Prostate-specific membrane antigen cars and methods of use thereof
The present disclosure provides modified immune cells (e.g., modified T cells) comprising a chimeric antigen receptor (CAR) having affinity for a prostate-specific membrane antigen (PSMA) (e.g., human PSMA). The present disclosure provides modified immune cells (e.g., modified T cells) comprising a CAR having affinity for PSMA and a dominant negative receptor and/or a switch receptor. The present disclosure provides modified immune cells (e.g., modified T cells) comprising a CAR having affinity for PSMA and a dominant negative receptor and/or a switch receptor, wherein the modified cell is capable of expressing and secreting a bispecific antibody.
1. A method of treating a cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a composition comprising a modified immune cell comprising:
(a) a chimeric antigen receptor (CAR) having affinity for a prostate specific membrane antigen (PSMA) on a target cell, wherein the CAR comprises the amino acid sequence set forth in SEQ ID NO:105; and
(b) a dominant negative receptor consisting of the amino acid sequence set forth in SEQ ID NO:115.
2. A method of treating a cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a composition comprising a modified immune cell, wherein the modified immune cell comprises:
(a) a chimeric antigen receptor (CAR) having affinity for a prostate specific membrane antigen (PSMA) on a target cell, wherein the CAR comprises an antigen binding domain, a transmembrane domain, and an intracellular domain, and wherein:
(i) the antigen binding domain comprises a heavy chain variable region and a light chain variable region selected from the group consisting of:
(a) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 21 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 16;
(b) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 28 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 33;
(c) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 40 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 45;
(d) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 52 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 57;
(e) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 64 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 69; or
(f) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 191 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 192; and
(ii) the intracellular domain comprises a 4-1BB signaling domain, an ICOS signaling domain, and a CD3 zeta signaling domain; and
(b) a dominant negative TGF-β receptor type II comprising the amino acid sequence set forth in SEQ ID NO:115.
3. The method of claim 2 , wherein the VH comprises the sequence of SEQ ID NO:191 and the VL comprises the sequence of SEQ ID NO:192.
4. A method of treating a cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a composition comprising a modified immune cell, wherein the modified immune cell comprises:
(a) a chimeric antigen receptor (CAR) having affinity for a prostate specific membrane antigen (PSMA) on a target cell, wherein the CAR comprises an antigen binding domain, a transmembrane domain, and an intracellular domain, and wherein the antigen binding domain comprises an antibody or an antigen-binding fragment thereof comprising:
(i) a heavy chain variable region (VH) that comprises the consensus sequence of SEQ ID NO:194, and/or
(ii) a light chain variable region (VL) that comprises the consensus sequence of SEQ ID NO: 195; and
(b) a dominant negative receptor and/or switch receptor.
5. The method of claim 4 , wherein:
(a) the VH comprises the sequence of SEQ ID NO: 198 and the VL comprises the sequence of SEQ ID NO: 199;
(b) the VH comprises the sequence of SEQ ID NO: 200 and the VL comprises the sequence of SEQ ID NO: 201;
(c) the VH comprises the sequence of SEQ ID NO: 202 and the VL comprises the sequence of SEQ ID NO: 199; and
(d) the VH comprises the sequence of SEQ ID NO: 202 and the VL comprises the sequence of SEQ ID NO: 201.
6. The method of claim 4 , wherein the antigen-binding fragment is selected from the group consisting of a Fab, a single-chain variable fragment (scFv), and a single-domain antibody.
7. The method of claim 4 , wherein the transmembrane domain comprises a transmembrane region derived from CD8.
8. The method of claim 7 , wherein the transmembrane region comprises the amino acid sequence of SEQ ID NO:88.
9. The method of claim 4 , wherein the transmembrane domain further comprises a hinge region derived from CD8.
10. The method of claim 9 , wherein the hinge region comprises the amino acid sequence of SEQ ID NO:86.
11. The method of claim 4 , wherein the intracellular domain comprises two signaling domains selected from the group consisting of a 4-1BB signaling domain, a CD3 zeta signaling domain, and an ICOS signaling domain.
12. The method of claim 4 , wherein the intracellular domain comprises:
(a) two signaling domains selected from the group consisting of a 4-1BB signaling domain comprising the amino acid sequence of SEQ ID NO:92,
(b) a CD3 zeta signaling domain comprising the amino acid sequence of SEQ ID NO:97 or SEQ ID NO:100, and
(c) an ICOS signaling domain comprising the amino acid sequence of SEQ ID NO:203.
13. The method of claim 4 , wherein the dominant negative receptor is a dominant negative TGF-β receptor type II.
14. The method of claim 13 , wherein the dominant negative TGF-β receptor type II comprises the amino acid sequence set forth in SEQ ID NO:115.
15. The method of claim 4 , wherein the modified immune cell is a T cell.
16. The method of claim 15 , wherein the T cell is an autologous cell or an allogeneic cell.
17. The method of claim 4 , wherein the modified cell is derived from a human cell.
18. The method of claim 4 , further comprising administering to the subject a lymphodepleting chemotherapy, wherein the lymphodepleting chemotherapy comprises administering to the subject a therapeutically effective amount of cyclophosphamide at about 200 mg/m 2 /day to about 2000 mg/m 2 /day, and/or fludarabine at about 20 mg/m 2 /day to about 900 mg/m 2 /day.
19. The method of claim 4 , wherein the cancer is a prostate cancer.
20. The method of claim 19 , wherein the prostate cancer is a castrate-resistant prostate cancer.
21. The method of claim 20 , wherein the castrate-resistant prostate cancer is selected from an advanced castrate-resistant prostate cancer, or a metastatic castrate-resistant prostate cancer.