Compositions and Methods Comprising Prostate Stem Cell Antigen (PSCA) Chimeric Antigen Receptors (CARs)

Abstract

The present disclosure provides modified immune cells or precursors thereof (e.g. T cells) comprising a chimeric antigen receptor (CAR) capable of binding human PSCA. CARs capable of binding human PSCA, and nucleic acids encoding the same are also provided. Provided herein are bispecific CARs capable of binding human PSCA and human PSMA, nucleic acids encoding the same, and modified immune cells comprising the same. Modified immune cells comprising a PSMA CAR and a PSCA CAR are also provided. Compositions and methods of treatment are also provided.

1 - 4 . (canceled) 
     
     
         5 . A nucleic acid comprising a polynucleotide sequence encoding a CAR comprising an antigen binding domain capable of binding prostate stem cell antigen (PSCA), a transmembrane domain, and an intracellular domain; wherein the antigen binding domain comprises:
 (a) a heavy chain variable region that comprises three heavy chain complementarity determining regions (HCDRs), wherein HCDR1 comprises the amino acid sequence DYYIH (SEQ ID NO: 1), HCDR2 comprises the amino acid sequence WIDPENGDTEFVPKF QG (SEQ ID NO: 2), and HCDR3 comprises the amino acid sequence TGGF (SEQ ID NO: 3); and   (b) a light chain variable region that comprises three light chain complementarity determining regions (LCDRs), wherein LCDR1 comprises the amino acid sequence SASSSVRFIHW (SEQ ID NO: 4), LCDR2 comprises the amino acid sequence DTSKLAS (SEQ ID NO: 5), and LCDR3 comprises the amino acid sequence QQWSSSPFT (SEQ ID NO: 6).   
     
     
         6 . The nucleic acid of  claim 5 , wherein the CAR comprises a polynucleotide sequence at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 20, 22, 24, or 26. 
     
     
         7 - 8 . (canceled) 
     
     
         9 . A vector comprising the nucleic acid of  claim 6 . 
     
     
         10 . (canceled) 
     
     
         11 . The vector of  claim 9 , wherein the vector is an expression vector. 
     
     
         12 - 16 . (canceled) 
     
     
         17 . A method of treating a disease in a subject in need thereof, comprising administering to the subject an effective amount of a modified cell comprising a chimeric antigen receptor (CAR),
 wherein the CAR comprises an antigen binding domain capable of binding prostate stem cell antigen (PSCA), a transmembrane domain, and an intracellular domain; and   wherein the antigen binding domain comprises:
 (a) a heavy chain variable region that comprises three heavy chain complementarity determining regions (HCDRs), wherein HCDR1 comprises the amino acid sequence DYYIH (SEQ ID NO: 1), HCDR2 comprises the amino acid sequence WIDPENGDTEFVPKF QG (SEQ ID NO: 2), and HCDR3 comprises the amino acid sequence TGGF (SEQ ID NO: 3); and 
 (b) a light chain variable region that comprises three light chain complementarity determining regions (LCDRs), wherein LCDR1 comprises the amino acid sequence SASSSVRFIHW (SEQ ID NO: 4), LCDR2 comprises the amino acid sequence DTSKLAS (SEQ ID NO: 5), and LCDR3 comprises the amino acid sequence QQWSSSPFT (SEQ ID NO: 6). 
   
     
     
         18 . The method of  claim 17 , wherein the disease is a cancer. 
     
     
         19 . The method of  claim 18 , wherein the disease is prostate cancer. 
     
     
         20 . The method of  claim 18 , wherein the subject is a human. 
     
     
         21 . The method of  claim 17 , wherein the CAR comprises:
 (a) the heavy chain variable region comprising an amino acid sequence at least 90% identical to SEQ ID NO: 7; and/or   (b) the light chain variable region comprising an amino acid sequence at least 90%, identical to SEQ ID NO: 8.   
     
     
         22 . The method of  claim 17 , wherein the antigen binding domain is a single-chain variable fragment (scFv) and comprises an amino acid sequence at least 90% identical to SEQ ID NO: 9. 
     
     
         23 . The method of  claim 17 , wherein the intracellular domain comprises:
 (a) a costimulatory domain of a protein selected from the group consisting of proteins in the TNFR superfamily, CD28, 4-1BB (CD137), OX40 (CD134), PD-1, CD7, LIGHT, CD83L, DAP10, DAP12, CD27, CD2, CD5, ICAM-1, LFA-1, Lck, TNFR-I, TNFR-II, Fas, CD30, CD40, ICOS, NKG2C, and B7-H3 (CD276), or a variant thereof, or an intracellular domain derived from a killer immunoglobulin-like receptor (KIR); and/or   (b) CD3 zeta intracellular domain.   
     
     
         24 . The method of  claim 17 , wherein the CAR comprises the amino acid sequence of SEQ ID NO: 21, 23, 25, or 27. 
     
     
         25 . The method of  claim 17 , wherein the CAR comprises the amino acid sequence of SEQ ID NO: 21. 
     
     
         26 . The method of  claim 17 , wherein the modified immune cell further comprises a CAR capable of binding prostate specific membrane antigen (PSMA), and wherein the PSMA-CAR comprises an antigen-binding domain, a transmembrane domain, and an intracellular domain, and the antigen binding domain of the PSMA-CAR comprises:
 (a) a heavy chain complementarity determining region (HCDR) 1 comprising the amino acid sequence of SEQ ID NO: 28, HCDR 2 comprising the amino acid sequence of SEQ ID NO: 29, and HCDR 3 comprising the amino acid sequence of SEQ ID NO: 30; and a light chain complementarity determining region (LCDR) 1 comprising the amino acid sequence of SEQ ID NO: 31, LCDR 2 comprising the amino acid sequence of SEQ ID NO: 32, and LCDR 3 comprising the amino acid sequence of SEQ ID NO: 33; or   (b) a heavy chain complementarity determining region (HCDR) 1 comprising the amino acid sequence of SEQ ID NO: 68, HCDR 2 comprising the amino acid sequence of SEQ ID NO: 69, and HCDR 3 comprising the amino acid sequence of SEQ ID NO: 70; and a light chain complementarity determining region (LCDR) 1 comprising the amino acid sequence of SEQ ID NO: 71, LCDR 2 comprising the amino acid sequence of SEQ ID NO: 72, and LCDR 3 comprising the amino acid sequence of SEQ ID NO: 73.   
     
     
         27 . The method of  claim 26 , wherein the PSMA CAR comprises an amino acid sequence at least about 90% identical to SEQ ID NO: 67, 87, 89, 91, 93, 95, or 97. 
     
     
         28 . The method of  claim 26 , wherein:
 (a) the PSCA CAR comprises the amino acid sequence of SEQ ID NO: 21 and the PSMA CAR comprises an amino acid sequence at least about about 90% identical to SEQ ID NO: 67; or   (b) the PSCA CAR comprises the amino acid sequence of SEQ ID NO: 21; and the PSMA CAR comprises an amino acid sequence at least about 90%, identical to SEQ ID NO: 87.   
     
     
         29 . The method of  claim 26 , wherein the intracellular domain of the PSCA CAR and the PSMA CAR each comprises:
 (a) a 4-1BB intracellular domain;   (b) a CD28 intracellular domain;   (c) an ICOS intracellular domain; or   (d) an ICOS(YMNM).   
     
     
         30 . The method of  claim 26 , wherein the costimulatory domain of the PSCA CAR is different from the costimulatory domain of the PSMA CAR. 
     
     
         31 . The method of  claim 26 , wherein the modified cell comprises the amino acid sequence of SEQ ID NO: 40, 42, 80, 81, or 82. 
     
     
         32 . A method of treating a disease in a subject in need thereof, comprising administering to the subject an effective amount of a modified cell comprising a vector comprising a nucleic acid sequence encoding a chimeric antigen receptor (CAR), wherein the CAR comprise an antigen binding domain capable of binding prostate stem cell antigen (PSCA), a transmembrane domain, and an intracellular domain, and wherein the vector comprises a nucleic acid sequence at least 85% identical to SEQ ID NO: 43 and a nucleic acid sequence at least 85% identical to SEQ ID NO: 44. 
     
     
         33 . The method of  claim 32 , wherein the vector comprises a nucleic acid sequence at least 85% identical to SEQ ID NO: 20, 22, 24, or 26. 
     
     
         34 . The method of  claim 32 , wherein the vector further comprises:
 (a) a nucleic acid sequence at least 85% identical to SEQ ID NO: 46 and a nucleic acid sequence at least 85% identical to SEQ ID NO: 47; or   (b) a nucleic acid sequence at least about 90% identical to SEQ ID NO: 88, 90, 92, 94, 96, or 98.   
     
     
         35 . The method of  claim 32 , wherein the vector comprises the nucleic acid sequence of SEQ ID NO: 39, 41, or 79. 
     
     
         36 . The method of  claim 31 , wherein:
 (a) the disease is a cancer;   (b) the disease is prostate cancer; or   (c) the subject is a human.