EP4619033A1 - Nouveau biomarqueur et cible pour le traitement de l'hypertension pulmonaire - Google Patents

Nouveau biomarqueur et cible pour le traitement de l'hypertension pulmonaire

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Publication number
EP4619033A1
EP4619033A1 EP23892308.0A EP23892308A EP4619033A1 EP 4619033 A1 EP4619033 A1 EP 4619033A1 EP 23892308 A EP23892308 A EP 23892308A EP 4619033 A1 EP4619033 A1 EP 4619033A1
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EP
European Patent Office
Prior art keywords
hfpef
receptor
microglobulin
beta
levels
Prior art date
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Pending
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EP23892308.0A
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German (de)
English (en)
Inventor
Yen-Chun Lai
Jia-Rong JHENG
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Indiana University
Indiana University Bloomington
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Indiana University
Indiana University Bloomington
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Application filed by Indiana University, Indiana University Bloomington filed Critical Indiana University
Publication of EP4619033A1 publication Critical patent/EP4619033A1/fr
Pending legal-status Critical Current

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/04—Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/24—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against cytokines, lymphokines or interferons
    • C07K16/244—Interleukins [IL]
    • C07K16/247—IL-4
    • 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
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
    • C12N15/1138—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing against receptors or cell surface proteins
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K2217/00—Genetically modified animals
    • A01K2217/07—Animals genetically altered by homologous recombination
    • A01K2217/075—Animals genetically altered by homologous recombination inducing loss of function, i.e. knock out
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K2227/00—Animals characterised by species
    • A01K2227/10—Mammal
    • A01K2227/105—Murine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
    • A61K48/005—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/76—Antagonist effect on antigen, e.g. neutralization or inhibition of binding
    • 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
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/10—Type of nucleic acid
    • C12N2310/14—Type of nucleic acid interfering nucleic acids [NA]
    • 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
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/10—Type of nucleic acid
    • C12N2310/20—Type of nucleic acid involving clustered regularly interspaced short palindromic repeats [CRISPR]

Definitions

  • the general field of the present disclosure are novel approaches to the treatment of pulmonary hypertension and methods for measuring or assessing the effectiveness of such treatments.
  • Pulmonary hypertension is a progressive illness often presenting with nonspecific symptoms including dyspnea, dizziness, lower extremity edema, and decreased exercise tolerance. At the cellular level, it is characterized by endothelial cell dysfunction and increased contractility of the small pulmonary arteries, which lead to abnormal intimal and smooth muscle proliferation together with resistance to apoptosis. Pulmonary vascular remodeling is a prominent feature of PH independent of the etiology. This remodeling increases pulmonary vascular resistance (PVR), which eventually leads to failure of the right ventricle (RV) due to rising afterload. Many symptoms of PH, including lower extremity edema and dyspnea, arise from RV failure. See Hensley et al., “Emerging therapeutics in pulmonary hypertension,” (2016) Am J Physiol Lung Cell Mol Physiol 314: L769-L781.
  • PH is defined by end-expiratory’ mean pulmonary artery pressure >20 mmHg and PVR >3 Wood units at rest. Id. PH is a nonspecific umbrella term, which covers elevated pulmonary artery pressure regardless of the etiology’. The initial clinical classification of PH has arisen from a World Health Organization-sponsored international meeting in 1973. PH has been subdivided into five groups based on the disease pathology’ and specific cause. Pulmonary arterial hypertension (PAH; Group 1 PH) specifically refers to disease processes, which result in vasoconstriction and stiffening of the small arteries in the lungs secondary to cell proliferation, fibrosis, as well as the development of in situ thrombi or plexiform lesions.
  • PAH Pulmonary arterial hypertension
  • PAH can be idiopathic, can be heritable, and can be associated with connective tissue disease, HIV, drug use, etc.
  • PH connective tissue disease
  • PH attributable to left heart disease or left ventricular diastolic dysfunction also referred to as PH associated with heart failure with preserved ejection fraction (PH-HFpEF)
  • PH-HFpEF PH associated with heart failure with preserved ejection fraction
  • Group 2 PH is a growing public health problem that is increasing in prevalence, affecting approximately 1.6 million patients in the United States alone.
  • Many patients with Group 1 PH due to pulmonary hypertension have many comorbidities including obesity’, hypertension, diabetes mellitus, and hypercholesterolemia, all clinical features of the metabolic syndrome (MS) which may itself predispose patients to PH.
  • MS metabolic syndrome
  • Robbins et al. “Association of the Metabolic Syndrome with Pulmonary’ Venous Hypertension,” (2009) CHEST 136: pp. 31-36.
  • no approved specific medication or consensus therapeutic strategy' for PH- HFpEF is available at present.
  • Sodium glucose cotransporter 2 (SGLT2) inhibitors have recently been approved for the treatment of HFpEF. See Anker et al., “Empagliflozin in Heart Failure with a Preserved Ejection Fraction,” (2021) N Engl J Med. 385: pp. 1451-1461. Although SGLT2 inhibitors have been reported to improve PH in SU5416-exposed obese ZSF1 (Ob-Su) rats, patients with type 2 diabetes, or HF, the search for effective therapies for PH-HFpEF remains.
  • B-type natriuretic peptide (BNP) and N-terminal proBNP (NT-proBNP) are at present the only guideline-recommended biomarkers for the diagnosis and risk stratification of HF and PH.
  • BNP B-type natriuretic peptide
  • NT-proBNP N-terminal proBNP
  • ESC/ERS Guidelines for the diagnosis and treatment of pulmonary hypertension The Joint Task Force for the Diagnosis and Treatment of Pulmonary Hypertension of the European Society of Cardiology (ESC) and the European Respiratory Society (ERS): Endorsed by: Association for European Paediatric and Congenital Cardiology (AEPC), International Society for Heart and Lung Transplantation (ISHLT),” (2015) Eur Respir J.
  • AEPC European Paediatric and Congenital Cardiology
  • ISHLT International Society for Heart and Lung Transplantation
  • B2M beta 2-microglobulin
  • MHC I major histocompatibility complex class I
  • Serum B2M concentration is positively correlated with patients with dialysis-related amyloidosis, chronic kidney disease (CKD), type 2 diabetes, COPD, acute coronary syndrome, atherosclerosis, multiple myeloma, and HIV. Circulating B2M levels are also associated with coronary heart disease (CHD) and all-cause mortality.
  • CHD coronary heart disease
  • CSF cerebrospinal fluid
  • B2M deletion improved glucose intolerance, reduced pulmonary vascular remodeling, lowered pulmonary hypertension, and attenuated RV hypertrophy in mice with HFD-induced PH-HFpEF.
  • the present disclosure provides novel approaches to the diagnosis/prognosis and treatment of PH-HFpEF.
  • PH-HFpEF is largely associated with metabolic syndrome, including diabetes, hypertension, kidney disfunction, and heart failure with preserved ejection fraction (HFpEF)
  • this invention provides potential diagnostic/prognostic value and treatment for metabolic syndrome and HFpEF.
  • current invention provides inhibitors targeting B2M, B2M receptors, and regulators of B2M receptors for the treatment of Group 1-5 PH.
  • the invention provides therapeutic approaches using the administration of inhibitors of B2M, B2M receptors, and regulators of B2M receptors for the treatment of Group 1-5 PH, metabolic syndrome, and HFpEF.
  • the current invention provides methods of treatment of PH- HFpEF by reducing B2M from the blood stream in a patient with elevated levels of B2M or other biomarkers.
  • the current invention provides methods of treatment of PH- HFpEF by administration of antagonists for B2M receptors to patients in need thereof. [0023] In some embodiments, the current invention provides methods of treatment of PH- HFpEF by various types of receptor inhibition therapy.
  • the current invention provides methods of treatment of PH- HFpEF skeletal muscle-specific gene therapy.
  • any embodiment of the invention is provided methods of determining the severity in PH-HFpEF patients by measuring the amount of B2M and related biomarker of PH-HFpEF severity' and prognosis.
  • the invention provides methods of depleting B2M in addition to one or more regulators of B2M gene expression or synthesis.
  • B2M can be inhibited, neutralized or depleted by the administration of and agent to the patient where the agent comprises an adeno-associated virus (AAV) or lentovirus-containing an a short-hairpin RNA (shRNA) against B2M.
  • AAV adeno-associated virus
  • shRNA short-hairpin RNA
  • the shRNA is commercially available and can be attached to or part of any vector known in the art including plasmids, viral vectors, bacteriophages, cosmids, and artificial chromosomes.
  • the agent comprises a monoclonal or polyclonal antibody directed against B2M or a B2M receptor. In yet other embodiments, the agent comprises a monoclonal or polyclonal antibody directed against B2M or a B2M receptor. In still other embodiments, the agent is an siRNA or antisense oligonucleotide that targets B2M or a B2M receptor.
  • the agent is an antagonist that binds to a B2M-mediated receptor and prevents the binding of B2M.
  • FIG. 1A-D depicts the changes in plasma protein abundance profiles in patients with PH-HFpEF.
  • FIG. 1A Schematic overview of mass spectrometry -based plasma proteomic analysis.
  • FIG. IB Volcano plot of the fold change and statistical significance for protein abundance levels.
  • FIG. 1C Changing protein abundances in PH-HFpEF [Age: 69.8 ⁇ 7.9; male sex: 8; BMI: 39.1 ⁇ 10.4; mean pulmonary artery pressure (mPAP): 39.4 ⁇ 8. 1 mmHg; pulmonary capillary wedge pressure (PCWP): 20.2 ⁇ 4.5 mmHg; WHO function class II: 1 (6%), III: 14 (88%), and IV: 1 (6%)].
  • FIG. ID Protein abundance ratio of B2M. Data are mean ⁇ SEM. P value was determined by Mann-Whitney U test.
  • FIG. 2A-F show the circulating levels of B2M are elevated in patients with PH-HFpEF. Circulating levels of B2M are elevated in patients with PH-HFpEF.
  • FIG. 2A Circulating levels of B2M were measured by ELISA in plasma of the validation cohort of control subjects, HFpEF patients without PH, and PH-HFpEF patients [confirmed diagnosis by RHC when the resting mPAP > 25 mmHg, PCWP > 15 mmHg, and transpulmonary pressure gradient (TPG) > 12 mmHg OR during exercise mPAP > 30 mmHg, PCWP > 20 mmHg, and total pulmonary resistance (TPR) > 3] .
  • TPG transpulmonary pressure gradient
  • FIG. 3A-F shows the skeletal muscle levels of B2M are increased in mice with skeletal muscle SIRT3 deficiency or HFD-induced PH-HFpEF.
  • FIG. 3A and 3B Skeletal muscle (FIG. 3A) and plasma (FIG. 3B) levels of B2M in wild-type (WT) and skeletal muscle-specific SIRT3 knockout (Sirt3 skn1 ’ 1 ’) mice.
  • FIG. 3C Plasma B2M levels correlate positively with right ventricular systolic pressures (RVSP) in S/rt3 skm '' mice.
  • FIG. 3D and 3E Skeletal muscle (FIG. 3D) and plasma (FIG.
  • FIG. 3E Plasma B2M levels correlate positively with RVSP in HFD-exposed mice. Spearman r is shown. Data are mean ⁇ SEM. P value was analyzed by Mann- Whitney U test.
  • FIG. 4A-C shows that muscle expression levels of B2M are increased in patients with PH-HFpEF.
  • FIG. 4A B2M expression levels were measured in muscle biopsies of PH-HFpEF patients and control subjects by Western blots.
  • FIG. 4B and 4C Correlation between skeletal muscle B2M expression with resting PCWP (FIG. 4B) or RAP (FIG. 4C). Spearman r is shown. Data are mean ⁇ SEM. P value w as analyzed by Mann-Whitney t/test.
  • FIG. 5A-D shows that treatment with B2M induces PAVECs migration/proliferation and promotes PAVSMCs proliferation.
  • FIG. 5A-D shows that treatment with B2M induces PAVECs migration/proliferation and promotes PAVSMCs proliferation.
  • FIG. 5 A and 5B Human PAVECs were administered with exogenous B2M (10 mg/ml) for 4 days. Representative images of cell migration and related quantitative data (FIG. 5A). Cell proliferation assessed by cell counts (FIG. 5B). FIG. 5C and 5D: Human PAVSMCs were exposed to B2M (10 mg/ml) for 5 days. Representative images of cell numbers and cell proliferation assessed by cell counts (FIG. 5C). Representative Western blots for PCNA protein expression levels (FIG. 5D). Data are mean ⁇ SEM. P value was analyzed by Student’s I test.
  • FIG. 6A-I sho s that B2M whole-body KO mice protects against metabolic syndrome- associated PH-HFpEF.
  • FIG. 6A 8-week-old WT and whole-body B2M knockout mice (82m 1 ') were fed a RD or HFD for 16 w eeks. At w eek 16, body w eights (FIG. 6B), glucose tolerant abilities (FIG. 6C), RVSP (FIG. 6D), and left ventricular end-diastolic pressure (LVEDP, FIG. 6G) were measured. Weights of RV (FIG. 6E) and LV+S (FIG. 6F) normalized to tibial length were used as index of ventricular hypertrophy.
  • FIG. 6E 8-week-old WT and whole-body B2M knockout mice (82m 1 ') were fed a RD or HFD for 16 w eeks. At w eek 16, body w eights (FIG. 6B), glucose
  • FIG. 6H Representative images of lung sections stained with a- smooth muscle actin (a-SMA) and quantification of wall thickness from the mean of 5-6 vessels per lung section from 3 mice/group. Scale bar, 30 jam.
  • FIG. 61 PCNA levels were analy zed by Western blot in PAVSMCs of 82m' 1 ' and WT mice. Data are mean ⁇ SEM. P value was analyzed by One-Way ANOVA followed by Tukey’s post hoc test. For glucose tolerance test, two-way ANOVA followed by Bonferroni’s post hoc test was performed.
  • a-SMA smooth muscle actin
  • FIG. 7A-B Correlation between circulating B2M levels and mPAP (FIG. 7A) or PCWP (FIG. 7B) measured during cardiopulmonary exercise testing in PH-HFpEF patients. Spearman r is shown.
  • FIG. 8A-D shows protein levels of B2M assessed by Western blots in kidney (FIG. 8A), LV (FIG. 8B), adipose tissue (FIG. 8C), and RV (FIG. 8D) of HFD-exposed mice. Data are mean ⁇ SEM. P value was analyzed by Mann-Whitney U test.
  • B2M beta 2-microglobulin
  • MHC I major histocompatibility complex class I
  • Serum B2M concentration is positively correlated with patients with dialysis-related amyloidosis, chronic kidney disease (CKD), type 2 diabetes, COPD, acute coronary syndrome, atherosclerosis, multiple myeloma, and HIV. Circulating B2M levels are also associated with coronary heart disease (CHD) and all-cause mortality.
  • CHD coronary heart disease
  • CSF cerebrospinal fluid
  • B2M has a significant positive correlation with disease severity in PH-HFpEF patients, demonstrating clinical relevance of B2M in PH-HFpEF pathogenesis and suggesting B2M as a relevant biomarker of PH-HFpEF severity and prognosis.
  • B2M-deficient mice are protected from metabolic syndrome-associated PH-HFpEF, further suggesting B2M as a clinically meaningful molecular target for the treatment of the disease.
  • B2M is regulated by sirtuin-3 (SIRT3), a mitochondria deacetylase, deficiency in skeletal muscle.
  • SIRT3 deficiency and its downstream reactive oxygen species (ROS) production have been associated with the development of pulmonary arterial hypertension (PAH, Group 1 PH), PH associated with hypoxia and lung disease (Group 3 PH), chronic thromboembolic PH (Group 4 PH), and PH associated with multifactorial causes (Group 5 PH).
  • this invention provides potential treatment for Group 1-5 PH.
  • the present disclosure provides:
  • [0052] methods of treatment of PH-HFpEF by various types of receptor inhibition therapy
  • [0053] methods of treatment of PH-HFpEF skeletal muscle-specific gene therapy
  • [0055] methods of inhibiting, treating, or preventing the effects of elevated B2M in patients comprising inhibiting, neutralizing or depleting B2M from the patient;
  • B2M can be inhibited, neutralized or depleted by the administration of and agent to the patient
  • the agent comprises an adeno-associated virus (AAV) or lentovirus-containing an a short-hairpin RNA (shRNA) against B2M
  • AAV adeno-associated virus
  • shRNA lentovirus-containing an a short-hairpin RNA
  • the shRNA is commercially available and can be attached to or part of any vector known in the art including plasmids, viral vectors, bacteriophages, cosmids, and artificial chromosomes;
  • the agent comprises a monoclonal or polyclonal antibody directed against B2M or a B2M receptor;
  • the agent comprises a monoclonal or polyclonal antibody directed against B2M or a B2M receptor;
  • agent is an siRNA or antisense oligonucleotide that targets B2M or a B2M receptor
  • the terms “treating” or “to treat” includes restraining, slowing, stopping, or reversing the progression or severity of an existing symptom or disorder.
  • the term “patient” refers to a human.
  • B2M can be neutralized or inhibited by several different non-limiting methods.
  • B2M can be neutralized or inhibited by administration of a therapeutically effective amount of an agent where the agent comprises an adeno-associated virus (AAV) or lentivirus-containing an a short-hairpin RNA (shRNA) against B2M (sh-“B2M”).
  • AAV adeno-associated virus
  • shRNA short-hairpin RNA
  • sh-“B2M” is commercially available and can be attached to or part of any vector known in the art including plasmids, viral vectors, bacteriophages, cosmids, and artificial chromosomes.
  • B2M can be neutralized or inhibited by administration of a therapeutically effective amount of an agent where the agent comprises an antibody, bivalent antibody or a monoclonal antibody directed against B2M, a B2M receptor or other B2M modulators.
  • B2M can be neutralized or inhibited by administration of a therapeutically effective amount of an agent where the agent comprises an siRNA or antisense oligonucleotide that targets B2M, a B2M receptor or other B2M modulators.
  • B2M, a B2M receptor or other B2M modulators can be neutralized or inhibited by administration of a therapeutically effective amount of an agent where the agent comprises an antagonist that binds to B2M, a B2M receptor or other B2M modulator and prevents the binding of B2M.
  • the target B2M, a B2M receptor or other B2M modulator inhibitor or a composition therein can be administered once per day, two or more times daily or once per week.
  • the target inhibitor or inhibitors or composition containing the same can occur by any conventional means including orally intramuscularly, intraperitoneally or intravenously into the subject. If injected, they can be injected at a single site per dose or multiple sites per dose.
  • a B2M inhibitor is an antibody directed against B2M, a B2M receptor or other B2M modulator as disclosed herein.
  • suitable antibodies directed against one or more targets are disclosed herein and known to those of skill in the art.
  • the B2M antibody can also include an antibody fragment or a bivalent antibody or fragment thereof, inhibiting one or more of B2M, a B2M receptor or other B2M modulator.
  • the B2M inhibitor may be part of a pharmaceutical composition where the composition may include either an antibody or fragment thereof for one or more of B2M, a B2M receptor or other B2M modulator.
  • anti-B2M antibodies described herein can be made or obtained by any means known in the art, including commercially. It is also contemplated that an antibody can be specifically reactive with a particular B2M protein or polypeptide may also be used as an antagonist.
  • An anti-B2M antibody herein may be an antibody or fragment thereof that binds to a B2M or a bivalent antibody that binds to B2M, a B2M receptor or other B2M modulator.
  • antibody refers to an immunoglobulin (Ig) whether natural or partly or wholly synthetically produced.
  • the term also covers any polypeptide or protein having a binding domain which is, or is homologous to, an antigen-binding domain.
  • the term further includes “antigen-binding fragments” and other interchangeable terms for similar binding fragments such as described below.
  • Native antibodies and native immunoglobulins are usually heterotetrameric glycoproteins of about 150,000 Daltons, composed of two identical light (L) chains and two identical heavy (H) chains.
  • Each light chain is typically linked to a heavy chain by one covalent disulfide bond, while the number of disulfide linkages varies among the heavy chains of different immunoglobulin isotypes.
  • Each heavy and light chain also has regularly spaced intrachain disulfide bridges.
  • Each heavy chain has at one end a variable domain (“VH” or “VH”) followed by a number of constant domains (“CH” or “CH”).
  • Each light chain has a variable domain at one end (“VL” or “VL”) and a constant domain (“CL” or “CL”) at its other end; the constant domain of the light chain is aligned with the first constant domain of the heavy chain, and the light-chain variable domain is aligned with the variable domain of the heavy chain.
  • Particular amino acid residues are believed to form an interface between the light- and heavy-chain variable domains.
  • the cytokine inhibitors as described herein can be a “synthetic polypeptide” derived from a “synthetic polynucleotide” derived from a “synthetic gene,” meaning that the corresponding polynucleotide sequence or portion thereof, or amino acid sequence or portion thereof, is derived, from a sequence that has been designed, or synthesized de novo, or modified, compared to an equivalent naturally occurring sequence.
  • Synthetic polynucleotides (antibodies or antigen binding fragments) or synthetic genes can be prepared by methods known in the art. including but not limited to, the chemical synthesis of nucleic acid or amino acid sequences.
  • Synthetic genes are typically different from naturally occurring genes, either at the amino acid, or polynucleotide level, (or both) and are typically located within the context of synthetic expression control sequences. Synthetic gene polynucleotide sequences, may not necessarily encode proteins with different amino acids, compared to the natural gene; for example, they can also encompass synthetic polynucleotide sequences that incorporate different codons but which encode the same amino acid (i.e., the nucleotide changes represent silent mutations at the amino acid level).
  • anti-B2M antibodies refers to the any of the B2M, a B2M receptor or other B2M modulator proteins disclosed herein, respectively or any fragment of the protein molecules thereof.
  • antigen-binding portion of an antibody refers to one or more fragments of an antibody that retain the ability to specifically bind to one or more of B2M, a B2M receptor or other B2M modulator cytokines.
  • the B2M antibodies may also include “diabodies” which refers to small antibody fragments with two antigen-binding sites, which fragments comprise a heavy chain variable domain (VH) connected to a light chain variable domain (VL) in the same polypeptide chain (VH-VL).
  • VH heavy chain variable domain
  • VL light chain variable domain
  • VH-VL polypeptide chain
  • cytokine antibodies may also include “chimeric” forms of non-human (e.g., murine) antibodies include chimeric antibodies which contain minimal sequence derived from a non-human Ig.
  • chimeric antibodies are murine antibodies in which at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin are inserted in place of the murine Fc.
  • Fc immunoglobulin constant region
  • the cytokine antibodies may also include a “monoclonal antibody” which refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations that may be present in minor amounts. Monoclonal antibodies are highly specific, being directed against a single antigenic site. Furthermore, in contrast to conventional (polyclonal) antibody preparations, which can include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen.
  • monoclonal indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies and is not to be construed as requiring production of the antibody by any particular method.
  • monoclonal antibodies can be made by a hybridoma method, recombinant DNA methods, or isolated from phage antibody.
  • binding refers to binding agents, antibodies or fragments thereof that are specific to a sequence of amino acid residues on a B2M, a B2M receptor or other B2M modulator protein (“binding site” or “epitope”), yet if are cross-reactive to other peptides/proteins, are not toxic at the levels at which they are formulated for administration to human use.
  • binding refers to a direct association between two molecules, due to, for example, covalent, electrostatic, hydrophobic, and ionic and/or hydrogen-bond interactions under physiological conditions and including interactions such as salt bridges and water bridges and any other conventional binding means.
  • affinity refers to the equilibrium constant for the reversible binding of two agents and is expressed as Kd.
  • Affinity of a binding protein to a ligand such as affinity of an antibody for an epitope can be, for example, from about 100 nanomolar (nM) to about 0.1 nM, from about 100 nM to about 1 picomolar (pM), or from about 100 nM to about 1 femtomolar (fM).
  • the term “avidity” refers to the resistance of a complex of two or more agents to dissociation after dilution. Apparent affinities can be determined by methods such as an enzyme linked immunosorbent assay (ELISA) or any other technique familiar to one of skill in the art. Avidities can be determined by methods such as a Scatchard analysis or any other technique familiar to one of skill in the art.
  • ELISA enzyme linked immunosorbent assay
  • '‘Epitope” refers to that portion of an antigen or other macromolecule capable of forming a binding interaction with the variable region binding pocket of an antibody.
  • the term “specific” refers to a situation in which an antibody will not show any significant binding to molecules other than the antigen containing the epitope recognized by the antibody.
  • the term is also applicable where, for example, an antigen binding domain is specific for a particular epitope which is carried by a number of antigens, in which case the antibody will be able to bind to the various antigens carry ing the epitope.
  • the terms “preferentially binds” or “specifically binds” mean that the antibodies bind to an epitope with greater affinity than it binds unrelated amino acid sequences, and, if cross-reactive to other polypeptides containing the epitope, are not toxic at the levels at which they are formulated for administration to human use.
  • binding refers to a direct association between two molecules, due to, for example, covalent, electrostatic, hydrophobic, and ionic and/or hydrogen-bond interactions under physiological conditions and includes interactions such as salt bridges and water bridges, as well as any other conventional means of binding.
  • RNA interference refers to the silencing or decreasing of gene expression by siRNAs. It is the process of sequence-specific, post-transcriptional gene silencing in animals and plants, initiated by siRNA that is homologous in its duplex region to the sequence of the silenced gene.
  • the gene may 7 be endogenous or exogenous to the organism, present integrated into a chromosome or present in a transfection vector that is not integrated into the genome. The expression of the gene is either completely or partially inhibited.
  • RNAi may also be considered to inhibit the function of a target RNA; the function of the target RNA may be complete or partial.
  • siRNAs refers to short interfering RNAs.
  • siRNAs comprise a duplex, or double-stranded region, of about 18-25 nucleotides long; often siRNAs contain from about two to four unpaired nucleotides at the 3' end of each strand. At least one strand of the duplex or double-stranded region of a siRNA is substantially homologous to or substantially complementary to a target RNA molecule.
  • siRNAs may also contain additional sequences; non-limiting examples of such sequences include linking sequences, or loops, as well as stem and other folded structures. siRNAs appear to function as key intermediaries in triggering RNA interference in invertebrates and in vertebrates, and in triggering sequence-specific RNA degradation during posttranscriptional gene silencing in plants.
  • any B2M, a B2M receptor or other B2M modulator gene can be silenced or “turned” off’ through the use of CRISPR technology as known to those of skill in the art.
  • De-identified plasma samples and muscle biopsies of PH-HFpEF subjects participating in clinical trials were collected at baseline within a month of confirmed diagnosis by RHC under protocols approved by the University of Pittsburgh Institutional Review Board and/or University' of California San Francisco Institutional Review Board.
  • De-identified plasma samples of PH-HFpEF subjects and HFpEF patients without PH were collected more than a month of RHC and/or echocardiogram under protocols approved by the University of Illinois at Chicago Institutional Review Board.
  • de-identified plasma samples of control subjects were collected under protocols approved by the Indiana University Institutional Review Board.
  • the search parameters included trypsin as the protease with maximum of two missed cleavages allowed; oxidation of methionine (+15.9949 Da), and deamidation of asparagine and glutamine (+0.9848 Da) were set as a dynamic modification while static modifications included carbamidomethyl (+57.0215 Da) at cysteine and TMT as a static modification of lysine residues and peptide N-termini (+229.1629 Da).
  • Precursor mass tolerance was set at 10 ppm and fragment mass tolerance w as set at 0.6 Da.
  • Peptide confidence w as estimated with the Percolator node. Peptides were filtered at q-value ⁇ 0.01 based on a decoy database search.
  • Reporter ions for TMT labeled peptides w ere quantified using the Reporter Ions Quantifier Node included a TMT 11 pl ex quantification method in Proteome Discoverer with a peak integration tolerance of 20 ppm and an integration method based on the most confident centroid peak at the MS3 level. Only unique peptides w ere used for quantification, with protein groups considered for peptide uniqueness. Peptides with an average reporter signal-to-noise ratio >10 were used for protein quantification. Correction for the isotopic impurity of reporter quantification values was applied. The normalization was performed in two steps. First, peptide reporter ion signal-to-noise (S/N) values were normalized to the total sum per channel.
  • S/N peptide reporter ion signal-to-noise
  • TMT batch effects were reduced by row -wise normalization based on median intensities. No imputation for missing values was performed. Once the normalization has been completed, the proteins.txt output file was imported into Perseus v. 1.6.13.0 software for further statistical analysis and data visualization.
  • the Perseus output was used for generating volcano plot.
  • B2m knockout (B2m' l ⁇ ) and wild-type (WT) mice were purchased from Jackson Laboratories (002087 and 000664, respectively; Bar Harbor, ME). Beginning at 8 weeks of age, B2in and WT mice were randomly assigned to high-fat diet (HFD; 60% lipids/kcal; Research Diets, New Brunsw ick. NJ) or regular diet (RD; 10% lipids/kcal) exposure for 16 weeks. As female mice are protective in experimental models of PH and HFpEF, 2021 only male mice were used in this study.
  • HFD high-fat diet
  • RD regular diet
  • RVSP right ventricular systolic pressure
  • LVEDP LV end-diastolic pressure
  • Plasma samples were collected using EDTA for anticoagulation and centrifuged for 15 minutes at 2500rpm. Plasma aliquots were immediately stored at -80°C. Plasma levels of human B2M (Abeam, #99977) and mouse B2M (Abeam, #223590) were quantified by commercially available ELISA kits according to manufacturer’s instructions. Blinded data analysis was performed.
  • Total protein extracts from tissues were homogenized (Bio-Gen 200 Homogenizer and 7X95 mm Saw-tooth Generator Probe; VWR International) in freshly prepared T-PER tissue protein extraction buffer (Thermo Fisher Scientific, Waltham, MA) with protease and phosphatase inhibitors (Thermo Scientific, Waltham, MA). Supernatants were separated by centrifugation at 14000g for 10 min at 4°C. Cells were washed with ice-cold PBS and lysed with universal nucleases-contained lysis buffer (Thermo Scientific. Waltham, MA) and protease/phosphatase inhibitor cocktail. Supernatants were separated by centrifugation at 13200rpm for 10 min at 4°C.
  • Primary human PAVECs were purchased from LONZA (Basel, Switzerland; 64-year- old male donor) and cultured in Endothelial Cell Basal Medium-2 (EBM-2, Lonza) supplemented with supplements and growth factors (EGM-2 MV; complete medium, Lonza).
  • Primary human PAV SMCs (LONZA; 51-year-old male donor) were cultured in Smooth Muscle Cell Growth Basal Medium (SmBM, Lonza) supplemented with Smooth Muscle Cell Growth Medium-2 supplements and growth factors (SmGM-2; complete medium, Lonza). Cells were maintained at 37°C in a humidified atmosphere of 5% CO2 and 95% air. Passages 4-9 were used in the study.
  • mice were fasted for 6 h before being subjected to intraperitoneal injection with 1.8 mg/g dextrose in 0.9% NaCl. Blood samples were taken at different time points as indicated, and blood glucose levels were measured with a portable glucose meter (ACCU-CHECK Aviva; Roche, Basel, Switzerland).
  • Mass spectrometry-based plasma proteomics identifies high protein abundance levels of B2M in patients with PH-HFpEF
  • the inventors used mass spectrometr -based bottom-up proteomics as a sensitive and comprehensive hypothesis-generating discovery’ technique to profile proteins in control subjects and patients with PH-HFpEF.
  • nLC-ESI-MS nano-capillary liquid chromatography electrospray ionization mass spectrometry
  • TMT tandem mass tag
  • Table 2 Clinical information of plasma samples used for proteomic analysis.
  • FIG. 1A-D depicts the changes in plasma protein abundance profiles in patients with PH-HFpEF.
  • FIG. 1A Schematic overview of mass spectrometry-based plasma proteomic analysis.
  • FIG. IB Volcano plot of the fold change and statistical significance for protein abundance levels.
  • FIG. ID Protein abundance ratio of B2M. Data are mean ⁇ SEM. P value was determined by Mann- Whitney U test.
  • Plasma B2M levels are significantly elevated in patients with PH-HFpEF and are associated with disease severity.
  • PH chronic kidney disease
  • cancers cancers
  • Table 3 Clinical information of plasma samples used for B2M measurement by ELISA
  • mPAP mean pulmonary' artery pressure
  • PCWP pulmonary capillary wedge pressure
  • TRV tricuspid regurgitation velocity
  • FIG. 2A-F show the circulating levels of B2M are elevated in patients with PH-HFpEF. Circulating levels of B2M are elevated in patients with PH-HFpEF.
  • FIG. 2A Circulating levels of B2M were measured by ELISA in plasma of the validation cohort of control subjects, HFpEF patients without PH, and PH-HFpEF patients [confirmed diagnosis by RHC when the resting mPAP > 25 mmHg, PCWP > 15 mmHg, and TPG > 12 mmHg OR during exercise mPAP > 30 mmHg, PCWP > 20 mmHg, and TPR > 3], Correlation between circulating levels of B2M and resting mPAP (FIG.
  • FIG. 7A and 7B Correlation between circulating B2M levels and mPAP (FIG. 7A) or PCWP (FIG. 7B) measured during cardiopulmonary exercise testing in PH-HFpEF patients. Spearman r is shown.
  • FIG. 3A-F shows the skeletal muscle levels of B2M are increased in mice with skeletal muscle SIRT3 deficiency or HFD-induced PH-HFpEF.
  • FIG. 3A and 3B Skeletal muscle (FIG. 3 A) and plasma (FIG. 3B) levels of B2M in WT and SirtS ⁇ '' 1 ' mice.
  • FIG. 3C Correlation between plasma B2M levels with RVSP in WT and SirtS 5 ⁇ ' mice.
  • FIG. 3D and 3E Skeletal muscle (FIG. 3D) and plasma (FIG. 3E) levels of B2M in mice fed a RD or HFDfor 16 weeks.
  • FIG. 3F Correlation between plasma B2M levels with RVSP in mice fed a RD or HFD. Spearman r is shown. Data are mean ⁇ SEM. P value was analyzed by Mann-Whitney U test.
  • FIG. 3D and 3E skeletal muscle expression and plasma concentration of B2M levels were increased in HFD-exposed mice.
  • a positive correlation betw een plasma B2M levels and RVSP was observed in HFD-exposed mice (FIG. 3F).
  • expression of B2M was found to be increased in kidney, but remained unchanged in LV, RV, and adipose tissue after 16 weeks of HFD exposure (FIG. 8A-D).
  • FIG. 8A-D shows protein levels of B2M assessed by Western blots in kidney (FIG. 8A), LV (FIG. 8B), adipose tissue (FIG. 8C), and RV (FIG.
  • Skeletal muscle expression levels of B2M are increased in PH-HFpEF patients and correlated significantly with resting PCWP and RAP.
  • Table 4 Clinical information of muscle biopsies used for B2M measurements.
  • mPAP mean pulmonary artery' pressure
  • PCWP pulmonary' capillary w edge pressure
  • BMI body mass index.
  • FIG. 4A-C shows that muscle expression levels of B2M are increased in patients with PH-HFpEF.
  • FIG. 4A B2M expression levels were measured in muscle biopsies of PH-HFpEF patients and control subjects by Western blots.
  • FIG. 4B and 4C Correlation between skeletal muscle B2M expression with resting PCWP (FIG. 4B) or RAP (FIG. 4C). Spearman r is shown. Data are mean ⁇ SEM. P value was analyzed by Mann-Whitney’ U test.
  • FIG. 4A muscle biopsies of PH-HFpEF patients had higher protein expression levels of B2M compared to that of control subjects.
  • B2M has been associated with pulmonary fibrosis, emphysema, cardiac fibroblast activation, atherosclerosis, and aging, it is currently unknown whether B2M can regulate pulmonary vascular remodeling.
  • B2M can regulate pulmonary vascular remodeling.
  • Beta2-microglobulin is a systemic pro-aging factor that impairs cognitive function and neurogenesis,’' (2015) Nat Med. 21: pp. 932-937; Amighi et al.. ‘“Beta 2 microglobulin and the risk for cardiovascular events in patients with asymptomatic carotid atherosclerosis,” (2011) Stroke 42: pp.
  • B2M pulmonary arterial vascular endothelial cells
  • PAVSMCs pulmonary arterial vascular smooth muscle cells
  • FIG. 5A-D shows that treatment with B2M induces PAVECs migration/proliferation and promotes PAVSMCs proliferation.
  • FIG. 5 A and 5B Human PAVECs were administered with exogenous B2M (10 mg/ml) for 4 days. Representative images of cell migration and related quantitative data (FIG. 5A). Cell proliferation assessed by cell counts (FIG. 5B).
  • FIG. 5C and 5D Human PAVSMCs were exposed to B2M (10 mg/ml) for 5 days. Representative images of cell numbers and cell proliferation assessed by cell counts (FIG. 5C). Representative Western blots for PCNA protein expression levels (FIG. 5D). Data are mean ⁇ SEM. P value was analyzed by Student’s / test
  • FIG. 6A-I shows that B2M whole-body KO mice protects against metabolic syndrome- associated PH-HFpEF.
  • FIG. 6A 8-week-old WT and whole-body B2M knockout mice (B2m' 1 ') were fed a RD or HFD for 1 weeks.
  • body weights FIG. 6B
  • glucose tolerant abilities FIG. 6C
  • RVSP FIG. 6D
  • LVEDP FIG. 6G
  • FIG. 6H Representative images of lung sections stained with a-smooth muscle actin (a-SMA) and quantification of wall thickness from the mean of 5-6 vessels per lung section from 3 mice/group. Scale bar, 30 pm.
  • FIG. 61 PCNA levels were analyzed by Western blot in PAVSMCs of B2m' 1 ' and WT mice. Data are mean ⁇ SEM. P value was analyzed by One-Way ANOVA followed by Tukey’s post hoc test. For glucose tolerance test, two-way ANOVA followed by Bonferroni’s post hoc test was performed.
  • a-SMA smooth muscle actin
  • HFD-exposed WT mice exhibited significantly higher body weights, glucose intolerance, RVSP, LVEDP, and bi-ventricular hypertrophy than RD-exposed WT mice (FIG. 6B-G). While no difference in body weights, LV hypertrophy, or LVEDP was observed, HFD-exposed B2m' l ⁇ mice exhibited significantly improved glucose intolerance, lowered RVSP, and reduced RV hypertrophy compared to WT mice fed with a HFD (FIG. 6B-G). In addition, HFD-exposed B2m' 1 ' mice had a lower percentage of wall thickness in comparison with HFD-exposed WT mice (FIG. 6H).
  • Plasma B2M levels will be measured using a human B2M ELISA kit in baseline plasma samples from a larger cohort of PH-HFpEF patients and random control subj ects. They will also examine the correlation between plasma B2M levels and markers of PH-HFpEF severity (e.g. hemodynamic parameters obtained by RHC at rest and during cardiopulmonary exercise testing, functional class, VO2 max, cardiac output). In addition, the inventors will compare plasma B2M levels in PH-HFpEF patients and HFpEF patients without PH to assess whether plasma B2M levels may be useful for stratifying patients into sub-populations.
  • markers of PH-HFpEF severity e.g. hemodynamic parameters obtained by RHC at rest and during cardiopulmonary exercise testing, functional class, VO2 max, cardiac output.
  • B2M inhibitors At present there are no commercially available B2M inhibitors. Therefore, the inventors will explore potential therapeutical approaches as following: a. designing peptides/small molecule antagonists for B2M receptors. B2M has been reported to interact with hemochromatosis protein (HFE) and transferrin receptor complex 1 (TFRC1) to regulate iron homeostasis and epithelial-mesenchymal transition of cancer cells. See Josson et al., “Beta2-microglobulin induces epithelial to mesenchymal transition and confers cancer lethality and bone metastasis in human cancer cells,” (2011) Cancer Res. 71: pp. 2600-2610.
  • HFE hemochromatosis protein
  • TFRC1 transferrin receptor complex 1
  • B2M has also been shown to impair neurogenesis and cognitive function via reduced cell surface expression of MHC I and transporter associated with antigen processing 1 (TAPI). See Smith et al. 2015. Additionally, B2M has been shown to activate monocyte via through transforming growth factor beta receptor 2 (TGFPR2) signaling. See Hilt et al., “Platelet-derived beta2M regulates monocyte inflammatory responses,” (2019) JCI Insight 4.
  • TGFPR2 transforming growth factor beta receptor 2
  • the inventors Using coimmunoprecipitation-coupled mass spectrometry, the inventors also identified cadherins desmoglein-1 (DSG1), desmoglein-2 (DSG2) and desmocollin- 1 (DSC 1), that may mediate B2M signaling through non-canonical junction-independent effects. Thus, in further studies, the inventors will determine whether HFE, TFRC1, MHC I, TAPI, TGF
  • DSG1 desmoglein-1
  • DSG2 desmoglein-2
  • DSC 1 desmocollin- 1
  • B2M is present at higher levels in patients with PH-HFpEF; 2) skeletal muscle SIRT3 deficiency is associated with induction and secretion of B2M in animal models and human subjects with PH-HFpEF; 3) circulating and skeletal muscle expression levels of B2M correlate with PH-HFpEF severity; 4) B2M increases PAVECs migration/proliferation and promotes PAVSMCs proliferation; and 5) loss of B2M improves metabolic syndrome-associated PH-HFpEF.
  • These findings not only reveal a previously unknown pathogenic role for B2M in PH-HFpEF, but also suggest the potential of using circulating and skeletal muscle B2M levels as biomarkers for PH-HFpEF.
  • B2M comprises the light chain of MHC I that forms an active part of the adaptive immune system. As it is noncovalently associated with the light chain and has no direct attachment to the cell membrane, free B2M circulates in the blood as a result of shedding and/or intracellular release. See Wilson et al., '‘Beta2-microglobulin as a biomarker in peripheral arterial disease: proteomic profiling and clinical studies,” (2007) Circulation 116: pp. 1396-1403. The net concentration of B2M is determined by its generation/secretion into circulation and its elimination by the kidneys. As such, in people without kidney disease, elevated B2M has been considered as a marker of altered cell proliferation. See Prizment et al. 2016.
  • B2M Increased circulating levels of B2M have been implicated in various cancers and associated with cancer progression with poor prognosis. See Id. and Zhang et al., "B2M overexpression correlates with malignancy and immune signatures in human gliomas,” (2021) Sci Rep. 11 : p. 5045. B2M has also been associated with increased colorectal cancer risk. See Prizment et al. 2016. Additionally, there is evidence to correlate elevated serum B2M concentrations with the development and progression of pulmonary fibrosis in patients with chronic obstructive pulmonary disease (COPD) and emphysema. See Wu et al. 2020; Molenaar et al. 2021.
  • COPD chronic obstructive pulmonary disease
  • B2M was also found to be associated with adverse cardiovascular outcomes in patients with asymptomatic carotid atherosclerosis. See Amighi et al. 2011. Furthermore, plasma proteome studies found B2M to be a risk marker for coronary heart disease in postmenopausal women and to correlate with disease severity in patients with peripheral arterial disease (PAD) or HF with reduced ejection fraction (HFrEF). See Wilson et al. 2007; Prentice et al., “Novel proteins associated with risk for coronary heart disease or stroke among postmenopausal women identified by in-depth plasma proteome profiling,” (2010) Genome Med. 2: p.
  • B2M is probably related to chronic pathophysiological processes and abnormalities in the left heart, pulmonary vasculature, and the right heart. Whether circulating B2M levels can be a useful non-invasive tool to facilitate screening, to define/ refine specific patient sub-phenotypes, and/or to identify patients in a high-throughput fashion requires further investigations.
  • B2M may act as an endocrine signaling molecule to induce pulmonary vascular remodeling via increasing PAVECs migration/proliferation and PAVSMCs proliferation. While the data show that skeletal muscle expression levels of B2M correlate with PH-HFpEF severity, little is known about the pathological mechanism underlying increased skeletal muscle B2M in PH-HFpEF. Upregulation and prolonged expression of MHC I in muscle cells is a hallmark of inflammatory myopathies, which have been recognized as one of an extra-cardiopulmonary source of inflammation in HFpEF and PH. See Nagaraju et al..
  • skeletal muscle B2M reflects and/or contributes to the chronic, systemic, metabolic inflammation in PH-HFpEF. Future studies are needed to determine whether skeletal muscle B2M contributes to PH-HFpEF pathogenesis and the interplay between immunological and metabolic processes, dependent or independent of MHC I, in this context.
  • SGLT2 inhibitors were very recently approved for the treatment of HFpEF. See Anker et al., “Empagliflozin in Heart Failure with a Preserved Ejection Fraction,” (2021) N Engl J Med. 385: pp. 1451-1461. SGLT2 inhibitors have been reported to improve exercise-induced PH in Ob-Su rats and type 2 diabetes patients and reduce PA pressure in HF patients. See Satoh et al. 2021; Kayano et al. 2020; Nassif et al. 2021.
  • HFE hemochromatosis gene
  • the current findings suggest the potential of using circulating and skeletal muscle B2M levels as biomarkers for PH-HFpEF to guide future research towards facilitation of screening, diagnosis, refinement of specific patient phenotype, and/or identification of patients in a more efficient manner.
  • the findings also provide new 7 insights into the mechanistic basis of B2M in the regulation of pulmonary 7 vascular proliferative remodeling and PH-HFpEF. From a translational perspective, the data indicate that B2M is an an important therapeutic target for the treatment of PH-HFpEF in the future.
  • this invention provides potential treatment for Group 1-5 PH.
  • [00191] inhibitors targeting B2M, B2M receptors, and regulators of B2M receptors for the treatment of Group 1-5 PH;
  • [00194] methods of treatment of PH-HFpEF by administration of antagonists for B2M receptors to patients in need thereof;
  • B2M can be inhibited, neutralized or depleted by the administration of and agent to the patient
  • the agent comprises an adeno-associated virus (AAV) or lentivirus-containing an a short-hairpin RNA (shRNA) against B2M
  • AAV adeno-associated virus
  • shRNA lentivirus-containing an a short-hairpin RNA
  • the shRNA is commercially available and can be attached to or part of any vector known in the art including plasmids, viral vectors, bacteriophages, cosmids, and artificial chromosomes;
  • the agent comprises a monoclonal or polyclonal antibody directed against B2M or a B2M receptor;
  • the agent comprises a monoclonal or polyclonal antibody directed against B2M or a B2M receptor;
  • agent is an siRNA or antisense oligonucleotide that targets B2M or a B2M receptor

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Abstract

L'invention concerne de nouvelles approches pour le traitement de l'hypertension pulmonaire et des procédés de mesure ou d'évaluation de l'efficacité de tels traitements. Plus spécifiquement, l'invention concerne de nouvelles approches pour le diagnostic/pronostic et le traitement de l'HP associé à une insuffisance cardiaque avec une fraction d'éjection préservée (HP-ICFEP) comprenant l'administration d'inhibiteurs de la bêta 2-microglobuline (B2M). L'invention concerne également des procédés de traitement de l'HP-ICFEP largement associé au syndrome métabolique, y compris le diabète, l'hypertension, l'insuffisance cardiaque et la dysfonction rénale.
EP23892308.0A 2022-11-17 2023-11-02 Nouveau biomarqueur et cible pour le traitement de l'hypertension pulmonaire Pending EP4619033A1 (fr)

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