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 method of treating a cancer in a subject comprising:
a. administering to the subject a first chimeric antigen receptor-effector cell (CAR-EC) switch, wherein the first CAR-EC switch comprises:
i. a first peptidic antigen comprising a yeast transcription factor GCN4 peptide that binds to and activates a 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 first targeting moiety that binds a cell surface molecule on a target cell; wherein the first targeting moiety comprises an antibody, or an antigen binding fragment of an antibody;
wherein the first peptidic antigen is grafted or fused to the first targeting moiety, and
b. administering to the subject a first chimeric antigen receptor-effector cell comprising an anti-GCN4 chimeric antigen receptor that binds to the first peptidic antigen of the first chimeric antigen receptor-effector cell switch;
wherein the effector cell is a T cell.
2. The method of claim 1 , wherein the first CAR-EC switch comprises two peptidic antigens.
3. The method of claim 1 , wherein the 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.
4. The method of claim 1 , wherein the first 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.
5. The method of claim 3 , wherein the first targeting moiety is an anti-CD19 antibody or an antigen binding fragments thereof.
6. The method of claim 1 , wherein the first 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.
7. The method of claim 1 , wherein the first 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; SEQ ID NOs: 27 and 29; 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.
8. The method of claim 1 , wherein the first peptidic antigen is fused to a region of the first targeting antibody or antigen binding fragment of the antibody selected from the group consisting of: (i) an N terminus of a light chain; (ii) an N terminus of a heavy chain; (iii) an N terminus of a VL domain of an IgG; (iv) an N terminus of a VH domain of an IgG; (vi) an N terminus of a VL domain of a Fab; and (vii) an N terminus of a VH domain of a Fab.
9. The method of claim 5 , wherein the first peptidic antigen is fused to a region of the first targeting antibody or antigen binding fragment of the antibody selected from the group consisting of: (i) an N terminus of a light chain; (ii) an N terminus of a heavy chain; (iii) an N terminus of a VL domain of an IgG; (iv) an N terminus of a VH domain of an IgG; (v) an N terminus of a VL domain of a Fab; and (vi) an N terminus of a VH domain of a Fab.
10. The method of claim 1 , wherein the first CAR-EC switch comprises a linker that links the first peptidic antigen and the first targeting moiety.
11. The method of claim 10 wherein (i) the linker comprises about 1 to about 20 amino acids; (ii) the linker comprises a sequence selected from SEQ ID NOs: 40-44; or (iii) the linker comprises about 1 to about 20 amino acids and the linker comprises a sequence selected from SEQ ID NOs: 40-44.
12. The method of claim 1 , wherein the cancer is selected from the group consisting of leukemia, lymphoma, breast cancer, pancreatic cancer, lung cancer, glioma, and glioblastoma.
13. The method of claim 1 , wherein the cell surface molecule is selected from the group consisting of: a tumor associated antigen; a cluster of differentiation protein; a receptor; an integral membrane protein and a glycoprotein.
14. A method of treating a cancer in a subject comprising:
a. administering to the subject a first chimeric antigen receptor-effector cell (CAR-EC) switch, wherein the first CAR-EC switch comprises:
i. a first peptidic antigen comprising a yeast transcription factor GCN4 peptide that binds to and activates a chimeric antigen receptor on an effector cell and that (i) comprises SEQ ID NO: 3; or (ii) comprises at least 12 amino acids of SEQ ID NO: 2; and
ii. a first targeting moiety that binds a cell surface molecule on a target cell; wherein the first targeting moiety comprises an antibody, or an antigen binding fragment of an antibody; wherein the first peptidic antigen is grafted or fused to the first targeting moiety, and
b. administering to the subject a first chimeric antigen receptor-effector cell comprising an anti-GCN4 chimeric antigen receptor that binds to the first peptidic antigen of the first chimeric antigen receptor-effector cell switch;
wherein the effector cell is a T cell.
15. The method of claim 14 , wherein the first targeting moiety comprises a targeting antibody or an antigen binding fragment of an antibody 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.
16. The method of claim 1 , further comprising administering one or more second CAR-EC switch to the subject, each second CAR-EC switch comprising:
a. a peptidic antigen that binds a chimeric antigen receptor on an effector cell; and
b. a second targeting moiety that binds a cell surface molecule on a target cell;
wherein the second targeting moiety comprises an antibody, or an antigen binding fragment of an antibody;
wherein the second targeting moiety of each second CAR-EC switch differs from the first targeting moiety comprised on the first CAR-EC switch; and
wherein the peptidic antigen comprised on the second CAR-EC switch is either
(i) the same as the first peptidic antigen comprised on the first CAR-EC switch or
(ii) a second peptidic antigen that differs from the first peptidic antigen comprised on the first CAR-EC switch;
provided that if the second CAR-EC switch comprises a second peptidic antigen, the method further comprises administering a second chimeric antigen receptor-effector cell that is a T cell comprising a chimeric antigen receptor that binds to the second peptidic antigen of the second CAR-EC switch.
17. The method of claim 16 , wherein
(i) the first targeting moiety comprises an anti-CD20 antibody or an antigen binding portion thereof and the second targeting moiety comprises an anti-CD19 antibody or an antigen binding portion thereof; or
(ii) the first targeting moiety comprises an anti-CD19 antibody or an antigen binding portion thereof and the second targeting moiety comprises an anti-CD20 antibody or an antigen binding portion thereof.
18. The method of claim 16 , wherein the second CAR-EC switch is administered to the subject after the first CAR-EC switch.
19. The method of claim 18 , wherein the second CAR-EC switch is administered to the subject after the subject has been diagnosed as having a modulated expression of the cell surface molecule to which the first targeting moiety binds.
20. The method of claim 19 , wherein
(i) the first targeting moiety comprises an anti-CD20 antibody or an antigen binding portion thereof and the second targeting moiety comprises an anti-CD19 antibody or an antigen binding portion thereof; or
(ii) the first targeting moiety comprises an anti-CD19 antibody or an antigen binding portion thereof and the second targeting moiety comprises an anti-CD20 antibody or an antigen binding portion thereof.