Glypican-3-specific antibody and uses thereof

Abstract

The present invention relates to compositions and methods for diagnosing and treating diseases, disorders or conditions associated with dysregulated expression of GPC3. The invention provides novel antibodies that specifically bind to glypican-3 (GPC3). The invention also relates to a fully human chimeric antigen receptor (CAR) wherein the CAR is able to target GPC3.

What is claimed: 
     
         1 . An isolated and substantially purified polypeptide comprising a single chain human anti-glypican-3 (GPC3) antibody fragment comprising a heavy chain and light chain, wherein the single chain antibody fragment comprises a heavy chain variable region and light chain variable region, wherein the amino acid sequence of the heavy chain variable region and the amino acid sequence of the light chain variable region are selected from the group consisting of: (a) SEQ ID NO: 12 and SEQ ID NO: 17; (b) SEQ ID NO: 13 and SEQ ID NO: 18; (c) SEQ ID NO: 14 and SEQ ID NO: 19; and, (d) SEQ ID NO: 15 and SEQ ID NO: 20. 
     
     
         2 . The isolated polypeptide of  claim 1 , wherein the heavy chain variable region and the light chain variable region are encoded by nucleic acid sequences selected from the group consisting of: (a) SEQ ID NO: 52 and SEQ ID NO: 57; (b) SEQ ID NO: 53 and SEQ ID NO: 58; (c) SEQ ID NO: 54 and SEQ ID NO: 59; and, (d) SEQ ID NO: 55 and SEQ ID NO: 60. 
     
     
         3 . A modified cell comprising a vector comprising a polynucleotide encoding the polypeptide of  claim 1 . 
     
     
         4 . The modified cell of  claim 3 , wherein the vector is a viral vector selected from the group consisting of an adenoviral vector, an adeno-associated virus vector, a retroviral vector, a poxvirus, a herpes simplex virus I, and a lentiviral vector. 
     
     
         5 . A method for diagnosing a condition associated with the expression of GPC3 in a cell in a subject, the method comprising a) contacting the cell with a human anti-GPC3 antibody fragment comprising a heavy chain and light chain, wherein the amino acid sequence of the heavy chain is selected from the group consisting of SEQ ID NOs: 12-16 and the amino acid sequence of the light chain is selected from the group consisting of SEQ ID NOs: 17-21; and b) detecting the presence of GPC3 wherein the presence of GPC3 diagnoses a condition associated with the expression of GPC3 in the subject. 
     
     
         6 . A method of diagnosing, prognosing, or determining risk of liver cancer in a mammal, the method comprising detecting the expression of GPC3 in a sample from the mammal by: a) contacting the sample with a human anti-GPC3 antibody fragment comprising a heavy chain and light chain, wherein the amino acid sequence of the heavy chain is selected from the group consisting of SEQ ID NOs: 12-16 and the amino acid sequence of the light chain is selected from the group consisting of SEQ ID NOs: 17-21; and b) detecting the presence of GPC3 wherein the presence of GPC3 diagnoses, prognoses or indicates the risk of liver cancer in the mammal. 
     
     
         7 . A method of inhibiting growth of a GPC3-expressing tumor cell, the method comprising contacting the tumor cell with a human anti-GPC3 antibody or a fragment thereof comprising a heavy chain and light chain, wherein the amino acid sequence of the heavy chain is selected from the group consisting of SEQ ID NOs: 12-16 and the amino acid sequence of the light chain is selected from the group consisting of SEQ ID NOs: 17-21. 
     
     
         8 . An isolated nucleic acid sequence encoding a chimeric antigen receptor (CAR), wherein the isolated nucleic acid sequence comprises the sequence of a human GPC3 binding domain and the sequence of a CD3 zeta signaling domain. 
     
     
         9 . The isolated nucleic acid sequence of  claim 8 , further comprising the sequence of a co-stimulatory signaling domain. 
     
     
         10 . The isolated nucleic acid sequence of  claim 9 , wherein the co-stimulatory signaling domain is selected from the group consisting of the CD28 signaling domain, the 4-1BB signaling domain, and any combination thereof. 
     
     
         11 . The isolated nucleic acid sequence of  claim 8 , wherein the human GPC3 binding domain is a human antibody or a fragment thereof is selected from the group consisting of an Fab fragment, an F(ab′) 2  fragment, an Fv fragment, and a single chain Fv (scFv). 
     
     
         12 . The isolated nucleic acid sequence of  claim 8 , wherein the antibody or a fragment thereof comprises a heavy chain and light chain, wherein the amino acid sequence of the heavy chain is selected from the group consisting of SEQ ID NOs: 12-16 and the amino acid sequence of the light chain is selected from the group consisting of SEQ ID NOs: 17-21. 
     
     
         13 . The isolated nucleic acid sequence  claim 8 , wherein the antibody or a fragment thereof comprises nucleic acid sequences for a heavy chain and light chain, wherein the nucleic acid sequence of the heavy chain is selected from the group consisting of SEQ ID NOs: 52-56 and the nucleic acid sequence of the light chain is selected from the group consisting of SEQ ID NOs: 57-61. 
     
     
         14 . An isolated chimeric antigen receptor (CAR) comprising a human GPC3 binding domain and a CD3 zeta signaling domain. 
     
     
         15 . The isolated CAR of  claim 14 , further comprising the sequence of a co-stimulatory signaling domain. 
     
     
         16 . The isolated CAR of  claim 15 , wherein the co-stimulatory signaling domain is selected from the group consisting of the CD28 signaling domain, the 4-1BB signaling domain, and any combination thereof. 
     
     
         17 . The isolated CAR of  claim 14 , wherein the human GPC3 binding domain is a human antibody or a fragment thereof is selected from the group consisting of an Fab fragment, an F(ab′) 2  fragment, an Fv fragment, and a single chain Fv (scFv). 
     
     
         18 . The isolated CAR of  claim 15 , wherein the antibody or a fragment thereof comprises a heavy chain and light chain, wherein the amino acid sequence of the heavy chain is selected from the group consisting of SEQ ID NOs: 12-16 and the amino acid sequence of the light chain is selected from the group consisting of SEQ ID NOs: 17-21. 
     
     
         19 . A method of providing an anti-tumor immunity in a mammal, the method comprising administering to the mammal an effective amount of a genetically modified cell comprising an isolated nucleic acid sequence encoding a chimeric antigen receptor (CAR), wherein the isolated nucleic acid sequence comprises the sequence of a human GPC3 binding domain and the nucleic acid sequence of a CD3 zeta signaling domain. 
     
     
         20 . The method of  claim 19 , wherein the cell is an autologous T cell. 
     
     
         21 . The method of  claim 19 , wherein the mammal is a human. 
     
     
         22 . A method of treating a mammal having a disease, disorder or condition associated with dysregulated expression of mesothelin, the method comprising administering to the mammal an effective amount of a genetically modified cell comprising an isolated nucleic acid sequence encoding a chimeric antigen receptor (CAR), wherein the isolated nucleic acid sequence comprises the sequence of a human GPC3 binding domain and the nucleic acid sequence of a CD3 zeta signaling domain. 
     
     
         23 . The method of  claim 22 , wherein the disease, disorder or condition associated with dysregulated expression of GPC3 is selected from the group consisting of liver cancer, pancreatic cancer, ovarian cancer, stomach cancer, lung cancer, endometrial cancer, hepatocellular carcinoma, and any combination thereof.