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 nucleic acid molecule encoding a chimeric antigen receptor (CAR) comprising:
a human signal sequence;
an antigen binding domain directed against B-cell maturation antigen (BCMA);
a human hinge domain;
a human transmembrane domain; and
at least one human intracellular T cell signaling domain.
2. The nucleic acid molecule 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 nucleic acid molecule 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 said variable light chain and said variable heavy chain are joined by a linker.
4. The nucleic acid molecule of claim 1 , wherein the antigen binding domain comprises a single chain variable fragment (scFv).
5. The nucleic acid molecule of claim 1 , wherein the transmembrane domain and at least one intracellular T-cell signaling domain are obtained from the same protein.
6. The nucleic acid molecule of claim 1 , wherein the transmembrane domain and at least one intracellular T-cell signaling domain are obtained from different proteins.
7. The nucleic acid molecule of claim 1 , wherein the CAR comprises at least two intracellular T-cell signaling domains.
8. The nucleic acid molecule of claim 7 , wherein the transmembrane domain and at least one intracellular T-cell signaling domain are obtained from the same protein.
9. The nucleic acid molecule of claim 1 , wherein the transmembrane domain comprises an amino acid sequence obtained from CD8α or CD28.
10. The nucleic acid molecule of claim 1 , wherein the transmembrane domain comprises an amino acid sequence obtained from CD8α.
11. The nucleic acid molecule 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 nucleic acid molecule 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 nucleic acid molecule of claim 1 , wherein the transmembrane domain and the hinge domain are obtained from the same protein.
14. The nucleic acid molecule of claim 1 , wherein the transmembrane domain and the hinge domain are obtained from CD8α.
15. The nucleic acid molecule 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 nucleic acid molecule 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 nucleic acid molecule 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 nucleic acid molecule 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 nucleic acid molecule 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 nucleic acid molecule of claim 19 , 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. A vector comprising the nucleic acid molecule of claim 1 .
22. The vector of claim 21 , wherein the vector is a viral vector.
23. The vector of claim 21 , wherein the vector is a retroviral vector.
24. The vector of claim 21 , wherein the vector is a lentiviral vector.
25. The vector of claim 21 , 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.
26. The vector of claim 25 , wherein the vector is a viral vector.
27. The vector of claim 25 , wherein the vector is a retroviral vector.
28. The vector of claim 25 , wherein the vector is a lentiviral vector.