BCMA-targeting engineered immune cell and use thereof
Provided are a BCMA-targeting engineered immune cell and the use thereof. In particular, a CAR specifically targeting BCMA is provided. In the CAR, an antigen binding domain contained therein is a J-derived scFv, having an antibody heavy-chain variable region shown in SEQ ID NO: 9, and an antibody light-chain variable region shown in SEQ ID NO: 10. Also provided are a CAR-T cell containing the CAR, and a duplex CAR and CAR T cells containing the J-derived scFv, and related uses thereof. Compared with CAR-T cells constructed by using other scFvs, the CAR-T cells constructed by the present invention have higher killing effects and tumor clearance ability.
1 . A bispecific chimeric antigen receptor (CAR), comprising:
a first CAR targeting CD19; and
a second CAR targeting B-Cell maturation antigen (BCMA); wherein the first CAR and the second CAR are linked by a 2A peptide,
wherein the structure of the first CAR is shown in Formula V:
L-scFv1′-H-TM-C-CD3ζ (V)
wherein the structure of the second CAR is shown in Formula (I):
L-scFv-H-TM-C-CD3ζ (I)
wherein,
each “-” is independently a linker peptide or a peptide bond;
L is none or a signal peptide sequence;
H is none or a hinge region;
TM is a transmembrane domain;
C is a co-stimulatory signal molecule;
CD3ζ is a cytoplasmic signal transduction sequence derived from CD3ζ;
scFv1′ is an antigen binding domain targeting CD19BCMA, comprising an antibody heavy chain variable region shown in SEQ ID NO: 11 and an antibody light chain variable region shown in SEQ ID NO: 12;
the structure of the antigen binding domain targeting CD19 is shown in Formula (C):
V L1 -V H1 (C)
wherein V H1 is the antibody heavy chain variable region
V L1 is the antibody light chain variable; and
“-” is a linker peptide or a peptide bond;
scFv is an antigen binding domain targeting BCMA, comprising an antibody heavy chain variable region shown in SEQ ID NO: 9 and an antibody light chain variable region shown in SEQ ID NO: 10;
the structure of the antigen binding domain targeting BCMA is shown in Formula (A):
V H -V L (A)
wherein V H is the antibody heavy chain variable region;
V L is the antibody light chain variable; and
“-” is a linker peptide or a peptide bond.
2 . A method for enhancing viability of immune cells in vivo or killing ability of immune cells to tumor cells with clonal proliferation ability, comprising expressing the bispecific CAR of claim 1 in the immune cells.
3 . The bispecific CAR of claim 1 , wherein the L is a signal peptide of a protein selected from the group consisting of: a CD8, CD28, GM-CSF, CD4, CD137, or a combination thereof.
4 . The bispecific CAR of claim 3 , wherein the L comprises a CD8 signal peptide.
5 . The bispecific CAR of claim 1 , wherein the H comprises a hinge region from CD8, CD28, CD137, or a combination thereof.
6 . The bispecific CAR of claim 5 , wherein at least one H comprises a hinge region from CD8.
7 . The bispecific CAR of claim 1 , wherein the TM comprises a transmembrane region of CD28, CD3 epsilon, CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, 4-1BB (CD137), CD154, or a combination thereof.
8 . The bispecific CAR of claim 7 , wherein each TM independently comprises a CD8 or CD28 transmembrane region.
9 . The bispecific CAR of claim 1 , wherein the C comprises a co-stimulatory signal molecule of OX40, CD2, CD7, CD27, CD28, CD30, CD40, CD70, CD134, 4-1BB (CD137), PD1, Dap10, CDS, ICAM-1, LFA-1 (CD11a/CD18), ICOS (CD278), NKG2D, GITR, TLR2, or a combination thereof.
10 . The bispecific CAR of claim 9 , wherein each C independently comprises a CD28 and/or 4-1BB co-stimulatory signal molecule.
11 . The bispecific CAR of claim 1 , wherein the CD32 comprises the amino acid of SEQ ID NO: 3.
12 . A nucleic acid molecule encoding the bispecific CAR of claim 1 .
13 . A vector comprising the nucleic acid molecule of claim 12 .
14 . An engineered immune cell comprising an exogenous nucleic acid molecule of claim 12 .
15 . An engineered immune cell comprising the vector of claim 13 .
16 . A method for enhancing viability of immune cells in vivo or killing ability of immune cells to tumor cells with clonal proliferation ability, comprising expressing the bispecific CAR of claim 6 in the immune cells.