Engineered immune cells targeting BCMA and their uses thereof
The present invention provides an engineered immune cell targeting BCMA and use thereof. In particular, the present invention provides a CAR specifically targeting BCMA, the CAR comprising an antigen-binding domain which is an S-derived scFv, an antibody heavy chain variable region as shown in SEQ ID NO: 9 and an antibody light chain variable region as shown in SEQ ID NO: 10. The present invention also provides a CAR-T cell comprising the CAR, a double CAR- and CAR-T cell comprising the S-derived scFv, and related use thereof. Compared to CAR-T cells constructed using other scFvs, the constructed CAR-T cell of the present invention has a better killing effect and tumor elimination capability.
1. A bispecific chimeric antigen receptor (CAR) polypeptide, comprising:
a first antigen binding domain exhibiting specific binding to CD19, comprising (i) a first heavy chain variable region (V H,CD19 ) and (ii) a first light chain variable region (V L,CD19 ); and
a second antigen binding domain exhibiting specific binding to BCMA, comprising (i) a second heavy chain variable region (V H,BCMA ) that comprises the polypeptide sequence of SEQ ID NO: 9 and (ii) a second light chain variable region (V L,BCMA ) that comprises the polypeptide sequence of SEQ ID NO: 10,
wherein the bispecific CAR polypeptide comprises a structure as shown in formula (I) or (II):
V L,CD19 -the second antigen binding domain-V H,CD19 (I),
V H,CD19 -the second antigen binding domain-V L,CD19 (II),
wherein “-” is each independently a linker peptide or a peptide bond.
2. The bispecific CAR polypeptide of claim 1 , wherein the V H,CD19 comprises the polypeptide sequence selected from the group consisting of SEQ ID NOs: 11 and 21-30, and wherein the V L,CD19 comprises the polypeptide sequence selected from the group consisting of SEQ ID NOs: 12 and 31-36.
3. The bispecific CAR polypeptide of claim 2 , wherein the V H,CD19 comprises the polypeptide sequence of SEQ ID NO: 11, and wherein the V L-CD19 comprises the polypeptide sequence of SEQ ID NO: 12.
4. The bispecific CAR polypeptide of claim 1 , wherein the bispecific CAR polypeptide comprises the structure as shown in formula (I).
5. The bispecific CAR polypeptide of claim 1 , wherein the bispecific CAR polypeptide comprises a structure as shown in formula III:
L-V L,CD19 -V H,BCMA -V L,BCMA -V H,CD19 -H-TM-C 2 -C 1 (III),
wherein:
“-” is each independently a linker peptide or a peptide bond;
L is absent or a signal domain;
H is absent or a hinge domain;
TM is a transmembrane domain;
C 2 is a costimulatory signaling domain; and
C 1 is a cytoplasmic signaling sequence derived from CD3ξ.
6. The bispecific CAR polypeptide of claim 5 , wherein:
L comprises the polypeptide sequence of SEQ ID NO: 16;
V L,CD19 comprises the polypeptide sequence of SEQ ID NO: 12;
V H,BCMA comprises the polypeptide sequence of SEQ ID NO: 9;
V L,BCMA comprises the polypeptide sequence of SEQ ID NO: 10;
V H,CD19 comprises the polypeptide sequence of SEQ ID NO: 11;
H comprises the polypeptide sequence of SEQ ID NO: 38;
TM comprises the polypeptide sequence of SEQ ID NO: 7;
C 2 comprises the polypeptide sequence of SEQ ID NO: 5; and
C 1 comprises the polypeptide sequence of SEQ ID NO: 3.
7. The bispecific CAR polypeptide of claim 6 , wherein:
(i) V L,CD19 and V H,BCMA are coupled via a linker peptide comprising the polypeptide sequence of SEQ ID NO: 40;
(ii) V H,BCMA and V L,BCMA are coupled via a linker peptide comprising the polypeptide sequence of SEQ ID NO: 19; and
(iii) V L,BCMA and V H,CD19 are coupled via a linker peptide comprising the polypeptide sequence of SEQ ID NO: 40.
8. The bispecific CAR polypeptide of claim 1 , wherein the bispecific CAR polypeptide comprises the structure as shown in formula (I), and wherein an engineered T cell expressing the bispecific CAR polypeptide is characterized by exhibiting greater A cytotoxicity against target cells expressing BCMA, as compared to a control engineered T cell expressing a CAR polypeptide that comprises the first antigen binding domain and the second antigen binding domain in tandem.
9. The bispecific CAR polypeptide of claim 1 , wherein the bispecific CAR polypeptide comprises the structure as shown in formula (I), and wherein an engineered T cell expressing the bispecific CAR polypeptide is characterized by exhibiting greater cytotoxicity against target cells expressing CD19, as compared to a control engineered T cell expressing a CAR polypeptide that comprises the first antigen binding domain and the second antigen binding domain in tandem.
10. An engineered immune cell comprising a bispecific chimeric antigen receptor (CAR) polypeptide, wherein the bispecific CAR comprises:
a first antigen binding domain exhibiting specific binding to CD19, comprising (i) a first heavy chain variable region (V H,CD19 ) and (ii) a first light chain variable region (V L,CD19 ); and
a second antigen binding domain exhibiting specific binding to BCMA, comprising (i) a second heavy chain variable region (V H,BCMA ) that comprises the polypeptide sequence of SEQ ID NO: 9 and (ii) a second light chain variable region (V L,BCMA ) that comprises the polypeptide sequence of SEQ ID NO: 10,
wherein the bispecific CAR polypeptide comprises a structure as shown in formula (I) or (II):
V L,CD19 -the second antigen binding domain-V H,CD19 (I),
V H,CD19 -the second antigen binding domain-V L,CD19 (II),
wherein “-” is each independently a linker peptide or a peptide bond.
11. The engineered immune cell of claim 10 , wherein, in the bispecific CAR, the V H,CD19 comprises the polypeptide sequence of SEQ ID NO: 11, and wherein the V L-CD19 comprises the polypeptide sequence of SEQ ID NO: 12.
12. The bispecific CAR polypeptide of claim 10 , wherein the bispecific CAR polypeptide comprises the structure as shown in formula (I).
13. The engineered immune cell of claim 10 , wherein the bispecific CAR polypeptide comprises a structure as shown in formula III:
L-V L,CD19 -V H,BCMA -V L,BCMA -V H,CD19 -H-TM-C 2 -C 1 (III),
wherein:
“-” is each independently a linker peptide or a peptide bond;
L is absent or a signal domain;
H is absent or a hinge domain;
TM is a transmembrane domain;
C 2 is a costimulatory signaling domain; and
C 1 is a cytoplasmic signaling sequence derived from CD3ξ.
14. The engineered immune cell of claim 13 , wherein:
L comprises the polypeptide sequence of SEQ ID NO: 16;
V L,CD19 comprises the polypeptide sequence of SEQ ID NO: 12;
V H,BCMA comprises the polypeptide sequence of SEQ ID NO: 9;
V L,BCMA comprises the polypeptide sequence of SEQ ID NO: 10;
V H,CD19 comprises the polypeptide sequence of SEQ ID NO: 11;
H comprises the polypeptide sequence of SEQ ID NO: 38;
TM comprises the polypeptide sequence of SEQ ID NO: 7;
C 2 comprises the polypeptide sequence of SEQ ID NO: 5; and
C 1 comprises the polypeptide sequence of SEQ ID NO: 3.
15. The engineered immune cell of claim 14 , wherein:
(i) V L,CD19 and V H,BCMA are coupled via a linker peptide comprising the polypeptide sequence of SEQ ID NO: 40;
(ii) V H,BCMA and V L,BCMA are coupled via a linker peptide comprising the polypeptide sequence of SEQ ID NO: 19; and
(iii) V L,BCMA and V H,CD19 are coupled via a linker peptide comprising the polypeptide sequence of SEQ ID NO: 40.
16. The engineered immune cell of claim 10 , wherein the engineered immune cell is an engineered T cell.
17. A method of treating a tumor or cancer of a subject in need thereof, the method comprising administering to the subject an engineered immune cell comprising a bispecific chimeric antigen receptor (CAR) polypeptide, wherein the bispecific CAR comprises:
a first antigen binding domain exhibiting specific binding to CD19, comprising (i) a first heavy chain variable region (V H,CD19 ) and (ii) a first light chain variable region (V L,CD19 ); and
a second antigen binding domain exhibiting specific binding to BCMA, comprising (i) a second heavy chain variable region (V H,BCMA ) that comprises the polypeptide sequence of SEQ ID NO: 9 and (ii) a second light chain variable region (V L,BCMA ) that comprises the polypeptide sequence of SEQ ID NO: 10,
wherein the bispecific CAR polypeptide comprises a structure as shown in formula (I) or (II):
V L,CD19 -the second antigen binding domain-V H,CD19 (I),
V H,CD19 -the second antigen binding domain-V L,CD19 (II),
wherein “-” is each independently a linker peptide or a peptide bond.
18. The method of claim 17 , wherein, in the bispecific CAR, the V H,CD19 comprises the polypeptide sequence of SEQ ID NO: 11, and wherein the V L -CD19 comprises the polypeptide sequence of SEQ ID NO: 12.
19. The bispecific CAR polypeptide of claim 17 , wherein the bispecific CAR polypeptide comprises the structure as shown in formula (I).
20. The method of claim 17 , wherein the bispecific CAR polypeptide comprises a structure as shown in formula III:
L-V L,CD19 -V H,BCMA -V L,BCMA -V H,CD19 -H-TM-C 2 -C 1 (III),
wherein:
“-” is each independently a linker peptide or a peptide bond;
L is absent or a signal domain;
H is absent or a hinge domain;
TM is a transmembrane domain;
C 2 is a costimulatory signaling domain; and
C 1 is a cytoplasmic signaling sequence derived from CD3ξ.
21. The method of claim 20 , wherein:
L comprises the polypeptide sequence of SEQ ID NO: 16;
V L,CD19 comprises the polypeptide sequence of SEQ ID NO: 12;
V H,BCMA comprises the polypeptide sequence of SEQ ID NO: 9;
V L,BCMA comprises the polypeptide sequence of SEQ ID NO: 10;
V H,CD19 comprises the polypeptide sequence of SEQ ID NO: 11;
H comprises the polypeptide sequence of SEQ ID NO: 38;
TM comprises the polypeptide sequence of SEQ ID NO: 7;
C 2 comprises the polypeptide sequence of SEQ ID NO: 5; and
C 1 comprises the polypeptide sequence of SEQ ID NO: 3.
22. The method of claim 21 , wherein:
(i) V L,CD19 and V H,BCMA are coupled via a linker peptide comprising the polypeptide sequence of SEQ ID NO: 40;
(ii) V H,BCMA and V L,BCMA are coupled via a linker peptide comprising the polypeptide sequence of SEQ ID NO: 19; and
(iii) V L,BCMA and V H,CD19 are coupled via a linker peptide comprising the polypeptide sequence of SEQ ID NO: 40.
23. The method of claim 17 , wherein the engineered immune cell is derived from an autologous cell of the subject.
24. The method of claim 17 , wherein the engineered immune cell is allogeneic to the subject.
25. The method of claim 17 , wherein the engineered immune cell is an engineered T cell.
26. The bispecific CAR polypeptide of claim 1 , wherein the bispecific CAR polypeptide comprises the structure as shown in formula (II).
27. The engineered immune cell of claim 10 , wherein the bispecific CAR polypeptide comprises the structure as shown in formula (II).
28. The method of claim 17 , wherein the bispecific CAR polypeptide comprises the structure as shown in formula (II).