BCMA chimeric antigen receptors and uses thereof

1. An isolated nucleic acid molecule encoding a chimeric antigen receptor (CAR), wherein the CAR comprises an anti-BCMA binding domain, a transmembrane domain, and an intracellular signaling domain comprising a primary signaling domain and a costimulatory signaling domain, wherein the anti-BCMA binding domain comprises a heavy chain variable region (VH) comprising a heavy chain complementarity determining region 1 (HC CDR1) of SEQ ID NO: 86, a heavy chain complementarity determining region 2 (HC CDR2) of SEQ ID NO: 87, and a heavy chain complementarity determining region 3 (HC CDR3) of SEQ ID NO: 88, and a light chain variable region (VL) comprising a light chain complementarity determining region 1 (LC CDR1) of SEQ ID NO: 95, a light chain complementarity determining region 2 (LC CDR2) of SEQ ID NO: 96, and a light chain complementarity determining region 3 (LC CDR3) of SEQ ID NO: 97, wherein:

(a) the primary signaling domain:

(i) comprises a functional signaling domain of CD3 zeta or FcR gamma;

(ii) comprises the amino acid sequence of SEQ ID NO: 9 or 10, or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto; or

(iii) is encoded by a nucleic acid molecule comprising the nucleic acid sequence of SEQ ID NO: 20, SEQ ID NO: 21, or SEQ ID NO: 256, or a nucleic acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto; and

(b) the costimulatory signaling domain:

(i) comprises a functional signaling domain of 4-1BB (CD137), OX40, CD2, CD27, CD28, ICAM-1, LFA-1, or ICOS (CD278);

(ii) comprises the amino acid sequence of SEQ ID NO: 7, or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto; or

(iii) is encoded by a nucleic acid molecule comprising the nucleic acid sequence of SEQ ID NO: 18 or SEQ ID NO: 255, or a nucleic acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto.

2. The isolated nucleic acid molecule of claim 1 , wherein the VH comprises the amino acid sequence of SEQ ID NO: 93, or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto.

3. The isolated nucleic acid molecule of claim 1 , comprising the nucleic acid sequence of SEQ ID NO: 260 or 94, or a nucleic acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto.

4. The isolated nucleic acid molecule of claim 1 , wherein the VL comprises the amino acid sequence of SEQ ID NO: 102, or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto.

5. The isolated nucleic acid molecule of claim 1 , comprising the nucleic acid sequence of SEQ ID NO: 261 or 103, or a nucleic acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto.

6. The isolated nucleic acid molecule of claim 1 , wherein the VH and VL comprise the amino acid sequences of

SEQ ID NOs: 93 and 102, respectively.

7. The isolated nucleic acid molecule of claim 1 , wherein the anti-BCMA binding domain comprises a single-chain fragment variable (scFv) comprising the amino acid sequence of SEQ ID NO: 105, or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto.

8. The isolated nucleic acid molecule of claim 1 comprising the nucleic acid sequence of SEQ ID NO: 253 or 106, or a nucleic acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto.

9. The isolated nucleic acid molecule of claim 1 , wherein the CAR comprises the amino acid sequence of SEQ ID NO: 107, or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto.

10. The isolated nucleic acid molecule of claim 1 , wherein the nucleic acid molecule comprises the nucleic acid sequence of SEQ ID NO: 259, 258, or 108, or a nucleic acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto.

11. The isolated nucleic acid molecule of claim 1 , wherein the VH and VL are connected by a linker comprising the amino acid sequence of SEQ ID NO: 63 or 104.

12. The isolated nucleic acid molecule of claim 1 , wherein:

(i) the transmembrane domain comprises a transmembrane domain of a protein chosen from the alpha, beta, or zeta chain of T-cell receptor, CD28, CD3 epsilon, CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137, or CD154;

(ii) the transmembrane domain comprises the amino acid sequence of SEQ ID NO: 6, or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto; or

(iii) the nucleic acid molecule comprises the nucleic acid sequence of SEQ ID NO: 17, or a nucleic acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto.

13. The isolated nucleic acid molecule of claim 1 , wherein the anti-BCMA binding domain is connected to the transmembrane domain by a hinge region, wherein:

(i) the hinge region comprises the amino acid sequence of SEQ ID NO: 2, 3, or 4, or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto; or

(ii) the nucleic acid molecule comprises the nucleic acid sequence of SEQ ID NO: 13, 14, or 15, or a nucleic acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto.

14. The isolated nucleic acid molecule of claim 1 , wherein the intracellular signaling domain comprises a costimulatory signaling domain comprising a functional signaling domain of 4-1BB and a primary signaling domain comprising a functional signaling domain of CD3 zeta.

15. The isolated nucleic acid molecule of claim 1 , wherein the CAR further comprises a leader sequence comprising the amino acid sequence of SEQ ID NO: 1.

16. The isolated nucleic acid molecule of claim 1 , wherein the CAR comprises one or more of the following properties:

(i) the CAR, when expressed in a cell, activates NFAT signaling in the cell in the presence of BCMA-expressing cells;

(ii) the CAR, when expressed in a cell, induces cytotoxicity of BCMA-expressing cells; and

(iii) the CAR, when expressed in a cell, induces expression of a cytokine in the cell in the presence of BCMA-expressing cells.

17. A vector comprising the nucleic acid molecule of claim 1 .

18. A cell comprising the nucleic acid molecule of claim 1 .

19. A method of making a cell comprising transducing a cell with the vector of claim 17 .

20. A method of making an RNA-engineered cell comprising introducing an in vitro transcribed RNA or synthetic RNA into a cell, wherein the RNA comprises the nucleic acid molecule of claim 1 .

21. A nucleic acid molecule encoding a chimeric antigen receptor (CAR) that binds to BCMA, wherein the CAR comprises, from N-terminus to C-terminus:

(i) leader sequence of SEQ ID NO: 1;

(ii) a HC CDR1 of SEQ ID NO: 86;

(iii) a HC CDR2 of SEQ ID NO: 87;

(iv) a HC CDR3 of SEQ ID NO: 88;

(v) a LC CDR1 of SEQ ID NO: 95;

(vi) a LC CDR2 of SEQ ID NO: 96;

(vii) a LC CDR3 of SEQ ID NO: 97;

(viii) a CD8 transmembrane domain and hinge of SEQ ID NO: 202;

(ix) a functional signaling domain of 4-1BB of SEQ ID NO: 7; and

(x) a functional signaling domain of CD3 zeta of SEQ ID NO: 10.

22. The nucleic acid molecule of claim 21 , wherein the CAR comprises a VH region of SEQ ID NO: 93 and a VL region of SEQ ID NO: 102 and/or a scFv of SEQ ID NO: 105.

23. The nucleic acid molecule of claim 21 , wherein the CAR an amino acid sequence of SEQ ID NO: 257 or SEQ ID NO: 107.

24. A cell comprising a chimeric antigen receptor (CAR) that binds to BCMA, wherein the CAR comprises, from N-terminus to C-terminus:

(i) leader sequence of SEQ ID NO: 1;

(ii) a HC CDR1 of SEQ ID NO: 86;

(iii) a HC CDR2 of SEQ ID NO: 87;

(iv) a HC CDR3 of SEQ ID NO: 88;

(v) a LC CDR1 of SEQ ID NO: 95;

(vi) a LC CDR2 of SEQ ID NO: 96;

(vii) a LC CDR3 of SEQ ID NO: 97;

(viii) a CD8 transmembrane domain and hinge of SEQ ID NO: 202;

(ix) a functional signaling domain of 4-1BB of SEQ ID NO: 7; and

(x) a functional signaling domain of CD3 zeta of SEQ ID NO: 10.

25. The cell of claim 24 , wherein the cell is a T cell.

26. The cell of claim 24 , wherein the cell is an NK cell.

27. The isolated nucleic acid of claim 14 , wherein:

(i) the costimulatory signaling domain comprises the amino acid sequence of SEQ ID NO: 7 (or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto) and the primary signaling domain comprises the amino acid sequence of SEQ ID NO: 9 or 10 (or an amino acid sequence having at least about 85%, 90%, 95%, or 99% sequence identity thereto); or

(ii) the costimulatory signaling domain comprises the amino acid sequence of SEQ ID NO: 7 and the primary signaling domain comprises the amino acid sequence of SEQ ID NO: 9 or 10.