Peptidic chimeric antigen receptor T cell switches and uses thereof
Disclosed herein are chimeric antigen receptor effector cells (CAR-ECs) and CAR-EC switches. The switchable CAR-ECs are generally T cells. The one or more chimeric antigen receptors may recognize a peptidic antigen on the CAR-EC switch. The CAR-ECs and switches may be used for the treatment of a condition in a subject in need thereof.
1. A kit comprising:
a. a chimeric antigen receptor-effector cell switch comprising:
i. a peptidic antigen comprising a yeast transcription factor GCN4 peptide that binds to and activates an anti-GCN4 chimeric antigen receptor on an effector cell; said GCN4 peptide comprising a portion of SEQ ID NO: 2 that is at least 12 amino acids; and
ii. a targeting moiety that binds a cell surface molecule on a target cell; and
b. a chimeric antigen receptor-effector cell comprising an anti-GNC4 chimeric antigen receptor that binds to the peptidic antigen of the chimeric antigen receptor-effector cell switch.
2. The kit of claim 1 , wherein the binding of the chimeric antigen receptor-effector cell switch to the chimeric antigen receptor on an effector cell induces an immune response that is cytotoxic to the target cell also bound to the chimeric antigen receptor-effector cell switch.
3. The kit of claim 1 , wherein the targeting moiety comprises a targeting peptide.
4. The kit of claim 1 , wherein the targeting moiety comprises a targeting antibody or an antigen binding fragment of an antibody.
5. The kit of claim 4 , wherein the targeting antibody or the antigen binding fragment of the antibody is selected from the group consisting of: an immunoglobulin, an Fc null immunoglobulin, a Fab, and antigen binding fragments thereof.
6. The kit of claim 4 , wherein the targeting moiety is selected from the group consisting of: an anti-EGFR antibody, an anti-Her2 antibody, an anti-EGFRvIII antibody, an anti-CD33 antibody, an anti-CLL-1 antibody, an anti-CEA antibody, an anti-CD19 antibody, an anti-CD22 antibody, an anti-BCMA antibody, and an anti-CS1 antibody, and antigen binding fragments thereof.
7. The kit of claim 4 , wherein the targeting antibody or the antigen binding fragment of the antibody comprises a light chain and a heavy chain pair, wherein the light chain and heavy chain are encoded by nucleic acid sequences comprising nucleic acid sequence pairs selected from the group consisting of: SEQ ID NOs: 8 and 9; SEQ ID NOs: 8 and 10; SEQ ID NOs: 11 and 12; SEQ ID NOs. 13 and 14; SEQ ID NOs: 15 and 16; SEQ ID NOs: 17 and 18; and SEQ ID NOs: 19 and 20.
8. The kit of claim 4 , wherein the targeting antibody or the antigen binding fragment of the antibody comprises a light chain and a heavy chain pair, wherein the light chain and heavy chain comprise amino acid sequence pairs selected from the group consisting of: SEQ ID NOs: 21 and 22; SEQ ID NOs: 23 and 24; SEQ ID NOs. 25 and 26; SEQ ID NOs: 27 and 28; and SEQ ID NOs: 27 and 29.
9. The kit of claim 4 , wherein the targeting antibody or the antigen binding fragment of the antibody comprises a light chain and a heavy chain pair, wherein the light chain and heavy chain comprise amino acid sequence pairs selected from the group consisting of: SEQ ID NOs: 30 and 29; SEQ ID NOs: 36 and 29; SEQ ID NOs: 31 and 28; SEQ ID NOs: 27 and 32; SEQ ID NOs: 27 and 33; SEQ ID NOs: 27 and 34; and SEQ ID NOs: 27 and 35.
10. The kit of claim 4 , wherein the peptidic antigen is fused to a terminus of the targeting antibody or the antigen binding fragment of the antibody.
11. The kit of claim 4 , wherein the peptidic antigen is fused to a region of the targeting antibody or antigen binding fragment of the antibody selected from the group consisting of: an N terminus of a light chain, a C terminus of a light chain, an N terminus of a heavy chain, a C terminus of a Fab heavy chain, and a C terminus of a heavy chain constant region.
12. The kit of claim 1 , further comprising a linker that links the peptidic antigen and the targeting moiety.
13. The kit of claim 12 , wherein the linker comprises about 1 to about 20 amino acids.
14. The kit of claim 12 , wherein the linker comprises a sequence selected from SEQ ID NOs: 38-42.
15. The kit of claim 1 , wherein the targeting moiety is selected from the group consisting of: an anti-Her2 antibody, an anti-BCMA antibody, an anti-CD19 antibody, an anti-CEA antibody, and antigen binding fragments thereof.
16. The kit of claim 1 , wherein the target cell is a cancer cell.
17. The kit of claim 1 , wherein the cell surface molecule is a tumor associated antigen.
18. The kit of claim 1 , wherein the cell surface molecule is selected from the group consisting of: a cluster of differentiation protein, a receptor, an integral membrane protein and a glycoprotein.
19. The kit of claim 4 , wherein the targeting antibody or antigen binding fragment of the antibody comprises a light chain and a heavy chain pair, wherein the light chain and heavy chain are encoded by nucleic acid sequences comprising nucleic acid sequence pairs selected from the group consisting of: SEQ ID NOs: 8 and 9; SEQ ID NOs: 8 and 10.
20. The kit of claim 4 , wherein the targeting antibody or the antigen binding fragment of the antibody comprises a light chain and a heavy chain pair, wherein the light chain and heavy chain comprise amino acid sequence pairs selected from the group consisting of: SEQ ID NOs: 27 and 28; and SEQ ID NOs: 27 and 29.
21. The kit of claim 1 , wherein the chimeric antigen receptor-effector cell switch comprises a light chain and a heavy chain pair, wherein the light chain and heavy chain comprise amino acid sequences of SEQ ID NOs: 36 and 29.
22. The kit of claim 1 , wherein the yeast transcription factor GCN4 peptide comprises a sequence that is at least 85% identical to SEQ ID NO: 3.
23. The kit of claim 1 , wherein the yeast transcription factor GCN4 peptide that binds to and activates the anti-GCN4 chimeric antigen receptor on an effector cell; said GCN4 peptide comprising a portion of SEQ ID NO: 2 that (i) is at least 12 amino acids (ii) is at least 85% identical to SEQ ID NO: 3; and (iii) that does not dimerize.
24. The kit of claim 23 , wherein the targeting moiety comprises a targeting antibody or an antigen binding fragment of an antibody.
25. The kit of claim 24 , wherein the targeting moiety is selected from the group consisting of: an anti-EGFR antibody, an anti-Her2 antibody, an anti-EGFRvIII antibody, an anti-CD33 antibody, an anti-CLL-1 antibody, an anti-CEA antibody, an anti-CD19 antibody, an anti-CD22 antibody, an anti-BCMA antibody, and an anti-CS1 antibody, and antigen binding fragments thereof.