Chimeric antigen receptors targeting B-cell maturation antigen
The invention provides an isolated and purified nucleic acid sequence encoding a chimeric antigen receptor (CAR) directed against B-cell Maturation Antigen (BCMA). The invention also provides host cells, such as T-cells or natural killer (NK) cells, expressing the CAR and methods for destroying multiple myeloma cells.
1. A therapeutically effective chimeric antigen receptor (CAR) comprising:
a human signal sequence;
an antigen binding domain directed against B-cell maturation antigen (BCMA);
a human transmembrane domain;
a hinge sequence positioned between the antigen binding domain and the transmembrane domain; and
at least one human intracellular T cell signaling domain.
2. The CAR of claim 1 , wherein
the antigen binding domain comprises an anti-BCMA antibody or antigen binding fragment thereof;
the transmembrane domain and the hinge domain are obtained from a first protein; and
wherein the CAR comprises at least two human intracellular T cell signaling domains, wherein one of the at least two human intracellular T cell signaling domains is a T cell signaling domain comprising an amino acid sequence obtained from a cytoplasmic portion of CD3ξ.
3. The CAR of claim 1 , wherein the antigen binding domain comprises at least a portion of a variable light chain and at least a portion of a variable heavy chain, and wherein the variable light chain and the variable heavy chain are joined by a linker.
4. The CAR of claim 1 , wherein the antigen binding domain comprises an anti-BCMA single chain variable fragment (scFv).
5. The CAR of claim 1 , wherein the transmembrane domain and at least one intracellular T cell signaling domain are obtained from the same protein.
6. The CAR of claim 1 , wherein the transmembrane domain and at least one intracellular T cell signaling domain are obtained from different proteins.
7. The CAR of claim 1 , wherein the CAR comprises at least two intracellular T cell signaling domains.
8. The CAR of claim 7 , wherein the transmembrane domain and at least one intracellular T cell signaling domain are obtained from the same protein.
9. The CAR of claim 1 , wherein the transmembrane domain comprises an amino acid sequence obtained from CD8α or CD28.
10. The CAR of claim 1 , wherein the transmembrane domain comprises an amino acid sequence obtained from CD8α.
11. The CAR of claim 1 , wherein at least one intracellular T cell signaling domain comprises an amino acid sequence obtained from a cytoplasmic portion of CD27, CD28, CD3ξ, OX40, or 4-1BB.
12. The CAR of claim 1 , wherein the signal sequence is a granulocyte-macrophage colony-stimulating factor (GM-CSF) receptor signal sequence or a CD8α signal sequence.
13. The CAR of claim 1 , wherein the transmembrane domain and the hinge domain are obtained from the same protein.
14. The CAR of claim 1 , wherein the transmembrane domain and the hinge domain are obtained from a CD8a protein.
15. The CAR of claim 1 , wherein the at least one intracellular T cell signaling domain comprises an amino acid sequence obtained from a cytoplasmic portion of CD3ξ.
16. The CAR of claim 15 , wherein the CAR further comprises a T cell signaling domain comprising an amino acid sequence obtained from a cytoplasmic portion of CD28.
17. The CAR of claim 15 , wherein the CAR further comprises a T cell signaling domain comprising an amino acid sequence obtained from a cytoplasmic portion of 4-1BB.
18. The CAR of claim 15 , wherein the CAR further comprises a T cell signaling domain comprising an amino acid sequence obtained from a cytoplasmic portion of OX40.
19. The CAR of claim 15 , wherein the CAR further comprises a T cell signaling domain comprising an amino acid sequence obtained from a cytoplasmic portion of CD27.
20. The CAR of claim 1 , wherein the transmembrane domain and the hinge domain are each obtained from CD8α, and the CAR further comprises a first intracellular T cell signaling domain from 4-1BB, and a second intracellular T cell signaling domain from CD3ξ.
21. The CAR of claim 20 , wherein the antigen binding domain comprises at least a portion of a variable light chain and at least a portion of a variable heavy chain, and wherein the variable light chain and the variable heavy chain are joined by a linker.
22. The CAR of claim 21 , wherein the antigen binding domain comprises an anti-BCMA scFv.
23. The CAR of claim 22 , wherein the signal sequence is a CD8α signal sequence.
24. The CAR of claim 22 , wherein the hinge domain is a CD8α hinge domain.