Compositions and methods for detecting or eliminating senescent cells to diagnose or treat disease

Abstract

Disclosed are agents (e.g., peptides, polypeptides, proteins, small molecules, antibodies, and antibody fragments that target senescent cells) and methods of their use for imaging senescent cells in vivo and for treating or preventing cancer, age-related disease, tobacco-related disease, or other diseases and disorders related to or caused by cellular senescence in a mammal. The methods include administering one or more of the agents of the invention to a mammal, e.g., a human. The agents, which specifically bind to senescent cells, can be labeled with a radioactive label or a therapeutic label, e.g., a cytotoxic agent.

1 . An agent comprising a peptide, polypeptide, antibody, or fragment thereof comprising an amino acid sequence set forth in any one of SEQ ID NOs:1-3 and 5-8 or a peptide, polypeptide, antibody, antibody fragment, or small molecule capable of specifically binding to an antigen comprising an amino acid sequence having at least 20 amino acids with at least 80% sequence identity to any one of the amino acid sequences set forth in SEQ ID NOS:11-23. 
     
     
         2 . The agent of  claim 1 , wherein said antigen comprises an amino acid sequence having at least 20 amino acids of any one of the amino acid sequences set forth in SEQ ID NOS:11-23. 
     
     
         3 . The agent of  claim 1 , wherein said antigen comprises any one of the amino acid sequences set forth in SEQ ID NOS:11-23. 
     
     
         4 . The agent of  claim 1 , further comprising one or more of a detectable label, a therapeutic agent, a chelating agent, or a linker moiety. 
     
     
         5 . The agent of  claim 4 , wherein said agent is indirectly attached to said detectable label. 
     
     
         6 . The agent of  claim 4 , wherein said agent is directly attached to said detectable label. 
     
     
         7 . The agent of  claim 4 , wherein said linker moiety comprises the amino acid sequence GGGC, GGGS, or GG. 
     
     
         8 . The agent of  claim 4 , wherein said detectable label is a radioactive agent, a fluorescent agent, a bioluminescent molecule, an epitope tag, or a heavy metal. 
     
     
         9 . The agent of  claim 8 , wherein said radioactive agent is an iodine, astatine, or bromine label that is attached to an amino acid of said agent. 
     
     
         10 . The agent of  claim 8 , wherein said radioactive agent is technetium-99m. 
     
     
         11 . The agent of  claim 8 , wherein said fluorescent agent is fluorescein isothiocyanate (FITC), allophycocyanin (APC), phycoerythrin (PE), rhodamine, tetramethyl rhodamine isothiocyanate (TRITC), fluorescent protein (GFP), enhanced GFP (eGFP), yellow fluorescent protein (YFP), cyan fluorescent protein (CFP), red fluorescent protein (RFP), or dsRed. 
     
     
         12 . The agent of  claim 8 , wherein said bioluminescent molecule is luciferase. 
     
     
         13 . The agent of  claim 8 , wherein said epitope tag is c-myc, hemagglutinin, or a histidine tag. 
     
     
         14 . The agent of  claim 4 , wherein said therapeutic agent is a cytotoxic agent. 
     
     
         15 . The agent of  claim 14 , wherein said cytotoxic agent is an alkylating agent, an antibiotic, an antineoplastic agent, an antimetabolic agent, a ribosomal activity inhibitor, an antiproliferative agent, a tubulin inhibitor, a topoisomerase I or II inhibitor, a growth factor, an hormonal agonist or antagonists, an apoptotic agent, an immunomodulator, a radioactive agent, a phospholipase, or a cytotoxic peptide or lysin. 
     
     
         16 . The agent of  claim 14 , wherein said cytotoxic agent is selected from the group consisting of the following compounds and their derivatives: ricin, doxorubicin, methotrexate, camptothecin, homocamptothecin, thiocolchicine, colchicine, combretastatin, combretastin A-4, podophyllotoxin, rhizoxin, rhizoxin-d, dolistatin, paclitaxel, CC 1065, ansamitocin p3, maytansinoid, streptolysin O, stoichactis toxin, phallolysin,  staphylococcus  alpha toxin, holothurin A, digitonin, melittin, lysolecithin, cardiotoxin, and cerebratulus A toxin. 
     
     
         17 . The agent of  claim 4 , wherein said chelating agent is an ininocarboxylic reactive group, a polyaminopolycarboxylic reactive group, diethylenetriaminepentaacetic acid (DTPA), or 1, 4,7,1 O-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA). 
     
     
         18 . The agent of  claim 1 , wherein said agent further comprises a pharmaceutically acceptable carrier. 
     
     
         19 . The agent of  claim 1 , wherein said agent specifically binds to a senescent cell. 
     
     
         20 . The agent of  claim 19 , wherein said senescent cell comprises a senescent lung, breast, colon, prostate, gastric, hepatic, ovarian, esophageal, or bronchial epithelial or stromal cell, a senescent skin epithelial or stromal cell; a senescent glial cell; or a senescent vascular endothelial or stromal cell. 
     
     
         21 . A method of imaging a senescent cell-containing region in a mammal in vivo, said method comprising the steps of:
 (a) administering to said mammal the agent of  claim 1  and a detectable label;   (b) allowing said agent to bind senescent cells and allowing unbound agent to be cleared from the body of said mammal; and   (c) obtaining an image of said senescent cell-containing region.   
     
     
         22 . The method of  claim 21 , wherein said mammal is a human. 
     
     
         23 . The method of  claim 21 , wherein said region is the breast, prostate, gastrointestinal tract, liver, lungs, intracranial space, nasopharynx, oropharynx, larynx, esophagus, mediastinum, abdomen and pelvis, any region of the body containing peripheral vasculature, or the entire body. 
     
     
         24 . The method of  claim 21 , wherein said image is obtained using scintigraphy. 
     
     
         25 . A method of predicting cancer risk in a mammal, said method comprising the steps of:
 (a) administering to said mammal the agent of  claim 1  and a detectable label;   (b) predicting an elevated cancer risk in said mammal by detecting binding of said agent to a senescent cell of said mammal.   
     
     
         26 . The method of  claim 25 , wherein said mammal is a human. 
     
     
         27 . The method of  claim 25 , wherein said cancer is prostate cancer, colon cancer, lung cancer, squamous cell cancer of the head and neck, esophageal cancer, hepatocellular carcinoma, gastric cancer, pancreatic cancer, ovarian cancer, or breast cancer. 
     
     
         28 . A method of treating or preventing disease in a mammal, said method comprising administering to said mammal the agent of  claim 14 . 
     
     
         29 . The method of  claim 28 , wherein said mammal is a human. 
     
     
         30 . The method of  claim 28 , wherein said disease is cancer, age-related disease, tobacco-related disease, or skin wrinkles. 
     
     
         31 . The method of  claim 30 , wherein said cancer is prostate cancer, colon cancer, lung cancer, squamous cell cancer of the head and neck, esophageal cancer, hepatocellular carcinoma, gastric cancer, pancreatic cancer, ovarian cancer, or breast cancer. 
     
     
         32 . The method of  claim 30 , wherein said age-related or tobacco-related disease is cardiovascular disease, cerebrovascular disease, peripheral vascular disease, Alzheimer's disease, osteoarthritis, cardiac diastolic dysfunction, benign prostatic hypertrophy, aortic aneurysm, or emphysema. 
     
     
         33 . A method of treating or preventing disease in a mammal, said method comprising administering to said mammal a nucleic acid molecule encoding a cytotoxic agent and the agent of  claim 1 . 
     
     
         34 . The method of  claim 33 , wherein said mammal is a human. 
     
     
         35 . The method of  claim 33 , wherein said disease is cancer, age-related disease, or tobacco-related disease. 
     
     
         36 . The method of  claim 31 , wherein said cancer is prostate cancer, colon cancer, lung cancer, squamous cell cancer of the head and neck, esophageal cancer, hepatocellular carcinoma, gastric cancer, pancreatic cancer, ovarian cancer, or breast cancer. 
     
     
         37 . The method of  claim 35 , wherein said age-related or tobacco-related disease is cardiovascular disease, cerebrovascular disease, peripheral vascular disease, Alzheimer's disease, osteoarthritis, cardiac diastolic dysfunction, benign prostatic hypertrophy, aortic aneurysm, or emphysema. 
     
     
         38 . The method of  claim 33 , wherein said nucleic acid molecule is administered in a vector. 
     
     
         39 . The method of  claim 38 , wherein said vector is an adenoviral vector. 
     
     
         40 . The method of  claim 33 , wherein said cytotoxic agent and said agent are expressed as a single polypeptide chain. 
     
     
         41 . A method of identifying a peptide or polypeptide capable of detecting senescent cells comprising the steps of:
 (a) culturing cells to produce senescent cells;   (b) exposing said senescent cells to a phage peptide library;   (c) recovering said senescent cells and phage from said phage peptide library bound to said senescent cells;   (d) eluting and amplifying said bound phage to produce amplified phage;   (e) repeating steps (a)-(d) one or more times; and   (f) recovering a peptide or polypeptide expressed by said amplified phage that is capable of detecting senescent cells.   
     
     
         42 . A method for identifying an antibody or antibody fragment capable of specifically binding to a senescent cell-specific antigen, said method comprising the steps of:
 (a) contacting an antibody or antibody fragment with a polypeptide comprising at least 20 amino acids comprising at least 80% amino acid sequence identity to any one of the amino acid sequences set forth in SEQ ID NOS:11-23; and   (b) identifying an antibody or antibody fragment that binds said polypeptide with a dissociation constant of less than 10 −7  M.   
     
     
         43 . A method for making an antibody or antibody fragment capable of specifically binding to a senescent cell-specific antigen, said method comprising the steps of:
 (a) administering a polypeptide comprising at least 20 amino acids comprising at least 80% amino acid sequence identity to any one of the amino acid sequences set forth in SEQ ID NOS:11-23 to a mammal;   (b) allowing said mammal to generate a humoral immune response to said polypeptide; and   (c) isolating from said mammal an antibody or antibody fragment that binds said polypeptide with a dissociation constant of less than 10 −7  M.   
     
     
         44 . The method of  claim 43 , wherein said mammal is a mouse, hamster, rat, guinea pig, chicken, goat, sheep, cow, horse, non-human primate, or human. 
     
     
         45 . A method for identifying a small molecule capable of specifically binding to a senescent cell-specific antigen, said method comprising the steps of:
 (a) contacting a small molecule with a polypeptide comprising at least 20 amino acids comprising at least 80% amino acid sequence identity to any one of the amino acid sequences set forth in SEQ ID NOS:11-23; and   (b) identifying a small molecule that binds said polypeptide with a dissociation constant of less than 10 −7  M.   
     
     
         46 . A method of making an antibody or antibody fragment, said method comprising recombinantly expressing a nucleic acid sequence that encodes an amino acid sequence comprising one or more of SEQ ID NOs:1-3 and 5-8, wherein said antibody or antibody fragment specifically binds a senescent cell. 
     
     
         47 . A method of cellular therapy comprising administering to a mammal in need thereof the agent of  claim 1  prior to, concurrent with, or following administration of a cellular therapeutic. 
     
     
         48 . The method of  claim 47 , wherein said mammal is a human. 
     
     
         49 . A method of cellular therapy comprising contacting the agent of  claim 1  to a donor cell, tissue, or organ prior to, concurrent with, or following administration of said cell, tissue, or organ to a mammal. 
     
     
         50 . The method of  claim 49 , wherein said mammal is a human. 
     
     
         51 . The method of  claim 49 , wherein said donor cell, tissue, or organ comprises an autologous, allogeneic, syngeneic, or xenogeneic cell, tissue, or organ. 
     
     
         52 . The method of  claim 49 , wherein said cell comprises a stem cell population. 
     
     
         53 . The method of  claim 52 , wherein said stem cell is selected from a hematopoietic, umbilical cord blood, totipotent, multipotent, or pluripotent stem cell. 
     
     
         54 . A kit comprising:
 (a) the agent of  claim 1 ; and   (b) one or more of a detectable label, a therapeutic agent, a chelating agent, or a linker moiety.   
     
     
         55 . A method of treating a disease related to or caused by cellular senescence in a mammal comprising administering to the mammal a pharmaceutical composition comprising: (a) a peptide, a polypeptide, or an antibody or antibody fragment thereof, wherein the peptide, the polypeptide, or the antibody or antibody fragment thereof specifically binds to a senescent cell; and (b) a pharmaceutically acceptable carrier. 
     
     
         56 . The method of  claim 55 , wherein the peptide, the polypeptide, or the antibody or antibody fragment thereof specifically binds to a senescent cell-specific peptide, polypeptide, or glycoprotein. 
     
     
         57 . The method of  claim 55 , wherein the peptide, the polypeptide, or the antibody or the antibody fragment thereof administered to the mammal is coupled to a therapeutic agent. 
     
     
         58 . The method of  claim 57 , wherein the therapeutic agent is a cytotoxic agent. 
     
     
         59 . The method of  claim 55 , wherein the disease is a cancer, an age-related disease, or a tobacco-related disease. 
     
     
         60 . A method of treating a disease related to or caused by cellular senescence in a mammal comprising administering to the mammal a pharmaceutical composition comprising: (a) a small molecule that specifically binds to a senescent cell; and (b) a pharmaceutically acceptable carrier. 
     
     
         61 . The method of  claim 60 , wherein the small molecule specifically binds to a senescent cell-specific peptide, polypeptide, or glycoprotein. 
     
     
         62 . The method of  claim 60 , wherein the disease is a cancer, an age-related disease, or a tobacco-related disease. 
     
     
         63 . A method for selectively killing a senescent cell in a mammal comprising administering to the mammal a pharmaceutical composition comprising (a) a peptide, a polypeptide, or an antibody or antibody fragment thereof, wherein the peptide, the polypeptide, or the antibody or antibody fragment thereof specifically binds to the senescent cell; and (b) a pharmaceutically acceptable carrier. 
     
     
         64 . The method of  claim 63 , wherein the peptide, the polypeptide, or the antibody or antibody fragment thereof specifically binds to a senescent cell-specific peptide, polypeptide, or glycoprotein. 
     
     
         65 . The method of  claim 63 , wherein the peptide, the polypeptide, or the antibody or the antibody fragment that is administered to the mammal is coupled to a therapeutic agent. 
     
     
         66 . The method of  claim 65 , wherein the therapeutic agent is a cytotoxic agent. 
     
     
         67 . The method of  claim 63 , wherein the senescent cell is a senescent lung cell, a senescent breast cell, a senescent colon cell, a senescent prostate cell, a senescent gastric cell, a senescent hepatic cell, a senescent ovarian cell, a senescent esophageal cell, a senescent bronchial epithelial or stromal cell, a senescent skin epithelial or stromal cell, a senescent glial cell, or a senescent vascular endothelial or stromal cell. 
     
     
         68 . The method of  claim 63 , wherein the pharmaceutical composition comprises the peptide, and wherein the peptide comprises (a) the amino acid sequence selected from any one of SEQ ID NOS:1-3; or (b) an amino acid sequence at least 90% identical to the amino acid sequence selected from any one of SEQ ID NOS:1-3. 
     
     
         69 . The method of  claim 63 , wherein the pharmaceutical composition comprises the antibody or antibody fragment thereof, and wherein the antibody or antibody fragment thereof comprises (a) the amino acid sequence selected from any one of SEQ ID NOS:1-3 in a complementarity determining region; or (b) an amino acid sequence at least 90% identical to the amino acid sequence selected from any one of SEQ ID NOS:1-3 in a complementarity determining region. 
     
     
         70 . The method of  claim 64 , wherein the senescent cell-specific peptide, polypeptide, or glycoprotein is a senescent cell-specific antigen comprising the amino acid sequence set forth in any one of SEQ ID NOS:11-23. 
     
     
         71 . A method for selectively killing a senescent cell in a mammal comprising administering to the mammal a pharmaceutical composition comprising (a) a small molecule that specifically binds to the senescent cell; and (b) a pharmaceutically acceptable carrier. 
     
     
         72 . The method of  claim 71 , wherein the small molecule specifically binds to a senescent cell-specific peptide, polypeptide, or glycoprotein. 
     
     
         73 . The method of  claim 71 , wherein the senescent cell is a senescent lung cell, a senescent breast cell, a senescent colon cell, a senescent prostate cell, a senescent gastric cell, a senescent hepatic cell, a senescent ovarian cell, a senescent esophageal cell, a senescent bronchial epithelial or stromal cell, a senescent skin epithelial or stromal cell, a senescent glial cell, or a senescent vascular endothelial or stromal cell. 
     
     
         74 . A method for depleting senescent cells from donor cells, a donor tissue, or a donor organ, said method comprising (a) contacting the donor cells, the donor tissue, or the donor organ with a peptide, a polypeptide, a small molecule, or an antibody or antibody fragment thereof, wherein the peptide, the polypeptide, the small molecule, or the antibody or antibody fragment thereof specifically binds to a senescent cell; and (b) administering the donor cells, the donor tissue, or the donor organ to the mammal prior to, concurrent with, or subsequent to the step of contacting. 
     
     
         75 . The method of  claim 74 , wherein the peptide, the polypeptide, small molecule, or the antibody or antibody fragment thereof is administered to the mammal prior to, concurrent with, or subsequent to administration of the donor cells, the donor tissue, or the donor organ to the mammal. 
     
     
         76 . The method of  claim 74 , wherein the peptide, the polypeptide, or the antibody or the antibody fragment thereof contacting the donor cells, tissue, or organ is coupled to a therapeutic agent. 
     
     
         77 . The method of  claim 76  wherein the therapeutic agent is a cytotoxic agent. 
     
     
         78 . A method of imaging a senescent cell-containing region in a mammal in vivo, said method comprising the steps of:
 (a) administering to the mammal a peptide, a polypeptide, a small molecule, or an antibody or antibody fragment thereof coupled to a detectable label, wherein the detectably labeled peptide, polypeptide, small molecule, or antibody or antibody fragment thereof specifically binds to a senescent cell;   (b) allowing the detectably labeled peptide, polypeptide, small molecule, or antibody or antibody fragment thereof to bind to the senescent cell and allowing unbound detectably labeled peptide, polypeptide, small molecule, or antibody or antibody fragment thereof to be cleared from the body of said mammal; and   (c) obtaining an image of the senescent cell-containing region.   
     
     
         79 . A method of predicting cancer risk in a mammal, said method comprising:
 (a) administering to the mammal a peptide, a polypeptide, a small molecule, or an antibody or antibody fragment thereof coupled to a detectable label, wherein the detectably labeled peptide, polypeptide, small molecule, or antibody or antibody fragment thereof specifically binds to a senescent cell; and   (b) detecting binding of the detectably labeled peptide, polypeptide, small molecule, or antibody or antibody fragment thereof to the senescent cell of the mammal, thereby predicting an elevated cancer risk in the mammal.   
     
     
         80 . A method for identifying a peptide or polypeptide that binds specifically to a senescent cell, said method comprising:
 (a) culturing cells to produce senescent cells;   (b) exposing the senescent cells to a phage peptide library;   (c) recovering the (i) senescent cells and (ii) the phage from the phage peptide library that is bound to the senescent cells;   (d) eluting and amplifying the bound phage to produce amplified phage;   (e) repeating steps (a)-(d) one or more times; and   (f) recovering a peptide or polypeptide expressed by the amplified phage, which peptide or polypeptide is capable of identifying senescent cells.   
     
     
         81 . A method for identifying a small molecule that specifically binds to a senescent cell, said method comprising (a) contacting a plurality of small molecule candidates with a senescent cell; and (b) determining specific binding of the small molecule candidates to the senescent cell, thereby identifying a small molecule that specifically binds to the senescent cell. 
     
     
         82 . A pharmaceutical composition comprising:
 (a) a peptide, a polypeptide, or an antibody or antibody fragment thereof coupled to a therapeutic agent, wherein the peptide, the polypeptide, or the antibody or antibody fragment thereof specifically binds to a senescent cell-specific peptide, polypeptide, or glycoprotein, and wherein the therapeutic agent is capable of killing a senescent cell; and   (b) a pharmaceutically acceptable carrier.   
     
     
         83 . The pharmaceutical composition of  claim 82 , wherein the composition comprises the peptide coupled to the therapeutic agent, and wherein the peptide comprises (a) the amino acid sequence selected from any one of SEQ ID NOS:1-3; or (b) an amino acid sequence at least 90% identical to the amino acid sequence selected from any one of SEQ ID NOS:1-3. 
     
     
         84 . The pharmaceutical composition of  claim 82 , wherein the composition comprises the antibody or antibody fragment thereof coupled to the therapeutic agent, and wherein the antibody or antibody fragment thereof comprises (a) the amino acid sequence selected from any one of SEQ ID NOS:1-3 in a complementarity determining region; or (b) an amino acid sequence at least 90% identical to the amino acid sequence selected from any one of SEQ ID NOS:1-3 in a complementarity determining region. 
     
     
         85 . The pharmaceutical composition of  claim 82 , wherein the senescent cell-specific peptide, polypeptide, or glycoprotein is a senescent cell-specific antigen comprising the amino acid sequence set forth in any one of SEQ ID NOS:11-23. 
     
     
         86 . A pharmaceutical composition comprising:
 (a) a small molecule that specifically binds to a senescent cell-specific peptide, polypeptide, or glycoprotein; and (b) a pharmaceutically acceptable carrier.