BISPECIFIC CHIMERIC ANTIGEN RECEPTORS TARGETING CD20 AND BCMA
The present disclosure provides bispecific chimeric antigen receptors targeting CD20 and BCMA. The CAR may comprise an scFv targeting CD20 and an scFv targeting BCMA, 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 nucleic acid encoding a bispecific chimeric antigen receptor (CAR), the bispecific 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 is located at the N-terminus of V H 1, wherein V L 1 and V H 1 comprise the amino acid sequences set forth in SEQ ID NO: 4 and SEQ ID NO: 8, 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 is located at the N-terminus of V H 2, wherein V L 2 and V H 2 comprise the amino acid sequences set forth in SEQ ID NO: 12 and SEQ ID NO: 16, respectively.
2 . The nucleic acid 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.
3 . The nucleic acid 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.
4 . The nucleic acid of claim 3 , 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.
5 . The nucleic acid of claim 3 , wherein the cytoplasmic signaling domain comprises a cytoplasmic signaling domain of CD3ζ.
6 . The nucleic acid of claim 3 , wherein the hinge region comprises a hinge region of IgG4, CD8, CD28, CD137, or combinations thereof.
7 . The nucleic acid of claim 3 , wherein the hinge region comprises a hinge region of CD8.
8 . The nucleic acid of claim 3 , 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.
9 . The nucleic acid of claim 3 , wherein the transmembrane domain comprises a transmembrane domain of CD8.
10 . The nucleic acid of claim 1 , wherein the anti-CD20 antigen-binding region is located at the N-terminus of the anti-BCMA antigen-binding region.
11 . A vector comprising the nucleic acid of claim 1 .
12 . A pharmaceutical composition, comprising the nucleic acid of claim 1 .
13 . A pharmaceutical composition, comprising the vector of claim 11 .