BISPECIFIC CHIMERIC ANTIGEN RECEPTORS TARGETING CD20 AND BCMA

1 . A bispecific chimeric antigen receptor (CAR), comprising:

(i) an anti-CD20 antigen-binding region which comprises a light chain variable region (V L 1) and a heavy chain variable region (V H 1), wherein V L 1 comprises three complementarity determining regions (CDRs), CDR1, CDR2 and CDR3, having amino acid sequences set forth in SEQ ID NO: 130, SEQ ID NO: 131, SEQ ID NO: 132, respectively, and wherein V H 1 comprises three CDRs, CDR1, CDR2 and CDR3, having amino acid sequences set forth in SEQ ID NO: 127, SEQ ID NO: 128, SEQ ID NO: 129, respectively; and

(ii) an anti-BCMA antigen-binding region which comprises a light chain variable region (V L 2) and a heavy chain variable region (V H 2), wherein V L 2 comprises three complementarity determining regions (CDRs), CDR1, CDR2 and CDR3, having amino acid sequences set forth in SEQ ID NO: 134, SEQ ID NO: 136, SEQ ID NO: 138, respectively, and wherein V H2 comprises three CDRs, CDR1, CDR2 and CDR3, having amino acid sequences set forth in SEQ ID NO: 141, SEQ ID NO: 143, SEQ ID NO: 145, respectively.

2 . The bispecific CAR of claim 1 , wherein V L 1 is located at the N-terminus of V H 1.

3 . The bispecific CAR of claim 1 , wherein V L 2 is located at the N-terminus of V H 2.

4 . The bispecific CAR of claim 1 , wherein V H 1 is located at the N-terminus of V L 1.

5 . The bispecific CAR of claim 1 , wherein V H 2 is located at the N-terminus of V L 2.

6 . The bispecific CAR of claim 1 , wherein V L 1 and V H 1 comprise amino acid sequences set forth in SEQ ID NO: 4 and SEQ ID NO: 8, respectively.

7 . The bispecific CAR of claim 1 , wherein V L 2 and V H 2 comprise amino acid sequences set forth in SEQ ID NO: 12 and SEQ ID NO: 16, respectively.

8 . The bispecific CAR of claim 1 , wherein the anti-CD20 antigen-binding region is a single-chain variable fragment (scFv) that specifically binds CD20, and wherein the anti-BCMA antigen-binding region is a scFv that specifically binds BCMA.

9 . The bispecific CAR of claim 1 , wherein the bispecific CAR further comprises one or more of the following:

(a) a signal peptide,

(b) a hinge region,

(c) a transmembrane domain,

(d) a co-stimulatory region, and

(e) a cytoplasmic signaling domain.

10 . The bispecific CAR of claim 9 , wherein the co-stimulatory region comprises a co-stimulatory region of 4-1BB (CD137), CD28, OX40, CD2, CD7, CD27, CD30, CD40, CD70, CD134, PD1, Dap10, CDS, ICAM-1, LFA-1 (CD11a/CD18), ICOS (CD278), NKG2D, GITR, TLR2, or combinations thereof.

11 . The bispecific CAR of claim 9 , wherein the cytoplasmic signaling domain comprises a cytoplasmic signaling domain of CD3ζ.

12 . The bispecific CAR of claim 9 , wherein the hinge region comprises a hinge region of IgG4, CD8, CD28, CD137, or combinations thereof.

13 . The bispecific CAR of claim 9 , wherein the transmembrane domain comprises a transmembrane domain of CD8, CD28, CD3ε, CD45, CD4, CD5, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137, CD154, or combinations thereof.

14 . The bispecific CAR of claim 1 , comprising an amino acid sequence about 80% to about 100% identical to the amino acid sequence set forth in SEQ ID NO:26, SEQ ID NO:40, SEQ ID NO:54, SEQ ID NO:68, SEQ ID NO:84, SEQ ID NO:98, SEQ ID NO:112, or SEQ ID NO:126.

15 . An immune cell expressing the bispecific CAR of claim 1 .

16 . The immune cell of claim 15 , wherein the immune cell is a T cell or a natural killer (NK) cell.

17 . A nucleic acid encoding the bispecific CAR of claim 1 .

18 . A vector comprising the nucleic acid of claim 17 .

19 . A pharmaceutical composition, comprising the bispecific CAR of claim 1 .

20 . A pharmaceutical composition, comprising the immune cell of claim 15 .