Chimeric antigen receptors targeting BCMA and methods of use thereof
Provided herein are single-domain antibodies targeting BCMA, and chimeric antigen receptors (such as monovalent CAR, and multivalent CAR including bi-epitope CAR) having one or more anti-BCMA single-domain antibodies. Further provided are engineered immune effector cells (such as T cells) having the chimeric antigen receptors. Pharmaceutical compositions, kits and methods of treating cancer are also provided.
1. An isolated nucleic acid comprising a nucleic acid sequence encoding a multivalent chimeric antigen receptor (CAR) comprising a polypeptide comprising:
(a) an extracellular antigen binding domain comprising a first anti-BCMA binding moiety and a second BCMA binding moiety, wherein each of the first and the second anti-BCMA binding moieties is a VHH domain;
(b) a transmembrane domain; and
(c) an intracellular signaling domain,
wherein the first BCMA binding moiety comprises
a CDR1 comprising the amino acid sequence of SEQ ID NO:15; a CDR2 comprising the amino acid sequence of SEQ ID NO:53; and a CDR3 comprising the amino acid sequence of SEQ ID NO:91, and the second BCMA binding moiety comprises
a CDR1 comprising the amino acid sequence of SEQ ID NO:18; a CDR2 comprising the amino acid sequence of SEQ ID NO:56; and a CDR3 comprising the amino acid sequence of SEQ ID NO:94.
2. The isolated nucleic acid of claim 1 , wherein the first BCMA binding moiety comprises the amino acid sequence of SEQ ID NO: 129 and the second BCMA binding moiety comprises the amino acid sequence of SEQ ID NO: 132.
3. A vector comprising the isolated nucleic acid of claim 1 .
4. The isolated nucleic acid of claim 1 , wherein the first BCMA binding moiety is located at the N-terminus of the second BCMA binding moiety.
5. The isolated nucleic acid of claim 1 , wherein the first BCMA binding moiety is located at the C-terminus of the second BCMA binding moiety.
6. The isolated nucleic acid of claim 1 , wherein the first BCMA binding moiety and the second BCMA binding moiety are fused to each other via a peptide linker.
7. The isolated nucleic acid of claim 6 , wherein the peptide linker is no more than about 50 amino acids long.
8. The isolated nucleic acid of claim 1 , wherein the transmembrane domain is derived from a molecule selected from the group consisting of CD8α, CD4, CD28, CD137, CD80, CD86, CD152 and PD1.
9. The isolated nucleic acid of claim 8 , wherein the transmembrane domain is derived from CD8α or CD28.
10. The isolated nucleic acid of claim 1 , wherein the intracellular signaling domain comprises a primary intracellular signaling domain of an immune effector cell.
11. The isolated nucleic acid of claim 10 , wherein the primary intracellular signaling domain is derived from CD3ζ.
12. The isolated nucleic acid of claim 1 , wherein the intracellular signaling domain comprises a co-stimulatory signaling domain.
13. The isolated nucleic acid of claim 12 , wherein the co-stimulatory signaling domain is derived from a co-stimulatory molecule selected from the group consisting of CD27, CD28, CD137, OX40, CD30, CD40, CD3, LFA-1, ICOS, CD2, CD7, LIGHT, NKG2C, B7-H3, ligands of CD83 and combinations thereof.
14. The isolated nucleic acid of claim 13 , wherein the co-stimulatory signaling domain comprises a cytoplasmic domain of CD28 and/or a cytoplasmic domain of CD137.
15. The isolated nucleic acid of claim 1 , wherein the multivalent CAR further comprises a hinge domain located between the C-terminus of the extracellular antigen binding domain and the N-terminus of the transmembrane domain.
16. The isolated nucleic acid of claim 15 , wherein the hinge domain is derived from CD8α.
17. The isolated nucleic acid of claim 16 , wherein the multivalent CAR further comprises a signal peptide located at the N-terminus of the polypeptide.
18. The isolated nucleic acid of claim 17 , wherein the signal peptide is derived from CD8α.
19. The isolated nucleic acid of claim 1 , comprising the nucleic acid sequence of SEQ ID NO: 356.
20. A multivalent CAR encoded by the isolated nucleic acid of claim 1 .
21. The multivalent CAR of claim 20 , comprising the amino acid sequence of SEQ ID NO: 318.
22. An engineered immune effector cell, comprising the multivalent CAR of claim 20 .
23. A pharmaceutical composition, comprising the engineered immune effector cell of claim 22 .
24. A method of treating cancer that expresses BCMA in an individual, comprising administering to the individual an effective amount of the pharmaceutical composition of claim 23 .
25. The method of claim 24 , wherein the cancer is multiple myeloma.
26. The method of claim 25 , wherein the cancer is refractory or relapsed multiple myeloma.