Bispecific chimeric antigen receptors, encoding polynucleotides and use of receptors thereof to treat cancer
The invention is directed to a bispecific chimeric antigen receptor, comprising: (a) at least two antigen-specific targeting regions; (b) an extracellular spacer domain; (c) a transmembrane domain; (d) at least one co-stimulatory domain; and (e) an intracellular signaling domain, wherein each antigen-specific targeting region comprises an antigen-specific single chain Fv (scFv) fragment, and binds a different antigen, and wherein the bispecific chimeric antigen receptor is co-expressed with a therapeutic control. The invention also provides methods and uses of the bispecific chimeric antigen receptors.
1. A bispecific chimeric antigen receptor (CAR), comprising:
a. two antigen-specific targeting regions;
b. a CD8a hinge extracellular spacer domain;
c. a CD8a transmembrane domain;
d. a 4-1BB co-stimulatory domain; and
e. CD3 zeta intracellular signaling domain wherein each antigen-specific targeting region comprises an antigen-specific single-domain antibody, a variable domain, a VH domain, or a full length heavy chain.
2. The bispecific CAR of claim 1 , wherein each antigen-specific targeting region binds a different antigen.
3. The bispecific CAR of claim 1 , wherein each antigen-specific targeting region comprises an antigen-specific single-domain antibody that is a variable domain of a heavy chain that is alone sufficient to confer antigen specificity.
4. The bispecific CAR of claim 1 , wherein each antigen-specific targeting region comprises an antigen-specific variable domain.
5. The bispecific CAR of claim 1 , wherein each antigen-specific targeting region comprises an antigen-specific VH domain.
6. The bispecific CAR of claim 3 , wherein the two antigen-specific targeting regions bind an antigen specific for cancer.
7. The bispecific CAR of claim 4 , wherein the two antigen-specific targeting regions bind an antigen specific for cancer.
8. The bispecific CAR of claim 5 , wherein the two antigen-specific targeting regions bind an antigen specific for cancer.
9. The bispecific CAR of claim 3 , wherein the two antigen-specific targeting regions bind to an antigen specifically expressed on cells associated with a hematological disease.
10. The bispecific CAR of claim 4 , wherein the two antigen-specific targeting regions bind to an antigen specifically expressed on cells associated with a hematological disease.
11. The bispecific CAR of claim 5 , wherein the two antigen-specific targeting regions bind to an antigen specifically expressed on cells associated with a hematological disease.
12. The bispecific CAR of claim 6 , wherein the two antigen-specific targeting regions bind to an antigen specifically expressed on B-cell lineage tumor cells.
13. The bispecific CAR of claim 7 , wherein the two antigen-specific targeting regions bind to an antigen specifically expressed on B-cell lineage tumor cells.
14. The bispecific CAR of claim 8 , wherein the two antigen-specific targeting regions bind to an antigen specifically expressed on B-cell lineage tumor cells.
15. The bispecific CAR of claim 3 , wherein the two antigen-specific targeting regions bind to an antigen specifically expressed on immune cells.
16. The bispecific CAR of claim 4 , wherein the two antigen-specific targeting regions bind to an antigen specifically expressed on immune cells.
17. The bispecific CAR of claim 5 , wherein the two antigen-specific targeting regions bind to an antigen specifically expressed on immune cells.
18. The bispecific CAR of claim 15 , wherein the two antigen-specific targeting regions bind to an antigen specifically expressed on plasma cells.
19. The bispecific CAR of claim 16 , wherein the two antigen-specific targeting regions bind to an antigen specifically expressed on plasma cells.
20. The bispecific CAR of claim 17 , wherein the two antigen-specific targeting regions bind to an antigen specifically expressed on plasma cells.
21. The bispecific CAR of claim 18 , wherein the two antigen-specific targeting regions bind to an antigen specifically expressed on cancerous plasma cells.
22. The bispecific CAR of claim 19 , wherein the two antigen-specific targeting regions bind to an antigen specifically expressed on cancerous plasma cells.
23. The bispecific CAR of claim 20 , wherein the two antigen-specific targeting regions bind to an antigen specifically expressed on cancerous plasma cells.
24. The bispecific CAR of claim 3 , wherein the two antigen-specific targeting regions are arranged in tandem and are separated by a linker peptide.
25. The bispecific CAR of claim 4 , wherein the two antigen-specific targeting regions are arranged in tandem and are separated by a linker peptide.
26. The bispecific CAR of claim 5 , wherein the two antigen-specific targeting regions are arranged in tandem and are separated by a linker peptide.
27. A polynucleotide encoding the bispecific CAR of claim 1 .
28. A genetically engineered cell derived from a T-lymphocyte (T-cell), wherein the genetically engineered cell expresses the bispecific CAR of claim 1 .
29. A method for treating a disease cancer in a subject in need thereof, comprising:
administering a therapeutically effective amount of a pharmaceutical composition comprising the genetically engineered cell of claim 28 and a pharmaceutically acceptable carrier to the subject so as to treat the cancer,
wherein the two antigen-specific targeting regions target an antigen associated with the cancer.
30. A method of producing T-cells expressing a CAR, comprising:
i. transfecting one or more T-cells with a vector comprising the polynucleotide of claim 27 ; and
ii. stimulating the one or more T-cells, whereby the T-cells proliferate so as to produce T-cells expressing a CAR.