Methods for treatment and diagnosis of cancer by targeting glycoprotein a repetitions predominant (garp) and for providing effective immunotherapy alone or in combination

Abstract

Isolated or recombinant monoclonal antibodies that bind to GARP are provided. In some cases, antibodies of the embodiments can be used for the detection, diagnosis and/or therapeutic treatment of human diseases, such as cancer. Further provided herein are methods and compositions for treating cancer in an individual comprising administering to the individual an effective amount of an anti-platelet agent and a T cell therapy.

What is claimed is: 
     
         1 . An isolated monoclonal antibody, wherein the antibody specifically binds to GARP and comprises:
 (I):
 (a) a first V H  CDR is identical to SEQ ID NO: 1; 
 (b) a second V H  CDR is identical to SEQ ID NO: 2; 
 (c) a third V H  CDR is identical to SEQ ID NO: 3; 
 (d) a first V L  CDR is identical to SEQ ID NO: 5; 
 (e) a second V L  CDR is identical to SEQ ID NO: 6; and 
 (f) a third V L  CDR is identical to SEQ ID NO: 7; or 
   (II):
 (a) a first V H  CDR is identical to SEQ ID NO: 9; 
 (b) a second V H  CDR is identical to SEQ ID NO: 10; 
 (c) a third V H  CDR is identical to SEQ ID NO: 11; 
 (d) a first V L  CDR is identical to SEQ ID NO: 13; 
 (e) a second V L  CDR is identical to SEQ ID NO: 14; and 
 (f) a third V L  CDR is identical to SEQ ID NO: 15. 
   
     
     
         2 . The antibody of  claim 1 , wherein the antibody comprises:
 (a) a first V H  CDR is identical to SEQ ID NO: 1;   (b) a second V H  CDR is identical to SEQ ID NO: 2;   (c) a third V H  CDR is identical to SEQ ID NO: 3;   (d) a first V L  CDR is identical to SEQ ID NO: 5;   (e) a second V L  CDR is identical to SEQ ID NO: 6; and   (f) a third V L  CDR is identical to SEQ ID NO: 7.   
     
     
         3 . The antibody of  claim 2 , wherein the antibody comprises a V H  domain at least about 80% identical to the V H  domain of 4d3 (SEQ ID NO: 4) and a V L  domain at least about 80% identical to the V L  domain of 4d3 (SEQ ID NO: 8). 
     
     
         4 . The antibody of  claim 3 , wherein the antibody comprises a V H  domain identical to the V H  domain of 4d3 (SEQ ID NO: 4) and a V L  domain identical to the V L  domain of 4d3 (SEQ ID NO: 8). 
     
     
         5 . The isolated antibody of  claim 1 , wherein the antibody comprises:
 (a) a first V H  CDR is identical to SEQ ID NO: 9;   (b) a second V H  CDR is identical to SEQ ID NO: 10;   (c) a third V H  CDR is identical to SEQ ID NO: 11;   (d) a first V L  CDR is identical to SEQ ID NO: 13;   (e) a second V L  CDR is identical to SEQ ID NO: 14; and   (f) a third V L  CDR is identical to SEQ ID NO: 15.   
     
     
         6 . The antibody of  claim 5 , wherein the antibody comprises a V H  domain at least about 80% identical to the V H  domain of 5c5 (SEQ ID NO: 12) and a V L  domain at least about 80% identical to the V L  domain of 5c5 (SEQ ID NO: 16). 
     
     
         7 . The antibody of  claim 3 . 1 , wherein the antibody comprises a V H  domain identical to the V H  domain of 5c5 (SEQ ID NO: 12) and a V L  domain identical to the V L  domain 5c5 (SEQ ID NO: 16). 
     
     
         8 . The antibody of any one of  claims 1 - 7 , wherein the antibody is recombinant. 
     
     
         9 . The antibody of  claim 1 , wherein the antibody is an IgG, IgM, IgA or an antigen binding fragment thereof. 
     
     
         10 . The antibody of any one of  claims 1 - 7 , wherein the antibody is a Fab′, a F(ab′)2, a F(ab′)3, a monovalent scFv, a bivalent scFv, or a single domain antibody. 
     
     
         11 . The antibody of any one of  claims 1 - 9 , wherein the antibody is a human, humanized antibody or de-immunized antibody. 
     
     
         12 . The antibody of any one of  claims 1 - 11 , wherein the antibody is fused or conjugated to a platelet binding agent. 
     
     
         13 . The antibody of any one of  claims 1 - 11 , wherein the antibody is conjugated to an imaging agent, a chemotherapeutic agent, a toxin or a radionuclide. 
     
     
         14 . A composition comprising an antibody of any one of  claims 1 - 13  in a pharmaceutically acceptable carrier. 
     
     
         15 . An isolated polynucleotide molecule comprising a nucleic acid sequence encoding an antibody of any one of  claims 1 - 11 . 
     
     
         16 . A recombinant polypeptide comprising an antibody V H  domain comprising CDRs 1-3 of the V H  domain of 4d3 (SEQ ID NOs: 1, 2, and 3); CDRs 1-3 of the V H  domain of 5c5 (SEQ ID NOs: 9, 10, and 11). 
     
     
         17 . A recombinant polypeptide comprising an antibody V L  domain comprising CDRs 1-3 of the V L  domain of 4d3 (SEQ ID NOs: 5, 6, and 7); or 5c5 (SEQ ID NOs: 13, 14, and 15). 
     
     
         18 . An isolated polynucleotide molecule comprising a nucleic acid sequence encoding a polypeptide of  claim 16  or  17 . 
     
     
         19 . A host cell comprising one or more polynucleotide molecule(s) encoding an antibody of any one of  claims 1 - 11  or a recombinant polypeptide of  claim 16  or  17 . 
     
     
         20 . The host cell of  claim 19 , wherein the host cell is a mammalian cell, a yeast cell, a bacterial cell, a ciliate cell or an insect cell. 
     
     
         21 . A method of manufacturing an antibody comprising:
 (a) expressing one or more polynucleotide molecule(s) encoding a V L  and V H  chain of an antibody of any one of  claims 1 - 11  in a cell; and   (b) purifying the antibody from the cell.   
     
     
         22 . A method for treating a subject having a cancer comprising administering an effective amount of an antibody of any one of  claims 1 - 10  to the subject. 
     
     
         23 . The method of  claim 22 , wherein the cancer is a breast cancer, lung cancer, head & neck cancer, prostate cancer, esophageal cancer, tracheal cancer, skin cancer brain cancer, liver cancer, bladder cancer, stomach cancer, pancreatic cancer, ovarian cancer, uterine cancer, cervical cancer, testicular cancer, colon cancer, rectal cancer, skin cancer or a hematological cancer. 
     
     
         24 . The method of  claim 22 , wherein the antibody is in a pharmaceutically acceptable composition. 
     
     
         25 . The method of  claim 22 , wherein the antibody is administered systemically. 
     
     
         26 . The method of  claim 22 , wherein the antibody is administered intravenously, intradermally, intratumorally, intramuscularly, intraperitoneally, subcutaneously, or locally. 
     
     
         27 . The method of  claim 22 , further comprising administering at least a second anticancer therapy to the subject. 
     
     
         28 . The method of  claim 27 , wherein the second anticancer therapy is a surgical therapy, chemotherapy, radiation therapy, cryotherapy, hormonal therapy, immunotherapy or cytokine therapy. 
     
     
         29 . The method of  claim 27 , wherein the second anticancer therapy comprises an adoptive T-cell therapy. 
     
     
         30 . A method for detecting a cancer in a subject comprising testing for the presence of elevated GARP relative to a control in a sample from the subject. 
     
     
         31 . The method of  claim 30 , further comprising testing for the presence of an elevated level of soluble GARP in the sample. 
     
     
         32 . The method of  claim 30 , further comprising testing for the presence of an elevated level GARP expressing cells in the sample. 
     
     
         33 . The method of  claim 30 , wherein the testing comprises contacting the sample with an antibody that binds to GARP. 
     
     
         34 . The method of  claim 30 , wherein the antibody that binds to GARP is an antibody according to any one of  claims 1 - 11 . 
     
     
         35 . The method of  claim 30 , further defined as an in vitro method. 
     
     
         36 . A method of treating cancer in a subject comprising administering an immunotherapy in combination with an anti-platelet agent. 
     
     
         37 . The method of  claim 36 , wherein the immunotherapy comprises a T-cell, NK-cell or NKT-cell therapy. 
     
     
         38 . The method of  claim 37 , wherein the T cell therapy comprises administration of tumor infiltrating lymphocytes (TILs), CD8 +  T cells and/or CD4 +  T cells. 
     
     
         39 . The method of  claim 37 , wherein the T cell therapy comprises administration of tumor-specific T cells. 
     
     
         40 . The method of  claim 39 , wherein the tumor-specific T cells are autologous. 
     
     
         41 . The method of  claim 39 , wherein the tumor-specific T cells are engineered to express a T cell receptor (TCR) or chimeric antigen receptor (CAR) receptor having antigenic specificity for a tumor antigen. 
     
     
         42 . The method of  claim 41 , wherein the tumor-antigen is selected from the group consisting of tEGFR, Her2, CD19, CD20, CD22, mesothelin, CEA, CD23, CD24, CD30, CD33, CD38, CD44, EGFR, EGP-2, EGP-4, EPHa2, ErbB2, FBP, MAGE-A1, MUC1, NY-ESO-1, and MART-1. 
     
     
         43 . The method of  claim 36 , wherein the immunotherapy comprises an immune checkpoint inhibitor. 
     
     
         44 . The method of  claim 43 , wherein the at least one immune checkpoint inhibitor is an anti-CTLA-4 antibody. 
     
     
         45 . The method of  claim 36 . 1 , the immune checkpoint inhibitor is a human programmed cell death 1 (PD-1) binding antagonist, a PDL1 binding antagonist ore a PDL2 binding antagonist. 
     
     
         46 . The method of  claim 45 , wherein the PD-1 binding antagonist is a monoclonal antibody or antigen binding fragment thereof. 
     
     
         47 . The method of  claim 45 , wherein the PD-1 binding antagonist is nivolumab, pembrolizumab, CT-011, BMS 936559, MPDL328OA or AMP-224. 
     
     
         48 . The method of  claim 36 , further comprising lymphodepletion of the subject prior to administration of the T cell therapy. 
     
     
         49 . The method of  claim 48 , wherein lymphodepletion comprises administration of cyclophosphamide and/or fludarabine. 
     
     
         50 . The method of  claim 41 , wherein the CAR comprises co-stimulatory molecule endodomains selected from the group consisting of CD28, CD27, 4-IBB, OX40 ICOS, and a combination thereof. 
     
     
         51 . The method of  claim 36 , wherein the anti-platelet agent is an anti-GARP antibody or fragment thereof. 
     
     
         52 . The method of  claim 36 , wherein the anti-GARP antibody is fused or conjugated to a platelet binding agent. 
     
     
         53 . The method of  claim 51 , wherein the anti-GARP antibody is a monoclonal antibody. 
     
     
         54 . The method of  claim 51 , wherein the anti-GARP antibody is a human, humanized antibody or de-immunized antibody. 
     
     
         55 . The method of  claim 51 , wherein the anti-GARP antibody comprises the CDR sequences of the 4d3 or 5c5 antibody. 
     
     
         56 . The method of  claim 36 , wherein the anti-platelet agent is not an anti-GARP antibody. 
     
     
         57 . The method of  claim 36 , wherein the anti-platelet agent is selected from the group consisting of a cyclooxygenase inhibitor, adenosine diphosphate (ADP) inhibitor, phosphodiesterase inhibitor, protease-activated receptor-1 (PAR-1) antagonist, glycoprotein IIB/IIIA inhibitor, adenosine reuptake inhibitor, and thromboxane inhibitor. 
     
     
         58 . The method of  claim 57 , wherein the ADP inhibitor is clopidogrel, prasugrel, or ticlopidine. 
     
     
         59 . The method of  claim 36 , wherein the method further comprises administering at least one additional therapeutic agent. 
     
     
         60 . The method of  claim 59 , wherein the at least one additional therapeutic agent is chemotherapy, immunotherapy, surgery, radiotherapy, or biotherapy. 
     
     
         61 . The method of  claim 59 , wherein the at least one additional therapeutic agent is a TGFβ inhibitor. 
     
     
         62 . The method of  claim 61 , wherein the TGFβ inhibitor is LY2157299, trabedersen, fresolimumab, LY2382770, lucanix, or PF-03446962. 
     
     
         63 . The method of  claim 36 , wherein the immunotherapy is administered before the anti-platelet agent, simultaneous with the anti-platelet agent, or after the anti-platelet agent. 
     
     
         64 . The method of  claim 36 , wherein the immunotherapy and anti-platelet agent are administered simultaneously. 
     
     
         65 . The method of  claim 36 , wherein the immunotherapy and anti-platelet agent are administered intravenously, intraperitoneally, intratracheally, intratumorally, intramuscularly, endoscopically, intralesionally, percutaneously, subcutaneously, regionally, or by direct injection or perfusion. 
     
     
         66 . The method of  claim 36 , wherein the cancer is bladder cancer, breast cancer, clear cell kidney cancer, head/neck squamous cell carcinoma, lung squamous cell carcinoma, melanoma, non-small-cell lung cancer (NSCLC), ovarian cancer, pancreatic cancer, prostate cancer, renal cell cancer, small-cell lung cancer (SCLC), triple negative breast cancer, acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CIVIL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma, Hodgkin's lymphoma (HL), mantle cell lymphoma (MCL), multiple myeloma (MM), myeloid cell leukemia-1 protein (Mcl-1), myelodysplastic syndrome (MDS), non-Hodgkin's lymphoma (NHL), or small lymphocytic lymphoma (SLL). 
     
     
         67 . The method of  claim 36 , wherein the cancer is breast cancer. 
     
     
         68 . The method of  claim 36 , wherein the cancer is melanoma. 
     
     
         69 . The method of  claim 36 , wherein the cancer is a GARP positive cancer. 
     
     
         70 . The method of  claim 36 , wherein the number of regulatory T cells (Tregs) is decreased in the subject relative to prior to administration of the therapy. 
     
     
         71 . The method of  claim 70 , wherein the Tregs are Foxp3 +  Tregs. 
     
     
         72 . The method of  claim 36 , wherein said subject is a human subject.