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 pharmaceutical composition comprising a plurality of T cells that comprise a nucleic acid sequence that encodes an anti-B-cell maturation antigen (BCMA) chimeric antigen receptor (CAR), the CAR comprising:
a human signal sequence;
an antigen binding domain directed against BCMA;
a human hinge domain;
a human transmembrane domain; and
at least one human intracellular T cell signaling domain;
wherein the T cells express the CAR and
wherein the pharmaceutical composition comprises a therapeutically effective amount of the T cells expressing the anti-BCMA CAR sufficient to treat multiple myeloma in a human patient.
2. The pharmaceutical composition 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 the same 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 CD3ζ.
3. The pharmaceutical composition 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 pharmaceutical composition of claim 1 , wherein the antigen binding domain comprises a single chain variable fragment (scFv).
5. The pharmaceutical composition of claim 1 , wherein the transmembrane domain and at least one intracellular T cell signaling domain are obtained from the same protein.
6. The pharmaceutical composition of claim 1 , wherein the transmembrane domain and at least one intracellular T cell signaling domain are obtained from different proteins.
7. The pharmaceutical composition of claim 1 , wherein the CAR comprises at least two human intracellular T cell signaling domains.
8. The pharmaceutical composition of claim 7 , wherein the transmembrane domain and at least one intracellular T cell signaling domain are obtained from the same protein.
9. The pharmaceutical composition of claim 1 , wherein the transmembrane domain comprises an amino acid sequence obtained from CD8α or CD28.
10. The pharmaceutical composition of claim 1 , wherein the transmembrane domain comprises an amino acid sequence obtained from CD8α.
11. The pharmaceutical composition 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, CD3ζ, OX40, or 4-1BB.
12. The pharmaceutical composition 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 pharmaceutical composition of claim 1 , wherein the transmembrane domain and the hinge domain are obtained from the same protein.
14. The pharmaceutical composition of claim 1 , wherein the transmembrane domain and the hinge domain are obtained from CD8α.
15. The pharmaceutical composition of claim 1 , wherein the CAR comprises an intracellular T cell signaling domain comprising an amino acid sequence obtained from a cytoplasmic portion of CD3ζ.
16. The pharmaceutical composition 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 pharmaceutical composition 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 pharmaceutical composition 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 pharmaceutical composition 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 pharmaceutical composition 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 CD3ζ.
21. The pharmaceutical composition of claim 1 , wherein the therapeutically effective amount comprises at least about 100 million T cells that express the anti-BCMA CAR.
22. The pharmaceutical composition of claim 1 , wherein the therapeutically effective amount comprises about 100 million to about 900 million T cells that express the anti-BCMA CAR.
23. The pharmaceutical composition of claim 1 , wherein the therapeutically effective amount comprises about 120 million to about 650 million T cells that express the anti-BCMA CAR.
24. The pharmaceutical composition of claim 1 , wherein the anti-BCMA CAR T cells proliferate and degranulate when contacted by BCMA-expressing cells.
25. The pharmaceutical composition of claim 1 , wherein the anti-BCMA CAR T cells are autologous or allogenic with respect to the patient.
26. The pharmaceutical composition of claim 25 , wherein the anti-BCMA CAR T cells are autologous and wherein the plurality of T cells is transduced with a retroviral vector comprising the nucleic acid sequence.
27. The pharmaceutical composition of claim 26 , wherein the retroviral vector is a lentiviral vector.