Materials and methods for treating senescence-related disorders

Abstract

Disclosed herein are materials and methods for the treatment of a senescence-associated condition.

Description

MATERIALS AND METHODS FOR TREATING SENESCENCE-RELATED DISORDERSFIELD OF THE INVENTION

[0001] The present disclosure is directed to the use of osteoprotegerin inhibitors for the treatment of senescence-associated conditions.INCORPORATION BY REFERENCE OF MATERIALS SUBMITTED ELECTRONICALLY

[0002] This application contains, as a separate part of the disclosure, a Sequence Listing in computer readable form (Filename: 70225_SeqListing.xml; Size: 67,134 bytes; Created: July 30, 2025), which is incorporated by reference in its entirety.BACKGROUND

[0003] Cellular senescence is a process wherein the cell cycle is arrested and cells enter a functional but non-dividing state. It is believed that this this cellular pathway evolved to protect against genetic instability and cancer in cells that have accumulated DNA- damaging insults, and thus senescence prevents unrestricted cell proliferation. There is also evidence that senescence acts against viral infections, inhibiting the replicative processes that are hijacked by viruses.

[0004] Senescent cells produce numerous cytokines and chemokines. Over short time scales, this response is thought to be adaptive, wherein the proinflammatory signals recruit immune elements to the afflicted area, enabling immune surveillance of potentially oncogenic or infected cells. In healthy and young individuals, it is believed that the senescent cells are efficiently removed by the immune system.

[0005] However, in diseased or aged individuals, it appears that immune system removal of senescent cells is impaired. Under such conditions, senescent cells accumulate and the proinflammatory action of these cells can become pathologic. Senescent cells accumulate in ever greater numbers over time, and the signaling molecules they secrete become deleterious to surrounding tissues, causing inflammation, remodeling tissue in aberrant ways, and potentially fostering the onset of cancer rather than suppressing it. The presence of senescent cells likely worsens the progression of many age-related diseases and processes. In some cases, an extended state of unresolved stress causes the senescent cells to exhibit what is known as the SASP, for senescence-associated secretory phenotype. SASP cells secrete a number of inflammatory signal molecules, including cytokines, chemokines, and other factors. This SASP secretome may profoundly affect neighboring, non-senescent cells, impairing their function and facilitating their transition to senescent cells as well.

[0006] Accumulation of senescent cells is thus believed to underlie a great number of disease conditions and age-related pathologies. For example, accumulation of senescent cells has been implicated in formation of lesions such as atherosclerotic plaques, in diabetes, in neurodegeneration, and other conditions.SUMMARY

[0007] In one aspect, the disclosure provides a method of treating a senescence- associated condition in a subject in need of treatment therefore comprising administering an osteoprotegerin inhibitor to the subject. In some embodiments, the senescence-associated condition is an inflammatory or autoimmune condition, diabetes or a diabetes-related condition, cardiovascular condition, neurodegenerative condition, or an age-related condition. In some embodiments, the subject does not have cancer.

[0008] In some embodiments, the osteoprotegerin inhibitor is an antibody that specifically binds osteoprotegerin.BRIEF DESCRIPTION OF THE FIGURES

[0009] Figure 1 A is a graph showing that TRAIL is upregulated on the cell surface of iNKT cells after stimulation with stimulation with a-GalCer. Figures 1 B and 1 C are graphs showing confirmation of cytokine release of IFNy and proliferation upon stimulation of the iNKT cells.

[0010] Figure 2 is a graph showing that senescent cells and tissues that senescent cells accumulate secrete osteoprotogerin (OPG), which acts as a decoy protein that binds TRAIL and inhibits its interaction with the TRAIL receptor DR5.

[0011] Figures 3A and 3B: (Figure 3A) Recombinant OPG protein inhibits cytotoxicity of iNKT cells. (Figure 3B) Blocking antibody of OPG can unmask the cytotoxic activity if iNKT cells.

[0012] Figures 4A-4C: Anti-OPG treatment of HFD clears senescent adipose progenitor cells and normalizes glucose control. Schematic of experiment (Figure 4A) reduction in senescent cells (Figure 4B) normalization of blood glucose (Figure 4C).DETAILED DESCRIPTION

[0013] The present disclosure is based, in part, on the discovery that osteoprotegerin (OPG) inhibitors are capable of stimulating iNKT cytotoxic activity.

[0014] In one aspect, described herein is a method of treating a senescence-associated condition in a subject in need of treatment therefore comprising administering an osteoprotegerin inhibitor to the subject.

[0015] In some embodiments, the osteoprotegerin inhibitor is an anti-OPG antibody. Exemplary anti-OPG antibodies include, but are not limited to, anti-OPG antibodies and binding compositions disclosed in United States Patent Application Publication Number 20190352413, and in “Anti-OPG Antibodies,” by De Arbeau Carvalho et aL; United States Patent Number 6,919,433, “Monoclonal antibodies that Bind OCIF,” by Goto et al.

[0016] Multiple commercially available anti-OPG antibodies are available, including, for example:, NB100-56505 and NBP1 -51670 (Novus Biochem); AM06539SU-N (Origene); GTX82749 (Genetex); LS-C169286 (Lifespan Biosciences); PA5-34946, PA5-86053, MAS- 15715, MA5-15960, MA5-34922, MA5-34923, and MA5-15726 (Invitrogen Antibodies); AM06539SU-N, SM7070P, AM06550SU-N, DM2005 and DM2018 (Acris Antibodies, GmbH); BF0156 (Affinity Biosciences); MAB10335, MAB6241 , MAB12971 , H00004982-K, and MAB3414 (Abnova Biosciences); OAAD00388 (Aviva Systems Biology); EM1701-98 and EM1701-99 (HUABIO Research); ALX-804-813-C100, ALX-804-813B-C100, ADI-AAM- 020-E, and ALX-804-532-C100 (Enzo Life Sciences), 139256 and 104289 (NovoPro Bioscience); 10-6001 (Abeomics); AO1471 a, AO1482a, and ALS12119 (Abgent); 3448-1 and ab124820 (RabMAbs); abx011830 and abx016009 (Abbexa); 030691 , 030692, and 08065-04 (United States Biological); MOB-1442z-S(P), CBMAB-00022-CQ, CBMAB- 00026- CQ, CBMAB-O0472-CQ, CBMAB-O0473-CQ, CBMAB-O0474-CQ, CBMAB-O0475- CQ, CBMAB-O0881-CQ, CBMAB-O0884-CQ, CBMAB-O0888-CQ, CBMAB-T3072-YJ, MOB- 1442z, MOB-1507CT, MOR-3609, MRO-1136-CN, NEUT-2118CQ, and ZG-0330C (Creative BioLabs); 10271-R340 (Sino Biologica); 98A1071 and 40938 (Active Motif); 119- 12729 (Ray Biotech); and APR08890G (Leading Antibody).

[0017] The term “antibody” refers to an intact immunoglobulin molecule (including polyclonal, monoclonal, chimeric, humanized, and / or human versions having full length heavy and / or light chains). The antibody may be any type of antibody, i.e., immunoglobulin, known in the art. In exemplary embodiments, the antibody is an antibody of class or isotype IgA, IgD, IgE, IgG, or IgM. In exemplary embodiments, the antibody described herein comprises one or more alpha, delta, epsilon, gamma, and / or mu heavy chains. In exemplary embodiments, the antibody described herein comprises one or more kappa or light chains. In exemplary aspects, the antibody is an IgG antibody and optionally is one of the four human subclasses: lgG1 , lgG2, lgG3 and lgG4.

[0018] “Specifically binds” as used herein means that the antibody (or antigen binding fragment) preferentially binds an antigen (osteoprotegerin peptide) over other proteins. In some embodiments, “specifically binds” means the antibody has a higher affinity for the antigen than for other proteins. Antibodies that specifically bind an antigen may have abinding affinity for the antigen of less than or equal to 1 x 1 O’7M, less than or equal to 2 x 1 O’7M, less than or equal to 3 x 10’7M, less than or equal to 4 x 10’7M, less than or equal to 5 x 10’7M, less than or equal to 6 x 10’7M, less than or equal to 7 x 10’7M, less than or equal to 8 x 10’7M, less than or equal to 9 x 10’7M, less than or equal to 1 x 10’8M, less than or equal to 2 x 10’8M, less than or equal to 3 x 10’8M, less than or equal to 4 x 10’8M, less than or equal to 5 x 10’8M, less than or equal to 6 x 10’8M, less than or equal to 7 x 10’8M, less than or equal to 8 x 10’8M, less than or equal to 9 x 10’8M, less than or equal to 1 x 1 O’9M, less than or equal to 2 x 10’9M, less than or equal to 3 x 10’9M, less than or equal to 4 x 10’9M, less than or equal to 5 x 10’9M, less than or equal to 6 x 10’9M, less than or equal to 7 x 10’9M, less than or equal to 8 x 10’9M, less than or equal to 9 x 10’9M, less than or equal to 1 x 1 O’10M, less than or equal to 2 x 1 O’10M, less than or equal to 3 x 1 O’10M, less than or equal to 4 x 10’10M, less than or equal to 5 x 10’10M, less than or equal to 6 x 10’10M, less than or equal to 7 x 1 O’10M, less than or equal to 8 x 1 O’10M, less than or equal to 9 x 1 O’10M, less than or equal to 1 x 10’11M, less than or equal to 2 x 10’11M, less than or equal to 3 x 10’11M, less than or equal to 4 x 10’11M, less than or equal to 5 x 10’11M, less than or equal to 6 x 10’11M, less than or equal to 7 x 10’11M, less than or equal to 8 x 10’11M, less than or equal to 9 x 10’11M, less than or equal to 1 x 10’12M, less than or equal to 2 x 10’12M, less than or equal to 3 x 10’12M, less than or equal to 4 x 10’12M, less than or equal to 5 x 10’12M, less than or equal to 6 x 10’12M, less than or equal to 7 x 10’12M, less than or equal to 8 x 10’12M, or less than or equal to 9 x 1012M. It will be appreciated that ranges having the values above as end points is contemplated in the context of the disclosure.

[0019] "CDR" refers to the complementarity determining region within antibody variable sequences. There are three CDRs in each of the variable regions of the heavy chain and the light chain, which are designated CDR1 , CDR2 and CDR3, for each of the variable regions. The term "set of six CDRs " as used herein refers to a group of three CDRs that occur in the light chain variable region and heavy chain variable region, which are capable of binding the antigen. The exact boundaries of CDRs have been defined differently according to different systems. The system described by Kabat (Kabat et al., Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, Md. (1987) and (1991 )) not only provides an unambiguous residue numbering system applicable to any variable region of an antibody, but also provides precise residue boundaries defining the three CDRs.These CDRs may be referred to as Kabat CDRs. Chothia and coworkers (Chothia & Lesk, J. Mol. Biol. 196:901 -917 (1987) and Chothia et al., Nature 342:877-883 (1989)) found that certain sub-portions within Kabat CDRs adopt nearly identical peptide backbone conformations, despite having great diversity at the level of amino acid sequence. Thesesub-portions were designated as L1 , L2 and L3 or H1 , H2 and H3 where the "L" and the "H" designates the light chain and the heavy chains regions, respectively. These regions may be referred to as Chothia CDRs, which have boundaries that overlap with Kabat CDRs. Other boundaries defining CDRs overlapping with the Kabat CDRs have been described by Padlan (FASEB J. 9:133-139 (1995)) and MacCallum (J Mol Biol 262(5)73245 (1996)). Still other CDR boundary definitions may not strictly follow one of the above systems, but will nonetheless overlap with the Kabat CDRs, although they may be shortened or lengthened in light of prediction or experimental findings that particular residues or groups of residues or even entire CDRs do not significantly impact antigen binding. The methods used herein may utilize CDRs defined according to any of these systems, although preferred embodiments use Kabat or Chothia defined CDRs.

[0020] CDRs are obtained by, e.g., constructing polynucleotides that encode the CDR of interest and expression in a suitable host cell. Such polynucleotides are prepared, for example, by using the polymerase chain reaction to synthesize the variable region using mRNA of antibody-producing cells as a template (see, for example, Larrick et al., Methods: A Companion to Methods in Enzymology, 2:106 (1991 ); Courtenay- Luck, "Genetic Manipulation of Monoclonal Antibodies," in Monoclonal Antibodies Production, Engineering and Clinical Application, Ritter et al. (eds.), page 166, Cambridge University Press (1995); and Ward et al., "Genetic Manipulation and Expression of Antibodies," in Monoclonal Antibodies: Principles and Applications, Birch et al., (eds.), page 137, Wiley-Liss, Inc. (1995)).

[0021] The exact definitional CDR boundaries and lengths are subject to different classification and numbering systems. CDRs may therefore be referred to by Kabat, Chothia, contact or any other boundary definitions, including the numbering system described herein. Despite differing boundaries, each of these systems has some degree of overlap in what constitutes the so-called “hypervariable regions” within the variable sequences. CDR definitions according to these systems may therefore differ in length and boundary areas with respect to the adjacent framework region. See for example Kabat (an approach based on cross-species sequence variability), Chothia (an approach based on crystallographic studies of antigen-antibody complexes), MacCallum, Honegger, and IMGT (Wu and Kabat, J. Exp. Med. 1970; 132:21 1 -250; Chothia et al., J. Mol. Biol, 1987, 196: 901 -917; MacCallum et al., J. Mol. Biol, 1996, 262: 732; Lefranc et al., Dev. Comp. Immunol. 2003; 27:55-77; Honegger et al., J. Mol. Bol. (2001 ); 309:657-670). Still another standard for characterizing the antigen binding side is the AbM definition used by Oxford Molecular’s AbM antibody modeling software. See, e.g., Protein Sequence and Structure Analysis of Antibody Variable Domains. In: Antibody Engineering Lab Manual (Ed.: Duebel, S. and Kontermann, R.,Springer-Verlag, Heidelberg). To the extent that two residue identification techniques define regions of overlapping, but not identical regions, they can be combined to define a hybrid CDR. In some embodiments, the CDRs disclosed herein are identified according to the Kabat numbering system. In some embodiments, the CDRs disclosed herein are identified according to the Chothia numbering system. In some embodiments, the CDRs disclosed herein are identified according to the IMGT approach. In some embodiments, the CDRs disclosed herein are identified according to the Honegger approach.

[0022] In some embodiments, the antibody (or antigen binding fragment) thereof comprises three heavy chain complementarity determining regions (CDRs) (CDR-H1 , CDR- H2, and CDR-H3) and three light chain complementarity determining regions (CDRs) (CDR- L1 , CDR-L2, and CDR-L3), wherein the CDR-H1 , CDR-H2, and CDR-H3 are present in a heavy chain variable domain (VH) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 1 , 16, 31 , 46, and 61 , and wherein the CDR-L1 , CDR-L2, and CDR-L3 are present in a light chain variable domain (VL) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 9. 24, 39, 54, and 69.

[0023] In various aspects, the antibody (or antigen binding fragment thereof) comprises at least one CDR sequence having at least 75% identity (e.g., at least 75%, 80%, 85%, 90%, 95% or 100% identity) to a CDR selected from CDR-H1 , CDR-H2, CDR-H3, CDR-L1 , CDR- L2, and CDR-L3 wherein CDR-H1 has the sequence given in any one of SEQ ID NOs: 2, 5, 17, 20, 32, 35, 47, 50, 62, and 65; CDR-H2 has the sequence given in any one of SEQ ID NOs: 3, 6, 18, 21 , 33, 36, 48, 51 , 63, and 66; CDR-H3 has the sequence given in any one of SEQ ID NOs: 4, 7, 19, 22, 34, 37, 49, 52, 64, and 67; CDR-L1 has the sequence given in any one of SEQ ID NO: 10, 13, 25, 28, 40, 43, 55, 58, 70, and 73; CDR-L2 has the sequence given in any one of SEQ ID NOs: 11 (DAS), 14, 26 (GAS), 29, 44, 59, and 74; and CDR-L3 has the sequence given in any one of SEQ ID NOs: 12, 27, 42, 57, and 72. The anti-OPG antibody, in various aspects, comprises two of the CDRs, three of the CDRs, four of the CDRs, five of the CDRs or all six of the CDRs.

[0024] In some embodiments, the anti-OPG antibody comprises a set of six CDRs as follows: CDR-H1 of SEQ ID NO: 2, CDR-H2 of SEQ ID NO: 3, CDR-H3 of SEQ ID NO: 4, CDR-L1 of SEQ ID NO: 10, CDR-L2 of SEQ ID NO: 11 and CDR-L3 of SEQ ID NO: 12.

[0025] In some embodiments, the anti-OPG antibody comprises a set of six CDRs as follows: CDR-H1 of SEQ ID NO: 5, CDR-H2 of SEQ ID NO: 6, CDR-H3 of SEQ ID NO: 7, CDR-L1 of SEQ ID NO: 13, CDR-L2 of SEQ ID NO: 14, and CDR-L3 of SEQ ID NO: 12.

[0026] In some embodiments, the anti-OPG antibody comprises a set of six CDRs as follows: CDR-H1 of SEQ ID NO: 17, CDR-H2 of SEQ ID NO: 18, CDR-H3 of SEQ ID NO: 19,CDR-L1 of SEQ ID NO: 23, CDR-L2 of SEQ ID NO: 26 (GAS), and CDR-L3 of SEQ ID NO: 27.

[0027] In some embodiments, the anti-OPG antibody comprises a set of six CDRs as follows: CDR-H1 of SEQ ID NO: 20, CDR-H2 of SEQ ID NO: 21 , CDR-H3 of SEQ ID NO: 22, CDR-L1 of SEQ ID NO: 28, CDR-L2 of SEQ ID NO: 29, and CDR-L3 of SEQ ID NO: 27.

[0028] In some embodiments, the anti-OPG antibody comprises a set of six CDRs as follows: CDR-H1 of SEQ ID NO: 32, CDR-H2 of SEQ ID NO: 33, CDR-H3 of SEQ ID NO: 34, CDR-L1 of SEQ ID NO: 40, CDR-L2 of SEQ ID NO: 41 (DAS), and CDR-L3 of SEQ ID NO: 42.

[0029] In some embodiments, the anti-OPG antibody comprises a set of six CDRs as follows: CDR-H1 of SEQ ID NO: 35, CDR-H2 of SEQ ID NO: 36, CDR-H3 of SEQ ID NO: 37, CDR-L1 of SEQ ID NO: 43, CDR-L2 of SEQ ID NO: 44, and CDR-L3 of SEQ ID NO: 42.

[0030] In some embodiments, the anti-OPG antibody comprises a set of six CDRs as follows: CDR-H1 of SEQ ID NO: 47, CDR-H2 of SEQ ID NO: 48, CDR-H3 of SEQ ID NO: 49, CDR-L1 of SEQ ID NO: 55, CDR-L2 of SEQ ID NO: 56 (GAS), and CDR-L3 of SEQ ID NO: 57.

[0031] In some embodiments, the anti-OPG antibody comprises a set of six CDRs as follows: CDR-H1 of SEQ ID NO: 50, CDR-H2 of SEQ ID NO: 51 , CDR-H3 of SEQ ID NO: 52, CDR-L1 of SEQ ID NO: 58, CDR-L2 of SEQ ID NO: 59, and CDR-L3 of SEQ ID NO: 57.

[0032] In some embodiments, the anti-OPG antibody comprises a set of six CDRs as follows: CDR-H1 of SEQ ID NO: 62, CDR-H2 of SEQ ID NO: 63, CDR-H3 of SEQ ID NO: 64, CDR-L1 of SEQ ID NO: 70, CDR-L2 of SEQ ID NO: 74, and CDR-L3 of SEQ ID NO: 72.

[0033] In some embodiments, the anti-OPG antibody comprises a set of six CDRs as follows: CDR-H1 of SEQ ID NO: 65, CDR-H2 of SEQ ID NO: 66, CDR-H3 of SEQ ID NO: 67, CDR-L1 of SEQ ID NO: 73, CDR-L2 of SEQ ID NO: 74, and CDR-L3 of SEQ ID NO: 72.

[0034] In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 75% identity (e.g., at least 75%, 80%, 85%, 90%, 95% or 100% identity) to an amino acid sequence set forth in any one of SEQ ID NOs: 1 , 16, 31 , 46, and 61 and / or a light chain variable region comprising an amino acid sequence having at least 75% identity (e.g., at least 75%, 80%, 85%, 90%, 95% or 100% identity) to an amino acid sequence set forth in any one of SEQ ID NOs: 9, 24, 39, 54, and

[0035] In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 1 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 9. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 16 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 24. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 31 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 39. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 46 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 54. In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 61 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 69.

[0036] In some embodiments, the antibody comprises a heavy chain amino acid sequence set forth in any one of SEQ ID NOs: 8, 23, 38, 53, and 68, and / or a light chain amino acid sequence set forth in any one of SEQ ID NOs: 15, 30, 45, 60, and 76.

[0037] In some embodiments, the antibody comprises a heavy chain amino acid sequence set forth in SEQ ID NO: 8 and a light chain amino acid sequence set forth in SEQ ID NO: 15. In some embodiments, the antibody comprises a heavy chain amino acid sequence set forth in SEQ ID NO: 23 and a light chain amino acid sequence set forth in SEQ ID NO: 30. In some embodiments, the antibody comprises a heavy chain amino acid sequence set forth in SEQ ID NO: 38 and a light chain amino acid sequence set forth in SEQ ID NO: 45. In some embodiments, the antibody comprises a heavy chain amino acid sequence set forth in SEQ ID NO: 53 and a light chain amino acid sequence set forth in SEQ ID NO: 60. In some embodiments, the antibody comprises a heavy chain amino acid sequence set forth in SEQ ID NO: 68 and a light chain amino acid sequence set forth in SEQ ID NO: 76.

[0038] Antigen binding fragments of the anti-OPG antibodies described herein are also contemplated. The antigen binding fragment can be any part of an antibody that has at least one antigen binding site, and the antigen binding fragment may be part of a larger structure (an “antibody product”) that retains the ability of the antigen binding fragment to recognize CRH. For ease of reference, these antibody products that include antigen binding fragments are included in the disclosure herein of “antigen binding fragment.” Examples of antigen binding fragments, include, but are not limited to, Fab, F(ab')2, a monospecific or bispecific Fab2, a trispecific Fabs, scFv, dsFv, scFv-Fc, bispecific diabodies, trispecific triabodies,minibodies, a fragment of IgNAR (e.g., V-NAR), a fragment of hcIgG (e.g., VhH), bis-scFvs, fragments expressed by a Fab expression library, and the like. In exemplary aspects, the antigen binding fragment is a domain antibody, VhH domain, V-NAR domain, VH domain, VL domain, or the like. Antibody fragments of the disclosure, however, are not limited to these exemplary types of antibody fragments. In exemplary aspects, antigen binding fragment is a Fab fragment. In exemplary aspects, the antigen binding fragment comprises two Fab fragments. In exemplary aspects, the antigen binding fragment comprises two Fab fragments connected via a linker. In exemplary aspects, the antigen binding fragment comprises or is a minibody comprising two Fab fragments. In exemplary aspects, the antigen binding fragment comprises, or is, a minibody comprising two Fab fragments joined via a linker. Minibodies are known in the art. See, e.g., Hu et al., Cancer Res 56: 3055- 3061 (1996). In exemplary aspects, the antigen binding fragment comprises or is a minibody comprising two Fab fragments joined via a linker, optionally, comprising an alkaline phosphatase domain.

[0039] A domain antibody comprises a functional binding unit of an antibody, and can correspond to the variable regions of either the heavy (VH) or light (VL) chains of antibodies. A domain antibody can have a molecular weight of approximately 13 kDa, or approximately one-tenth of a full antibody. Domain antibodies may be derived from full antibodies such as those described herein.

[0040] In some embodiments, the scFv is attached to a human Fc domain. In some embodiments, the Fc domain does not activate Fc effector functions.

[0041] In some embodiments, the OPG inhibitor is a soluble receptor that specifically binds OPG. In some embodiments, the OPG inhibitor is a soluble receptor that blocks OPG activity (either directly or indirectly). Zhao et al., Frontiers Pharmacol., 14:1171293, 2023, the disclosure of which is incorporated herein by reference in its entirety.Methods of Antibody or Antigen Binding Fragment Production

[0042] Suitable methods of making antibodies are known in the art. For instance, standard hybridoma methods are described in, e.g., Harlow and Lane (eds.), Antibodies: A Laboratory Manual, CSH Press (1988), and CA. Janeway et al. (eds.), Immunobiology, 5th Ed., Garland Publishing, New York, NY (2001)). Monoclonal antibodies for use in the methods of the disclosure may be prepared using any technique which provides for the production of antibody molecules by continuous cell lines in culture. These include but are not limited to the hybridoma technique originally described by Koehler and Milstein (Nature 256: 495-497, 1975), the human B-cell hybridoma technique (Kosbor et al., Immunol Today 4:72, 1983; Cote et al., Proc Natl Acad Sci 80: 2026-2030, 1983) and the EBV-hybridomatechnique (Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R Liss Inc, New York N.Y., pp 77-96, (1985). Alternatively, other methods, such as EBV-hybridoma methods (Haskard and Archer, J. Immunol. Methods, 74(2), 361-67 (1984), and Roder et al., Methods Enzymol., 121 , 140-67 (1986)), and bacteriophage vector expression systems (see, e.g., Huse et al., Science, 246, 1275-81 (1989)) are known in the art. Further, methods of producing antibodies in non-human animals are described in, e.g., U.S. Patents 5,545,806, 5,569,825, and 5,714,352, and U.S. Patent Application Publication No. 2002 / 0197266 Al). Antibodies may also be produced by inducing in vivo production in the lymphocyte population or by screening recombinant immunoglobulin libraries or panels of highly specific binding reagents as disclosed in Orlandi et al (Proc Natl Acad Sci 86: 3833-3837; 1989), and Winter G and Milstein C (Nature 349: 293-299, 1991). If the full sequence of the antibody or antigen-binding fragment is known, then methods of producing recombinant proteins may be employed. See, e.g., “Protein production and purification” Nat Methods 5(2): 135-146 (2008). In some embodiments, the antibodies (or antigen binding fragments) are isolated from cell culture or a biological sample if generated in vivo.

[0043] Phage display also can be used to generate the antibodies described herein. In this regard, phage libraries encoding antigen-binding variable (V) domains of antibodies can be generated using standard molecular biology and recombinant DNA techniques (see, e.g., Sambrook et al. (eds.), Molecular Cloning, A Laboratory Manual, 3rd Edition, Cold Spring Harbor Laboratory Press, New York (2001)). Phage encoding a variable region with the desired specificity are selected for specific binding to the desired antigen, and a complete or partial antibody is reconstituted comprising the selected variable domain. Nucleic acid sequences encoding the reconstituted antibody are introduced into a suitable cell line, such as a myeloma cell used for hybridoma production, such that antibodies having the characteristics of monoclonal antibodies are secreted by the cell (see, e.g., Janeway et al., supra, Huse et al., supra, and U.S. Patent 6,265,150). Related methods also are described in U.S. Patent No. 5,403,484; U.S. Patent No. 5,571 ,698; U.S. Patent No. 5,837,500; U.S. Patent No. 5,702,892. The techniques described in U.S. Patent No. 5,780,279; U.S. Patent No. 5,821 ,047; U.S. Patent No. 5,824,520; U.S. Patent No. 5,855,885; U.S. Patent No. 5,858,657; U.S. Patent No. 5,871 ,907; U.S. Patent No. 5,969,108; U.S. Patent No.6,057,098; and U.S. Patent No. 6,225,447.

[0044] Antibodies can be produced by transgenic mice that are transgenic for specific heavy and light chain immunoglobulin genes. Such methods are known in the art and described in, for example U.S. Patent Nos. 5,545,806 and 5,569,825, and Janeway et al., supra.

[0045] Methods for generating humanized antibodies are well known in the art and are described in detail in, for example, Janeway et al., supra, U.S. Patent Nos. 5,225,539, 5,585,089 and 5,693,761 , European Patent No. 0239400 Bl, and United Kingdom Patent No. 2188638. Humanized antibodies can also be generated using the antibody resurfacing technology described in U.S. Patent No. 5,639,641 and Pedersen et al., J. Mol. Biol, 235, 959-973 (1994). A preferred chimeric or humanized antibody has a human constant region, while the variable region, or at least a CDR, of the antibody is derived from a non-human species. Methods for humanizing non-human antibodies are well known in the art. (See U.S. Patent Nos. 5,585,089, and 5,693,762.)

[0046] Techniques developed for the production of “chimeric antibodies,” e.g., the splicing of mouse antibody genes to human antibody genes to obtain a molecule with appropriate antigen specificity and biological activity, can be used (Morrison et al., Proc Natl Acad Sci 81 : 6851-6855 (1984); Neuberger et al., Nature 312: 604-608 (1984); Takeda et al., Nature 314: 452-454 (1985)). Alternatively, techniques described for the production of single chain antibodies (U.S. Patent No. 4,946,778) can be adapted to produce CRH-specific single chain antibodies.

[0047] Likewise, using techniques known in the art to isolate CDRs, compositions comprising CDRs are generated. Compositions comprising one, two, and / or three CDRs of a heavy chain variable region or a light chain variable region of a monoclonal antibody can be generated. The CDRs of exemplary antibodies are provided herein as SEQ ID NOs: 4-9 and 12-17. Techniques for cloning and expressing nucleotide and polypeptide sequences are well-established in the art (see, e.g., Sambrook et al., Molecular Cloning: A Laboratory Manual, 2nd Edition, Cold Spring Harbor, New York (1989)). The amplified CDR sequences are ligated into an appropriate expression vector. The vector comprising one, two, three, four, five and / or six cloned CDRs optionally contains additional polypeptide encoding regions linked to the CDR.

[0048] Chemically constructed bispecific antibodies may be prepared by chemically crosslinking heterologous Fab or F(ab')2 fragments by means of chemicals such as heterobifunctional reagent succinimidyl-3-(2-pyridyldithiol)-propionate (SPDP, Pierce Chemicals, Rockford, III.). The Fab and F(ab')2 fragments can be obtained from intact antibody by digesting it with papain or pepsin, respectively (Karpovsky et al., J. Exp. Med. 160:1686-701 (1984); Titus et al., J. Immunol., 138:4018-22 (1987)).

[0049] Methods of testing antibodies for the ability to bind to an epitope of CRH, regardless of how the antibodies are produced, are known in the art and include, e.g., radioimmunoassay (RIA), ELISA, Western blot, immunoprecipitation, surface plasmonresonance (e.g., BIAcore), and competitive inhibition assays (see, e.g., Janeway et al., infra, and U.S. Patent Application Publication No. 2002 / 0197266).

[0050] Antibody fragments that contain the antigen binding, or idiotype, of the antibody molecule may be generated by techniques known in the art. For example, a F(ab')2 fragment may be produced by pepsin digestion of the antibody molecule; Fab' fragments may be generated by reducing the disulfide bridges of the F(ab')2 fragment; and two Fab' fragments which may be generated by treating the antibody molecule with papain and a reducing agent. The disclosure is not limited to enzymatic methods of generating antigen binding fragments; the antigen binding fragment may be a recombinant antigen binding fragment produced by expressing a polynucleotide encoding the fragment in a suitable host cell.

[0051] A single-chain variable region fragments (scFv), which consists of a truncated Fab fragment comprising the variable (V) domain of an antibody heavy chain linked to a V domain of an antibody light chain via a synthetic peptide, can be generated using routine recombinant DNA technology techniques (see, e.g., Janeway et al., supra). Similarly, disulfide-stabilized variable region fragments (dsFv) can be prepared by recombinant DNA technology (see, e.g., Reiter et al., Protein Engineering, 7, 697-704 (1994)).

[0052] Recombinant antibody fragments, e.g., scFvs, can also be engineered to assemble into stable multimeric oligomers of high binding avidity and specificity to different target antigens. Such diabodies (dimers), triabodies (trimers) or tetrabodies (tetramers) are well known in the art, see e.g., Kortt et al., Biomol Eng. 2001 18:95-108, (2001) and Todorovska et al., J Immunol Methods. 248:47-66, (2001).

[0053] Therapeutic Methods

[0054] The antibodies or antigen binding fragments thereof described herein are useful for treating senescence-associated conditions. A “senescence-associated condition” refers to any state or condition associated with the accumulation of senescent cells in a subject. In some embodiments, the subject does not have cancer.

[0055] “Treating” or “treatment” refers to achieving any preventative or therapeutic effect. In some embodiments, the therapeutic effect is a reduction in the abundance of senescent cells in the subject, for example, wherein a therapeutic effect comprises a reduction in the number of senescent cells in a selected compartment. The reduction may be an absolute reduction in cell numbers, or a proportional reduction in the ratio of senescent cells to nonsenescent cells. In various embodiments, a therapeutic effect is defined as a reduction in senescent cells in the selected compartment of at least 10%, at least 20%, at least 30%, atleast 40%, at least 50%, at least 60%, at least 70%, at least 80% or at least 90%, compared to the target prior to treatment, or compared to like, untreated targets.

[0056] In some embodiments, the therapeutic effect is a reduction in the abundance of senescence-associated secretory phenotype (SASP) cells or SASP factors in the selected compartment. The reduction in SASP cells or SASP factors may be any reduction in the absolute number of SASP cells or factors in the selected compartment, for example, a reduction of at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80% or at least 90%, compared to the target prior to treatment or compared to like, untreated targets.

[0057] In some embodiments, the senescence-associated condition is an inflammatory or autoimmune condition. Exemplary inflammatory or autoimmune conditions include, but are not limited to, acute and chronic lung inflammation, lung fibrosis (including idiopathic lung fibrosis), ankylosing spondylarthritis, arthritis (including psoriatic arthritis, osteoarthritis, reactive arthritis, juvenile, and rheumatoid arthritis), arthropathy, asthma, Ataxia Telangiestasi, cachexia, chronic bronchitis, chronic pulmonary obstructive diseases, Crohn's disease, cystic fibrosis, Ehlers-Danlos Syndrome, endotoxic shock, fibromyalgia, gout, hypermobility syndrome, inflammatory bowel disease (Crohn's disease and ulcerative colitis), ischemia induced inflammation, ischemia-reperfusion injury, kidney ischemia, kyphosis, limb ischemia, liver fibrosis, local and systemic inflammation, lupus, metabolic acidosis, multiple sclerosis, osteoarthritis, psoriasis, sarcopenia, scleroderma and vasculitis, sepsis or septic shock, and ulcerative colitis.

[0058] In various embodiments, the senescence-associated condition is a condition caused by metabolic dysfunction associated with the accumulation of senescent cells. For example, in various embodiments, the condition is metabolic dysfunction associated with the accumulation of senescent preadipocytes in epidydmal white adipose tissue, for example, as occurs in Type II diabetes.

[0059] In some embodiments, the senescence-associated condition is diabetes or a diabetes- related condition. For example, in various embodiments, the condition is diabetes Type I, diabetes Type II diabetes- associated inflammation, diabetic ulcer, diabetic nephropathy, or diabetic adhesive capsulitis, metabolic dysfunction associated with the accumulation of senescent preadipocytes in epidydmal white adipose tissue, glucose intolerance, glucose dysregulation and other symptoms of diabetes.

[0060] In some embodiments, the senescence-associated condition is a cardiovascular condition. Exemplary cardiovascular conditions include, but are not limited to, angina, aortic aneurysm, arrhythmia, atherosclerosis, brain aneurysm, cardiac diastolic dysfunction,cardiac fibrosis, cardiac stress resistance, cardiomyopathy, carotid artery disease, chronic obstructive pulmonary disease, congestive heart failure, coronary artery disease, coronary thrombosis, endocarditis, hypertension, hypercholesterolemia, hyperlipidemia, idiopathic pulmonary fibrosis, mitral valve prolapse, myocardial infarction, peripheral vascular disease, and stroke.

[0061] In some embodiments, the senescence associated condition is a neurodegenerative condition. Exemplary neurodegenerative conditions include, but are not limited to, Alzheimer’s disease, Parkinson’s disease, multiple sclerosis, amyotrophic lateral sclerosis, Huntington’s disease, epilepsy, chronic traumatic encephalopathy, frontotemporal dementia, dementia, and motor neuron dysfunction.

[0062] In various embodiments, the senescence associated condition is an age-related condition. Exemplary age-related conditions include, but are not limited to, loss of pulmonary function, macular degeneration, renal failure, frailty, muscle fatigue, liver fibrosis, pancreatic fibrosis, oral submucosa fibrosis, progressive muscle loss, decreased bone-density, age- associated memory impairment, age-related hearing loss, sarcopenia, skin atrophy, brain atrophy, arteriosclerosis, pulmonary emphysema, immunologic incompetence, cataracts, skin wrinkling, and graying of hair.Pharmaceutical Compositions

[0063] Pharmaceutical compositions comprising an anti-OPG antibody or antigen-binding fragment thereof described herein are also contemplated. In some embodiments, the pharmaceutical composition contains formulation materials for modifying, maintaining or preserving, for example, the pH, osmolarity, viscosity, clarity, color, isotonicity, odor, sterility, stability, rate of dissolution or release, adsorption or penetration of the composition. In such embodiments, suitable formulation materials include, but are not limited to, amino acids (such as glycine, glutamine, asparagine, arginine, proline, methionine or lysine); antimicrobials; antioxidants (such as reducing agents, oxygen / free-radical scavengers, and chelating agents (e.g., ascorbic acid, EDTA, sodium sulfite or sodium hydrogen-sulfite)); buffers (such as borate, bicarbonate, Tris-HCI, citrates, phosphates or other organic acids); bulking agents (such as mannitol or glycine); chelating agents (such as ethylenediamine tetraacetic acid (EDTA)); complexing agents (such as caffeine, polyvinylpyrrolidone, betacyclodextrin or hydroxypropyl-beta-cyclodextrin); fillers; monosaccharides; disaccharides; and other carbohydrates (such as glucose, mannose or dextrins); proteins (such as serum albumin, gelatin or immunoglobulins); coloring, flavoring and diluting agents; emulsifying agents; hydrophilic polymers (such as polyvinylpyrrolidone); low molecular weight polypeptides; salt-forming counter-ions (such as sodium); preservatives (such asbenzalkonium chloride, benzoic acid, salicylic acid, thimerosal, phenethyl alcohol, methylparaben, propylparaben, chlorhexidine, sorbic acid or hydrogen peroxide); solvents (such as glycerin, propylene glycol or polyethylene glycol); sugar alcohols (such as mannitol or sorbitol); suspending agents; surfactants or wetting agents (such as pluronics, PEG, sorbitan esters, polysorbates such as polysorbate 20, polysorbate, triton, tromethamine, lecithin, cholesterol, tyloxapal); stability enhancing agents (such as sucrose or sorbitol); tonicity enhancing agents (such as alkali metal halides, preferably sodium or potassium chloride, mannitol sorbitol); delivery vehicles; diluents; excipients and / or pharmaceutical adjuvants. See, REMINGTON'S PHARMACEUTICAL SCIENCES, 18" Edition, (A. R. Genrmo, ed.), 1990, Mack Publishing Company.

[0064] Selection of the particular formulation materials described herein may be driven by, for example, the intended route of administration, delivery format and desired dosage. See, for example, REMINGTON'S PHARMACEUTICAL SCIENCES, supra. The primary vehicle or carrier in a pharmaceutical composition may be either aqueous or non-aqueous in nature. For example, a suitable vehicle or carrier may be water for injection, physiological saline solution or artificial cerebrospinal fluid, possibly supplemented with other materials common in compositions for parenteral administration. Neutral buffered saline or saline mixed with serum albumin are further exemplary vehicles. In specific embodiments, pharmaceutical compositions comprise Tris buffer of about pH 7.0-8.5, or acetate buffer of about pH 4.0-5.5, and may further include sorbitol or a suitable substitute therefor. In certain embodiments, the composition may be prepared for storage by mixing the selected composition having the desired degree of purity with optional formulation agents (REMINGTON'S PHARMACEUTICAL SCIENCES, supra) in the form of a lyophilized cake or an aqueous solution. Further, in some embodiments, the antibody or (antigen binding fragment thereof) may be formulated as a lyophilizate using appropriate excipients such as sucrose.

[0065] The pharmaceutical compositions described herein can be selected for parenteral delivery. Alternatively, the compositions may be selected for inhalation or for delivery through the digestive tract, such as orally. Preparation of such pharmaceutically acceptable compositions is within the skill of the art. The formulation components are present preferably in concentrations that are acceptable to the site of administration. In certain embodiments, buffers are used to maintain the composition at physiological pH or at a slightly lower pH, typically within a pH range of from about 5 to about 8.

[0066] When parenteral administration is contemplated, the composition may be provided in the form of a pyrogen-free, parenterally acceptable aqueous solution comprising the desired antibody or fragment in a pharmaceutically acceptable vehicle. A particularly suitable vehicle for parenteral injection is sterile distilled water in which the antibody orfragment is formulated as a sterile, isotonic solution, properly preserved. In certain embodiments, implantable drug delivery devices may be used to introduce the desired antibody (or antigen binding fragment thereof).

[0067] Additional pharmaceutical compositions will be evident to those skilled in the art, including formulations involving antigen binding proteins in sustained- or controlled-delivery formulations. Techniques for formulating a variety of other sustained- or controlled-delivery means, such as liposome carriers, bio-erodible microparticles or porous beads and depot injections, are also known to those skilled in the art. See, for example, International Patent Application No. PCT / US93 / 00829, which is incorporated by reference and describes controlled release of porous polymeric microparticles for delivery of pharmaceutical compositions. Sustained-release preparations may include semipermeable polymer matrices in the form of shaped articles, e.g., films, or microcapsules. Sustained release matrices may include polyesters, hydrogels, polylactides (as disclosed in U.S. Pat. No. 3773919 and European Patent Application Publication No. EP058481 , each of which is incorporated by reference), copolymers of L-glutamic acid and gamma ethyl-L-glutamate (Sidman et al., 1983, Biopolymers 2:547-556), poly (2-hydroxyethyl-methacrylate) (Langer et al., 1981 , J. Biomed. Mater. Res. 15:167-277 and Langer, 1982, Chem. Tech. 12:98-105), ethylene vinyl acetate (Langer et al., 1981 , supra) or poly-D(-)-3-hydroxybutyric acid (European Patent Application Publication No. EP133988). Sustained release compositions may also include liposomes that can be prepared by any of several methods known in the art. See, e.g., Eppstein et al., 1985, Proc. Natl. Acad. Sci. U.S.A. 82:3688-3692; European Patent Application Publication Nos. EP036676; EP088046 and EP143949, incorporated by reference.

[0068] Administration of the compositions described herein will be via any common route so long as the target tissue is available via that route. The pharmaceutical compositions may be introduced into the subject by any conventional method, e.g., by intravenous, intradermal, intramusclar, intramammary, intraperitoneal, intrathecal, intraocular, retrobulbar, intrapulmonary (e.g., term release); by oral, sublingual, nasal, anal, vaginal, or transdermal delivery, or by surgical implantation at a particular site, e.g., embedded under the splenic capsule, brain, or in the cornea. The treatment may consist of a single dose or a plurality of doses over a period of time. In some embodiments, the composition is delivered via local administration to the brain.

[0069] The following Examples are provided to further illustrate aspects of the disclosure and are not meant to constrain the disclosure to any particular application or theory of operation.EXAMPLESExample 1 - TNF-related apoptosis-inducing ligand (TRAIL) is upregulated on the surface of iNKT cells

[0070] TRAIL is known to acts as a signaling molecule that can induce apoptosis (programmed cell death) in target cells. As shown in Figure 1 , stimulation with lipid antigen a-GalCer upregulated TRAIL (Fig 1 A). Cytokine release of IFN-g and proliferation (Fig 2 B,C) confirms the stimulation of iNKT cells.Example 2 - Senescent cells secrete osteoprotogerin (OPG)

[0071] Next, we identified that senescent cells and tissues that accumulate senescent cells secrete osteoprotogerin (OPG), which acts as a decoy protein that binds TRAIL and inhibits its interaction with the TRAIL receptor DR5 (Figure 2).Example 3 - OPG interferes with INKT cytotoxic activity

[0072] To extend the role of OPG in regulating lymphocyte cytotoxicity, mouse iNKT hybridoma cells were used as effector cells (DN32.D3). To generate target senescent cells, adipocyte-derived mesenchymal progenitor (ADMP) cells treated with etoposide were used to induce a senescent phenotype. iNKT hybridoma cells were activated by co-culturing with bone marrow-derived dendritic cells (BMDC) pulsed with 100 ng / mL a-GalCer. While no cell death was observed in BMDC pulsed with vehicle formulation, significant target cell death was observed when BMDC pulsed with a-GalCer were used to stimulate iNKT hybridomas (Fig 3A). This increase in cytotoxicity of a-GalCer-stimulated iNKT cells could be mitigated by the addition of recombinant OPG protein (Fig 3A). The ability of recombinant OPG to dampen the cytotoxic function of both CD8+ T cells and iNKT cells suggests that secreted OPG could effectively inhibit TRAIL-mediated Teff cytotoxicity. To directly test whether secreted OPG inhibited Teff cytotoxicity, we next reduced the amount of a-GalCer such that low-dose (10 ng / mL) a-GalCer-loaded BMDC / iNKT cells did not lead to any appreciable cytotoxicity of target cells, however addition of an antibody against OPG (aOPG) resulted in dose-dependent increase in target cell death (Fig 3B). Together, these results support the notion that OPG from senescent-like stromal cells can directly inhibit cytotoxic T-cell effector function.Example 4 - Effect of anti-OPG antibody on removal of senescent cells in vivo

[0073] The following Example was performed to assess whether blocking OPG with an anti-OPG antibody would lead to removal of senescent cells in vivo. As shown Figure 4A, chow, HFD and HFD mice treated with anti-OPG antibody were assessed for blood glucose control and insulin resistance two weeks after treatment. The adipose tissue from thesemice was isolated and the SVF fraction was assessed for senescent cells as described previously by our lab. As shown in Figures 4C and 4D, treatment of HFD mice clearly reduced the senescent cell burden to chow levels. Correspondingly, GTT assay showed improvement in blood glucose control and concomitant with a reduction in insulin resistance.

Claims

What is claimed is:1 . A method of treating a senescence-associated condition in a subject in need of treatment therefore comprising administering an osteoprotegerin inhibitor to the subject.

2. The method of claim 1 , wherein the senescence-associated condition is an inflammatory or autoimmune condition, diabetes or a diabetes-related condition, cardiovascular condition, neurodegenerative condition, or an age-related condition.

3. The method of claim 2, wherein the inflammatory or autoimmune condition is acute and chronic lung inflammation, lung fibrosis (including idiopathic lung fibrosis), ankylosing spondylarthritis, arthritis (including psoriatic arthritis, osteoarthritis, reactive arthritis, juvenile, and rheumatoid arthritis), arthropathy, asthma, Ataxia Telangiestasi, cachexia, chronic bronchitis, chronic pulmonary obstructive diseases, Crohn's disease, cystic fibrosis, Ehlers-Danlos Syndrome, endotoxic shock, fibromyalgia, gout, hypermobility syndrome, inflammatory bowel disease (Crohn's disease and ulcerative colitis), ischemia induced inflammation, ischemia-reperfusion injury, kidney ischemia, kyphosis, limb ischemia, liver fibrosis, local and systemic inflammation, lupus, metabolic acidosis, multiple sclerosis, osteoarthritis, psoriasis, sarcopenia, scleroderma and vasculitis, sepsis or septic shock, and ulcerative colitis.

4. The method of claim 2, wherein the diabetes or diabetes-related condition is diabetes Type I, diabetes Type II diabetes- associated inflammation, diabetic ulcer, diabetic nephropathy, or diabetic adhesive capsulitis, metabolic dysfunction associated with the accumulation of senescent preadipocytes in epidydmal white adipose tissue, glucose intolerance, glucose dysregulation and other symptoms of diabetes.

5. The method of claim 2, wherein the cardiovascular condition is angina, aortic aneurysm, arrhythmia, atherosclerosis, brain aneurysm, cardiac diastolic dysfunction, cardiac fibrosis, cardiac stress resistance, cardiomyopathy, carotid artery disease, chronic obstructive pulmonary disease, congestive heart failure, coronary artery disease, coronary thrombosis, endocarditis, hypertension, hypercholesterolemia, hyperlipidemia, idiopathic pulmonary fibrosis, mitral valve prolapse, myocardial infarction, peripheral vascular disease, or stroke.

6. The method of claim 2, wherein the neurodegenerative condition is Alzheimer’s disease, Parkinson’s disease, multiple sclerosis, amyotrophic lateral sclerosis, Huntington’s disease, epilepsy, chronic traumatic encephalopathy, frontotemporal dementia, dementia, or motor neuron dysfunction.

7. The method of claim 2, wherein the age-related condition is loss of pulmonary function, macular degeneration, renal failure, frailty, muscle fatigue, liver fibrosis, pancreatic fibrosis, oral submucosa fibrosis, progressive muscle loss, decreased bone-density, age- associated memory impairment, age-related hearing loss, sarcopenia, skin atrophy, brain atrophy, arteriosclerosis, pulmonary emphysema, immunologic incompetence, cataracts, skin wrinkling, and graying of hair.

8. The method of claim 1 , wherein the senescence-associated condition is fibrosis.

9. The method of claim 1 , wherein the senescence-associated condition is diabetes.

10. The method of any one of claims 1-9, wherein the osteoprotegerin inhibitor is an antibody or antigen binding fragment thereof that specifically binds osteoprotegerin.11 . The method of claim 10, wherein the antibody or antigen binding fragment thereof comprises three heavy chain complementarity determining regions (CDRs) (CDR- H1 , CDR-H2, and CDR-H3) and three light chain complementarity determining regions (CDRs) (CDR-L1 , CDR-L2, and CDR-L3), wherein the CDR-H1 , CDR-H2, and CDR-H3 are present in a heavy chain variable domain (VH) comprising the amino acid sequence set forth in any one of SEQ ID NOs: 1 , 16, 31 , 46, and 61 , and wherein the CDR-L1 , CDR-L2, and CDR-L3 are present in a light chain variable domain (VL) comprising the amino acid sequence set forth in any one of SEQ ID NOs:

9. 24, 39, 54, and 69.

12. The isolated antibody or antigen binding fragment of claim 11 , wherein: the CDR-H1 , CDR-H2, and CDR-H3, and the CDR-L1 , CDR-L2, and CDR-L3 are defined according to Kabat; the CDR-H1 , CDR-H2, and CDR-H3, and the CDR-L1 , CDR-L2, and CDR-L3 are defined according to Chothia; the CDR-H1 , CDR-H2, and CDR-H3, and the CDR-L1 , CDR-L2, and CDR-L3 are defined according to IMGT; the CDR-H1 , CDR-H2, and CDR-H3, and the CDR-L1 , CDR-L2, and CDR-L3 are defined according to AbM; the CDR-H1 , CDR-H2, and CDR-H3, and the CDR-L1 , CDR-L2, and CDR-L3 are defined according to Contact; or the CDR-H1 , CDR-H2, and CDR-H3, and the CDR-L1 , CDR-L2, and CDR-L3 are defined according to Honneger (AHo).

13. The method of any one of claims 1-12, wherein the subject does not have cancer.

14. The method of claim 13, wherein the cancer is breast cancer, bladder cancer, brain cancer, cervical cancer, colorectal cancer, esophageal cancer, gastric cancer, head and neck cancer, kidney cancer, lung cancer, leukemia, lymphoma, myeloma, ovarian cancer, pancreatic cancer, prostate cancer, sarcoma, or skin cancer.

15. The method of claim 13, wherein the cancer is a stromagenic cancer.

Images