Chimeric antigen receptor
The present invention provides a chimeric antigen receptor (CAR) which binds human CD22, having an antigen-binding domain which comprises a) a heavy chain variable region (VH) having complementarity determining regions (CDRs) with the following sequences: CDR1—NYWIH (SEQ ID No. 1); CDR2—GINPGNNYATYRRKFQG (SEQ ID No. 2) CDR3—EGYGNYGAWFAY (SEQ ID No. 3); and b) a light chain variable region (VL) having CDRs with the following sequences: CDR1—RSSQSLANSYGNTFLS (SEQ ID No. 4); CDR2—GISNRFS (SEQ ID No. 5) CDR3—LQGTHQPYT (SEQ ID No. 6). The present invention also provides a cell comprising such a CAR and the use of such a cell to treat cancer.
1. A chimeric antigen receptor (CAR) which comprises a CD22-binding single-chain variable fragment (scFv) domain which comprises
a) a heavy chain variable region (VH) having complementarity determining regions (CDRs) with the following sequences:
CDR1
(SEQ ID No. 1)
NYWIH;
CDR2
(SEQ ID No. 2)
GINPGNNYATYRRKFQG
CDR3
(SEQ ID No. 3)
EGYGNYGAWFAY;
and
b) a light chain variable region (VL) having CDRs with the following sequences:
CDR1
(SEQ ID No. 4)
RSSQSLANSYGNTFLS;
CDR2
(SEQ ID No. 5)
GISNRFS
CDR3
(SEQ ID No. 6)
LQGTHQPYT.
2. A CAR according to claim 1 , wherein the CD22-binding domain comprises a VH domain having the sequence shown as SEQ ID No. 7; or a VL domain having the sequence shown as SEQ ID No 8 or a variant thereof having at least 90% sequence identity which retains the capacity to bind CD22.
3. A CAR according to claim 1 , which comprises a spacer domain selected from the following: a human IgG1 Fc domain; an IgG1 hinge; an IgG1 hinge-CD8 stalk; a CD8 stalk; or a coiled-coil spacer domain.
4. A nucleic acid sequence which encodes a CAR according to claim 1 .
5. A nucleic acid construct which comprises a first nucleic acid sequence according to claim 4 and a second nucleic acid sequence encoding another CAR or a suicide gene.
6. A vector which comprises a nucleic acid sequence according to claim 4 .
7. A vector which comprises a nucleic acid construct according to claim 5 .
8. A cell which expresses a CAR according to claim 1 .
9. A method for making a cell according to claim 8 , which comprises the step of introducing into a cell a nucleic acid sequence which encodes a chimeric antigen receptor (CAR) which comprises a CD22-binding single-chain variable fragment (scFv) domain which comprises
a) a heavy chain variable region (VH) having complementarity determining regions (CDRs) with the following sequences:
CDR1 - NYWIH
(SEQ ID No. 1);
CDR2 - GINPGNNYATYRRKFQG
(SEQ ID No. 2)
CDR3 - EGYGNYGAWFAY
(SEQ ID No. 3);
and
b) a light chain variable region (VL) having CDRs with the following sequences:
CDR1 - RSSQSLANSYGNTFLS
(SEQ ID No. 4);
CDR2 - GISNRFS
(SEQ ID No. 5)
CDR3 - LQGTHQPYT
(SEQ ID No. 6).
10. A pharmaceutical composition which comprises a plurality of cells according to claim 8 .
11. A method for treating a B-cell malignancy which comprises the step of administering a pharmaceutical composition according to claim 10 to a subject.
12. A method for making a cell according to claim 8 , which comprises the step of introducing into a cell a nucleic acid construct which encodes a chimeric antigen receptor (CAR) which comprises a CD22-binding single-chain variable fragment (scFv) domain which comprises
a) a heavy chain variable region (VH) having complementarity determining regions (CDRs) with the following sequences:
CDR1 - NYWIH;
(SEQ ID No. 1);
CDR2 - GINPGNNYATYRRKFQG
(SEQ ID No. 2)
CDR3 - EGYGNYGAWFAY;
(SEQ ID No. 3);
and
b) a light chain variable region (VL) having CDRs with the following sequences:
CDR1 - RSSQSLANSYGNTFLS
(SEQ ID No. 4);
CDR2 - GISNRFS
(SEQ ID No. 5)
CDR3 - LQGTHQPYT
(SEQ ID No. 6).