolytic virus.
  • the method comprises administering a first and second dose of DDM to the individual prior to administering the oncolytic virus, wherein the oncolytic virus is administered directly after the second dose of DDM (e.g., without intravesical administration of a saline wash after the second dose of DDM and before administration of the oncolytic virus).
  • the method comprises administration of one or more additional therapeutic agents (e.g., an immune checkpoint modulator or chemotherapeutic agent described herein).
  • the additional therapeutic agent is administered based on a determination of GDB (e.g., a GDB value associated with intermediate or high risk) in a urine sample from the subject as described herein.
  • the method comprises an induction phase comprising administering the oncolytic virus to the individual once per week for six weeks.
  • the method comprises a maintenance phase subsequent to the induction phase, wherein the maintenance phase comprises administering the oncolytic virus to the individual weekly for three weeks every three or six months.
  • the start of the induction phase and the start of the maintenance phase are separated by about three months or about six months.
  • the oncolytic virus is administered at a dose of about 1 x 10 8 to about 1 x 10 14 viral particles (e.g., about 1 x 10 12 viral particles).
  • NMIBC high-risk non-muscle invasive bladder cancer
  • the BCG induction course comprises 5 or 6 weekly intravesical instillations of BCG.
  • the BCG maintenance course comprises 3 weekly intravesical instillations of BCG.
  • the oncolytic virus comprises a viral vector comprising a tumor cell-specific promoter (e.g. , a human E2F- 1 promoter) operably linked to a viral gene essential for replication of the oncolytic virus, and a heterologous gene encoding an immune-related molecule (e.g., GM-CSF).
  • the oncolytic virus is an adenovirus serotype 5, wherein the endogenous Ela promoter of a native adenovirus serotype 5 is replaced by the human E2F-1 promoter, and the endogenous E3 19kD coding region of the native adenovirus serotype 5 is replaced by a nucleic acid encoding human GM-CSF.
  • the oncolytic virus is cretostimogene.
  • the method comprises intravesically administering at least one dose ofN-Dodecyl-P-D-maltoside (DDM) to the individual prior to administering the oncolytic virus (e.g., prior to each intravesical instillation of the oncolytic virus).
  • DDM N-Dodecyl-P-D-maltoside
  • the method comprises intravesically administering a first and second dose of DDM to the individual, and intravesically administering a saline wash after administering the first and second dose of DDM to the individual and prior to administering the oncolytic virus.
  • the method comprises intravesically administering a single dose of DDM to the individual directly prior to each instillation of the oncolytic virus, without administration of a saline wash between the single dose of DDM and the administration of the oncolytic virus.
  • the method comprises administering a first and second dose of DDM to the individual prior to administering the oncolytic virus, wherein the oncolytic virus is administered directly after the second dose of DDM (e.g., without intravesical administration of a saline wash after the second dose of DDM and before administration of the oncolytic virus).
  • the method comprises administration of one or more additional therapeutic agents (e.g., an immune checkpoint modulator or chemotherapeutic agent described herein).
  • the additional therapeutic agent is administered based on a determination of GDB (e.g., a GDB value associated with intermediate or high risk) in a urine sample from the subject as described herein.
  • the method comprises an induction phase comprising administering the oncolytic virus to the individual once per week for six weeks.
  • the method comprises a maintenance phase subsequent to the induction phase, wherein the maintenance phase comprises administering the oncolytic virus to the individual weekly for three weeks every three or six months.
  • the start of the induction phase and the start of the maintenance phase are separated by about three months or about six months.
  • the oncolytic virus is administered at a dose of about 1 x 10 8 to about 1 x 10 14 viral particles (e.g., about 1 x 10 12 viral particles).
  • the individual is a human individual.
  • the individual being treated for bladder cancer has been identified as having one or more of the conditions described herein. Identification of the conditions as described herein by a skilled physician is routine in the art (e.g., via blood tests, X-rays, ultrasound, CT scans, PET scans, PET/CT scans, MRI scans, PET/MRI scans, nuclear medicine radioisotope scans, endoscopy, biopsy, angiography, CT-angiography, etc.) and may also be suspected by the individual or others, for example, due to tumor growth, hemorrhage, ulceration, pain, enlarged lymph nodes, cough, jaundice, swelling, weight loss, cachexia, sweating, anemia, paraneoplastic phenomena, thrombosis, etc.
  • the individual is selected for any one of the treatment methods described herein based on any one or more of a number of risk factors and/or diagnostic approaches appreciated by the skilled artisan, including, but not limited to, genetic profiling, family history, medical history (e.g., appearance of related conditions and viral infection history), lifestyle or habits.
  • risk factors and/or diagnostic approaches appreciated by the skilled artisan, including, but not limited to, genetic profiling, family history, medical history (e.g., appearance of related conditions and viral infection history), lifestyle or habits.
  • the method prevents progression of the bladder cancer (e.g., NMIBC, e.g., high-risk NMIBC) in the individual.
  • the method prevents progression of NMIBC to muscle invasive bladder cancer by at least about any one of 3, 6, 9, 12, 18, 24, 30, 36, 42, 48, 54, 60 or more months.
  • the method inhibits growth or reduces the size of one or more bladder tumors (e.g. , one or more bladder tumors associated with NMIBC, such as high-risk NMIBC).
  • the size of the bladder tumor is reduced for at least about 10% (including for example at least about any of 20%, 30%, 40%, 60%, 70%, 80%, 90%, or 100%).
  • the method causes bladder cancer remission (partial or complete) in the individual.
  • the individual has bladder cancer remission for at least about any one of 2, 3, 4, 5, 6, 12, 24, or more months.
  • the method inhibits metastasis of bladder tumors in the individual.
  • at least about 10% including for example at least about any of 20%, 30%, 40%, 60%, 70%, 80%, 90%, or 100%
  • metastasis is inhibited.
  • a method of inhibiting metastasis of bladder tumors to lymph nodes or other organs is provided.
  • Metastasis can be assessed by any known methods in the art, such as by blood tests, bone scans, x-ray scans, CT scans, PET scans, and biopsy.
  • the method prolongs survival (such as disease-free survival, progression-free survival, recurrence-free survival, or cystectomy-free survival) in the individual.
  • survival is prolonged for at least about 2, 3, 4, 5, 6, 9, 12, 15, 18, 24, 30, 36 or more months.
  • the method improves quality of life in the individual.
  • the individual does not need a cystectomy (such as partial cystectomy or radical cystectomy) after receiving the therapies described herein for at least about any one of 3, 6, 9, 12, 18, 24, 30, 36, 42, 48, 54, 60 or more months.
  • a cystectomy such as partial cystectomy or radical cystectomy
  • the methods described herein are performed without subjecting the individual to cystectomy (such as radical cystectomy).
  • the methods described herein are bladder-sparing (e.g, prevent the need for a cystectomy for at least about any one of 3, 6, 9, 12, 18, 24, 30, 36, 42, 48, 54, 60 or more months).
  • the intravesical administration of the oncolytic virus in the methods described herein provide a unique opportunity of a relatively convenient yet effective intravesical tumor exposure to the oncolytic virus, as well as a potentially reduced toxicity to other tissues.
  • Suitable dosages for the oncolytic virus depend on factors such as the nature of the oncolytic virus, type of the bladder carcinoma in situ being treated, and routes of administration.
  • particles as related to an oncolytic virus mean the collective number of physical singular units of the oncolytic virus. This number can be converted to, or is equivalent to, another number meaning infectious titer units, e.g., plaque forming unit (pfu) or international unit, by infectivity assays as known in the art.
  • the oncolytic virus is administered at a dose of about any one of IxlO 5 particles, IxlO 6 particles, IxlO 7 particles, IxlO 8 particles, IxlO 9 particles, IxlO 10 particles, 2xlO 10 particles, 5xlO 10 particles, IxlO 11 particles, 2xlO n particles, 5xlO n particles, IxlO 12 particles, 2xl0 12 particles, 5xl0 12 particles, IxlO 13 particles, 2xl0 13 particles, 5xl0 13 particles, IxlO 14 particles, or IxlO 15 particles.
  • the oncolytic virus is administered at a dose of any one of about IxlO 5 particles to about IxlO 6 particles, about IxlO 6 particles to about IxlO 7 particles, about IxlO 7 particles to about IxlO 8 particles, about IxlO 8 particles to about IxlO 9 particles, about IxlO 9 particles to about IxlO 10 particles, about IxlO 10 particles to about IxlO 11 particles, about IxlO 11 particles to about 5xl0 n particles, about 5xl0 n particles to about IxlO 12 particles, about IxlO 12 particles to about 2xl0 12 particles, about 2xl0 12 particles to about 5xl0 12 particles, about 5xl0 12 particles to about IxlO 13 particles, about IxlO 13 particles to about IxlO 14 particles, or about IxlO 14 particles to about IxlO 15 particles.
  • the oncolytic virus is administered daily. In some embodiments, the oncolytic virus is administered is administered at least about any one of lx, 2x, 3x, 4x, 5x, 6x, or 7x (i.e., daily) a week. In some embodiments, the oncolytic virus is administered weekly. In some embodiments, the oncolytic virus is administered weekly without break; weekly, two out of three weeks; weekly three out of four weeks; once every two weeks; once every 3 weeks; once every 4 weeks; once every 6 weeks; once every 8 weeks, monthly, or every two to 12 months.
  • the intervals between each administration are less than about any one of 6 months, 3 months, 1 month, 20 days, 15, days, 12 days, 10 days, 9 days, 8 days, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, or 1 day. In some embodiments, the intervals between each administration are more than about any one of 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 8 months, or 12 months. In some embodiments, there is no break in the dosing schedule. In some embodiments, the interval between each administration is no more than about a week.
  • the administration of the oncolytic virus can be over an extended period of time, such as from about a month up to about seven years.
  • the oncolytic virus is administered over a period of at least about any one of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 30, 36, 48, 60, 72, or 84 months.
  • the oncolytic virus is administered over a period of between 1 and 6 weeks.
  • the oncolytic virus is administered weekly for 6 weeks.
  • the oncolytic virus is administered weekly for 3 weeks every 3 or 6 months.
  • the oncolytic vims is administered in two or more treatment courses, for example, one or more induction courses and/or one or more maintenance courses.
  • each treatment course comprises weekly administration of the oncolytic vims for about 1 to 6 weeks. In some embodiments, each treatment course comprises weekly administration of the oncolytic vims for about 3 weeks or about 6 weeks. In some embodiments, the interval between each treatment course about any one of 1, 2, 3, 4, 5, 6, or more months (e.g., about 3 months or 6 months). In some embodiments, the same treatment course (e.g., the induction course or the maintenance course) is repeated at least once, such as about any of 1, 2, 3, 4, 5, 6, 7, 8, or more times. In some embodiments, the individual is treated with an induction course of the oncolytic vims, followed by a maintenance course. In some embodiments, the induction course and the maintenance course have the same dose and schedule. In some embodiments, the initial course and the maintenance course have different doses and/or schedules.
  • the individual receives no more than 21 doses of an oncolytic vims. In some embodiments, the individual receives no more than 30, 25, 20, 15, 14, 13, 12, 11, 10, 9, 8, 7, or 6 doses of an oncolytic vims.
  • the oncolytic vims is administered during an induction phase and a maintenance phase.
  • the induction phase comprises administering the oncolytic vims once per week for six weeks.
  • the maintenance phase comprises administering the oncolytic vims once per week for three weeks every three or six months.
  • the oncolytic vims is administered once per week for six weeks, followed by once per week for three weeks every three or six months.
  • administering the oncolytic vims comprises administering the oncolytic vims once per week for six weeks, followed by administering the oncolytic vims once per week for one or three weeks every three or six months.
  • the induction phase comprises intravesical administration of the oncolytic vims once per week for six weeks, starting on month 0 of a treatment regimen.
  • the individual is re-evaluated at month 3 of the treatment regimen.
  • the method comprises administering a second induction phase treatment course of intravesical administration of the oncolytic vims once per week for six weeks.
  • the induction phase comprises administering the oncolytic vims once per week for six weeks.
  • the maintenance phase comprises administering the oncolytic virus once per week for three weeks about every 12 weeks or 24 weeks.
  • the oncolytic virus is administered once per week for six weeks, followed by once per week for three weeks about every 12 weeks or about every 24 weeks.
  • administering the oncolytic virus comprises administering the oncolytic virus once per week for six weeks, followed by administering the oncolytic virus once per week for one or three weeks about every 12 weeks or about every 24 weeks.
  • the induction phase comprises intravesical administration of the oncolytic virus once per week for six weeks, starting on week 0 of a treatment regimen.
  • the individual is re-evaluated at week 13 of the treatment regimen.
  • the method comprises intravesically administering the oncolytic virus once per week for 3 weeks every 12 or 24 weeks starting at week 13.
  • the method further comprises intravesical administration of the oncolytic virus once per week for three weeks starting at weeks 13, 25, 37, 49, and 73.
  • the method comprises administering a second induction phase treatment course of intravesical administration of the oncolytic virus once per week for six weeks starting at week 13. In certain embodiments, if the individual has not experienced a complete response (absence of detectable bladder tumor) at week 13, the method further comprises a) intravesical administration of the oncolytic virus once per week for six weeks starting at week 13, and b) intravesical administration of the oncolytic virus once per week for three weeks starting at weeks 25, 37, 49, and 73.
  • the maintenance phase comprises administering an oncolytic virus once per week for three weeks, following an induction phase. In some embodiments, the maintenance phase comprises delivering the oncolytic virus once per week for three weeks every three or six months. In some embodiments, the maintenance phase comprises administering the oncolytic virus weekly for three weeks every 3 or 6 months 2, 3, 4, 5, 6, 7, or 8 times, or as needed. In some embodiments, the maintenance phase comprises administering an oncolytic virus weekly for one or three weeks on months 6, 12, and 18, where month 0 begins at the first induction phase administration. In some embodiments, the maintenance phase comprises administering an oncolytic virus weekly for one or three weeks on months 3, 6, 12, and 18, where month 0 begins at the first induction phase administration.
  • the dosage schedule can be modified based upon the individual’s response to the oncolytic virus.
  • an individual is administered an oncolytic virus once per week for six weeks on month 0 or month 1 and is reevaluated three months later (e.g., at month 3 or month 4).
  • individuals who have a complete response when reevaluated begin a maintenance phase comprising administration of an oncolytic virus once per week for 1 to 6 weeks (e.g, 3 weeks) every 3 or 6 months.
  • individuals who do not have a complete response at 3 months receive a second induction dose of oncolytic virus once per week for 6 weeks.
  • the oncolytic virus is administered by instillation as a solution via a catheter.
  • the total volume of the solution used for the intravesical installation is about any of 1 mb, 10 mb, 50 mb, 75 mb, 100 mb, 125 mb, 150 mb, 200 mb, 250 mb, 300 mb, 400 mb, or 500 mb.
  • the total volume of the solution used for the intravesical installation is any of about 1 mb to about 10 mb, about 10 mb to about 50 mb, about 50 mb to about 75 mb, about 50 mb to about 100 mb, about 75 mb to about 100 mb, about 85 mb to about 100 mb, about lOOmL to about 125 mb, about 75 mb to about 125 mb, about 100 mb to about 150 mb, about 150 mb to about 200 mb, about 200 mb to about 300 mb, about 300 mb to about 400 mb, about 400 mb to about 500 mb, about 50 mb to about 500 mb, about 50 mb to about 250 mb, or about 100 mb to about 250 mb.
  • the oncolytic virus is administered in a volume of between about 1 mb and about 10 mb, between about 10 mb and about 50 mb, between about 50 mb and about 75 mb, between about 50 mb and about 100 mb, between about 75 mb and about 100 mb, between about lOOmL and about 125 mb, between about 75 mb and about 125 mb, between about 100 mb and about 150 mb, between about 150 mb and about 200 mb, between about 200 mb and about 300 mb, between about 300 mb and about 400 mb, between about 400 mb and about 500 mb, between about 50 mb and about 500 mb, between about 50 mb and about 250 mb, or between about 100 mb and about 250 mb.
  • the oncolytic virus is administered at a dose of about IxlO 8 to about IxlO 15 particles (such as about IxlO 11 to about IxlO 14 particles, for example about 1x10 12 particles). In some embodiments, the oncolytic virus is administered at a volume of about 50 to about 500mL (such as about 50 mb, about 100 mb, or about 150mL) by instillation. In some embodiments, the oncolytic virus is administered at a dose of about IxlO 12 in about 50 mb. [0252] In some embodiments of the methods described herein, the method comprises one or more intravesical instillations of the oncolytic virus.
  • the intravesical instillation of the oncolytic virus comprises administering the oncolytic virus into the bladder and allowing the oncolytic virus to dwell in the bladder for a dwell time of at least about 5 minutes before draining the bladder. In some embodiments, the intravesical instillation of the oncolytic virus comprises allowing the oncolytic virus to dwell in the bladder for a dwell time of about 5, 10, 15, 20, 25, 30, 45, or 60 minutes. In certain embodiments, the intravesical instillation of the oncolytic virus comprises allowing the oncolytic virus to dwell in the bladder for a dwell time of about 60 minutes, or between about 55 minutes and about 65 minutes. In certain embodiments, the duration of the dwell time begins after completion of the administration of the full volume of the oncolytic virus into the bladder.
  • the solution of the oncolytic virus may be retained in the bladder for a certain amount of time before voiding, in order to achieve uniform distribution or sufficient exposure of the oncolytic virus among the bladder tumor cells.
  • the solution is retained in the bladder of the individual for at least about any of 5 minutes, 10 minutes, 15 minutes, 20 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, or more.
  • the solution is retained in the bladder of the individual for any of about 5 minutes to aboutlO minutes, about 10 minutes to about 15 minutes, about 10 minutes to about 20 minutes, about 20 minutes to about 30 minutes, about 30 minutes to about 45 minutes, about 45 minutes to about 50 minutes, about 50 minutes to about 1 hour, about 5 minutes to about 15 minutes, about 10 minutes to about 30 minutes, about 30 minutes to about 1 hour, or about 1 hour to about 2 hours.
  • the oncolytic virus e.g., cretostimogene
  • the efficiency of the intravesical administration of the oncolytic virus is further enhanced by a pretreatment comprising intravesical administration of an effective amount of a transduction enhancing agent, such as DDM.
  • the methods described herein may further comprise a step of intravesically administering to the individual a pretreatment composition prior to the administration of the oncolytic virus.
  • the pretreatment composition comprises a transduction enhancing agent, such as N-Dodecyl-[3-D-maltoside (DDM).
  • DDM is a nonionic surfactant comprised of a maltose derivatized with a single twelve-carbon chain, and acts as a mild detergent and solubilizing agent. It has been used as a food additive and is known to enhance mucosal surface permeation in rodents, probably due to its effect on membrane associated GAG and tight junctions.
  • the pretreatment composition is administered intravesically.
  • the pretreatment composition comprises a solution of the transduction enhancing agent (such as DDM).
  • Suitable concentration of the pretreatment composition includes, but are not limited to, about any one of 0.01%, 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 1%, 2%, 3%, 4%, or 5% of the transducing enhancing agent (such as DDM).
  • the pretreatment composition comprises any of about 0.01% to about 0.05%, about 0.05% to about 0.1%, about 0.1% to about 0.5%, about 0.5% to about 1%, about 1% to about 2%, about 2% to about 3%, about 3% to about 4%, about 4% to about 5%, about 0.01% to about 1%, about 0.05% to about 2%, about 1% to about 5%, or about 0.1% to about 5% of the transduction enhancing agent (such as DDM).
  • the transduction enhancing agent such as DDM
  • each administration of the oncolytic virus is administered as a part of a two-step instillation regimen, wherein each two- step instillation regimen comprises 1) intravesically administering a pretreatment composition comprising a transduction-enhancing agent to the individual; and subsequently 2) intravesically administering the oncolytic virus to the individual directly after administering the pretreatment composition, wherein there is no intervening bladder wash between the administration of the pretreatment composition and the administration of the oncolytic virus.
  • the methods described herein may comprise administering the two-step instillation regimen as a part of any treatment regimen described herein.
  • weekly administration of the oncolytic virus comprises weekly administration of the two-step instillation regimen.
  • Suitable dosages for the pretreatment composition include, but are not limited to, about any of 0.1 mg/kg, 0.5 mg/kg, 1 mg/kg, 1.5mg/kg, 2 mg/kg, 2.5 mg/kg, 5mg/kg, 10 mg/kg, 25 mg/kg, 50 mg/kg, 100 mg/kg, 150 mg/kg, 200 mg/kg, 250 mg/kg, 300 mg/kg, 400 mg/kg, 500 mg/kg, 0.
  • 1 mg/kg to 0.5 mg/kg 0.5 mg/kg to 1 mg/kg, 0.5 mg/kg to 1 mg/kg, 1 mg/kg to 2 mg/kg, 2 mg/kg to 5mg/kg, 5mg/kg to 10 mg/kg, 10 mg/kg to 25 mg/kg, 25 mg/kg to 50 mg/kg, 50 mg/kg to 100 mg/kg, 100 mg/kg to 150 mg/kg, 150 mg/kg to 200 mg/kg, 200 mg/kg to 250 mg/kg, 250 mg/kg to 500 mg/kg, or 0.5 mg/kg to about 5 mg/kg.
  • a suitable dosage for the pretreatment composition is about any one of 0.1 g, 0.2 g, 0.5g, 0.75 g, 1 g, 1.5 g, 2 g, 2.5 g, 5 g, or 10 g of the transduction enhancing agent (such as DDM).
  • the transduction enhancing agent such as DDM
  • the pretreatment step is carried out by contacting the luminal surface of the bladder in the individual with the pretreatment composition prior to the administration of the oncolytic virus.
  • the pretreatment composition may comprise about 0.01% to about 0.5% (such as about 0.05 to about 0.2%, for example about 0.1%) of the transduction enhancing agent (such as DDM).
  • the total volume of the pretreatment composition (such as DDM) is about 10 mL to about 1000 mL (such as about 10 mL to about 100 mL, about 100 mL to about 500 mL, or about 500 mL to about 1000 mL).
  • the pretreatment composition comprises about 0.1% DDM.
  • a suitable dosage for the pretreatment composition is about any one of 0.1 g, 0.2 g, 0.5g, 0.75 g, 1 g, 1.5 g, 2 g, 2.5 g, 5 g, or 10 g of the transduction enhancing agent (such as DDM).
  • the effective amount of the pretreatment composition is about 1g of DDM (e.g., 100 mL of 0. 1% DDM solution).
  • the pretreatment step comprises intravesically instilling 75 mL or 100 mL of a pretreatment composition (such as a DDM solution).
  • the pretreatment composition (such as DDM) is administered immediately (such as no more than 5 minutes) prior to the administration of the oncolytic virus.
  • the oncolytic virus is administered without a saline wash following DDM administration.
  • the pretreatment composition (such as DDM) is administered no more than about any of 5 minutes, 10 minutes, 15 minutes, 20 minutes, 30 minutes, 45 minutes, 1 hour, 90 minutes, 2 hours, 3 hours, or 4 hours before the administration of the oncolytic virus.
  • the pretreatment composition (such as DDM) is administered no more than about 2 hours before the administration of the oncolytic virus.
  • the pretreatment composition (such as DDM solution) is retained in the bladder for at least about any one of 5 minutes, 10 minutes, 15 minutes, or 20 minutes. In some embodiments, the pretreatment composition (such as DDM solution) is retained in the bladder for any of about 5 minutes to about 10 minutes, about 10 minutes to about 15 minutes, about 12 minutes to about 15 minutes, about 15 minutes to about 20 minutes, or about 10 minutes to about 20 minutes. In some embodiments, the pretreatment composition (such as DDM solution) is retained in the bladder for about 12 minutes to about 15 minutes.
  • the pretreatment step is carried out by contacting the luminal surface of the bladder in the individual with the pretreatment composition prior to the administration of the oncolytic virus. In some embodiments, the method further comprises washing the luminal surface of the bladder contact with the pretreatment composition. In some embodiments, the method further comprises washing the luminal surface of the bladder after contacting the bladder with the pretreatment composition prior to the administration of the oncolytic virus.
  • the individual is a human individual. In some embodiments, the individual has been identified as having one or more of the bladder cancer conditions described herein.
  • Identification of the conditions as described herein by a skilled physician is routine in the art (e.g., via blood tests, X-rays, ultrasound, CT scans, PET scans, PET/CT scans, MRI scans, PET/MRI scans, nuclear medicine radioisotope scans, endoscopy, biopsy, angiography, CT-angiography, etc.) and may also be suspected by the individual or others, for example, due to tumor growth, hemorrhage, ulceration, pain, enlarged lymph nodes, cough, jaundice, swelling, weight loss, cachexia, sweating, anemia, paraneoplastic phenomena, thrombosis, etc.
  • the individual is selected for any one of the treatment methods described herein based on any one or more of a number of risk factors and/or diagnostic approaches appreciated by the skilled artisan, including, but not limited to, genetic profiling, family history, medical history (e.g., appearance of related conditions and viral infection history), lifestyle or habits.
  • risk factors and/or diagnostic approaches appreciated by the skilled artisan, including, but not limited to, genetic profiling, family history, medical history (e.g., appearance of related conditions and viral infection history), lifestyle or habits.
  • Suitable dosages for the additional therapeutic agents described herein depend on factors such as the nature of the additional therapeutic agent or combination of additional therapeutic agent and the routes of administration.
  • Exemplary doses of the additional therapeutic agent include, but are not limited to, about any one of 1 mg/m 2 , 5 mg/m 2 , 10 mg/m 2 , 20 mg/m 2 , 50 mg/m 2 , 100 mg/m 2 , 200 mg/m 2 , 300 mg/m 2 , 400 mg/m 2 , 500 mg/m 2 , 750 mg/m 2 , 1000 mg/m 2 , or more.
  • the dose of the additional therapeutic agent is included in any one of the following ranges: about 1 to about 5 mg/m 2 , about 5 to about 10 mg/m 2 , about 10 to about 20 mg/m 2 , about 20 to about 50 mg/m 2 , about 50 to about 100 mg/m 2 , about 100 mg/m 2 to about 200 mg/m 2 , about 200 to about 300 mg/m 2 , about 300 to about 400 mg/m 2 , about 400 to about 500 mg/m 2 , about 500 to about 750 mg/m 2 , or about 750 to about 1000 mg/m 2 .
  • the additional therapeutic agent e.g., immune checkpoint modulator or chemotherapeutic agent
  • the dose of the additional therapeutic agent is about any one of 1 pg/kg, 2 pg/kg, 5 pg/kg, 10 pg/kg, 20 pg/kg, 50 pg/kg, 0.1 mg/kg, 0.2 mg/kg, 0.3 mg/kg, 0.4 mg/kg, 0.5 mg/kg, 1 mg/kg, 2 mg/kg, 5 mg/kg, 10 mg/kg, 20 mg/kg, 50 mg/kg, 100 mg/kg, or more.
  • the additional therapeutic agent e.g, immune checkpoint modulator or chemotherapeutic agent
  • the dose of the additional therapeutic agent is any one of about 1 pg/kg to about 5 pg/kg, about 5 pg/kg to about 10 pg/kg, about 10 pg/kg to about 50 pg/kg, about 50 pg/kg to about 0.
  • the additional therapeutic agent e.g., immune checkpoint modulator or chemotherapeutic agent
  • 1 mg/kg about 0.1 mg/kg to about 0.2 mg/kg, about 0.2 mg/kg to about 0.3 mg/kg, about 0.3 mg/kg to about 0.4 mg/kg, about 0.4 mg/kg to about 0.5 mg/kg, about 0.5 mg/kg to about 1 mg/kg, about 1 mg/kg to about 5 mg/kg, about 5 mg/kg to about 10 mg/kg, about 10 mg/kg to about 20 mg/kg, about 20 mg/kg to about 50 mg/kg, about 50 mg/kg to about 100 mg/kg, or about 1 mg/kg to about 100 mg/kg.
  • the dose of the additional therapeutic agent is about any one of 1 pg, 10 pg, 50 pg, 100 pg, 500 pg, 1 mg, 2 mg, 4 mg, 6 mg, 12 mg, 18 mg, 24 mg, 50 mg, 100 mg, 500 mg or 1000 mg.
  • the dose of the additional therapeutic agent is any one of about 1 pg to about 10 pg, about 10 pg to about 50 10 pg, about 50 pg to about 100 pg, about 100 pg to about 500 pg, about 500 pg to about 1 mg, about 1 mg to about 5 mg, about 5 mg to about 10 mg, about 10 mg to about 25 mg, about 25 mg to about 50 mg, about 50 mg to about 100 mg, about 100 mg to about 500 mg, about 500 mg to about 1000 mg, about 1 pg to about 1 mg, about 1 mg to about 1000 mg, or about 1 pg to about 1000 mg.
  • the additional therapeutic agent e.g., immune checkpoint modulator or chemotherapeutic agent
  • the dose of the additional therapeutic agent (e.g., immune checkpoint modulator or chemotherapeutic agent) administered per tumor site is no more than about any of 10 pg, 50 pg, 100 pg, 500 pg, 1 mg, 2 mg, 4 mg, 6 mg, 12 mg, 18 mg, 24 mg, 50 mg, or 100 mg.
  • the dose of the additional therapeutic agent (e.g., immune checkpoint modulator or chemotherapeutic agent) administered locally (e.g., intravesically) is any one of about 10 pg to about 50 pg, about 50 pg to about 100 pg, about 100 pg to about 500 pg, about 100 pg to about 1 mg, about 1 mg to about 2 mg, about 2 mg to about 5 mg, about 5 mg to about 10 mg, about 10 mg to about 15 mg, about 10 mg to about 25 mg, about 25 mg to about 50 mg, about 50 mg to about 100 mg, about 1 mg to about 50 mg, or about 100 pg to about 10 mg.
  • the dose of the additional therapeutic agent (e.g., immune checkpoint modulator or chemotherapeutic agent) administered locally (e.g., intravesically) is based on the size of the tumor.
  • the additional therapeutic agent (e.g., immune checkpoint modulator or chemotherapeutic agent)is administered daily.
  • the additional therapeutic agent e.g., immune checkpoint modulator or chemotherapeutic agent
  • is administered is administered at least about any one of lx, 2x, 3x, 4x, 5x, 6x, or 7x (i.e., daily) a week.
  • the additional therapeutic agent e.g., immune checkpoint modulator or chemotherapeutic agent
  • the intervals between each administration are less than about any one of 6 months, 3 months, 1 month, 20 days, 15, days, 12 days, 10 days, 9 days, 8 days, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, or 1 day. In some embodiments, the intervals between each administration are more than about any one of 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 8 months, or 12 months. In some embodiments, there is no break in the dosing schedule. In some embodiments, the interval between each administration is no more than about a week.
  • the additional therapeutic agent e.g., immune checkpoint modulator or chemotherapeutic agent
  • the additional therapeutic agent is administered with the same dosing schedule as the oncolytic virus. In some embodiments, the additional therapeutic agent (e.g., immune checkpoint modulator or chemotherapeutic agent) is administered with a different dosing schedule as the oncolytic virus. In some embodiments, the oncolytic virus is administered weekly for four weeks.
  • the administration of the additional therapeutic agent can be over an extended period of time, such as from about a month up to about seven years.
  • the additional therapeutic agent e.g., immune checkpoint modulator or chemotherapeutic agent
  • the additional therapeutic agent e.g., immune checkpoint modulator or chemotherapeutic agent
  • the additional therapeutic agent e.g., the immune checkpoint modulator or chemotherapeutic agent
  • the additional therapeutic agent is administered about every six weeks, about every three months, or about every six months.
  • the additional therapeutic agent e.g., the immune checkpoint modulator or chemotherapeutic agent
  • the additional therapeutic agent is an immune checkpoint inhibitor and is administered systemically (e.g., systemically) about once every six weeks for about six months, and subsequently about once every three months for about six months, and further subsequently about once every six months for about twelve months.
  • the additional therapeutic agent is an immune checkpoint inhibitor and is administered systemically (e.g., systemically) about once every six weeks for about 24 weeks, and subsequently about once every 12 weeks for about 24 weeks, and further subsequently about once every 24 weeks for about 48 weeks.
  • the immune checkpoint inhibitor is administered weekly at weeks 0, 7, 13, 19, 25, 37, 49, 73, 79, and 97 of a treatment regimen.
  • the immune checkpoint inhibitor is an inhibitor of PD-1 (e.g., an anti-PD-1 antibody, e.g., pembrolizumab or nivolumab) or an inhibitor of PD-L1 (e.g., an anti-PD-Ll antibody).
  • the immune checkpoint inhibitor is administered intravenously.
  • Exemplary routes of administration of the oncolytic virus, the additional therapeutic agent (e.g., immune checkpoint modulator or chemotherapeutic agent), prior therapy, and/or the pretreatment compositions include, but are not limited to, intratumoral, intravesical, intramuscular, intraperitoneal, intravenous, intra-arterial, intracranial, intrapleural, subcutaneous, and epidermal routes, or be delivered into lymph glands, body spaces, organs or tissues known to contain such live cancer cells (such as intrahepatic or intrapancreatic injections).
  • the local administration is carried out by direct injection of the agent(s) into the tumor.
  • the local administration is carried out by direct injection of the agent(s) to a site close to the tumor cells.
  • the local administration is carried out intravesically.
  • the systemic administration is via intravenous infusion.
  • GDB Genomic Disease Burden
  • kits for treating bladder cancer in an individual comprising intravesically administering to the individual an effective amount of an oncolytic virus and, based on a determined genomic disease burden (GDB) value, a determined VAF value, an assigned minimum residual disease (MRD) status, and/or assigned risk status, in a first urine sample from the individual, administering to the individual one or more additional therapeutic agents.
  • GDB genomic disease burden
  • VAF genomic disease burden
  • MRD assigned minimum residual disease
  • risk status in a first urine sample from the individual
  • GDB tumor mutational burden
  • VAF percentile of variant allele frequency
  • the VAF from the urine samples may be determined using methods known in the art to be suitable for detecting variant alleles in a sample, for example, nucleic acid sequencing methods, such as next-generation DNA or RNA sequencing or probe-based detection of mutations.
  • VAF may be determined using whole-genome sequencing or amplicon sequencing of nucleic acids in the urine sample.
  • the VAF may be calculated for any number of genes, such as for a specific set of genes associated with bladder cancer (e.g., for about 10, 20, 30, 40, 50, 60, 70, 80, 90, or 100 or more genes).
  • UroAmp® Convergent Genomics
  • Methods of determining GDB are known in the art, including, for example in Salari et al. (2023. "Development and Multicenter Case-Control Validation of Urinary Comprehensive Genomic Profiling for Urothelial Carcinoma Diagnosis, Surveillance, and Risk-Prediction.” Clinical Cancer Research 29.18: 3668-3680), Bicocca et al. (2022.
  • the methods described herein comprise intravesically administering to the individual an effective amount of an oncolytic virus; and, based on a determined GDB value, a determined VAF value, an assigned MRD status, and/or assigned risk status, in a urine sample from the individual, administering one or more additional therapeutic agents to the individual.
  • the method comprises determining the GDB value, the VAF value, or both.
  • one or more additional therapeutic agents are selected from the group consisting of a chemotherapeutic agent, a targeted therapy agent, an immune checkpoint modulator, and an immunomodulatory agent.
  • the additional therapeutic agent is administered based on a determination that the GDB value or VAF value exceeds a predetermined threshold. In some embodiments, the additional therapeutic agent is administered based on an assigned risk status of intermediate risk or high risk. In some embodiments, the additional therapeutic agent is administered based on an assigned MRD status of MRD-positive.
  • the GDB value (e.g, as determined by UroAmp®) is a percentile-based metric that quantifies the extent of genomic alterations in a patient's urine-derived DNA, comparing it to a reference population of bladder cancer patients. Higher GDB scores indicate a greater burden of disease and are associated with an increased risk of recurrence and progression in non-muscle invasive bladder cancer (NMIBC).
  • NMIBC non-muscle invasive bladder cancer
  • GDB values are presented as numerical values corresponding to percentile. For example, a GDB value of 20 corresponds to the 20 th percentile; a GDB value of 42 correspond to the 42 nd percentile; a GDB value of 80 corresponds to the 80 th percentile; and the like.
  • the percentile ranking of GDB scores can be used to stratify patients into “low,” “intermediate,” and “high” risk groups. “Low GDB” or “low risk” patients have GDB scores in the lower percentiles, often associated with minimal genomic alterations and a favorable prognosis. “Intermediate GDB” or “intermediate risk” patients have GDB scores in the mid-range percentiles, indicating a moderate level of genomic alterations and an intermediate risk of disease recurrence or progression. “High GDB” or “High-risk” patient have GDB scores in the upper percentiles, reflecting a significant burden of genomic alterations and a high risk of adverse outcomes.
  • Percentile cutoffs for categorizing GDB scores as low, intermediate, or high may vary depending on the specific study, tumor context, and clinical setting. Any percentile cutoff or range described herein may be used in the methods described herein to assign a risk status of low, intermediate, or high risk to an individual having bladder cancer.
  • the methods described herein comprise an initial therapy comprising intravesically administering to the individual an effective amount of an oncolytic virus, wherein the initial therapy is administered based on a determination of a first genomic disease burden (GDB) value in a first urine sample (e.g. at a first timepoint, e.g., at baseline) from the individual, wherein the individual is assigned a first risk status of low risk, intermediate risk, or high risk based on the first GDB value.
  • GDB genomic disease burden
  • the initial therapy further comprises administration of one or more additional therapeutic agents selected from the group consisting of a chemotherapeutic agent, a targeted therapy agent, an immune checkpoint modulator, and an immunomodulatory agent.
  • the initial therapy does not comprise administration of the additional therapeutic agent.
  • the method further comprises: subsequent to the initial therapy, administering a subsequent therapy to the individual, wherein the subsequent therapy is administered based on a determination of a second GDB value in a second urine sample from the individual (e.g., at a second, later timepoint (e.g. 3 months or 6 months) after the initial therapy, the treatment decided by the first GDB value), wherein the individual is assigned a second risk status of low risk, intermediate risk, or high risk based on the second GDB value.
  • the subsequent therapy comprises intravesical administration of an effective amount of the oncolytic virus and administration of one or more additional therapeutic agents (e.g., one or more additional therapeutic agents that was not administered during the initial therapy).
  • the method comprises determining the first GDB value, the second GDB value, or both.
  • the method comprises assigning the first risk status, the second risk status, or both to the individual based on the first GDB value, the second GDB value, or both, respectively.
  • one or more additional therapeutic agents are selected from the group consisting of a chemotherapeutic agent, a targeted therapy agent, an immune checkpoint modulator, and an immunomodulatory agent.
  • the methods described herein comprise administering an initial therapy comprising intravesically administering to the individual an effective amount of an oncolytic virus, wherein the initial therapy is administered based on a determination of an initial minimum residual disease (MRD) status of MRD-positive or MRD-negative, wherein the individual is assigned an initial MRD status of MRD-positive or MRD-negative based on a first variant allele frequency (VAF) value in a first urine sample from the individual at a first timepoint.
  • MRD initial minimum residual disease
  • VAF variant allele frequency
  • the initial therapy further comprises administration of one or more additional therapeutic agents selected from the group consisting of a chemotherapeutic agent, a targeted therapy agent, an immune checkpoint modulator, and an immunomodulatory agent.
  • the initial therapy does not comprise administration of the additional therapeutic agent.
  • the method further comprises: subsequent to the initial therapy, administering a subsequent therapy to the individual, wherein the subsequent therapy is administered based on a determination of an updated MRD status in a second urine sample from the individual (e.g., at a second, later timepoint (e.g.
  • the method comprises determining the first VAF value, the second VAF value, or both. In some embodiments, the method comprises assigning the initial MRD status, the updated MRD status, or both to the individual based on the first VAF value, the second VAF value, or both, respectively.
  • the method comprises determining the first genomic disease burden (GDB) value, the second GDB value, or both. In some embodiments, the method comprises assigning the initial MRD status, the updated MRD status, or both to the individual based on the first GDB value, the second GDB value, or both, respectively. In some embodiments, one or more additional therapeutic agents are selected from the group consisting of a chemotherapeutic agent, a targeted therapy agent, an immune checkpoint modulator, and an immunomodulatory agent.
  • MRD status is assigned to an individual based on a VAF value in a urine sample from the subject.
  • an MRD status of MRD-negative is assigned to the individual if the VAF value is less than a certain threshold, e.g., less than 20%, less than 15%, less than 11%, less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, or less than 1%.
  • a certain threshold e.g., less than 20%, less than 15%, less than 11%, less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, or less than 1%.
  • an MRD status of MRD-negative is assigned to the individual if the VAF is less than 8%.
  • an MRD status of MRD-positive is assigned to the individual if the VAF is at least a certain threshold, e.g., at least 20%, at least 15%, at least 11%, at least 10%, at least 9%, at least 8%, at least 7%, at least 6%, at least 5%, at least 4%, at least 3%, at least 2%, or at least 1%. In certain embodiments, an MRD status of MRD-positive is assigned to the individual if the VAF is at least 8%.
  • MRD status is assigned to an individual based on a VAF value and a GDB value in a urine sample from the subject.
  • an MRD status of MRD-negative is assigned to the individual if the VAF value is less than a certain threshold (e.g., less than 20%, less than 15%, less than 11%, less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, or less than 1%) and the GDB value is less than a certain threshold (e.g., less than 50, less than 45, less than 44, less than 43, less than 42, less than 41, less than 40, or less than 35).
  • a certain threshold e.g., less than 50, less than 45, less than 44, less than 43, less than 42, less than 41, less than 40, or less than 35.
  • an MRD status of MRD-negative is assigned to the individual if the VAF is less than 8% and the GDB is less than 42. In some embodiments, an MRD status of MRD-positive is assigned to the individual if the GDB value is at least a certain threshold (e.g., at least 50, at least 45, at least 44, at least 43, at least 42, at least 41, at least 40, or at least 35). In certain embodiments, an MRD status of MRD-positive is assigned to the individual if the GDB value is at least a certain threshold (e.g., at least 50, at least 45, at least 44, at least 43, at least 42, at least 41, at least 40, or at least 35) independent of the VAF value.
  • a certain threshold e.g., at least 50, at least 45, at least 44, at least 43, at least 42, at least 41, at least 40, or at least 35
  • an MRD status of MRD-positive is assigned to the individual if the GDB value is at least 42, independent of the VAF value.
  • the methods described herein comprise administering an initial therapy comprising intravesical administration of an oncolytic virus based on a determination of a first genomic disease burden (GDB) value in a first urine sample from the individual at a first timepoint, and administering a subsequent therapy based on the difference between the first GDB value and a second GDB value determined in a second urine sample from the individual at a second timepoint subsequent to the initial therapy.
  • the subsequent therapy is administered based on the difference between the first GDB value and the second GDB value.
  • the subsequent therapy comprises an escalated therapy, such as administration of the oncolytic virus in combination with one or more additional therapeutic agents (e.g., one or more immune checkpoint modulators or chemotherapeutic agents).
  • the subsequent therapy is the same as the initial therapy or is a de-escalated therapy, such as administration of the oncolytic virus without administration of one or more additional therapeutic agents.
  • the individual is assigned a first risk status of low risk, intermediate risk, or high risk based on the first GDB value.
  • the initial therapy comprises intravesical administration of an oncolytic virus and administration of one or more additional therapeutic agents selected from the group consisting of a chemotherapeutic agent, a targeted therapy agent, an immune checkpoint modulator, and an immunomodulatory agent.
  • the initial therapy comprises intravesical administration of the oncolytic virus without administration of the additional therapeutic agent.
  • the individual is assigned a first risk status of high risk based on a determination that the first GDB value exceeds a predetermined threshold.
  • the subsequent therapy is administered based on a difference between the first risk status and a second risk status of low risk, intermediate risk, or high risk assigned based on a second GDB value determined in a second urine sample from the individual at a second timepoint subsequent to the initial therapy.
  • the subsequent therapy is an escalated therapy comprising administration of the oncolytic virus and administration of the additional therapeutic agent, and, optionally, administration of one or more further additional therapeutic agents selected from the group consisting of a chemotherapeutic agent, a targeted therapy agent, an immune checkpoint modulator, and an immunomodulatory agent.
  • the subsequent therapy is a de-escalated therapy comprising administration of the oncolytic virus without administration of the additional therapeutic agent.
  • the subsequent therapy is an escalated therapy comprising administration of the oncolytic virus and administration of the additional therapeutic agent, and, optionally, administration of one or more further additional therapeutic agents selected from the group consisting of a chemotherapeutic agent, a targeted therapy agent, an immune checkpoint modulator, and an immunomodulatory agent.
  • the subsequent therapy is a de-escalated therapy comprising administration of the oncolytic virus without administration of the additional therapeutic agent.
  • the initial therapy comprises intravesical administration of the oncolytic virus without administration of the additional therapeutic agent
  • the escalated therapy comprises intravesical administration of the oncolytic virus and administration of the additional therapeutic agent.
  • the initial therapy comprises intravesical administration of the oncolytic virus and administration of the additional therapeutic agent
  • the escalated therapy comprises intravesical administration of the oncolytic virus, the additional therapeutic agent, and one or more further additional therapeutic agents selected from the group consisting of a chemotherapeutic agent, a targeted therapy agent, an immune checkpoint modulator, and an immunomodulatory agent.
  • the initial therapy comprises intravesical administration of the oncolytic virus and administration of the additional therapeutic agent
  • the de-escalated therapy comprises intravesical administration of the oncolytic virus without administration of the additional therapeutic agent.
  • the methods described herein comprise administering an initial therapy comprising intravesical administration of an oncolytic virus based on a determination of a first variant allele frequency (VAF) value (e.g., a first maximum VAF value) in a first urine sample from the individual at a first timepoint, and administering a subsequent therapy based on the difference between the first VAF value (e.g., first maximum VAF value) and a second VAF value (e.g., a second maximum VAF value) determined in a second urine sample from the individual at a second timepoint subsequent to the initial therapy.
  • the subsequent therapy is administered based on the difference between the first VAF value and the second VAF value.
  • the subsequent therapy comprises an escalated therapy, such as administration of the oncolytic virus in combination with one or more additional therapeutic agents (e.g., one or more immune checkpoint modulators or chemotherapeutic agents).
  • the subsequent therapy is the same as the initial therapy or is a de-escalated therapy, such as administration of the oncolytic virus without administration of one or more additional therapeutic agents.
  • the individual is assigned a first GDB value based on the first VAF value.
  • the individual is assigned a first risk status of low risk, intermediate risk, or high risk based on the for VAF value or the first GDB value.
  • the initial therapy comprises intravesical administration of an oncolytic virus and administration of one or more additional therapeutic agents selected from the group consisting of a chemotherapeutic agent, a targeted therapy agent, an immune checkpoint modulator, and an immunomodulatory agent.
  • the initial therapy comprises intravesical administration of the oncolytic virus without administration of the additional therapeutic agent.
  • the individual is assigned a first risk status of high risk based on a determination that the first VAF value or the first GDB value exceeds a predetermined threshold. In some embodiments, the individual is assigned a second GDB value based on the second VAF value. In some embodiments, the subsequent therapy is administered based on a difference between the first risk status and a second risk status of low risk, intermediate risk, or high risk assigned based on the second VAF value or the second GDB value.
  • the subsequent therapy is an escalated therapy comprising administration of the oncolytic virus and administration of the additional therapeutic agent, and, optionally, administration of one or more further additional therapeutic agents selected from the group consisting of a chemotherapeutic agent, a targeted therapy agent, an immune checkpoint modulator, and an immunomodulatory agent.
  • the subsequent therapy is a de-escalated therapy comprising administration of the oncolytic virus without administration of the additional therapeutic agent.
  • the subsequent therapy is an escalated therapy comprising administration of the oncolytic virus and administration of the additional therapeutic agent, and, optionally, administration of one or more further additional therapeutic agents selected from the group consisting of a chemotherapeutic agent, a targeted therapy agent, an immune checkpoint modulator, and an immunomodulatory agent.
  • the subsequent therapy is a de-escalated therapy comprising administration of the oncolytic virus without administration of the additional therapeutic agent.
  • the initial therapy comprises intravesical administration of the oncolytic virus without administration of the additional therapeutic agent
  • the escalated therapy comprises intravesical administration of the oncolytic virus and administration of the additional therapeutic agent.
  • the initial therapy comprises intravesical administration of the oncolytic virus and administration of the additional therapeutic agent
  • the escalated therapy comprises intravesical administration of the oncolytic virus, the additional therapeutic agent, and one or more further additional therapeutic agents selected from the group consisting of a chemotherapeutic agent, a targeted therapy agent, an immune checkpoint modulator, and an immunomodulatory agent.
  • the initial therapy comprises intravesical administration of the oncolytic virus and administration of the additional therapeutic agent
  • the de-escalated therapy comprises intravesical administration of the oncolytic virus without administration of the additional therapeutic agent.
  • the methods described herein comprise administering an initial therapy comprising intravesical administration of an oncolytic virus based on a determination of a first genomic disease burden (GDB) value in a first urine sample from the individual at a first timepoint, and administering a subsequent therapy based on the difference between the first risk status assigned by the first GDB value and a second risk status assigned by a second GDB value determined in a second urine sample from the individual at a second timepoint subsequent to the initial therapy.
  • the subsequent therapy is administered based on the difference between the first GDB risk status and the second GDB risk status.
  • the subsequent therapy comprises an escalated therapy, such as administration of the oncolytic virus in combination with one or more additional therapeutic agents (e.g., one or more immune checkpoint modulators or chemotherapeutic agents).
  • the subsequent therapy is the same as the initial therapy or is a de-escalated therapy, such as administration of the oncolytic virus without administration of one or more additional therapeutic agents.
  • the individual is assigned a first risk status of low risk, intermediate risk, or high risk based on the first GDB value.
  • the initial therapy comprises intravesical administration of an oncolytic virus and administration of one or more additional therapeutic agents selected from the group consisting of a chemotherapeutic agent, a targeted therapy agent, an immune checkpoint modulator, and an immunomodulatory agent.
  • the initial therapy comprises intravesical administration of the oncolytic virus without administration of the additional therapeutic agent.
  • the individual is assigned a first risk status of high risk based on a determination that the first GDB value exceeds a predetermined threshold.
  • the subsequent therapy is an escalated therapy comprising administration of the oncolytic virus and administration of the additional therapeutic agent, and, optionally, administration of one or more further additional therapeutic agents selected from the group consisting of a chemotherapeutic agent, a targeted therapy agent, an immune checkpoint modulator, and an immunomodulatory agent.
  • the subsequent therapy is a de-escalated therapy comprising administration of the oncolytic virus without administration of the additional therapeutic agent.
  • the initial therapy comprises intravesical administration of the oncolytic virus without administration of the additional therapeutic agent
  • the escalated therapy comprises intravesical administration of the oncolytic virus and administration of the additional therapeutic agent.
  • the initial therapy comprises intravesical administration of the oncolytic virus and administration of the additional therapeutic agent
  • the escalated therapy comprises intravesical administration of the oncolytic virus, the additional therapeutic agent, and one or more further additional therapeutic agents selected from the group consisting of a chemotherapeutic agent, a targeted therapy agent, an immune checkpoint modulator, and an immunomodulatory agent.
  • the initial therapy comprises intravesical administration of the oncolytic virus and administration of the additional therapeutic agent
  • the de-escalated therapy comprises intravesical administration of the oncolytic virus without administration of the additional therapeutic agent.
  • the methods described herein comprise administering an initial therapy comprising intravesical administration of an oncolytic virus based on an assignment of an initial minimum residual disease (MRD) status of MRD-negative or MRD- positive to the individual based on a first variant allele frequency (VAF) value in a first urine sample from the individual at a first timepoint, and administering a subsequent therapy based on the difference between the initial MRD status assigned by the first VAF value and an updated MRD status assigned by a second VAF value determined in a second urine sample from the individual at a second timepoint subsequent to the initial therapy.
  • the subsequent therapy is administered based on the difference between the initial MRD status and the updated MRD status.
  • the subsequent therapy comprises an escalated therapy, such as administration of the oncolytic virus in combination with one or more additional therapeutic agents (e.g., one or more immune checkpoint modulators or chemotherapeutic agents).
  • the subsequent therapy is the same as the initial therapy or is a de-escalated therapy, such as administration of the oncolytic virus without administration of one or more additional therapeutic agents.
  • the individual is assigned an initial MRD status of MRD-negative or MRD-positive based on the first VAF value.
  • the initial therapy comprises intravesical administration of an oncolytic virus and may further include administration of one or more additional therapeutic agents selected from the group consisting of a chemotherapeutic agent, a targeted therapy agent, an immune checkpoint modulator, and an immunomodulatory agent.
  • the initial therapy comprises intravesical administration of the oncolytic virus without administration of the additional therapeutic agent.
  • the individual is assigned an initial MRD status of MRD-positive based on a determination that the first VAF value exceeds a predetermined threshold.
  • the subsequent therapy is an escalated therapy comprising administration of the oncolytic virus and administration of the additional therapeutic agent, and, optionally, administration of one or more further additional therapeutic agents selected from the group consisting of a chemotherapeutic agent, a targeted therapy agent, an immune checkpoint modulator, and an immunomodulatory agent.
  • the subsequent therapy is a de-escalated therapy comprising administration of the oncolytic virus without administration of the additional therapeutic agent.
  • the initial MRD status is based on the first VAF value and a first genomic disease burden (GDB) value in a first urine sample from the individual at a first timepoint
  • the updated MRD status is based on the second VAF value and a second GDB value in a second urine sample from the individual at a second timepoint.
  • the individual is assigned an MRD status of MRD-negative if the VAF value is less than about 5-10% (e.g., less than about 8%), and/or is assigned an MRD status of MRD-positive if the VAF value is at least about 5-10% (e.g., at least about 8%).
  • the individual is assigned an MRD status of MRD-negative if the VAF value is less than about 5-10% (e.g., less than about 8%) and the GDB value is less than about 40-45 (e.g, less than about 42), and/or the individual is assigned an MRD status of MRD-positive if the GDB value is at least about 40-45 (e.g., less than about 42) regardless of the VAF value.
  • the initial therapy comprises intravesical administration of the oncolytic virus without administration of the additional therapeutic agent
  • the escalated therapy comprises intravesical administration of the oncolytic virus and administration of the additional therapeutic agent.
  • the initial therapy comprises intravesical administration of the oncolytic virus and administration of the additional therapeutic agent
  • the escalated therapy comprises intravesical administration of the oncolytic virus, the additional therapeutic agent, and one or more further additional therapeutic agents selected from the group consisting of a chemotherapeutic agent, a targeted therapy agent, an immune checkpoint modulator, and an immunomodulatory agent.
  • the initial therapy comprises intravesical administration of the oncolytic virus and administration of the additional therapeutic agent
  • the de-escalated therapy comprises intravesical administration of the oncolytic virus without administration of the additional therapeutic agent.
  • the methods described herein comprise identifying an individual having bladder cancer as a candidate for either a) combination therapy comprising intravesical administration of an oncolytic virus and administration of one or more additional therapeutic agents; or b) therapy comprising intravesical administration of an oncolytic virus without administration of the additional therapeutic agent.
  • the individual is identified as a candidate for either a) combination therapy comprising intravesical administration of an oncolytic virus and administration of one or more additional therapeutic agents; or b) therapy comprising intravesical administration of an oncolytic virus without administration of the additional therapeutic agent, based on a determination of a GDB value, VAF value, MRD status, and/or a risk status assigned based on a GDB value, in a urine sample from the individual.
  • the method comprises determining the GDB value.
  • the method comprises assigning to the individual a risk status of low risk, intermediate risk, or high risk based on the determined GDB value.
  • the individual if the risk status is intermediate or low risk, the individual is identified as a candidate for therapy comprising intravesical administration of an oncolytic virus without administration of the additional therapeutic agent. In some embodiments, if the risk status is high risk, the individual is identified as a candidate for therapy comprising intravesical administration of an oncolytic virus and administration of one or more additional therapeutic agents. In some embodiments, one or more additional therapeutic agents are selected from the group consisting of a chemotherapeutic agent, a targeted therapy agent, an immune checkpoint modulator, and an immunomodulatory agent.
  • determining a GDB value comprises collecting a urine sample from the individual, sequencing nucleic acids (e.g., DNA) from the urine sample to generate urine nucleic acid sequence data, and calculating the GDB value based on the urine nucleic acid sequence data.
  • methods of sequencing nucleic acids in urine samples include, for example, next generation sequencing (such as Illumina sequencing), single-molecule sequencing (such as PacBio sequencing), shotgun sequencing, and the like.
  • Different modes of sequencing may be used for determining a first GDB value, for example, whole genome sequencing, amplicon sequencing, RNA sequencing (e.g., whole transcriptome sequencing or amplicon RNA sequencing), and the like.
  • determining a GDB value comprises collecting a urine sample from an individual, extracting DNA from the sample, subjecting the extracted DNA to whole genome sequencing to generate whole genome sequence data, and calculating the GDB value or the second GDB value based on the whole genome sequence data.
  • determining a GDB value comprises collecting a urine sample from an individual, extracting DNA from the sample, subjecting the extracted DNA to amplicon sequencing targeting a plurality of genes associated with bladder cancer to generate amplicon sequence data, and calculating the GDB value or the second GDB value based on the amplicon sequence data.
  • the amplicon sequencing targets at least 1, at least 2, at least 3, at least 4, at least 5, at least 10, at least 15, at least 20, at least 25, at least 30, at least 35, at least 40, at least 45, at least 50, at least 60, at least 70, at least 80, at least 90, at least 100, or more genes associated with bladder cancer. In certain embodiments, the amplicon sequencing targets about 10, about 15, about 20, about 25, about 30, about 35, about 40, about 45, about 50, about 60, about 70, about 80, about 90, or about 100 genes associated with bladder cancer.
  • the amplicon sequencing targets between about 5 and about 100, between about 10 and about 100, between about 20 and about 100, between about 30 and about 100, between about 40 and about 100, between about 50 and about 100, between about 60 and about 100, between about 70 and about 100, between about 80 and about 100, between about 90 and about 100, between about 5 and about 90, between about 10 and about 90, between about 20 and about 90, between about 30 and about 90, between about 40 and about 90, between about 50 and about 90, between about 60 and about 90, between about 70 and about 90, between about 80 and about 90, between about 5 and about 80, between about 10 and about 80, between about 20 and about 80, between about 30 and about 80, between about 40 and about 80, between about 50 and about 80, between about 60 and about 80, between about 70 and about 80, between about 5 and about 70, between about 10 and about 70, between about 20 and about 70, between about 30 and about 70, between about 40 and about 70, between about 50 and about 70, between about 60 and about 80, between about 70 and about 80, between about 5
  • the calculating a GDB comprises: determining the variant allele frequency (VAF) of the urine nucleic acid sequence data and calculating the GDB value as a percentile ranking of the VAF of the urine nucleic acid sequence data relative to a training dataset.
  • the method comprises determining the VAF of each of a set of genes associated with bladder cancer in the urine nucleic acid sequence data and calculating the GDB value as a percentile ranking of the VAF across one or more of the genes associated with bladder cancer.
  • the training dataset comprises a plurality of VAFs of urine nucleic acid sequence data from a plurality of urine samples.
  • the training dataset comprises a plurality of VAF of nucleic acid sequence data from a plurality of urine samples from plurality of individuals who have been diagnosed with bladder cancer.
  • the training set comprises VAF of each of a set of genes associated with bladder cancer in a plurality of individuals diagnosed with bladder cancer.
  • the training set comprises VAF of one or more (or each) of the same set of genes that are determined in the individual to be treated or identified for treatment using the methods described herein.
  • the method comprises determining the VAF of at least 1, at least 2, at least 3, at least 4, at least 5, at least 10, at least 15, at least 20, at least 25, at least 30, at least 35, at least 40, at least 45, at least 50, at least 60, at least 70, at least 80, at least 90, at least 100, or more genes associated with bladder cancer. In certain embodiments, the method comprises determining the VAF of about 10, about 15, about 20, about 25, about 30, about 35, about 40, about 45, about 50, about 60, about 70, about 80, about 90, or about 100 genes associated with bladder cancer.
  • the amplicon sequencing targets between about 5 and about 100, between about 10 and about 100, between about 20 and about 100, between about 30 and about 100, between about 40 and about 100, between about 50 and about 100, between about 60 and about 100, between about 70 and about 100, between about 80 and about 100, between about 90 and about 100, between about 5 and about 90, between about 10 and about 90, between about 20 and about 90, between about 30 and about 90, between about 40 and about 90, between about 50 and about 90, between about 60 and about 90, between about 70 and about 90, between about 80 and about 90, between about 5 and about 80, between about 10 and about 80, between about 20 and about 80, between about 30 and about 80, between about 40 and about 80, between about 50 and about 80, between about 60 and about 80, between about 70 and about 80, between about 5 and about 70, between about 10 and about 70, between about 20 and about 70, between about 30 and about 70, between about 40 and about 70, between about 50 and about 70, between about 60 and about 80, between about 70 and about 80, between about 5
  • the methods described herein comprise assigning to the individual a first risk status of low risk, intermediate risk, or high risk based on a GDB value determined in a urine sample from the individual.
  • the individual is assigned a risk status of intermediate or high risk if the GDB value exceeds a predetermined threshold.
  • the individual is assigned a risk status of intermediate if the GDB value exceeds a first predetermined threshold and does not exceed a second predetermined threshold.
  • the individual is assigned a risk status of high risk if the GDB value exceeds the second predetermined threshold.
  • the assigned risk status corresponds to a relative risk of progression of bladder cancer despite intravesical administration of the oncolytic virus (e.g., in the absence of additional therapeutic agent, such as a chemotherapeutic agent or an immune checkpoint modulator). In some embodiments, the assigned risk status corresponds to a relative risk of non-response to intravesical administration of the oncolytic virus (e.g., in the absence of additional therapeutic agent, such as a chemotherapeutic agent or an immune checkpoint modulator).
  • the assigned risk status corresponds to a relative risk of recurrence of bladder cancer after intravesical administration of the oncolytic virus (e.g., in the absence of additional therapeutic agent, such as a chemotherapeutic agent or an immune checkpoint modulator).
  • the individual is assigned a risk status (e.g., a first risk status or a second risk status) of low risk if the GDB value (e.g., the first GDB value or the second GDB value, respectively) is less than about 50 (less than about the 50 th percentile).
  • the individual is assigned a risk status (e.g., a first risk status or a second risk status) of low risk if the GDB value (e.g.
  • the first GDB value or the second GDB value, respectively is less than about 50, 45, 40, 35, 30, 25, 20, 15, 10, 5, or 1 (less than about the 50 th , 45 th , 40 th , 35 th , 30 th , 25 th , 20 th , 15 th , 10 th , 5 th or 1 st percentile).
  • the individual is assigned a risk status (e.g., a first risk status or a second risk status) of low risk if the GDB value (e.g., the first GDB value or the second GDB value, respectively) is less than about 20 (the 20 th percentile).
  • the individual is assigned a risk status (e.g. , a first risk status or a second risk status) of low risk if the GDB value (e.g., the first GDB value or the second GDB value, respectively) is less than a maximum threshold GDB value between 1 and 50 (between the 1 st and 50 th percentiles).
  • the individual is assigned a risk status (e.g. , a first risk status or a second risk status) of low risk if the GDB value (e.g. , the first GDB value or the second GDB value, respectively) is less than a maximum threshold GDB value between 10 and 30 (between the 10 th and 30 th percentiles).
  • the individual is assigned a risk status (e.g., a first risk status or a second risk status) of low risk if the GDB value (e.g. , the first GDB value or the second GDB value, respectively) is less than a maximum threshold GDB value between 15 and 25 (between the 15 th and 25 th percentiles).
  • the individual is assigned a risk status (e.g., a first risk status or a second risk status) of low risk if the GDB value (e.g.
  • the first GDB value or the second GDB value, respectively) is less than a maximum threshold GDB value between 1 and 5 (between the 1 st and 5 th percentiles), between 1 and 10 (between the 1 st and 10 th percentiles), between 1 and 15 (between the 1 st and 15 th percentiles), between 1 and 20 (between the 1 st and 20 th percentiles), between 1 and 25 (between the 1 st and 25 th percentiles), between 1 and 30 (between the 1 st and 30 th percentiles), between 1 and 35 (between the 1 st and 35 th percentiles), between 1 and 40 (between the 1 st and 40 th percentiles), between 1 and 45 (between the 1 st and 45 th percentiles), between 1 and 50 (between the 1 st and 50 th percentiles), between 5 and 10 (between the 5 th and 10 th percentiles), between 5 and 15 (between the 5 th and 15 th percentiles), between 5 and
  • the individual is assigned a risk status (e.g. , a first risk status or a second risk status) of high risk if the GDB value (e.g., the first GDB value or the second GDB value, respectively) is greater than about 50 (greater than about the 50 th percentile).
  • the individual is assigned a risk status (e.g., a first risk status or a second risk status) of high risk if the GDB value (e.g.
  • the first GDB value or the second GDB value, respectively is greater than about 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 99 (less than about the 50 th , 55 th , 60 th , 65 th , 70 th , 75 th , 80 th , 85 th , 90 th , 95 th or 99 th percentile).
  • the individual is assigned a risk status (e.g., a first risk status or a second risk status) of high risk if the GDB value (e.g. , the first GDB value or the second GDB value, respectively) is greater than about 80 (the 80 th percentile).
  • the individual is assigned a risk status (e.g. , a first risk status or a second risk status) of high risk if the GDB value (e.g., the first GDB value or the second GDB value, respectively) is greater than a minimum threshold GDB value between 50 and 99 (between the 50 th and 99 th percentiles).
  • the individual is assigned a risk status (e.g., a first risk status or a second risk status) of high risk if the GDB value (e.g., the first GDB value or the second GDB value, respectively) is greater than a minimum threshold GDB value between 70 and 90 (between the 70 th and 90 th percentiles).
  • the individual is assigned a risk status (e.g., a first risk status or a second risk status) of high risk if the GDB value (e.g., the first GDB value or the second GDB value, respectively) is greater than a minimum threshold GDB value between 75 and 85 (between the 75 th and 85 th percentiles).
  • the individual is assigned a risk status (e.g. , a first risk status or a second risk status) of high risk if the GDB value (e.g.
  • the first GDB value or the second GDB value, respectively) is greater than a minimum threshold GDB value between 50 and 55 (between the 50 th and 55 th percentiles), between 50 and 60 (between the 50 th and 60 th percentiles), between 50 and 65 (between the 50 th and 65 th percentiles), between 50 and 70 (between the 50 th and 70 th percentiles), between 50 and 75 (between the 50 th and 75 th percentiles), between 50 and 80 (between the 50 th and 80 th percentiles), between 50 and 85 (between the 50 th and 85 th percentiles), between 50 and 90 (between the 50 th and 90 th percentiles), between 50 and 95 (between the 50 th and 95 th percentiles), between 50 and 99 (between the 50 th and 99 th percentiles), between 55 and 60 (between the 55 th and 60 th percentiles), between 55 and 65 (between the 55 th and 65 th percentiles), between 55 and 70 (between the 55 th and 70 th
  • the individual is assigned a risk status of intermediate risk if they do not meet the criteria for being assigned a risk status of low risk or high risk.
  • the individual is assigned a risk status (e.g., a first risk status or a second risk status) of intermediate risk if the GDB value (e.g. , the first GDB value or the second GDB value, respectively) is greater than a maximum threshold GDB value described herein and less than a minimum threshold GDB value described herein.
  • the individual is assigned a risk status (e.g., a first risk status or a second risk status) of intermediate risk if the GDB value (e.g., the first GDB value or the second GDB value, respectively) is between 1 and 99 (between the 1 st and 99 th percentiles).
  • the individual is assigned a risk status (e.g., a first risk status or a second risk status) of intermediate risk if the GDB value (e.g. , the first GDB value or the second GDB value, respectively) is between 20 and 80 (between the 20 th and 80 th percentiles).
  • the individual is assigned a risk status (e.g., a first risk status or a second risk status) of intermediate risk if the GDB value (e.g. , the first GDB value or the second GDB value, respectively) is between 1 and 5 (between the 1 st and 5 th percentiles), between 1 and 10 (between the 1 st and 10 th percentiles), between 1 and 15 (between the 1 st and 15 th percentiles), between 1 and 20 (between the 1 st and 20 th percentiles), between 1 and 25 (between the 1 st and 25 th percentiles), between 1 and 30 (between the 1 st and 30 th percentiles), between 1 and 35 (between the 1 st and 35 th percentiles), between 1 and 40 (between the 1 st and 40 th percentiles), between 1 and 45 (between the 1 st and 45 th percentiles), between 1 and 50 (between the 1 st and 50 th percentiles), between 5 and a risk status
  • the methods described herein comprise assigning to the individual an initial minimum residual disease (MRD) status of MRD-positive or MRD- negative based on a VAF value determined in a urine sample from the individual.
  • MRD initial minimum residual disease
  • the individual is assigned an MRD status of MRD-positive if the VAF value exceeds a predetermined threshold.
  • the individual is assigned an MRD status of MRD-negative of the VAF value is less than a predetermined threshold.
  • the initial MRD status is based on the first VAF value and a first genomic disease burden (GDB) value in a first urine sample from the individual at a first timepoint
  • the updated MRD status is based on the second VAF value and a second GDB value in a second urine sample from the individual at a second timepoint.
  • the individual is assigned an MRD status of MRD-negative if the VAF value is less than about 5- 10% (e.g., less than about 8%), and/or is assigned an MRD status of MRD-positive if the VAF value is at least about 5-10% (e.g., at least about 8%).
  • the individual is assigned an MRD status of MRD-negative if the VAF value is less than about 5- 10% (e.g., less than about 8%) and the GDB value is less than about 40-45 (e.g., less than about 42), and/or the individual is assigned an MRD status of MRD-positive if the GDB value is at least about 40-45 (e.g., less than about 42) regardless of the VAF value.
  • the assigned MRD status corresponds to a relative risk of progression of bladder cancer despite intravesical administration of the oncolytic virus (e.g., in the absence of additional therapeutic agent, such as a chemotherapeutic agent or an immune checkpoint modulator).
  • the assigned MRD status corresponds to a relative risk of non-response to intravesical administration of the oncolytic virus (e.g., in the absence of additional therapeutic agent, such as a chemotherapeutic agent or an immune checkpoint modulator). In some embodiments, the assigned MRD status corresponds to a relative risk of recurrence of bladder cancer after intravesical administration of the oncolytic virus (e.g., in the absence of additional therapeutic agent, such as a chemotherapeutic agent or an immune checkpoint modulator).
  • kits, unit dosages, and articles of manufacture useful for any one of the methods described herein.
  • a pharmaceutical composition comprising an oncolytic virus (such as cretostimogene) and a pharmaceutically acceptable carrier, wherein the oncolytic virus comprises a viral vector comprising a tumor cell-specific promoter operably linked to a viral gene essential for replication of the oncolytic virus, and a heterologous gene encoding an immune-related molecule.
  • the pharmaceutical composition may be used for treating bladder cancer (such as high-risk NMIBC, e.g., BCG-naive high-risk NMIBC or high-risk NMIBC after prior therapy with BCG) according to any one of the methods described herein.
  • Suitable pharmaceutical carriers include sterile water; saline, dextrose; dextrose in water or saline; condensation products of castor oil and ethylene oxide combining about 30 to about 35 moles of ethylene oxide per mole of castor oil; liquid acid; lower alkanols; oils such as com oil; peanut oil, sesame oil and the like, with emulsifiers such as mono- or di-glyceride of a fatty acid, or a phosphatide, e.g., lecithin, and the like; glycols; polyalkylene glycols; aqueous media in the presence of a suspending agent, for example, sodium carboxymethylcellulose; sodium alginate; poly(vinylpyrolidone); and the like, alone, or with suitable dispensing agents such as lecithin; polyoxyethylene stearate; and the like.
  • a suspending agent for example, sodium carboxymethylcellulose; sodium alginate; poly(vinylpyrolidone); and the
  • the carrier may also contain adjuvants such as preserving stabilizing, wetting, emulsifying agents and the like together with the penetration enhancer.
  • the final form may be sterile and may also be able to pass readily through an injection device such as a hollow needle.
  • the proper viscosity may be achieved and maintained by the proper choice of solvents or excipients.
  • the pharmaceutical carrier may include active or passive excipients for drug delivery, such as polymer and non-polymer systems. Other pharmaceutically acceptable carriers and their formulation are described in standard formulation treatises, e.g., Remington's Pharmaceutical Sciences by E. W. Martin. [0293]
  • the pharmaceutical compositions described herein may include other agents, excipients, or stabilizers to improve properties of the composition.
  • excipients and diluents include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, saline solution, syrup, methylcellulose, methyl- and propylhydroxybenzoates, talc, magnesium stearate and mineral oil.
  • the pharmaceutical composition is formulated to have a pH in the range of about 4.5 to about 9.0, including for example pH ranges of about any one of 5.0 to about 8.0, about 6.5 to about 7.5, or about 6.5 to about 7.0.
  • the pharmaceutical composition can also be made to be isotonic with blood by the addition of a suitable tonicity modifier, such as glycerol.
  • kits for treating bladder cancer such as high-risk NMIBC, e.g., BCG-naive high-risk NMIBC or high-risk NMIBC after prior therapy with BCG
  • an oncolytic virus such as cretostimogene
  • the oncolytic virus comprises a viral vector comprising a tumor cell-specific promoter operably linked to a viral gene essential for replication of the oncolytic virus, and a heterologous gene encoding an immune-related molecule.
  • the kit further comprises a pretreatment composition comprising a transduction enhancing agent, such as N-Dodecyl-P-D-maltoside (DDM).
  • DDM N-Dodecyl-P-D-maltoside
  • the kit further comprises devices, materials, and/or instructions for carrying out any one of the methods described above.
  • the kit comprises a device for intravesical administration of the oncolytic virus.
  • Medical devices for intravesical administration may include a catheter, for example, a Rusch 173430 Foley Catheter & BARD LUBRI-SIL Foley Catheter #70516SI.
  • the instructions relating to the use of the oncolytic virus generally include information as to dosage, dosing schedule, and route of administration for the intended treatment.
  • the containers may be unit doses, bulk packages (e.g., multi-dose packages) or sub-unit doses.
  • kits may be provided that contain sufficient dosages of the oncolytic virus as disclosed herein to provide effective treatment of an individual for an extended period, such as any of a week, 2 weeks, 3 weeks, 4 weeks, 6 weeks, 8 weeks, 3 months, 4 months, 5 months, 7 months, 8 months, 9 months, 12 months or more.
  • kits may be provided that contain between one and six doses of the oncolytic virus (e.g., one, two, three, four, five, or six unit doses). Kits may also include multiple unit doses of the oncolytic virus and instructions for use, packaged in quantities sufficient for storage and use in pharmacies, for example, hospital pharmacies and compounding pharmacies.
  • kits of the invention are in suitable packaging.
  • suitable packaging includes, but is not limited to, vials, bottles, jars, flexible packaging (e.g., sealed Mylar or plastic bags), and the like. Kits may optionally provide additional components such as buffers and interpretative information.
  • the present application thus also provides articles of manufacture, which include vials (such as sealed vials), bottles, jars, flexible packaging, and the like.
  • kits for treating bladder cancer in an individual comprising: a) one or more agents for determining a genomic disease burden (GDB) value in a urine sample from an individual; b) an oncolytic virus; and c) a device for intravesically administering the oncolytic virus.
  • the kit further comprises instructions for determining a GDB value in the urine sample, e.g., using a method described herein.
  • the kit further comprises instructions for determining a risk value for the individual based on the GDB value, e.g. , using a method described herein.
  • the kit further comprises one or more additional therapeutic agents selected from the group consisting of an immune checkpoint modulator or a chemotherapeutic agent.
  • the kit further comprises instructions for administering the additional therapeutic agent based on the determination of the GDB value and/or the determination of the risk value.
  • the kit further comprises a device for administering the additional therapeutic agent.
  • the kit further comprises a device for systemic (e.g., intravenous) administration of the additional therapeutic agent.
  • the kit further comprises a device for local (e.g., intravesical) administration of the additional therapeutic agent.
  • the oncolytic virus comprises a viral vector comprising a tumor cell-specific promoter (e.g.
  • the oncolytic virus is an adenovirus serotype 5, wherein the endogenous Ela promoter of a native adenovirus serotype 5 is replaced by the human E2F-1 promoter, and the endogenous E3 19kD coding region of the native adenovirus serotype 5 is replaced by a nucleic acid encoding human GM-CSF.
  • the oncolytic virus is cretostimogene.
  • the kit further comprises at least one dose of N-Dodecyl-P-D-maltoside (DDM), instructions for administering DDM as a pretreatment composition prior to intravesical administration of the oncolytic virus, and/or a device for intravesical administration of DDM.
  • the additional therapeutic agent is an immune checkpoint modulator (e.g., an immune checkpoint inhibitor).
  • the immune checkpoint modulator comprises an inhibitor of (e.g., an antibody that binds to) an immune checkpoint molecule selected from the group consisting of CTLA-4, PD-1, PD-L1, PD-L2, TIM3, B7-H3, B7-H4, LAG-3, KIR, and ligands thereof.
  • the immune checkpoint modulator comprises an anti -PD-1 antibody (e.g., nivolumab, pidilizumab, pembrolizumab, BMS-936559, atezolizumab, lambrolizumab, MK- 3475, AMP -224, AMP-514, STI-A1110, TSR-042, or any combination thereof).
  • the additional therapeutic agent is a chemotherapeutic agent.
  • the chemotherapeutic agent comprises gemcitabine, cisplatin, carboplatin, paclitaxel, docetaxel, ifosfamide, doxorubicin, methotrexate, vinblastine, mitomycin, 5- fluorouracil (5-FU), or any combination thereof.
  • the chemotherapeutic agent comprises gemcitabine.
  • the cancer is nonmuscle invasive bladder cancer (NMIBC). In certain embodiments, the cancer is high-risk NMIBC.
  • kits for treating bladder cancer in an individual comprising: a) one or more agents for determining a variant allele frequency (VAF) value in a urine sample from an individual; b) an oncolytic virus; and c) a device for intravesically administering the oncolytic virus.
  • the kit further comprises instructions for determining a VAF value in the urine sample, e.g., using a method described herein.
  • the kit further comprises instructions for determining a genomic disease burden (GDB) value, a minimum residual disease (MRD) status, and/or a risk status for the individual based on the VAF value, e.g., using a method described herein.
  • GDB genomic disease burden
  • MRD minimum residual disease
  • the kit further comprises one or more additional therapeutic agents selected from the group consisting of an immune checkpoint modulator or a chemotherapeutic agent.
  • the kit further comprises instructions for administering the additional therapeutic agent based on the determination of the VAF value, the determination of the GDB value, and/or the determination of the risk value.
  • the kit further comprises a device for administering the additional therapeutic agent.
  • the kit further comprises a device for systemic (e.g., intravenous) administration of the additional therapeutic agent.
  • the kit further comprises a device for local (e.g., intravesical) administration of the additional therapeutic agent.
  • the oncolytic virus comprises a viral vector comprising a tumor cell-specific promoter (e.g. , a human E2F-1 promoter) operably linked to a viral gene essential for replication of the oncolytic virus, and a heterologous gene encoding an immune-related molecule (e.g., GM-CSF).
  • a tumor cell-specific promoter e.g. , a human E2F-1 promoter
  • a heterologous gene encoding an immune-related molecule e.g., GM-CSF
  • the oncolytic virus is an adenovirus serotype 5, wherein the endogenous Ela promoter of a native adenovirus serotype 5 is replaced by the human E2F-1 promoter, and the endogenous E3 19kD coding region of the native adenovirus serotype 5 is replaced by a nucleic acid encoding human GM-CSF.
  • the oncolytic virus is cretostimogene.
  • the kit further comprises at least one dose of N-Dodecyl-P-D-maltoside (DDM), instructions for administering DDM as a pretreatment composition prior to intravesical administration of the oncolytic virus, and/or a device for intravesical administration of DDM.
  • the additional therapeutic agent is an immune checkpoint modulator (e.g., an immune checkpoint inhibitor).
  • the immune checkpoint modulator comprises an inhibitor of (e.g., an antibody that binds to) an immune checkpoint molecule selected from the group consisting of CTLA-4, PD-1, PD-L1, PD-L2, TIM3, B7-H3, B7-H4, LAG-3, KIR, and ligands thereof.
  • the immune checkpoint modulator comprises an anti-PD-1 antibody (e.g., nivolumab, pidilizumab, pembrolizumab, BMS- 936559, atezolizumab, lambrolizumab, MK- 3475, AMP -224, AMP-514, STI-Al l 10, TSR- 042, or any combination thereof).
  • the additional therapeutic agent is a chemotherapeutic agent.
  • the chemotherapeutic agent comprises gemcitabine, cisplatin, carboplatin, paclitaxel, docetaxel, ifosfamide, doxorubicin, methotrexate, vinblastine, mitomycin, 5 -fluorouracil (5-FU), or any combination thereof.
  • the chemotherapeutic agent comprises gemcitabine.
  • the cancer is non-muscle invasive bladder cancer (NMIBC). In certain embodiments, the cancer is high-risk NMIBC.
  • kits for treating bladder cancer in an individual comprising: a) one or more agents for determining a minimum residual disease (MRD) status of MRD-positive or MRD-negative based on a genomic disease burden (GDB) value in a urine sample from an individual; b) an oncolytic virus; and c) a device for intravesically administering the oncolytic virus.
  • the kit further comprises instructions for determining a MRD status based on a GDB value in the urine sample, e.g., using a method described herein.
  • the kit further comprises one or more additional therapeutic agents selected from the group consisting of an immune checkpoint modulator or a chemotherapeutic agent.
  • the kit further comprises instructions for administering the additional therapeutic agent based on the determination of the MRD status and/or the determination of the GDB value.
  • the kit further comprises a device for administering the additional therapeutic agent.
  • the kit further comprises a device for systemic (e.g., intravenous) administration of the additional therapeutic agent.
  • the kit further comprises a device for local (e.g., intravesical) administration of the additional therapeutic agent.
  • the oncolytic virus comprises a viral vector comprising a tumor cell-specific promoter (e.g.
  • the oncolytic virus is an adenovirus serotype 5, wherein the endogenous Ela promoter of a native adenovirus serotype 5 is replaced by the human E2F-1 promoter, and the endogenous E3 19kD coding region of the native adenovirus serotype 5 is replaced by a nucleic acid encoding human GM-CSF.
  • the oncolytic virus is cretostimogene.
  • the kit further comprises at least one dose of N-Dodecyl-P-D-maltoside (DDM), instructions for administering DDM as a pretreatment composition prior to intravesical administration of the oncolytic virus, and/or a device for intravesical administration of DDM.
  • the additional therapeutic agent is an immune checkpoint modulator (e.g., an immune checkpoint inhibitor).
  • the immune checkpoint modulator comprises an inhibitor of (e.g., an antibody that binds to) an immune checkpoint molecule selected from the group consisting of CTLA-4, PD-1, PD-L1, PD-L2, TIM3, B7-H3, B7-H4, LAG-3, KIR, and ligands thereof.
  • the immune checkpoint modulator comprises an anti-PD-1 antibody (e.g., nivolumab, pidilizumab, pembrolizumab, BMS- 936559, atezolizumab, lambrolizumab, MK- 3475, AMP -224, AMP-514, STI-Al l 10, TSR- 042, or any combination thereof).
  • the additional therapeutic agent is a chemotherapeutic agent.
  • the chemotherapeutic agent comprises gemcitabine, cisplatin, carboplatin, paclitaxel, docetaxel, ifosfamide, doxorubicin, methotrexate, vinblastine, mitomycin, 5 -fluorouracil (5-FU), or any combination thereof.
  • the chemotherapeutic agent comprises gemcitabine.
  • the cancer is non-muscle invasive bladder cancer (NMIBC). In certain embodiments, the cancer is high-risk NMIBC.
  • kits for treating high-risk non-muscle invasive bladder cancer comprising: a) an oncolytic adenovirus; b) gemcitabine; and c) a device for intravesically administering the oncolytic adenovirus, gemcitabine, or both the oncolytic adenovirus and gemcitabine; wherein the oncolytic virus comprises a viral vector comprising a tumor cell-specific promoter operably linked to a viral gene essential for replication of the oncolytic virus, and a heterologous gene encoding an immune-related molecule.
  • the oncolytic virus preferentially replicates in a cancer cell.
  • the cancer cell is defective in the Rb pathway.
  • the tumor cell-specific promoter is an E2F-1 promoter.
  • the E2F-1 promoter comprises the nucleotide sequence set forth in SEQ ID NO: 1.
  • the immune-related molecule is selected from the group consisting ofGM-CSF, IL-2, IL-12, interferon, CCL4, CCL19, CCL21, CXCL13, TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, TLR10, RIG-I, MDA5, LGP2, and LTap.
  • the immune-related molecule is GM-CSF.
  • the heterologous gene is operably linked to a viral promoter.
  • the viral gene essential for replication of the oncolytic virus is selected from the group consisting of E1A, E1B, and E4.
  • the heterologous gene is operably linked to an El promoter or an E3 promoter.
  • the oncolytic virus is an adenovirus serotype 5, wherein the endogenous Ela promoter of a native adenovirus serotype 5 is replaced by the human E2F-1 promoter, and the endogenous E3 19kD coding region of the native adenovirus serotype 5 is replaced by a nucleic acid encoding human GM-CSF.
  • the oncolytic adenovirus is cretostimogene.
  • the kit further comprises instructions for a method of administering the oncolytic adenovirus, gemcitabine, or both the oncolytic adenovirus and gemcitabine.
  • the method comprises administering the gemcitabine in combination with the oncolytic adenovirus, wherein the method comprises a 6-week induction phase comprising administering the oncolytic adenovirus and gemcitabine to the individual on weeks 1, 2, 3, 4, 5, and 6.
  • the method comprises a maintenance phase subsequent to the induction phase, wherein the maintenance phase comprises a 3-week treatment comprising administering the oncolytic adenovirus and gemcitabine to the individual every three or six months.
  • the 3-week treatment comprises administering the oncolytic adenovirus and gemcitabine weekly on weeks 1, 2, and 3.
  • the start of the induction phase and the start of the maintenance phase are separated by about three or six months.
  • the method comprises reevaluating the individual at month three or around week 11 following the start of the induction phase. In some embodiments, if the individual experiences a complete or partial response at month three, the method comprises beginning the maintenance phase at month three or around week 13 following the start of the induction phase.
  • the maintenance phase comprises administering the 3-week treatment every three months for nine months, followed by administering the 3-week treatment every six months.
  • the method comprises administering a second induction dose at month three or around week 13 following the start of the induction phase.
  • the maintenance phase comprises administering the 3 -week treatment every three months for six months, followed by administering the 3 -week treatment every six months.
  • the method comprises intravesical administration of the oncolytic adenovirus followed by intravesical administration of gemcitabine on the same day of treatment.
  • the method comprises intravesical administration of the oncolytic adenovirus followed immediately by intravesical administration of gemcitabine.
  • the method comprises a 6-week induction phase comprising: i) administering the oncolytic adenovirus to the individual on weeks 1, 2, 4 and 5; and ii) administering gemcitabine to the individual on weeks 3 and 6.
  • the method comprises a maintenance phase subsequent to the induction phase, wherein the maintenance phase comprises a 3 -week treatment comprising administering the oncolytic adenovirus and gemcitabine to the individual every three or six months.
  • the 3 -week treatment comprises administering the oncolytic adenovirus weekly on weeks 1 and 2 followed by administering gemcitabine in week 3.
  • the method comprises reevaluating the individual at month three or around week 11 following the start of the induction phase. In some embodiments, if the individual experiences a complete or partial response at month three or around week 11 following the start of the induction phase, the method comprises beginning the maintenance phase at month three or around week 13 following the start of the induction phase. In some embodiments, the maintenance phase comprises administering the 3 -week treatment every three months for nine months, followed by administering the 3 -week treatment every six months. In some embodiments, if the individual experiences a complete or partial response at month three or around week 11 following the start of the induction phase, the method comprises administering a second induction dose at month three or around week 13 following the start of the induction phase.
  • the maintenance phase comprises administering the 3-week treatment every three months for six months, followed by administering the 3-week treatment every six months.
  • the method comprises administering the gemcitabine is administered at a dose of around 1 g or around 2 g. In some embodiments, the method comprises administering the gemcitabine in 50 to 100 mL saline. In some embodiments, the kit comprises a dosage unit of gemcitabine in an amount of around 1 g or around 2 g. In some embodiments, the kit comprises a dosage unit of the oncolytic adenovirus in an amount of between about 1 x 10 8 and about 1 x 10 14 viral particles.
  • the article of manufacture can comprise a container and a label or package insert on or associated with the container.
  • Suitable containers include, for example, bottles, vials, syringes, etc.
  • the containers may be formed from a variety of materials such as glass or plastic.
  • the container holds a composition which is effective for treating bladder cancer (e.g, high-risk NMIBC) as described herein, and may have a sterile access port.
  • the label or package insert indicates that the composition is used for treating the particular condition in an individual.
  • the label or package insert will further comprise instructions for administering the composition to the individual.
  • Articles of manufacture and kits comprising combination therapies described herein are also contemplated.
  • Package insert refers to instructions customarily included in commercial packages of therapeutic products that contain information about the indications, usage, dosage, administration, contraindications and/or warnings concerning the use of such therapeutic products.
  • the package insert indicates that the composition is used for treating bladder cancer (e.g., high-grade NMIBC).
  • the article of manufacture may further comprise a second container comprising a pharmaceutically-acceptable buffer, such as bacteriostatic water for injection (BWFI), phosphate-buffered saline, Ringer's solution and dextrose solution. It may further include other materials desirable from a commercial and user standpoint, including other buffers, diluents, fdters, needles, and syringes.
  • BWFI bacteriostatic water for injection
  • phosphate-buffered saline such as bacteriostatic water for injection (BWFI), phosphate-buffered saline, Ringer's solution and dextrose solution.
  • BWFI bacteriostatic water for injection
  • Example 1 Clinical Trial of Cretostimogene for Treating BCG-Naive and BCG- Exposed High-Risk Non-Muscle Invasive Bladder Cancer (NMIBC)
  • This example describes an open-label, single-arm, multicenter study of the safety and efficacy of intravesical cretostimogene monotherapy in patients with high-risk nonmuscle invasive bladder cancer (NMIBC) who are either BCG-naive or BCG-exposed.
  • BCG- naive patients are those with high-risk NMIBC who have not received prior treatment with Bacillus Calmette-Guerin (BCG), z.e., are completely BCG-naive.
  • BCG-exposed patients encompass three patient subpopulations:
  • BCG-resistant NMIBC Patients with persistent or recurrent high-grade Ta or CIS at 3 -month evaluation that follows at least 5 of 6 doses of induction BCG.
  • Cretostimogene is a replication selective oncolytic adenovirus that destroys bladder tumor cells through their defective retinoblastoma (Rb) pathway.
  • Rb retinoblastoma
  • CIS-containing high-risk NMIBC e.g., CIS only or CIS with concurrent Ta or T1 tumors
  • patients with CIS z.e., Ta or T1 tumors only
  • CIS-containing high-risk NMIBC e.g., CIS only or CIS with concurrent Ta or T1 tumors
  • CIS z.e., Ta or T1 tumors only
  • BCG-exposed patients meeting the criteria for 1. BCG-resistant NMIBC, 2. Delayed relapse after inadequate BCG, and 3. Delayed relapse after adequate BCG, as outlined above, are included in the trial. Patients will be stratified by the definitions of “BCG-Resistant” and by “Late Recurrences”, acknowledging the clinical and likely biologic heterogeneity inherent in these two populations. Patients will be treated for a 2-year treatment duration.
  • each patient will receive intravesical cretostimogene at a dose of IxlO 12 viral particles weekly for six weeks. Treatment will be performed in a classic induction course followed by maintenance therapy. Cretostimogene induction will be administered as 6 weekly intravesical instillations followed by 3 weekly maintenance doses at months 3, 6, 9, 12, 18, and 24. Given the strength of response for re-induction with cretostimogene based on prior studies, those patients in Non-Response at the first disease evaluation would be offered the opportunity for cretostimogene re-induction.
  • cretostimogene has shown strong efficacy in the treatment ofNMIBC as a monotherapy. Additionally, cretostimogene (also known as CG0070) has also shown remarkable efficacy in clinical trials in combination with immune checkpoint inhibitors (see, e.g., Li, Roger, et al.
  • biomarker-guided combination therapy comprising intravesical administration of cretostimogene and administration of one or more additional therapeutic agents having bladder cancer, such as high-risk non-muscle invasive bladder cancer (NMIBC).
  • NMIBC high-risk non-muscle invasive bladder cancer
  • the biomarker evaluated in this trial is genomic disease burden (GDB), a measure of tumor mutation burden measured from urine-derived DNA.
  • GDB genomic disease burden
  • urinary GDB will be evaluated in patients using UroAmp®, and each patient will be assigned a first risk status of low risk, intermediate risk, or high risk based on the GDB value. Patients assigned a first risk status of low risk will be treated with cretostimogene monotherapy and evaluated as described in Example 1.
  • Patients assigned a first risk status of high risk will be treated with cretostimogene as described in Example 1 in combination with one or more additional therapeutic agents, such as an immune checkpoint modulator or chemotherapeutic agent described herein, and evaluated as described in Example 1.
  • Patients assigned a first risk status of intermediate risk will be treated with cretostimogene monotherapy and evaluated as described in Example 1 and, at 6 months, will be evaluated a second time for GDB and assigned a second risk status of low risk or intermediate/high risk.
  • Patients having a second risk status of low risk will continue with cretostimogene monotherapy maintenance therapy.
  • a variety of additional therapeutic agents are administered individually or in combination to the patients.
  • Any one or more of the immune checkpoint inhibitors or chemotherapeutic agents described herein may be tested in the clinical studies.
  • the routes of administration, doses, dosing frequencies, duration, and/or maintenance dosing scheme for administration of the additional therapeutic agents may be any of those described herein.
  • the routes of administration, doses, dosing frequencies, duration, and/or maintenance dosing scheme can be the same or different from those listed in Table 1.
  • Table 1 is presented here for examples only, immune checkpoint inhibitors outside the scope of Table 1 can also be evaluated in the clinical studies.
  • Example 3 Phase 2 Clinical Trial of Cretostimogene for Treating High-Risk NonMuscle Invasive Bladder Cancer (NMIBC)
  • This example describes a phase 2, open-label, multi-arm, multicenter study of the safety and efficacy of intravesical cretostimogene in patients with high-risk non-muscle invasive bladder cancer (NMIBC) who are either BCG-naive, BCG-exposed, or BCG- unresponsive.
  • NMIBC non-muscle invasive bladder cancer
  • Cretostimogene is a replication selective oncolytic adenovirus that destroys bladder tumor cells through their defective retinoblastoma (Rb) pathway.
  • Rb retinoblastoma
  • Cretostimogene in conjunction with DDM has been administered to 265 patients across 3 completed and 3 ongoing clinical trials. Doses explored ranged from 1 x 10 12 to 3 x 10 13 vp given IVE as part of single and multi-dose regimens on monthly or weekly schedules. Prior to cretostimogene instillation, 75-100 m of 0.1% DDM was administered via IVE as a preparative agent. Across these studies, the most common toxicities were those localized to the bladder including pollakiuria, dysuria, bladder spasm, hematuria, urinary urgency, and nocturia. No Grade 4 or higher adverse events (AEs) related to cretostimogene or DDM have been reported thus far across the program (Tyson, M.
  • AEs adverse events
  • the pivotal Phase 3 BOND-003 study is an ongoing, open-label, single-arm trial, entitled, “A Phase 3 Study of Cretostimogene in Patients with non-Muscle-Invasive Bladder Cancer (NMIBC) Unresponsive to Bacillus Calmette -Guerin (BCG),” in which Cohort C enrolled patients with BCG-unresponsive CIS. All Cohort C patients received weekly induction cretostimogene for 6 weeks (with a reinduction course for non-responders at 3 months), followed by maintenance cretostimogene every 3 months for the first year and then every 6 months for up to 36 months. Preliminary data reported in 2024 on efficacy as measured by Complete Response (CR) were available for 110 patients in Cohort C.
  • CR Complete Response
  • Cretostimogene will be administered intravesically to patients having high-risk nonmuscle-invasive bladder cancer (NMIBC) in three cohorts. Cohorts A and B will receive intravesical cretostimogene as a monotherapy. Patients in Cohort CX will also receive gemcitabine, a deoxycytidine analog that has direct apoptotic effects on tumor cells via DNA replication interference and is used in a wide range of tumor types, including bladder cancer.
  • NMIBC nonmuscle-invasive bladder cancer
  • DDM n-dodecyl
  • FIG. 4 A summary of the cohorts to be evaluated in the study is depicted in FIG. 4.
  • NMIBC tumor risk stratification will be determined by 2024 American Urological Association (AU A) guidelines (Holzbeierlein 2024) and for the purposes of this study will include patients with a current diagnosis of: CIS (with or without concomitant papillary Ta/Tl disease); high-grade (HG) Tl; or HG Ta that is recurrent, multi-focal, >3cm, or has recurred post-BCG.
  • CIS with or without concomitant papillary Ta/Tl disease
  • HG high-grade
  • HG Ta that is recurrent, multi-focal, >3cm, or has recurred post-BCG.
  • All pathology specimens must be predominantly urothelial (transitional cell) and have less than 50% histologic subtype/variant histology (e.g., micropapillary, nested, plasmacytoid, neuroendocrine, sarcomatoid, squamous, or glandular differentiation).
  • histologic subtype/variant histology e.g., micropapillary, nested, plasmacytoid, neuroendocrine, sarcomatoid, squamous, or glandular differentiation.
  • BCG-naive NMIBC will be defined as the following:
  • BCG-exposed NMIBC will be defined as the following:
  • BCG-resistant NMIBC Persistent or recurrent HG Ta or CIS at the first evaluation that follows at least 5 of 6 doses of induction BCG;
  • Adequate BCG is defined as at least 5 of 6 doses of induction BCG and at least 2 doses of 1 course of either reinduction or maintenance BCG. Inadequate BCG is defined as 3-6 doses.
  • HG NMIBC i.e., CIS and/or HG T1 and/or HG Ta
  • HGT1 disease at the first evaluation following at least 5 of 6 doses of induction BCG meet the current FDA definition of BCG-unresponsive disease (FDA 2018; FDA 2024) and do not meet the definition of BCG-exposed disease. Participants who have in the past met the FDA definition of BCG-Unresponsive NMIBC are not classified as BCG-exposed.
  • BCG-unresponsive NMIBC will be defined as the following based upon current FDA guidelines (FDA 2018; FDA 2024):
  • HGT1 disease at the first evaluation following at least 5 of 6 doses of induction BCG [0336] Participants with BCG-unresponsive disease must have had a prior diagnosis of any HG NMIBC (i.e., CIS and/or HG T1 and/or HG Ta) before receiving BCG therapy.
  • Adequate BCG is defined as at least 5 of 6 doses of induction BCG and at least 2 doses of 1 course of either reinduction or maintenance BCG with the minimum (5+2) completed within 12 months of initial induction dose. Any prior combination BCG treatment will be considered as additional BCG therapy.
  • BCG-unresponsive disease can be current or historical.
  • Study accrual is expected to occur over a maximum of approximately 12-18 months. Participant participation will be up to 48 months.
  • Cohort A consists of 3 treatment arms. Arms 1 and 2 will enroll up to 50 patients with pathologically confirmed carcinoma in situ (QS)-containing, high-risk NMIBC (i.e., CIS with or without concomitant Ta/Tl) who are naive to Bacillus Calmette -Guerin (BCG) treatment. Arm 3 will enroll up to 75 patients with pathologically confirmed, papillary-only, high risk, high-grade (HG) NMIBC (i.e., HG Ta/Tl papillary-only disease without CIS) who are naive to BCG treatment.
  • QS carcinoma in situ
  • HG high risk, high-grade NMIBC
  • Arms 1 and 2 will enroll up to 50 participants with pathologically confirmed, CIS containing, high-risk NMIBC (i.e., CIS with or without concomitant Ta/Tl) who are naive to BCG treatment.
  • Arm 3 (US and Canada) will enroll up to 75 participants with pathologically confirmed, papillary-only, high risk, HG NMIBC (i.e., HG Ta/Tl papillary-only disease without CIS) who are naive to BCG treatment.
  • Participants will be randomized 1 : 1 to receive DDM and cretostimogene as a 5 step instillation procedure (Arm 1) or a modified 2-step instillation procedure (Arm 2). Participants in Arm 3 will not be randomized and will have treatment allocated to receive cretostimogene using the modified 2 step instillation procedure.
  • Treatment will be performed in a classic induction course followed by maintenance therapy.
  • the induction and maintenance therapy schedules for all arms of this cohort are depicted in FIG. 5A.
  • cretostimogene will be administered as an induction course every week for 6 treatments on Weeks 1, 2, 3, 4, 5, and 6. If the participant has persistent CIS and/or HG Ta disease at Week 11, the participant may receive a reinduction cycle of 6 weekly treatments on Weeks 13, 14, 15, 16, 17, and 18. Participants who have HG T1 disease at the Week 11 assessment should discontinue the treatment phase.
  • efficacy assessments will be performed at 6-month intervals for up to an additional 2 years (i.e., up to 4 years from Week 1 Day 1 of treatment) or until disease recurrence, initiation of new anti-cancer therapy, documented withdrawal of consent by the participant, participant is lost to follow-up, or the study is terminated.
  • efficacy assessment procedures should be performed 2 weeks prior to the first dose of the next treatment cycle with an allowable window ranging from up to 3 weeks before dosing to the day of dosing.
  • Participants with recurrent disease will be discontinued from study treatment and followed for survival every 6 months from the date of recurrence until 4 years from Week 1 Day 1 of treatment or until documented withdrawal of consent by the participants, participant is lost to follow-up, or the study is terminated. Any new cancer therapy (including radical cystectomy/radiotherapy) and stage of disease at time of new therapy will also be documented.
  • participant To be eligible for participation in this trial, participants must meet all the following entry criteria. The participant must:
  • INR International normalized ratio
  • PT prothrombin time
  • the participant must be excluded from participating in the trial if the participant: [0361] 1. Has a current or past history of muscle invasive (T2 or higher stage), locally advanced (T3/T4, any N) or metastatic bladder cancer.
  • HG, non-muscle-invasive urothelial carcinoma in the upper genitourinary tract (kidneys, renal collecting systems, ureters) or urethra (including CIS of the prostatic urethra) within 24 months prior to randomization or treatment allocation
  • history of T2 urothelial carcinoma in the upper genitourinary tract in the past 48 months OR any history of T3 or higher urothelial carcinoma in the upper genitourinary tract.
  • Patients with a history of ⁇ T2 urothelial carcinoma of the upper genitourinary tract must have had curative treatment with no evidence of recurrence, nodal disease or metastasis since treatment.
  • HIV testing is not mandatory unless required by local health authorities. HIV without viral load or stable infection may be considered in consultation with Medical Monitor.
  • systemic immunosuppressive medication including high-dose corticosteroids (e.g., systemic corticosteroids >10 mg per day prednisone or equivalent), within 4 weeks prior to randomization or treatment allocation unless it is determined that the medication will not interfere with cretostimogene ’s mechanism of action or place the participant at undue risk.
  • Participants with contrast dye allergies may be given 1-3 dose(s) of a systemic steroid to complete a contrast-enhanced CT urogram) for trial purposes.
  • a steroid dose may also be administered as part of general anesthesia premedication, if required.
  • a 14 day minimum separation of the steroid dose and cretostimogene dose must occur.
  • live vaccines include, but are not limited to, the following: measles, mumps, rubella, varicella/zoster (chicken pox), yellow fever, rabies, intradermal BCG, and typhoid vaccine.
  • Seasonal influenza vaccines for injection are generally killed virus vaccines and are allowed; however, intranasal influenza vaccines (e.g., FluMist®) are live, attenuated vaccines and are not allowed. COVID-19 vaccination is permitted at least 7 days after cretostimogene treatment.
  • risk of metastasis or death e.g., risk of death or metastasis ⁇ 5% at 5 years
  • Cretostimogene will be administered at a dose of 1 x 10 12 vp intravesically following instillation of 0. 1% DDM.
  • Arm 1 Initial bladder wash with 100 mb normal saline then drain; Second bladder wash with 75 mb 0.1% DDM then drain; Instillation of 100 mb 0.1% DDM with 15 minutes dwell time ( ⁇ 5 minutes) then drain; Third bladder wash with 100 mb of normal saline then drain; Instillation of cretostimogene in up to 100 mb normal saline with 60 minutes dwell time ( ⁇ 5 minutes) then drain.
  • Arm 2 Instillation of 100 mb 0.1% DDM with 15 minutes dwell time ( ⁇ 5 minutes) then drain; Instillation of cretostimogene in up to 100 mb normal saline with 60 minutes dwell time ( ⁇ 5 minutes) then drain.
  • Arm 3 Instillation of 100 mb 0.1% DDM with 15 minutes dwell time ( ⁇ 5 minutes) then drain. Instillation of cretostimogene in up to 100 mb normal saline with 60 minutes dwell time ( ⁇ 5 minutes) then drain. [0386] The optimal instillation volume of DDM and cretostimogene is 85-100 mL. For participants with known limited bladder capacity, 55 mL ( ⁇ 10%) is the minimum target volume of cretostimogene allowed. Provided the entire volume of diluted cretostimogene is instilled, a reduction in normal saline flush volume will not be considered a protocol deviation.
  • Safety assessments will include physical examination, vital signs, assessment of treatment-emergent AEs, safety laboratory assessments.
  • Treatment-emergent AEs and serious adverse events will be captured from Treatment Day 1 through 30 days after the last study treatment; in addition, SAEs that are caused by a protocol-required screening procedure will be captured. Any other abnormal findings noted at Screening, including clinically relevant physical examination or laboratory findings, will be recorded as medical history.
  • Cystoscopy with directed TURBT/biopsy as indicated will occur every 3 months in Year 1 and Year 2, and every 6 months in Year 3 and Year 4.
  • Urine cytology will occur every 3 months in Year 1 and Year 2, and every 6 months in Year 3 and Year 4.
  • CT urogram/MR Urogram will occur every 6 months in all participants and as part of additional workup of an isolated positive urine cytology.
  • For-cause bladder mapping trigone, bladder dome, right, left, and posterior bladder walls
  • prostatic urethra biopsy in all males regardless of prior documented involvement
  • bilateral direct upper tract additional evaluation will occur as part of additional workup of an isolated positive urine cytology. Number of Participants
  • Arms 1 and 2 will enroll up to 50 participants with BCG-naive, CIS containing high risk NMIBC (i.e., CIS with or without Ta/Tl disease).
  • Arm 3 will enroll up to 75 patients with pathologically confirmed, papillary-only, high-risk, HG Ta/Tl NMIBC (i.e., HG Ta/Tl papillary-only disease without CIS) who are naive to BCG treatment.
  • Analysis for this cohort will utilize descriptive statistics (such as mean, median, standard deviation, ranges for continuous data, and percentages for catel data). These statistics will be used to summarize participant characteristics, treatment administration/compliance, efficacy, safety, and laboratory or exploratory parameters. For time-to-event variables (RFS, EFS, PFS, Overall Survival, etc.) survival analysis methods including Kaplan-Meier analyses will be used. Data will be displayed graphically, where appropriate.
  • Arms 1 and 2 will randomize up to 50 participants with BCG-Naive, CIS-containing high risk NMIBC in a 1: 1 ratio to Arm 1 and Arm 2.
  • the percentage of participants with CR at any time will be presented by randomized treatment arm and overall, with 95% confidence interval estimates. Included within analysis of secondary endpoints will be DoR, presented by randomized treatment arm and overall as well as presentation of cretostimogene genome and GM-CSF levels by randomized treatment arm and overall.
  • Arm 3 will enroll up to 75 participants with BCG-naive, papillary-only high-risk HG NMIBC.
  • HG EFS For the primary endpoint of HG EFS, a time-to-event analysis from time from first treatment on study to the first HG event (e.g., HG disease recurrence, progression, cystectomy or death) will be presented.
  • first HG event e.g., HG disease recurrence, progression, cystectomy or death
  • Cohort B will enroll up to 150 participants with pathologically confirmed high-risk HG NMIBC (i.e., CIS with or without concomitant HG Ta or T1 disease (Arm 1) OR HG Ta/Tl disease without CIS (Arm 2)) who have previously been exposed to BCG treatment [0402] Primary Objectives:
  • Cohort B will enroll up to 150 participants with pathologically confirmed high-risk HG NMIBC (i.e., CIS with or without concomitant HG Ta or T1 disease OR HG Ta/Tl disease without CIS) who have previously been exposed to BCG treatment.
  • HG NMIBC i.e., CIS with or without concomitant HG Ta or T1 disease OR HG Ta/Tl disease without CIS
  • cretostimogene will be administered as an induction course every week for 6 treatments on Weeks 1, 2, 3, 4, 5, and 6. If the participant has persistent CIS and/or HG Ta at Week 11, the participant may receive a re induction cycle of 6 weekly treatments on Weeks 13, 14, 15, 16, 17, and 18. Participants who have HG T1 disease at the Week 11 assessment should discontinue the treatment phase.
  • efficacy assessments will be performed at 6 month intervals for up to an additional 2 years (i.e., up to 4 years from Week 1 Day 1 of treatment) or until disease recurrence, initiation of new anti -cancer therapy, documented withdrawal of consent by the participant, participant is lost to follow-up, or the study is terminated.
  • efficacy assessment procedures should be performed 2 weeks prior to the first dose of the next treatment cycle with an allowable window ranging from up to 3 weeks before dosing to the day of dosing.
  • participant To be eligible for participation in this trial, participants must meet all the following entry criteria. The participant must:
  • HG NMIBC i.e., CIS with or without Ta/Tl disease or HG Ta/Tl papillary-only disease without CIS
  • INR or PT ⁇ 1.5 xULN unless receiving anticoagulation therapy and INR or PT is within the therapeutic range of intended use of anticoagulants. Participants must be able to suspend, based on local practice, anticoagulant and antiplatelet therapy for study-specific biopsies and procedures
  • the participant must be excluded from participating in the trial if the participant: [0426] 1. Has a current or past history of muscle invasive (T2 or higher stage), locally advanced (T3/T4, any N) or metastatic bladder cancer.
  • HG, non-muscle-invasive urothelial carcinoma in the upper genitourinary tract (kidneys, renal collecting systems, ureters) or urethra (including CIS of the prostatic urethra) within 24 months prior to randomization or treatment allocation
  • history of T2 urothelial carcinoma in the upper genitourinary tract in the past 48 months OR any history of T3 or higher urothelial carcinoma in the upper genitourinary tract.
  • Patients with a history of : T2 urothelial carcinoma of the upper genitourinary tract must have had curative treatment with no evidence of recurrence, nodal disease or metastasis since treatment.
  • HIV testing is not mandatory unless required by local health authorities. HIV without viral load or stable infection may be considered in consultation with Medical Monitor.
  • systemic immunosuppressive medication including high-dose corticosteroids (e.g., systemic corticosteroids >10 mg per day prednisone or equivalent), within 4 weeks prior to randomization or treatment allocation unless it is determined that the medication will not interfere with cretostimogene ’s mechanism of action or place the participant at undue risk.
  • Participants with contrast dye allergies may be given 1-3 dose(s) of a systemic steroid to complete a contrast-enhanced CT urogram) for trial purposes.
  • a steroid dose may also be administered as part of general anesthesia premedication, if required.
  • a 14 day minimum separation of the steroid dose and cretostimogene dose must occur.
  • live vaccines include, but are not limited to, the following: measles, mumps, rubella, varicella/zoster (chicken pox), yellow fever, rabies, intradermal BCG, and typhoid vaccine.
  • Seasonal influenza vaccines for injection are generally killed virus vaccines and are allowed; however, intranasal influenza vaccines (e.g., FluMist®) are live, attenuated vaccines and are not allowed. COVID-19 vaccination is permitted at least 7 days after cretostimogene treatment.
  • intranasal influenza vaccines e.g., FluMist®
  • Hepatitis B has known active Hepatitis B (defined as HBsAg reactive) or known active Hepatitis C virus (defined as HCV RNA [qualitative] is detected) infection.
  • Participants with active acute or active chronic Hepatitis B or C, who take medications with anti-Hepatitis B or C activity are ineligible.
  • Participants who have evidence of hepatic decompensation or cirrhosis on physical exam, laboratory, pathologic or radiographic testing also are ineligible.
  • Participants with inactive chronic Hepatitis B infection (asymptomatic healthy carriers) who meet the trial-defined laboratory parameters are eligible for trial entry. No testing for Hepatitis B and Hepatitis C is required unless mandated by a local health authority.
  • Cretostimogene will be administered at a dose of 1 x 10 12 vp intravesically following instillation of 0. 1% DDM.
  • a summary of the instillation procedure is provided: [0448] Instillation of 100 m 0.1% DDM with 15 minutes dwell time ( ⁇ 5 minutes) then drain; Instillation of cretostimogene in up to 100 m normal saline with 60 minutes dwell time ( ⁇ 5 minutes) then drain.
  • the optimal instillation volume of DDM and cretostimogene is 85-100 mb. Eor participants with known limited bladder capacity, 55 mb ( ⁇ 10%) is the minimum target volume of cretostimogene allowed. Provided the entire volume of diluted cretostimogene is instilled, a reduction in normal saline flush volume will not be considered a protocol deviation.
  • Safety assessments will include physical examination, vital signs, assessment of treatment-emergent AEs, safety laboratory assessments.
  • Treatment-emergent AEs and serious adverse events will be captured from Treatment Day 1 through 30 days after the last study treatment; in addition, SAEs that are caused by a protocol-required screening procedure will be captured. Any other abnormal findings noted at Screening, including clinically relevant physical examination or laboratory findings, will be recorded as medical history.
  • All AEs will be coded using the Medical Dictionary for Regulatory Activities (MedDRA) and graded according to the Common Terminology Criteria for Adverse Events (CTCAE) version 5.0.
  • MedDRA Medical Dictionary for Regulatory Activities
  • CCAE Common Terminology Criteria for Adverse Events
  • Cystoscopy with directed TURBT/biopsy as indicated will occur every 3 months in Year 1 and Year 2 and every 6 months in Year 3 and Year 4.
  • Urine cytology will occur every 3 months in Year 1 and Year 2 and every 6 months in Year 3 and Year 4.
  • CT urogram/ MR urogram will occur every 6 months in all participants and as part of additional workup of an isolated positive urine cytology.
  • Cohort B will enroll up to 150 participants with BCG-exposed, high-risk, HG NMIBC (i.e., CIS with or without Ta/Tl disease [Arm 1] or HG Ta/Tl papillary disease without CIS [Arm 2]).
  • HG NMIBC i.e., CIS with or without Ta/Tl disease [Arm 1] or HG Ta/Tl papillary disease without CIS [Arm 2]).
  • Cohort B will allocate treatment for up to 150 participants with BCG-exposed High Risk NMIBC, ⁇ 75 participants with CIS-containing High-Risk NMIBC (Arm 1) and ⁇ 75 participants with papillary-only High-Risk NMIBC (Arm 2).
  • Analysis for this cohort will utilize descriptive statistics (such as mean, median, standard deviation, ranges for continuous data, and percentages for categoril data). These statistics will be used to summarize participant characteristics, treatment administration/compliance, efficacy, safety, and laboratory or exploratory parameters. For time-to-event variables (RFS, EFS, PFS, Overall Survival, etc.) survival analysis methods including Kaplan-Meier analyses will be used. Data will be displayed graphically, where appropriate.
  • HG EFS Primary endpoint in Arm 2
  • Cohort CX will enroll up to 50 participants with pathologically confirmed, high-risk HG BCG unresponsive or BCG exposed NMIBC (i.e., CIS with or without Ta/Tl disease OR HG Ta/Tl papillary-only disease without CIS).
  • Cohort CX will enroll up to 50 participants with pathologically confirmed, high-risk HG BCG-unresponsive or BCG-exposed NMIBC (i.e., CIS with or without Ta/Tl disease or HG Ta/Tl papillary-only disease without CIS). Participants will be randomized 1: 1 to receive either DDM and cretostimogene with concurrent intravesical gemcitabine (Arm 1) or DDM and cretostimogene with sequential intravesical gemcitabine (Arm 2).
  • Treatment will be performed in a classic induction course followed by maintenance therapy.
  • the induction and maintenance therapy schedules for Arm 1 of Cohort CX are depicted in FIG. 5A.
  • the induction and maintenance therapy schedules for Arm 2 of Cohort CX are depicted in FIG. 5B.
  • cretostimogene and gemcitabine will be administered in combination (DDM and cretostimogene instilled first, followed by gemcitabine) as an induction course every week for 6 treatments on Weeks 1, 2, 3, 4, 5, and 6. If the participant has persistent CIS and/or HG Ta disease at Week 11, the participant may receive a reinduction cycle of 6 weekly treatments of cretostimogene and gemcitabine on Weeks 13, 14, 15, 16, 17, and 18. Participants who have HG T1 disease at the Week 11 assessment should discontinue the treatment phase.
  • cretostimogene will be administered as an induction course on Weeks 1, 2, 4 and 5.
  • gemcitabine will be administered. If the participant has persistent CIS and/or HG Ta disease at Week 11, the participant may receive a reinduction cycle of cretostimogene on Weeks 13, 14, 16 and 17 and gemcitabine on Weeks 15 and 18. Participants who have HG T1 disease at the Week 11 assessment should discontinue the treatment phase.
  • efficacy assessments will be performed at 6 month intervals for up to an additional 2 years (i.e., up to 4 years from Week 1 Day 1 of treatment) or until disease recurrence, initiation of new anti -cancer therapy, documented withdrawal of consent by the participant, participant is lost to follow-up, or the study is terminated.
  • efficacy assessment procedures should be performed 2 weeks prior to the first dose of the next treatment cycle with an allowable window ranging from up to 3 weeks before dosing to the day of dosing.
  • participant [0479] To be eligible for participation in this trial, participants must meet all the following entry criteria. The participant must: [0480] 1. Be >18 years of age (or legal age of majority in the jurisdiction) on day of signing informed consent.
  • HG BCG-unresponsive or BCG-exposed NMIBC i.e., CIS with or without Ta/Tl disease or HG Ta/Tl papillary-only disease without CIS.
  • the participant must be excluded from participating in the trial if the participant: [0489] 1. Has a current or past history of muscle invasive (T2 or higher stage), locally advanced (T3/T4, any N) or metastatic bladder cancer. [0490] 2. Any HG, non-muscle-invasive urothelial carcinoma (Tl, HG Ta, or CIS) in the upper genitourinary tract (kidneys, renal collecting systems, ureters) or urethra (including CIS of the prostatic urethra) within 24 months prior to randomization or treatment allocation OR history of T2 urothelial carcinoma in the upper genitourinary tract in the past 48 months OR any history of T3 or higher urothelial carcinoma in the upper genitourinary tract. Patients with a history of : T2 urothelial carcinoma of the upper genitourinary tract must have had curative treatment with no evidence of recurrence, nodal disease or metastasis since treatment.
  • HIV testing is not mandatory unless required by local health authorities. HIV without viral load or stable infection may be considered in consultation with Medical Monitor.
  • systemic immunosuppressive medication including high-dose corticosteroids (e.g., systemic corticosteroids >10 mg per day prednisone or equivalent), within 4 weeks prior to randomization or treatment allocation unless it is determined that the medication will not interfere with cretostimogene’s mechanism of action or place the participant at undue risk.
  • Participants with contrast dye allergies may be given 1-3 dose(s) of a systemic steroid to complete a contrast-enhanced CT urogram) for trial purposes.
  • a steroid dose may also be administered as part of general anesthesia premedication, if required.
  • a 14 day minimum separation of the steroid dose and cretostimogene dose must occur.
  • live vaccines include, but are not limited to, the following: measles, mumps, rubella, varicella/zoster (chicken pox), yellow fever, rabies, intradermal BCG, and typhoid vaccine.
  • Seasonal influenza vaccines for injection are generally killed virus vaccines and are allowed; however, intranasal influenza vaccines (e.g., FluMist®) are live, attenuated vaccines and are not allowed. COVID-19 vaccination is permitted at least 7 days after cretostimogene treatment.
  • [0501] 13 Has known active Hepatitis B (defined as HBsAg reactive) or known active Hepatitis C virus (defined as HCV RNA [qualitative] is detected) infection. Participants with active acute or active chronic Hepatitis B or C, who take medications with anti-Hepatitis B or C activity are ineligible. Participants who have evidence of hepatic decompensation or cirrhosis on physical exam, laboratory, pathologic or radiographic testing also are ineligible. Participants with inactive chronic Hepatitis B infection (asymptomatic healthy carriers) who meet the trial-defined laboratory parameters are eligible fortrial entry. No testing for Hepatitis B and Hepatitis C is required unless mandated by a local health authority.
  • [0505] Has not recovered (i.e., to Grade I or to Baseline status) from adverse events (AEs) due to a previously administered agent or therapy. Participants with Grade : 2 neuropathy or participants having any alopecia are an exception to this criterion and may qualify for the study. If the participant received major surgery, they must be at least 4 weeks from surgery and have recovered adequately from the toxicity and/or complications from the surgery prior to randomization or treatment allocation. TURBT or biopsy is not considered major surgery. Participants with irreversible but not clinically significant toxicity are eligible. [0506] 18. Has an active infection requiring systemic therapy (e.g., active infection that requires long-term intravenous or oral antibiotics).
  • systemic therapy e.g., active infection that requires long-term intravenous or oral antibiotics.
  • Cretostimogene will be administered at a dose of 1 x 10 12 vp intravesically following instillation of 0.1% DDM.
  • Arm 1 gemcitabine will be administered at a dose of 1g intravesically.
  • Arm 2 gemcitabine will be administered at a dose of 2g intravesically
  • a summary of the instillation procedure is provided: [0511] Arm 1: Instillation of 100 mL 0.1% DDM with 15 minutes dwell time ( ⁇ 5 minutes) then drain. Then, instillation of cretostimogene in up to 100 mL normal saline with 60 minutes dwell time ( ⁇ 5 minutes) then drain. Then, instillation of 1g gemcitabine in 50-100ml normal saline with 60 minutes dwell time ( ⁇ 5 minutes) then drain.
  • the optimal instillation volume of DDM and cretostimogene is 85-100 mL.
  • 55 mL ( ⁇ 10%) is the minimum target volume of cretostimogene allowed.
  • a reduction in normal saline flush volume will not be considered a protocol deviation.
  • Safety assessments will include physical examination, vital signs, assessment of treatment-emergent AEs, safety laboratory assessments.
  • Treatment-emergent AEs and serious adverse events will be captured from Treatment Day 1 through 30 days after the last study treatment; in addition, SAEs that are caused by a protocol-required screening procedure will be captured. Any other abnormal findings noted at Screening, including clinically relevant physical examination or laboratory findings, will be recorded as medical history.
  • All AEs will be coded using the Medical Dictionary for Regulatory Activities (MedDRA) and graded according to the Common Terminology Criteria for Adverse Events (CTCAE) version 5.0.
  • MedDRA Medical Dictionary for Regulatory Activities
  • CCAE Common Terminology Criteria for Adverse Events
  • Cystoscopy with directed TURBT/biopsy as indicated will occur every 3 months in Year 1 and Year 2, and every 6 months in Year 3 and Year 4.
  • Urine cytology will occur every 3 months in Year 1 and Year 2, and every 6 months in Year 3 and Year 4.
  • CT urogram/MR Urogram will occur every 6 months in all participants and as part of additional workup of an isolated positive urine cytology.
  • For-cause bladder mapping trigone, bladder dome, right, left, and posterior bladder walls
  • prostatic urethra biopsy in all males regardless of prior documented involvement
  • bilateral direct upper tract additional evaluation will occur as part of additional workup of an isolated positive urine cytology
  • Cohort CX will enroll up to 50 participants with pathologically confirmed, high-risk HG BCG-unresponsive or BCG-exposed NMIBC (i.e., CIS with or without Ta/Tl disease or HG Ta/Tl papillary-only disease without CIS.
  • HG BCG-unresponsive or BCG-exposed NMIBC i.e., CIS with or without Ta/Tl disease or HG Ta/Tl papillary-only disease without CIS.
  • Cohort CX will randomize up to 50 participants with high-risk BCG-unresponsive or BCG-exposed NMIBC in a 1 : 1 ratio to concurrent (Arm 1) or sequential (Arm 2) cretostimogene and gemcitabine.
  • Analysis for this cohort will utilize descriptive statistics (such as mean, median, standard deviation, ranges for continuous data, and percentages for categoril data). These statistics will be used to summarize participant characteristics, treatment administration/compliance, efficacy, safety, and laboratory or exploratory parameters.
  • time-to-event variables RFS, EFS, PFS, Overall Survival, etc.
  • survival analysis methods including Kaplan-Meier analyses will be used. Data will be displayed graphically, where appropriate.
  • HG EFS For the primary efficacy endpoint (HG EFS), a time-to-event analysis from time from first treatment on study to the first HG event (e.g., HG disease recurrence, progression, cystectomy or death) will be presented by randomized treatment arm and overall.
  • MRD minimal residual disease
  • VAF variant allele frequency
  • GDB cretostimogene monotherapy or cretostimogene in combination with pembrolizumab.
  • RFS durable 12-month regression-free survival
  • VAF, GDB, and MRD can serve as useful prognostic tools for oncolytic virotherapy.
  • BOND-003 is an ongoing phase 3 open-label, single-arm trial evaluating cretostimogene monotherapy in conjunction with DDM bladder pre-treatment.
  • BOND-003 is registered on ClinicalTrials.gov under the identifier NCT04452591 and is described in Example 3 and in Tyson 2024 (Topline Results from BOND-003: A Phase 3, Study of Intravesical Cretostimogene Grenadenorepvec for the Treatment of BCG-Unresponsive, High-Risk NMIBC with CIS. Presented at: Society of Urologic Oncology Annual
  • CORE-001 was a phase 2, single-arm study to evaluate the efficacy and safety of intravesical administration of cretostimogene and intravenous pembrolizumab the intravenous PD-1 inhibitor pembrolizumab in BCG-unresponsive CIS.
  • Cretostimogene was administered in conjunction with a DDM pre-treatment composition using the same dosage and treatment schedule as the BOND-003 trial, with all patients receiving weekly induction cretostimogene for 6 weeks (with a reinduction course for non-responders at 3 months), followed by maintenance cretostimogene every 3 months for the first year and then every 6 months for up to 36 months.
  • Pembrolizumab was administered intravenously every 6 weeks for up to 24 months.
  • GDB LJroAmp -Derived Genomic Disease Burden
  • MRD Minimal Residual Disease
  • UroAmp® Convergent Genomics
  • utDNA urine tumor DNA
  • Genomic Disease Burden (GDB): Definition and Calculation. GDB is a quantitative metric that reflects the overall molecular tumor burden in a patient's urine sample. It is calculated by comparing the total variant allele frequency (VAF) profile of a given sample against an empirical distribution derived from a reference cohort of bladder cancer patients tested using the UroAmp® platform. The GDB score is expressed as a percentile ranking — the higher the GDB percentile, the greater the molecular tumor load. GDB provides a continuous measure of disease burden, making it valuable for baseline risk stratification, longitudinal monitoring, and treatment response assessment.
  • VAF total variant allele frequency
  • GDB is calculated from genomic alterations identified through UroAmp®’s urinebased comprehensive genomic profiling (uCGP), which uses next-generation sequencing (NGS) to detect:
  • MSI Microsatellite Instability
  • GDB GDB
  • MRD Minimum Residual Disease
  • VAF Variant Allele Frequency
  • Validated MRD Algorithm Uses a defined VAF threshold to classify samples.
  • MRD-negative patients (VAF ⁇ 8%) had a 12-month RFS of 56%, compared to 22% in MRD-positive patients (VAF > 8%)
  • VAF > 8% MRD-positive patients
  • GDB-Enhanced MRD Algorithm Improves sensitivity in detecting residual disease by incorporating GDB as an override mechanism. Specifically, patients who are initially classified as MRD-negative based on VAF may be reclassified as MRD-positive if their GDB score is > 42, allowing the algorithm to capture additional high-risk cases that may otherwise go undetected through VAF alone. In the same Phase 2 nadofaragene firadenovec study, the authors evaluated an enhanced MRD algorithm that incorporated Genomic Disease Burden (GDB) as an override criterion. Specifically, samples initially classified as MRD-negative were reclassified as MRD-positive if the GDB score was > 42.
  • GDB Genomic Disease Burden
  • MRD is tracked over time, enabling classification of patients into:
  • GDB vs. MRD Relationship and Distinction: While both GDB and MRD are derived from the same uCGP data, they serve distinct purposes:
  • MRD detects low-level disease associated with recurrence risk using VAF thresholds.
  • GDB is not part of the core MRD algorithm but can enhance sensitivity when used as an override in the GDB-enhanced MRD model.
  • GDB and MRD represent independent yet synergistic tools for guiding clinical decisions.
  • MRD status is influenced by VAF thresholds, while GDB aggregates all mutation data.
  • VAF% > 8% provided a robust and consistent threshold for MRD classification across studies.
  • GDB reflects total disease burden, useful for monitoring and trend analysis.
  • MRD offers actionable insight into residual disease and recurrence risk.
  • the GDB-enhanced MRD algorithm combines the strengths of both, improving sensitivity for high-risk detection.
  • FIGS. 6A-6B show the molecular response to cretostimogene monotherapy in BOND-003 trial patients, including mutation count, GDB, and MRD status. There is a dramatic loss of aneuploidy in responders at the 6-month timepoint (FIG. 6B) relative to the pre-treatment timepoint (FIG. 6A), as well as an overall reduction in GDB. This demonstrates that loss of aneuploidy is associated with reduced risk of progression.
  • FIGS. 7A-7B show the molecular response to the combination of cretostimogene and pembrolizumab administered in the CORE-001 trial patients. Similar to BOND-003, there is a dramatic reduction in aneuploidy and in overall GDB at the 6-month timepoint (FIG. 7B) relative to the pretreatment timepoint (FIG. 7A), further confirming that loss of aneuploidy is associated with reduced risk of progression. Additionally, at the 6-month timepoint (FIG.
  • FIG. 8 displays the most frequently mutated genes identified in baseline urine samples from BCG-unresponsive NMIBC patients enrolled in the BOND-003 and CORE-001 trials, highlighting TERT (over 50% of patients) and TP53 (over 40% of patients) as frequently-altered genes in this patient population.
  • RBI variants were identified in over 10% of the cohort, which is notable as Rb pathway genes are key regulators of cell cycle control and major factors for cretostimogene replication (FIG. 8). In addition, nearly 20% of patients were found to have aneuploidy (FIG. 8), confirming the results shown in FIG. 6A.
  • MRD status was determined based on variant allele frequency (VAF), the patients having a VAF of at least 8% classified as MRD-positive, and the patients having a VAF of less than 8% classified as MRD-negative.
  • VAF variant allele frequency
  • a p-value of ⁇ 0.001 indicates a statistically significant difference in survival outcomes between the two groups.
  • FIG. 10 presents a series of box plots stratified by response category in BOND-003 trial:
  • Each plot tracks maximum variant allele frequency (VAF%) over time: W1 (baseline), W13 ( ⁇ 3 months after the 1st dose), W25 ( ⁇ 6 months after the 1st dose), W37 ( ⁇ 9 months after the 1st dose), W49 ( ⁇ 12 months after the 1st dose).
  • the number of patients (population size) contributing data at each timepoint is also shown.
  • VAF NR to CR patients start with relatively high VAF at Wl, which remains modestly elevated at Wl 3 (after the first induction cycle), but notably declines by W25-W49 following a second induction course (FIG. 10).
  • FIGS. 6A-6B and 7A-7B demonstrate that a loss of aneuploidy is associated with reduced risk of progression.
  • FIG. 9 and FIG. 10 demonstrate that MRD status and VAF, respectively, can be used as a prognostic for recurrence.
  • This Example describes Cohort P of BOND-003, a Phase 3, single-arm, open-label clinical trial designed to evaluate intravesical cretostimogene in patients with BCG- unresponsive NMIBC.
  • Cohort P is designed to evaluate the activity of IVE cretostimogene in patients with tissue pathology-confirmed high-grade Ta (HG Ta) or high-grade T1 (HG Tl) NMIBC without carcinoma in situ (CIS).
  • HG EFS high-grade event free survival

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  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

La présente divulgation concerne des méthodes pour des individus atteints d'un cancer de la vessie comprenant l'administration intravésicale à l'individu d'un virus oncolytique, tel que le crétostimogène grenadenorepvec, éventuellement en combinaison avec un ou plusieurs agents thérapeutiques supplémentaires, tels qu'un modulateur de point de contrôle immunitaire ou un agent chimiothérapeutique. L'invention concerne également des compositions pharmaceutiques et des kits pour le traitement du cancer de la vessie.
PCT/US2025/031754 2024-05-31 2025-05-30 Méthodes de traitement du cancer de la vessie à l'aide d'un virus oncolytique et d'un agent améliorant la transduction Pending WO2025251009A1 (fr)

Applications Claiming Priority (8)

Application Number Priority Date Filing Date Title
US202463654584P 2024-05-31 2024-05-31
US63/654,584 2024-05-31
US202463714668P 2024-10-31 2024-10-31
US63/714,668 2024-10-31
US202563790586P 2025-04-17 2025-04-17
US63/790,586 2025-04-17
US202563794976P 2025-04-25 2025-04-25
US63/794,976 2025-04-25

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WO2025251009A1 true WO2025251009A1 (fr) 2025-12-04

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7459154B2 (en) * 2002-12-26 2008-12-02 Cell Genesys, Inc. Methods and reagents for the enhancement of virus transduction in the bladder epithelium
US11596660B2 (en) * 2017-04-14 2023-03-07 Cg Oncology, Inc. Methods of treating bladder cancer
US12090183B2 (en) * 2016-03-10 2024-09-17 Cg Oncology, Inc. Methods of treating solid or lymphatic tumors by combination therapy

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7459154B2 (en) * 2002-12-26 2008-12-02 Cell Genesys, Inc. Methods and reagents for the enhancement of virus transduction in the bladder epithelium
US12090183B2 (en) * 2016-03-10 2024-09-17 Cg Oncology, Inc. Methods of treating solid or lymphatic tumors by combination therapy
US11596660B2 (en) * 2017-04-14 2023-03-07 Cg Oncology, Inc. Methods of treating bladder cancer

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