Chimeric antigen receptor (CAR) comprising a CD19-binding domain
There is provided a chimeric antigen receptor (CAR) comprising a CD19-binding domain which comprises a) a heavy chain variable region (VH) having complementarity determining regions (CDRs) with the following sequences: CDR1—GY-AFSSS (SEQ ID No. 1); CDR2—YPGDED (SEQ ID No. 2) CDR3—SLLYGDYLDY (SEQ ID No. 3); and b) a light chain variable region (VL) having CDRs with the following sequences: CDR1—SASSSVSYMH (SEQ ID No. 4); CDR2—DTSKLAS (SEQ ID No. 5) CDR3—QQWNINPLT (SEQ ID No. 6). There is also provided a cell comprising such a CAR, and the use of such a cell in the treatment of cancer, in particular a B cell malignancy.
1. A method of causing cell-mediated killing of CD19-expressing cells in a subject, the method comprising a step of administering a composition to the subject, wherein the composition comprises a plurality of cells expressing a chimeric antigen receptor (CAR), said CAR comprising a CD19-binding domain which comprises:
a) a heavy chain variable region (VH) having complementarity determining regions (CDRs) with the following sequences:
CDR1-
(SEQ ID NO: 1)
GYAFSSS;
CDR2-
(SEQ ID NO: 2)
YPGDED; and
CDR3-
(SEQ ID NO: 3)
SLLYGDYLDY;
b) a light chain variable region (VL) having CDRs with the following sequences:
CDR1-
(SEQ ID NO: 4)
SASSSVSYMH;
CDR2-
(SEQ ID NO: 5)
DTSKLAS; and
CDR3-
(SEQ ID NO: 6)
QQWNINPLT.
2. The method according to claim 1 , wherein the CD19-binding domain comprises a VH domain having the sequence shown as SEQ ID NO: 7.
3. The method according to claim 2 , wherein the CD19-binding domain comprises a VL domain having the sequence shown as SEQ ID NO: 8.
4. The method according to claim 1 , wherein the CD19-binding domain comprises a VL domain having the sequence shown as SEQ ID NO: 8.
5. The method according to claim 1 , wherein the CD19-binding domain comprises an scFv in the orientation VH-VL.
6. The method according to claim 5 , wherein the CD19-binding domain comprises the sequence shown as SEQ ID NO: 9.
7. The method according to claim 1 , wherein the CAR further comprises a transmembrane domain and an intracellular T-cell signalling domain.
8. The method according to claim 7 , wherein the CAR further comprises a spacer connecting the CD19-binding domain to the transmembrane domain.
9. The method according to claim 8 , wherein the spacer comprises one of the following: a human an IgG1 Fc domain; an IgG1 hinge; or a CD8 stalk.
10. The method according to claim 8 , wherein the intracellular T-cell signalling domain comprises one or more of the following endodomains: a CD28 endodomain; a 41BB endodomain, an OX40 endodomain and a CD3-Zeta endodomain.
11. The method according to claim 10 , wherein the intracellular T-cell signalling domain comprises the 41BB endodomain and the CD3-Zeta endodomain.
12. The method according to claim 10 , wherein the intracellular T-cell signalling domain comprises the OX40 endodomain and the CD3-Zeta endodomain.
13. The method according to claim 10 , wherein the intracellular T-cell signalling domain comprises the CD28 endodomain; the OX40 endodmain; and the CD3-Zeta endodomain.
14. The method according to claim 1 , wherein the CAR comprises the sequence shown as any of SEQ ID NO: 10 to 15.
15. The method according to claim 1 , wherein the CDRs are grafted on to a human or humanised framework.
16. The method according to claim 1 , wherein the composition comprises a plurality of T cells that express the CAR.
17. The method according to claim 1 , wherein the composition comprises a plurality of natural killer (NK) cells that express the CAR.
18. The method according to claim 1 , wherein the composition further comprises a pharmaceutically acceptable carrier, diluent or excipient.
19. The method according to claim 1 , further comprising steps of transducing or transfecting cells from the subject ex vivo with a vector containing a nucleic acid that encodes the CAR, then administering transfected cells back to the subject.
20. The method according to claim 1 , wherein the subject has a cancerous disease associated with CD19 expression.
21. A chimeric antigen receptor (CAR) comprising a CD19-binding domain which comprises:
a) a heavy chain variable region (VH) having complementarity determining regions (CDRs) with the following sequences:
CDR1-
(SEQ ID No. 1)
GYAFSSS;
CDR2-
(SEQ ID No. 2)
YPGDED; and
CDR3-
(SEQ ID No. 3)
SLLYGDYLDY;
b) a light chain variable region (VL) having CDRs with the following sequences:
CDR1-
(SEQ ID No. 4)
SASSSVSYMH;
CDR2-
(SEQ ID No. 5)
DTSKLAS; and
CDR3-
(SEQ ID No. 6)
QQWNINPLT;
and
wherein the CAR further comprises a transmembrane domain and an intracellular T-cell signalling domain, wherein the intracellular T-cell signalling domain comprises a 41BB endodomain and a CD3-Zeta endodomain.