BCMA-targeted chimeric antigen receptor as well as preparation method therefor and application thereof
The present invention provides a BCMA-targeted chimeric antigen receptor (CAR) as well as a preparation method therefor and an application thereof. Specifically, the present invention provides the BCMA-targeted CAR, which comprises a BCMA-targeted scFv, a hinge region, a transmembrane region, and an intracellular signal structure domain. The present invention provides a nucleic acid molecule for coding the CAR and a corresponding expression vector as well as CAR-T cells and application thereof. The CAR of the present invention targets BCMA-positive cells, and can be used for treating BCMA-positive B-cell lymphoma, multiple myeloma and plasma cell leukemia.
1. A chimeric antigen receptor (CAR), comprising an anti-BCMA antigen binding region which comprises: (i) a light chain variable region (V L ) having an amino acid sequence set forth in SEQ ID NO: 1, and (ii) a heavy chain variable region (V H ) having an amino acid sequence set forth in SEQ ID NO: 2;
wherein V L is located at the N-terminus of V H ,
wherein the anti-BCMA antigen-binding region is a single-chain variable fragment (scFv) that specifically binds BCMA,
wherein the CAR further comprises:
(a) a signal peptide having an amino acid sequence set forth in SEQ ID NO: 9,
(b) a hinge region having an amino acid sequence set forth in SEQ ID NO: 12,
(c) a transmembrane domain having an amino acid sequence set forth in SEQ ID NO: 13,
(d) a co-stimulatory region having an amino acid sequence set forth in SEQ ID NO: 14, and
(e) a cytoplasmic signaling domain having an amino acid sequence set forth in SEQ ID NO: 15.
2. The CAR of claim 1 , comprising from N-terminus to C-terminus: the signal peptide, V L , V H , the hinge region, the transmembrane domain, the co-stimulatory region, and the cytoplasmic signaling domain.
3. An engineered T cell expressing a chimeric antigen receptor (CAR), wherein the CAR comprises an anti-BCMA antigen binding region which comprises: (i) a light chain variable region (V L ) having an amino acid sequence set forth in SEQ ID NO: 1, and (ii) a heavy chain variable region (V H ) having an amino acid sequence set forth in SEQ ID NO: 2;
wherein V L is located at the N-terminus of V H ,
wherein the anti-BCMA antigen-binding region is a single-chain variable fragment (scFv) that specifically binds BCMA,
wherein the CAR further comprises:
(a) a signal peptide having an amino acid sequence set forth in SEQ ID NO: 9,
(b) a hinge region having an amino acid sequence set forth in SEQ ID NO: 12,
(c) a transmembrane domain having an amino acid sequence set forth in SEQ ID NO: 13,
(d) a co-stimulatory region having an amino acid sequence set forth in SEQ ID NO: 14, and
(e) a cytoplasmic signaling domain having an amino acid sequence set forth in SEQ ID NO: 15.
4. A pharmaceutical composition comprising an engineered T cell expressing a chimeric antigen receptor (CAR), wherein the CAR comprises an anti-BCMA antigen binding region which comprises: (i) a light chain variable region (V L ) having an amino acid sequence set forth in SEQ ID NO: 1, and (ii) a heavy chain variable region (V H ) having an amino acid sequence set forth in SEQ ID NO: 2;
wherein V L is located at the N-terminus of V H ,
wherein the anti-BCMA antigen-binding region is a single-chain variable fragment (scFv) that specifically binds BCMA,
wherein the CAR further comprises:
(a) a signal peptide having an amino acid sequence set forth in SEQ ID NO: 9,
(b) a hinge region having an amino acid sequence set forth in SEQ ID NO: 12,
(c) a transmembrane domain having an amino acid sequence set forth in SEQ ID NO: 13,
(d) a co-stimulatory region having an amino acid sequence set forth in SEQ ID NO: 14, and
(e) a cytoplasmic signaling domain having an amino acid sequence set forth in SEQ ID NO: 15.