WO2024250002A1 - Méthodes de prédiction de survie sans progression et de survie globale chez des sujets atteints d'un cancer à l'aide de cellules hybrides de macrophages tumoraux - Google Patents

Méthodes de prédiction de survie sans progression et de survie globale chez des sujets atteints d'un cancer à l'aide de cellules hybrides de macrophages tumoraux Download PDF

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WO2024250002A1
WO2024250002A1 PCT/US2024/032257 US2024032257W WO2024250002A1 WO 2024250002 A1 WO2024250002 A1 WO 2024250002A1 US 2024032257 W US2024032257 W US 2024032257W WO 2024250002 A1 WO2024250002 A1 WO 2024250002A1
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cancer
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tmhcs
cells
biological sample
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Daniel L. Adams
Cha-Mei Tang
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Creatv Microtech Inc
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D43/00—Separating particles from liquids, or liquids from solids, otherwise than by sedimentation or filtration
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/575—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D2239/00—Aspects relating to filtering material for liquid or gaseous fluids
    • B01D2239/06—Filter cloth, e.g. knitted, woven non-woven; self-supported material
    • B01D2239/065—More than one layer present in the filtering material
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D2239/00—Aspects relating to filtering material for liquid or gaseous fluids
    • B01D2239/12—Special parameters characterising the filtering material
    • B01D2239/1216—Pore size
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/52—Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis

Definitions

  • the present invention generally relates to the use of biomarkers in the blood and other bodily fluids to make predictions regarding overall survival and progression free survival in subjects having cancer, such as solid tumors.
  • CTCs circulating tumor cells
  • CTCs are not consistently associated with the development and/or presence of cancer in a subject, even in stage IV cancer patients. While CTCs are found most often in stage IV of breast, prostate and colorectal cancers, they are rare in early stages of the same cancer. CTCs are also rare in other cancers.
  • Circulating Cancer Associated Macrophage-Like cells are another cancer- related cell type that is found in the blood of subjects having cancer.
  • CAMLs are associated with all solid tumors tested and all stages of cancer.
  • CAMLs are polyploid and very large in size, ⁇ 20 pm to -300 pm in size. These polyploid cells can be either CD45(-) or CD45(+) and express CD11c, CD 14 and CD31, which confirms their origin as a myeloid lineage. They are often found in the process of engulfing CTCs and cell debris I 14 - 19 !.
  • the examination of protein expression levels in CAMLs can also assist clinicians in making informed treatment decisions.
  • Assays associated with additional types of cancer-associated circulating cells, in blood and other body fluids could be used to provide further important prognostic and treatment information for a subject having cancer. The present invention is directed to discovery of such cells and other important goals.
  • the present invention is directed prognostic methods that make use of Tumor Macrophage Hybrid Cells (TMHCs), a unique group of cell types that include fusions and/or clusters of different cells that are found in the blood of subjects having solid tumors, including carcinoma, sarcoma, neuroblastoma and melanoma. These circulating cells have been shown to be associated with the presence of solid tumors in a subject having cancer. TMHCs have been found in peripheral blood of subjects having solid tumors via microfdtration using precision microfilters.
  • TMHCs Tumor Macrophage Hybrid Cells
  • TMHCs are an indicator of more aggressive disease with worse clinical outcomes.
  • medical applications associated with TMHCs include, but are not limited to, use of the cells themselves as biomarkers to provide early detection of cancer and diagnosis of cancer, in particular, in the early detection and diagnosis of cancer relapse or recurrence, and in the determination of cancer mutation.
  • the present invention is directed to a method for predicting overall survival (OS) and/or progression free survival (PFS) of a subject having cancer.
  • the method comprises determining the presence of TMHCs in a biological sample from a subject having cancer, wherein the presence of TMHCs predicts lower OS and/or PFS than a subject having the same cancer without the presence of TMHCs.
  • the invention is directed to a method for predicting presence of metastatic spread and/or metastatic progression in a subject having cancer.
  • the method comprises determining the presence of TMHCs in a biological sample from a subject having cancer, wherein the presence of TMHCs predicts presence of metastatic spread and/or metastatic progression in the subject.
  • the invention is directed to a method for predicting cancer progression in a subject having cancer.
  • the method comprises determining the presence TMHCs in a first biological sample and a second biological sample at a later date, and optional additional biological samples, obtained from a subject having cancer, wherein when TMHCs are present in the second and/or additional biological sample but not present in the first biological sample, the cancer is predicted to progress in the subject.
  • the invention is directed to a method for predicting response to treatment in a subject having cancer.
  • the method comprises determining the presence TMHCs in a first biological sample and a second biological sample at a later date, and optional additional biological samples, obtained from a subject having cancer, wherein the first sample is obtained from the subject prior to or during cancer treatment, wherein the second sample and optional additional samples are obtained from the subject after at least one cancer treatment, wherein when TMHCs are present in the first biological sample but not present in the second and/or additional biological sample, the subject is predicted to respond to the treatment.
  • the TMHCs comprise one or more of the following cell types:
  • CTC circulating tumor cell
  • EMT CTCs epithelial-mesenchymal transition circulating tumor cell
  • OS and/or PFS is over a period of at least 12 months. In other aspects of the first embodiment, OS and/or PFS is over a period of at least 24 months.
  • the biological sample is one or more of blood (such as peripheral blood), lymph node, bone marrow, cerebral spinal fluid, and urine.
  • the biological sample may be antecubital-vein blood, inferior-vena-cava blood, femoral vein blood, portal vein blood, or jugular-vein blood.
  • the size of the biological sample is between 5 and 15 mb.
  • the cancer in each of the embodiments of the invention is a Stage I cancer, Stage II cancer, Stage III cancer, Stage IV cancer, carcinoma, sarcoma, neuroblastoma, melanoma, epithelial cell cancer, lung cancer, breast cancer, prostate cancer, pancreatic cancer, bladder cancer, kidney cancer, head and neck cancer, colorectal cancer, liver cancer, ovarian cancer, osteosarcoma, esophageal, brain & ONS, larynx, bronchus, oral cavity and pharynx, stomach, testis, thyroid, uterine cervix, uterine corpus cancer or other solid tumor cancers, or a blood cancer.
  • the TMHCs may be isolated from the biological samples for the determining steps using one or more means selected from the group consisting of size exclusion methodology, immunocapture, red blood cell lysis, white blood cell depletion, a high-molecular weight polysaccharide such as FICOLL®, electrophoresis, dielectrophoresis, flow cytometry, magnetic levitation, and various microfluidic chips, slits, channels, hydrodynamic size-based sorting, grouping, trapping, concentrating large cells, eliminating small cells, or a combination thereof.
  • the TMHCs may be isolated from the biological samples using size exclusion methodology that comprises using a microfilter.
  • the microfdter may have a pore size ranging from about 5 microns to about 20 microns. Additionally, the pores of the microfdter may have a round, race-track shape, oval, square and rectangular pore shape.
  • the microfdter may also have precision pore geometry and uniform pore distribution.
  • the TMHCs may be isolated using a microfluidic chip via physical size-based sorting, hydrodynamic size-based sorting, grouping, trapping, immunocapture, concentrating large cells, or eliminating small cells based on size.
  • the TMHCs may be isolated from the biological samples for the determining steps using a microfdtration assay.
  • the treatment is one or more of chemotherapy, single drug, combination of drugs, immunotherapy, radiation therapy, chemoradiation, radiation combined with single or multiple drugs, chemoradiation combined with single or multiple drugs, cancer vaccine, and cell therapy.
  • the treatment may be a cancer vaccine and the subject expresses at least one HLA allele.
  • the subject may be treated with one or more of chemotherapy, single drug, combination of drugs, immunotherapy, radiation therapy, chemoradiation, radiation combined with single or multiple drugs, chemoradiation combined with single or multiple drugs, cancer vaccine, and cell therapy.
  • the immunotherapy may be PD-L1 immunotherapy.
  • TMHCs may be used independently as cancer markers, or in combination with biomarker expression by other circulating cells, such as CAMLs and other circulating tumor cells (CTCs).
  • CTCs circulating tumor cells
  • Suitable CTC subtypes include, but are not limited to, pathologically definable CTCs (PDCTCs), apoptotic CTCs, circulating cancer associated vascular endothelial cells (CAVEs).
  • PDCTCs pathologically definable CTCs
  • CAVEs circulating cancer associated vascular endothelial cells
  • the presence of these additional cell types themselves and also cell-free DNA (cfDNA), circulating tumor DNA (ctDNA), methylated DNA, proteomic, metabolomic, lipidomic and other biomarkers may provide a more complete understanding of a patient’s disease.
  • the methods further comprise administering a therapeutically effective amount of a cancer treatment to the subject.
  • the subject may be a subject in which the OS and PFS is predicted to be lower or shorter than the OS and PFS of another subject.
  • FIG. l is a diagram of the different TMFC subtypes and images of TMFCs from metastatic breast cancer patient blood.
  • FIG. 2 is a diagram of the different CTC cluster subtypes and images of CTC clusters from metastatic breast cancer patient blood.
  • FIG. 3 shows survival times.
  • A. shows progression free survival (PFS) for mBC patients with any TMFCs (types (1 )-(4)) detected in blood.
  • B. shows overall survival (OS) for mBC patients with any TMFCs (types (l)-(4)) detected in blood.
  • PFS progression free survival
  • OS overall survival
  • FIG. 4 shows Cox Proportional Survival Analysis of PFS (A.) and OS (B.) for no CTCs (Green Line), with CTCs (Black Line) or with TMFCs (not including hyperploidy fusion cells (CAMLs)) (Red Line) at BL.
  • FIG. 5 shows PFS of Homotypic vs Heterotypic CTC Clusters.
  • FIG. 6 shows PFS of EMT Clusters vs > 50 pm CAMLs.
  • Cancer is one of the most feared illness in the world, affecting all populations and ethnicities in all countries. Approximately 40% of both men and women will develop cancer in their lifetime. In the United States alone, at any given time there are more than 12 million cancer patients, with 1.7 million new cancer cases and more than 0.6 million deaths estimated yearly. Cancer death worldwide is estimated to be about 8 million annually, of which 3 million occur in developed countries where patients have access to treatment.
  • Liquid biopsies provide real-time, sequential tracking of diagnostically -important circulating cells isolated from a subject having cancer.
  • Cells such as circulating tumor cells (CTCs) are found in the peripheral blood of cancer patients and previous work has shown that CTC-based assays can be used as a substitute to tissue biopsies [1 ' 4] .
  • CAML cancer associated macrophage-like cell
  • CAMLs have been identified in the blood using a non-affinity microfiltration based method which captures both CTCs and CAMLs, and allows for singular or parallel analysis of these cancer specific circulating cell subtypes [1, 6 ' 16] .
  • CAMLs are a recently defined circulating myeloid derived stromal cell, found in all the stages of invasive malignancy and in various solid malignancies (e.g. breast, prostate, non-small cell lung carcinoma (NSCLC), and pancreatic) [lt 13, 14, 17] .
  • NSCLC non-small cell lung carcinoma
  • CAMLs are specialized myeloid polyploid cells in the blood in all stages of solid tumors. They are easy to identify by their large size (greater than 20 pm), polyploid nucleus and morphologies: round, rod shaped, with one tail or two tails 180 degrees apart.
  • CAMLs typically express CD31, CD 14, CD45 and cytokeratin, and can also express EpCAM, CD 146, CD11c and tie2 [11, 13, 14, 17] .
  • TMHCs Tumor Macrophage Hybrid Cells
  • TMHCs have been found in peripheral blood of subjects having solid tumors via microfdtration using precision microfilters.
  • TMHCs The presence of TMHCs in the blood of a subject having cancer is an indicator of more aggressive disease with worse clinical outcomes.
  • medical applications associated with TMHCs include, but are not limited to, use of the cells themselves as biomarkers to provide early detection of cancer and diagnosis of cancer, in particular, in the early detection and diagnosis of cancer relapse or recurrence, and in the determination of cancer mutation.
  • the simple presence of one or more of these circulating cell types in a blood sample obtained from a subject having cancer provides diagnostic information.
  • THCs Tumor Macrophage Hybrid Cells
  • TMFCs tumor macrophage fusion cells
  • CTCs circulating tumor cell clusters
  • TMHCs Tumor Macrophage Hybrid Cells
  • TMHCs Tumor Macrophage Hybrid Cells
  • EMT CTCs epithelial-mesenchymal transition circulating tumor cell
  • Partial tumor macrophage fusion cells are cellular fusions between tumor cells and macrophages where there is some membrane interaction between the cells and both cells retaining their original phenotypes (FIG. 1).
  • Homodimeric tumor macrophage fusion cells are cellular fusions between tumor cells and macrophages where the two cells have a fused membrane and shared cytoplasm (FIG. 1).
  • Cannibalistic tumor macrophage fusion cells are cellular fusions between tumor cells and macrophages where the tumor cells has been taken up by the macrophage and both cells retaining their original phenotypes (FIG. 1).
  • Binucleated tumor macrophage fusion cells are cellular fusions between tumor cells and macrophages where both cells completely merge to become one cell with dual expression of phenotypes (FIG. 1).
  • Heterotypic CTC clusters are clusters of CTCs and immune/stromal white blood cells (WBCs) (FIG. 2).
  • Homotypic CTC clusters are clusters of only CTCs (FIG. 2).
  • Epithelial-mesenchymal transition circulating tumor cell (EMT CTCs) clusters are clusters of CTCs undergoing epithelial-mesenchymal transition (EMT), a process whereby tumor cells downregulate epithelial traits and upregulate mesenchymal traits (FIG. 2).
  • CTCs associated with carcinomas express a number of cytokeratins (CKs).
  • CK 8, 18, & 19 are the cytokeratins most commonly expressed and used in diagnostics, but surveying need not be limited to these markers alone.
  • the surface of solid tumor CTCs usually express epithelial cell adhesion molecule (EpCAM). However, this expression is not uniform or consistent.
  • CTCs do not express any CD45 because it is a white blood cell marker. In assays to identify tumor-associated cells, such as CTCs and CAMLs, it is sufficient to use antibodies against markers associated with the solid tumor such as CK 8, 18 and 19, or antibodies against CD45 or DAPI.
  • CTCs upregulate and/or down regulate phenotypes and marker expression in relation to tumor progression, tumor spread, and in response to tumor treatments. Therefore, assessing CTCs and the markers expressed by such cells in the peripheral blood can provide important information regarding the status of the cancer in the subject.
  • CTCs that can be used as cancer diagnostics, as well as means for screening and monitoring treatment and determining the susceptibility of a tumor in a particular subject to a particular treatment, can be divided into three subgroups.
  • the first subgroup is pathologically definable CTCs (PDCTCs) [6 ’ 101 .
  • PDCTCs can be characterized by: a “cancer-like” nucleus stained by DAPI; cytokeratin having a filamentous pattern; expression of one or more of CK 8, 18 and 19 (CTCs from epithelial cancers usually express at least CK 8, 18 and 19); lack of CD45 expression.
  • the second subgroup is apoptotic CTCs.
  • the cytokeratin pattern degrades into dots. Therefore, early apoptotic CTC have some cytokeratin dots and later apoptotic CTCs have all of the cellular cytokeratin degraded into dots.
  • Apoptotic CTCs can also be characterized as expressing CK 8, 18 and 19; a degrading nuclei; lack of CD45 expression.
  • EMTCTCs epithelial to mesenchymal transition
  • EMT epithelial to mesenchymal transition
  • EMTCTCs encompass cells in various stages of transition [6] .
  • EMTCTCs can be generally described by the down regulation of epithelial proteins, e.g. EpCAM and CK, and the upregulation of mesenchymal stem cell proteins, e.g. vimentin and CD34 [13] .
  • EMTCTC subtyping is typically performed using non-proteomic methods, i.e. mRNA expression or DNA analysis [13] .
  • CAVEs express cytokeratin and various subtypes endothelial cell markers such as CD31, CD146, CD144, CD105, but do not express CD14 or CD45 [18 b [0052] Combining staining techniques with morphology, pathologically-definable CTCs (PDCTC), apoptotic CTCs and CAMLs can be identified [6] .
  • PDCTC pathologically-definable CTCs
  • CAMLs can be identified [6] .
  • TMHCs unique characteristics make them well-suited for use in clinical methodology including methods of screening and diagnosis diseases such as cancer, monitoring treatment, monitoring of disease progression and recurrence. It has been shown that the presence of TMHCs in the blood of a subject is an indicator of more aggressive disease with worse clinical outcomes.
  • the present invention is directed to a method for predicting overall survival (OS) and/or progression free survival (PFS) of a subject having cancer.
  • the method comprises determining the presence of TMHCs in a biological sample from a subject having cancer, wherein the presence of TMHCs predicts lower OS and/or PFS than a subject having the same cancer without the presence of TMHCs.
  • the invention is directed to a method for predicting presence of metastatic spread and/or metastatic progression in a subject having cancer.
  • the method comprises determining the presence of TMHCs in a biological sample from a subject having cancer, wherein the presence of TMHCs predicts presence of metastatic spread and/or metastatic progression in the subject.
  • the invention is directed to a method for predicting cancer progression in a subject having cancer.
  • the method comprises determining the presence TMHCs in a first biological sample and a second biological sample at a later date, and optional additional biological samples, obtained from a subject having cancer, wherein when TMHCs are present in the second and/or additional biological sample but not present in the first biological sample, the cancer is predicted to progress in the subject.
  • the invention is directed to a method for predicting response to treatment in a subject having cancer.
  • the method comprises determining the presence TMHCs in a first biological sample and a second biological sample at a later date, and optional additional biological samples, obtained from a subject having cancer, wherein the first sample is obtained from the subject prior to or during cancer treatment, wherein the second sample and optional additional samples are obtained from the subject after at least one cancer treatment, wherein when TMHCs are present in the first biological sample but not present in the second and/or additional biological sample, the subject is predicted to respond to the treatment.
  • OS overall survival
  • progression free survival means the length of time survived by a subject having cancer from a selected date, such as the date on which treatment began or the date on which blood is drawn to assess cancer progression, and where the cancer has not worsened or progressed.
  • OS or PFS, or both is over a period of at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 months, or more. In one aspect of the invention, OS or PFS, or both, is over a period of at least about 12 months or at least about 24 months.
  • the amount of the biological sample in which the circulating cells can vary.
  • the biological sample should generally be at least about 2.5 mL.
  • the amount of biological sample may also be at least about 3, 4, 5, 6, 7, 7.5, 8, 9, 10, 11, 12, 12.5, 13, 14, 15, 16, 17, 17.5, 18, 19, 20, 21, 22, 22.5, 23, 24, 25, 26, 27, 27.5, 28, 29, or 30 mL, or more.
  • the amount of biological sample may also be between about 2.5 and 20 mL, between about 5 and 15 mL, or between about 5 and 10 mL. In one aspect of the invention, the biological sample is about 7.5 mL.
  • the source of the biological sample may be, but is not limited to, one or more of blood (such as peripheral blood), lymph node, bone marrow, cerebral spinal fluid, and urine.
  • blood such as peripheral blood
  • lymph node such as lymph node
  • bone marrow such as lymph node
  • cerebral spinal fluid such as urine
  • urine such as urine
  • the biological sample is blood
  • the blood may be antecubital-vein blood, inferior-vena-cava blood femoral vein blood, portal vein blood, or jugular-vein blood, for example.
  • the sample may be a fresh sample or a cryo-preserved sample that is thawed [14] .
  • the cancer may be a Stage I cancer, Stage II cancer, Stage III cancer, Stage IV cancer, carcinoma, sarcoma, neuroblastoma, melanoma, epithelial cell cancer, lung cancer, breast cancer, prostate cancer, pancreatic cancer, bladder cancer, kidney cancer, head and neck cancer, colorectal cancer, liver cancer, ovarian cancer, osteosarcoma, esophageal, brain & ONS, larynx, bronchus, oral cavity and pharynx, stomach, testis, thyroid, uterine cervix, uterine corpus cancer or other solid tumor cancers.
  • TMHCs are isolated from the biological samples for the determining steps using one or more means selected from size exclusion methodology, immunocapture, red blood cell lysis, white blood cell depletion, a high- molecular weight polysaccharide such as FICOLL®, electrophoresis, dielectrophoresis, flow cytometry, magnetic levitation, and various microfluidic chips, slits, channels, hydrodynamic size-based sorting, grouping, trapping, concentrating large cells, eliminating small cells, or a combination thereof.
  • the size exclusion methodology comprises use of a microfilter.
  • TMHCs are isolated from the biological samples using size exclusion methodology that comprises using a microfilter.
  • Suitable microfilters can have a variety of pore sizes and shapes.
  • the microfilter may have a pore size ranging from about 5 microns to about 20 microns. In certain aspects of the invention, the pore size is between about 5 and 10 microns; in other aspects, the pore size is between about 7 and 8 microns. The larger pore sizes will eliminate most of the WBC contamination on the filter.
  • the pores of the microfilter may have any shape, with acceptable shapes including round, race-track shape, oval, slit, square, rectangular and/or other shapes.
  • the microfilter may have precision pore geometry, uniform pore distribution, more than one pore geometry, and/or non-uniform distribution.
  • the microfilter may be single layer, or multi-layers with different shapes on different layers.
  • TMHCs are isolated from the biological samples using a microfluidic chip via physical size-based sorting, slits, channels, hydrodynamic sizebased sorting, grouping, trapping, immunocapture, concentrating large cells, or eliminating small cells based on size.
  • the circulating cell capture efficiency can vary depending on the collection method.
  • the size of a circulating cell that can be captured on different platforms can also vary depending, for example, on the identity of the cell. Collection of circulating cells using CELLSIEVE 1M microfilters provides 100% capture efficiency and high quality cells.
  • the sample when the biological sample is peripheral blood or blood, the sample may be collected from a subject using a blood collection tube.
  • CELLSAVETM blood collection tubes (Menarini Silicon Biosystems Inc., San Diego, CA), for example, provide stable cell morphology and size.
  • TMHCs are isolated from the biological samples using a CELLSIEVETM microfilter low-pressure microfiltration assay.
  • TMHCs may be used independently as cancer markers, or in combination with biomarker expression by other circulating cells, such as circulating tumor cells (CTCs).
  • CTCs circulating tumor cells
  • Suitable CTC subtypes include, but are not limited to, pathologically definable CTCs (PDCTCs), and apoptotic CTCs.
  • Other suitable circulating cell types include CAMLs and circulating cancer associated vascular endothelial cells (CAVEs). The presence of these additional cell types themselves and also cell-free DNA (cfDNA), circulating tumor DNA (ctDNA), methylated DNA, proteomic, metabolomic, lipidomic and other biomarkers may provide a more complete understanding of a patient’s disease.
  • the method optionally further comprises administering a therapeutically effective amount of a cancer treatment to the subject.
  • the subject may be a subject in which the OS and PFS is predicted to be lower or shorter than the OS and PFS of another subject.
  • cancer treatment will correspond to the particular type of cancer being treated.
  • suitable cancer treatments include chemotherapy, single drugs, combination of drugs, immunotherapy, radiation therapy, chemoradiation, chemoradiation combined with single or multiple drug, chemoradiation combined with single or multiple drugs (such as immunotherapy drugs), cell therapy, and other therapies.
  • Additional cancer treatments include, but are not limited to, immunotherapeutic agents, chemotherapeutic agents, radiotherapeutic agents, existing cancer drugs, CCR5 antagonists and CXCR4 antagonists.
  • cancer treatments include, but are not limited to, one or more of antibodies or antagonists that block the activity of CCL3, CCL5 (RANTES), CCL7 or CCL8; Leronlimab (PRO 140); T-VEC, AM-0010, CXCR4 antagonist, TGF-beta kinase inhibitor galunisertib, anti-CSF-lR monoclonal antibody, Abemaciclib, Faslodex, necitumumab, AZD9291, Cyramza (ramucirumab), TPIV 200, Galunisertib, cancer vaccines, cytokines, cellbased therapies, bi- and multi-specific antibodies, tumor-targeting mAbs, Rituximab, oncolytic viruses, reovirus, Blinatumomab, Sipuleucel-T, T-Vec, IL-2, IFN-a, Trastuzumab, Celuximab, bevacizumab, Tim-3, BTLA,
  • the subjects mentioned in the methods of the present invention will be a human, a nonhuman primate, bird, horse, cow, goat, sheep, a companion animal, such as a dog, cat or rodent, or other mammal.
  • the subject having cancer may be undergoing treatment for the cancer. Such treatments include, but are not limited to targeted agents, chemotherapy, and radiation therapy.
  • a companion or complementary diagnostic is a diagnostic test that can be used in combination with a therapeutic drug for a selected disease or condition.
  • the companion diagnostic determines the suitability of the drug for treatment of the disease or condition in a particular patient, i.e. the companion diagnostic can help to predict whether the subject will be a responder or non-responder to the effects of the drug on the disease or condition in the subject.
  • the companion diagnostic provides information that is used in treatment decisions and that is essential for the safe and effective use of a corresponding drug or biological product.
  • TFCs Tumor Macrophage Fusion Cells
  • TMFCs Tumor Macrophage Fusion Cells
  • TMFCs & CTCs were compared to a patient’s progression free survival (PFS) and overall survival (OS) hazard ratios (HRs), analyzed by censored univariate analysis based on RECIST vl.l over 24 months.
  • PFS progression free survival
  • OS overall survival
  • HRs hazard ratios
  • CTCs were found in 39% of patients, Partial TMFCs in 25% of patients, Homodimeric TMFCs in 6% of patients, Cannibalistic TMFCs in 0% of patients, Binucleated TMFCs in 2% of patients, Hyperploidy CAMLs in 96% of patients.
  • TMFCs were detected and described in the blood of mBC patients, demonstrating an association with poor clinical outcomes. These data suggest a TMFC involvement in the pathogenesis of cancer.
  • TMFCs Tumor Macrophage Fusion Cells
  • TMFCs in this analysis did not include Hyperploidy Fusion cells (i.e. CAMLs). TMFCs & CTCs were isolated using a CellSieveTM microfilter and differentiated by staining for CK, CD45, CD14, and DAPI.
  • TMFCs were detected in 26% of patient samples at BL and 27% of patient samples at FU. TMFCs were significantly prognostic for worse PFS and OS at both BL and FU. Any type of CTCs were detected in 39% at BL and 38% at FU. Any type of CTCs were significantly prognostic for worse PFS but not OS at BL. At FU, CTCs were significantly prognostic for worse PFS and OS. At BL, patients without CTCs or TMFCs had the best survival outcomes (Fig. 4A and 4B), followed by patients with just CTC, and patients with TMFCs having the worst outcomes. TMFCs at BL and FU were the only significant independent parameters for PFS and OS (Table 3).
  • TMFCs appears to represent a subgroup of CTCs with additional prognostic value not previously analyzed. Presence of TMFCs correlates with significantly worse PFS and OS versus patients without CTCs, or versus patients with normal CTCs. Patients with drops in TMFC populations after induction of new treatments had better outcomes, indicating responses to specific treatment types.
  • C. Circulating Tumor Cell Clusters (CTCCs) C. Circulating Tumor Cell Clusters (CTCCs)
  • Circulating tumor cell clusters are aggregated groups of tumor cells that detached from primary tumors and circulate in the bloodstream.
  • CTCs Circulating Tumor Cells
  • Fig. 2 various subtypes exist (Fig. 2), including homotypic clusters made of only tumor cells and heterotypic CTCCs made of CTCs attached to immune/stromal white blood cells (WBCs).
  • CTCs can undergo Epithelial-Mesenchymal Transition (EMT), a process where tumor cells downregulate epithelial traits and upregulate mesenchymal traits, and also form clustered EMTs (CEMTs). Further, CTCs can fuse with macrophages forming Cancer- Associated Macrophage-Like cells (CAMLs) when in circulation.
  • EMT Epithelial-Mesenchymal Transition
  • CTCs can fuse with macrophages forming Cancer- Associated Macrophage-Like cells (CAMLs) when in circulation.
  • AMLs Cancer- Associated Macrophage-Like cells
  • Single CTCs, EMTs, and CAMLs were enumerated, as well as homotypic CTCCs, heterotypic CTCCs and EMT CTCCs from the blood of metastatic breast cancer (mBC) patients to quantify these CTC populations and assess their clinical utility by median progression free survival (mPFS) and median overall survival (mOS) over 24 months
  • CTCs were defined as having an intact DAPI nucleus and strong filamentous CK.
  • Homotypic CTCCs were defined as >2 CTCs attached together.
  • Heterotypic CTCCs were defined as >1 CTC attached to >1 WBC.
  • EMTs were defined as having DAPI nuclei and weak non-filamentous CK.
  • EMT CTCCS were defined as >2 EMTs.
  • CAMLs were defined as having an enlarged polynucleated DAPI, and positive for CD45/CD14 or non- filamentous CK.
  • CTC Clusters appear to represent an array of subtypes with different biological and clinical meanings.
  • the CTC cluster subtypes from the blood of mBC patients were stratified and enumerated, and compared to clinical outcomes.
  • CAML hyperploidy and CTC clustering appears to indicate poor prognosis.

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Abstract

L'invention concerne des méthodes de prédiction de la survie globale (OS) et de la survie sans progression (PFS) de sujets atteints d'un cancer, sur la base de la présence de cellules hybrides de macrophages tumoraux (TMHC), un groupe unique de types de cellules qui comprennent des fusions et/ou des agrégats de différentes cellules trouvées dans le sang de sujets ayant des tumeurs solides.
PCT/US2024/032257 2023-06-02 2024-06-03 Méthodes de prédiction de survie sans progression et de survie globale chez des sujets atteints d'un cancer à l'aide de cellules hybrides de macrophages tumoraux Ceased WO2024250002A1 (fr)

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Citations (3)

* Cited by examiner, † Cited by third party
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US20190128892A1 (en) * 2016-04-14 2019-05-02 Creatv Microtech, Inc. Methods of using pd-l1 expression in treatment decisions for cancer therapy
WO2019178226A1 (fr) * 2018-03-13 2019-09-19 Creatv Microtech, Inc. Procédés de surveillance de la réponse au traitement et de la progression d'une maladie chez des sujets à l'aide de cellules circulantes
US20200049713A1 (en) * 2017-03-31 2020-02-13 Creatv Microtech, Inc. Methods of using giant cell nucleic acid characterization in cancer screening, diagnostics, treatment and recurrence

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20190128892A1 (en) * 2016-04-14 2019-05-02 Creatv Microtech, Inc. Methods of using pd-l1 expression in treatment decisions for cancer therapy
US20200049713A1 (en) * 2017-03-31 2020-02-13 Creatv Microtech, Inc. Methods of using giant cell nucleic acid characterization in cancer screening, diagnostics, treatment and recurrence
WO2019178226A1 (fr) * 2018-03-13 2019-09-19 Creatv Microtech, Inc. Procédés de surveillance de la réponse au traitement et de la progression d'une maladie chez des sujets à l'aide de cellules circulantes

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