Chimeric antigen receptor (CAR) binding to BCMA, and uses thereof
The invention provides a chimeric antigen receptor (CAR) which can specifically bind to a BCMA protein comprising a BCMA binding structural domain, a transmembrane domain, a costimulatory domain, and an intracellular signaling domain. The invention also provides uses of the CAR in treating diseases or conditions linked to the expression of BCMA.
1. A chimeric antigen receptor (CAR), wherein the CAR comprises a B-cell maturation antigen (BCMA)-binding domain, a transmembrane domain, a costimulatory domain and an intracellular signal transduction domain, the BCMA-binding domain comprises an antibody or a fragment thereof capable of specifically binding a BCMA protein, wherein the antibody or the fragment, comprises a heavy chain complementary determining region 1 (HCDR1), a heavy chain complementary determining region 2 (HCDR2) and a heavy chain complementary determining region 3 (HCDR3), wherein the HCDR1 comprises the amino acid sequence as set forth in SEQ ID NO: 9, the HCDR2 comprises the amino acid sequence as set forth in SEQ ID NO: 10, and the HCDR3 comprises the amino acid sequence as set forth in SEQ ID NO: 11, and wherein the antibody or the fragment, comprises a light chain complementary determining region 1 (LCDR1), a light chain complementary determining region 2 (LCDR2) and a light chain complementary determining region 3 (LCDR3), and wherein the LCDR1 comprises the amino acid sequence as set forth in SEQ ID NO: 17, the LCDR2 comprises the amino acid sequence as set forth in SEQ ID NO: 18, and the LCDR3 comprises the amino acid sequence as set forth in SEQ ID NO: 19.
2. The CAR of claim 1 , wherein the antibody comprises a heavy chain variable region, and the heavy chain variable region comprises the amino acid sequence as set forth in SEQ ID NO: 7.
3. The CAR of claim 1 , wherein the antibody comprises a light chain variable region, and the light chain variable region comprises the amino acid sequence as set forth in SEQ ID NO: 15.
4. The CAR of claim 1 , wherein the antibody is a single-chain antibody fragment.
5. The CAR of claim 1 , wherein the antibody comprises the amino acid sequence shown as SEQ ID NO: 43.
6. The CAR of claim 1 , wherein the transmembrane domain comprises a transmembrane domain derived from a protein selected from a group consisting of α, β or ζ chain of the T cell receptor, CD28, CD3e, CD45, CD4, CD5, CD8a, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137 and CD154.
7. The CAR of claim 1 , wherein the transmembrane domain comprises the amino acid sequence as set forth in SEQ ID NO: 27.
8. The CAR of claim 1 , wherein the costimulatory domain comprises a costimulatory domain derived from a protein selected from a group consisting of CD28, 4-1BB, OX-40 and ICOS.
9. The CAR of claim 1 , wherein the costimulatory domain comprises the amino acid sequence as set forth in SEQ ID NO: 29 or SEQ ID NO: 31.
10. The CAR of claim 1 , wherein the intracellular signal transduction domain comprises a signal transduction domain derived from CD3ζ.
11. The CAR of claim 1 , wherein the intracellular signal transduction domain comprises the amino acid sequence as set forth in SEQ ID NO: 33.
12. The CAR of claim 1 , wherein the CAR further comprises a hinge region linking the BCMA-binding domain to the transmembrane domain.
13. The CAR of claim 12 , wherein the hinge region comprises the amino acid sequence as set forth in SEQ ID NO: 25.
14. The CAR of claim 1 , wherein the CAR is further linked to a signal peptide.
15. The CAR of claim 14 , wherein the signal peptide comprises the amino acid sequence as set forth in SEQ ID NO: 3.
16. The CAR of claim 1 , wherein the CAR is further linked to a cleaving peptide.
17. The CAR of claim 16 , wherein the cleaving peptide comprises an amino acid sequence derived from a T2A peptide.
18. The CAR of claim 16 , wherein the cleaving peptide comprises the amino acid sequence as set forth in SEQ ID NO: 35.
19. The CAR of claim 1 , comprising the amino acid sequence as set forth in SEQ ID NO: 49 or SEQ ID NO: 51.
20. An isolated nucleic acid molecule, encoding the CAR of claim 1 .
21. An isolated nucleic acid molecule encoding CAR, comprising the nucleotide sequence as set forth in SEQ ID NO: 50 or SEQ ID NO: 52.
22. A vector, comprising the nucleic acid molecule of claim 20 .
23. The vector of claim 22 , wherein the vector is selected from a plasmid, a retroviral vector and a lentiviral vector.
24. An immune effector cell, comprising the CAR of claim 1 .
25. The cell of claim 24 , wherein the immune effector cell is selected from a T lymphocyte and a natural killer (NK) cell.
26. A method of preparing an immune effector cell, comprising introducing the vector of claim 22 into the immune effector cell.
27. A composition, comprising the immune effector cell of claim 24 .
28. A method for treating a mammalian subject having a plasmocyte malignant disease or a B-cell malignant disease, the method comprises administering to said mammalian subject an effective amount of the immune effector cell of claim 24 .
29. The method of claim 28 , wherein the plasmocyte malignant disease is multiple myeloma.
30. The method of claim 28 , wherein the B-cell malignant disease is Hodgkin's lymphoma or non-Hodgkin's lymphoma.