GPRC5D chimeric antigen receptors and cells expressing the same
The present disclosure provides for chimeric antigen receptors (CARs) that specifically target a G-protein coupled receptor, G-protein coupled receptor family C group 5 member D (GPRC5D), and immunoresponsive cells comprising such CARs, for the treatment of cancer.
1. A chimeric antigen receptor (CAR), comprising an extracellular antigen-binding domain, a transmembrane domain and an intracellular signaling domain, wherein the extracellular antigen-binding domain comprises:
a heavy chain CDR1 having the amino acid sequence of SEQ ID NO: 66, a heavy chain CDR2 having the amino acid sequence of SEQ ID NO: 67, and a heavy chain CDR3 having the amino acid sequence of SEQ ID NO: 68, and further comprises a light chain CDR1 having the amino acid sequence of SEQ ID NO: 69, a light chain CDR2 having the amino acid sequence of SEQ ID NO: 70, and a light chain CDR3 having the amino acid sequence of SEQ ID NO: 71; or
a heavy chain CDR1 having the amino acid sequence of SEQ ID NO: 58, a heavy chain CDR2 having the amino acid sequence of SEQ ID NO: 59, and a heavy chain CDR3 having the amino acid sequence of SEQ ID NO: 60, and further comprises a light chain CDR1 having the amino acid sequence of SEQ ID NO: 61, a light chain CDR2 having the amino acid sequence of SEQ ID NO: 62, and a light chain CDR3 having the amino acid sequence of SEQ ID NO: 63;
wherein the extracellular antigen-binding domain binds the anti-G protein receptor family C group 5 member D (GPRC5D) antigen.
2. The CAR of claim 1 , wherein the extracellular antigen-binding domain comprises:
a light chain variable region comprising an amino acid sequence of SEQ ID NO: 73 and a heavy chain variable region comprising an amino acid sequence of SEQ ID NO: 72; or
a light chain variable region comprising an amino acid sequence of SEQ ID NO: 65 and a heavy chain variable region comprising an amino acid sequence of SEQ ID NO: 64.
3. The CAR of claim 1 , wherein the extracellular antigen-binding domain comprises a single-chain variable fragment (scFv).
4. The CAR of claim 3 , wherein the scFv comprises a linker polypeptide between the light chain variable region and the heavy chain variable region.
5. The CAR of claim 4 , wherein the linker polypeptide comprises an amino acid sequence of SEQ ID NO: 7.
6. The CAR of claim 3 , wherein the scFv comprises an amino acid sequence selected from the group consisting of SEQ ID NOS: 78, 77, 75, and 76.
7. The CAR of claim 1 , wherein the extracellular antigen-binding domain comprises a signal polypeptide.
8. The CAR of claim 7 , wherein the signal polypeptide comprises an amino acid sequence of SEQ ID NO: 11.
9. The CAR of claim 1 , wherein the intracellular signaling domain comprises a polypeptide component selected from the group consisting of a TNF receptor superfamily member 9 (CD137) component, a T-cell surface glycoprotein CD3 zeta chain (CD3z) component, a cluster of differentiation (CD27) component, a cluster of differentiation superfamily member component, and a combination thereof.
10. The CAR of claim 9 , wherein the CD137 component comprises an amino acid sequence of SEQ ID NO: 12.
11. The CAR of claim 9 , wherein the CD3z component comprises an amino acid sequence of SEQ ID NO: 13.
12. The CAR of claim 9 , wherein the intracellular signaling domain comprises an amino acid sequence of SEQ ID NO: 14.
13. The CAR of claim 1 , wherein the transmembrane domain comprises a CD8a transmembrane region (CD8a-TM) polypeptide.
14. The CAR of claim 13 , wherein the CD8a-TM polypeptide comprises an amino acid sequence of SEQ ID NO: 15.
15. The CAR of claim 1 , further comprising a hinge region linking the transmembrane domain to the extracellular antigen-binding domain.
16. The CAR of claim 15 , wherein the hinge region is a CD8a-hinge region.
17. The CAR of claim 16 , wherein the CD8a-hinge region comprises an amino acid sequence of SEQ ID NO: 16.
18. The CAR of claim 1 , wherein the extracellular antigen-binding domain comprises an amino acid sequence selected from the group consisting of SEQ ID NOS: 82, 81, 80, and 79.
19. The CAR of claim 1 , wherein the CAR comprises an amino acid sequence selected from the group consisting of SEQ ID NOS: 86, 85, 83, and 84.
20. An isolated lymphocyte expressing the CAR of claim 1 .
21. The isolated lymphocyte of claim 20 , wherein the lymphocyte is a T lymphocyte.
22. The isolated lymphocyte of claim 21 , wherein the T lymphocyte is a naïve T cell.
23. The isolated lymphocyte of claim 21 , wherein the T lymphocyte is a memory stem T cell.
24. The isolated lymphocyte of claim 21 , wherein the T lymphocyte is a central memory T cell.
25. The isolated lymphocyte of claim 21 , wherein the T lymphocyte is CD4+.
26. The isolated lymphocyte of claim 21 , wherein the T lymphocyte is CD8+.
27. The isolated lymphocyte of claim 21 , wherein the T lymphocyte is CD4+ and CD8+.
28. A pharmaceutical composition, comprising an effective amount of the lymphocyte of claim 20 .
29. A pharmaceutical composition, comprising
an effective amount of the lymphocyte of claim 20 and a pharmaceutically acceptable excipient.
30. A method of treating a subject having a GPRC5D-expressing cancer, the method comprising: administering a therapeutically effective amount of the lymphocyte of claim 20 to a subject in need thereof, whereby the lymphocyte induces killing of GPRC5D-expressing cancer cells in the subject.
31. The method of claim 30 , wherein the cancer is selected from the group consisting of a lung cancer, a gastric cancer, a colon cancer, a hepatocellular carcinoma, a renal cell carcinoma, a bladder urothelial carcinoma, a metastatic melanoma, a breast cancer, an ovarian cancer, a cervical cancer, a head and neck cancer, a pancreatic cancer, a glioma, a glioblastoma, and a non-Hodgkin's lymphoma (NHL), an acute lymphocytic leukemia (ALL), a chronic lymphocytic leukemia (CLL), a chronic myelogenous leukemia (CML), a smoldering multiple myeloma (SMM), a multiple myeloma (MM), an acute myeloid leukemia (AML), and combinations thereof.
32. The method of claim 31 , wherein the cancer is multiple myeloma.
33. A method of targeted killing of a GPRC5D-expressing cancer cell, the method comprising: contacting the GPRC5D-expressing cancer cell with the lymphocyte of claim 20 , whereby the lymphocyte induces killing of the GPRC5D-expressing cancer cell.
34. The method of claim 33 , wherein the cancer cell is selected from the group consisting of a lung cancer, a gastric cancer, a colon cancer, a hepatocellular carcinoma, a renal cell carcinoma, a bladder urothelial carcinoma, a metastatic melanoma, a breast cancer, an ovarian cancer, a cervical cancer, a head and neck cancer, a pancreatic cancer, a glioma, a glioblastoma, and a non-Hodgkin's lymphoma (NHL), an acute lymphocytic leukemia (ALL), a chronic lymphocytic leukemia (CLL), a chronic myelogenous leukemia (CML), a smoldering multiple myeloma (SMM), a multiple myeloma (MM), an acute myeloid leukemia (AML), and combinations thereof.
35. The method of claim 33 , wherein the cancer cell is a multiple myeloma cell.
36. A method of detecting the presence of a GPRC5D-expressing cancer in a subject, comprising: (a) contacting a cell sample obtained from the subject with the CAR of claim 1 , thereby forming a CAR-cell complex, and (b) detecting the complex, wherein detection of the complex is indicative of the presence of a GPRC5D-expressing cancer in the subject.