BISPECIFIC CHIMERIC ANTIGEN RECEPTORS TARGETING BCMA AND CD19
The present disclosure provides bispecific chimeric antigen receptors targeting BCMA and CD19. The CAR may comprise an scFv targeting BCMA and an scFv targeting CD19, a hinge region, a transmembrane domain, a co-stimulatory region, and a cytoplasmic signaling domain. The chimeric antigen receptors can be used to treat autoimmune disorders or cancer.
1 . A bispecific chimeric antigen receptor (CAR), comprising:
(i) an anti-BCMA 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 about 80% to about 100% identical to the amino acid sequences set forth in SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, respectively, and wherein V H 1 comprises three CDRs, CDR1, CDR2 and CDR3, having amino acid sequences about 80% to about 100% identical to the amino acid sequences set forth in SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 13, respectively; and
(ii) an anti-CD19 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 about 80% to about 100% identical to the amino acid sequences set forth in SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, respectively, and wherein V H 2 comprises three CDRs, CDR1, CDR2 and CDR3, having amino acid sequences about 80% to about 100% identical to the amino acid sequences set forth in SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, 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 I.
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 have amino acid sequences about 80% to about 100% identical to amino acid sequences set forth in SEQ ID NO: 24 and SEQ ID NO: 28, respectively.
7 . The bispecific CAR of claim 1 , wherein V L 2 and V H 2 have amino acid sequences about 80% to about 100% identical to amino acid sequences set forth in SEQ ID NO: 32 and SEQ ID NO: 36, respectively.
8 . The bispecific CAR of claim 1 , wherein the anti-BCMA antigen-binding region is a single-chain variable fragment (scFv) that specifically binds BCMA, and wherein the anti-CD19 antigen-binding region is a scFv that specifically binds CD19.
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 9 , comprising an amino acid sequence about 80% to about 100% identical to the amino acid sequence set forth in SEQ ID NO:46, SEQ ID NO:60, SEQ ID NO:74, SEQ ID NO:88, SEQ ID NO:104, SEQ ID NO:121, SEQ ID NO:138, or SEQ ID NO:155.
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 method of treating an autoimmune disorder, the method comprising administering the immune cell of claim 15 to a subject in need thereof.
19 . The method of claim 18 , wherein the autoimmune disorder is systemic lupus erythematosus (SLE), systemic sclerosis, inflammatory myopathy, systemic scleroderma, multiple sclerosis, myasthenia gravis, a myositis autoantibody-driven disease, or neuromyelitis optica.
20 . The method of claim 19 , wherein the inflammatory myopathy is polymyositis, dermatomyositis, or inclusion-body myositis.
21 . The method of claim 19 , wherein the subject has lupus nephritis.
22 . The method of claim 18 , wherein the immune cell is allogeneic or autologous.