BISPECIFIC OR-GATE CHIMERIC ANTIGEN RECEPTOR RESPONSIVE TO CD19 AND CD20
A CD19-OR-CD20 chimeric antigen receptor (CAR) protein construct is provided. Also provided are nucleic acids encoding the CD19-OR-CD20 CAR; and methods of use, e.g. in the treatment of B cell malignancies. The CD19-OR-CD20 CAR of the invention is a bispecific CAR that can trigger T-cell activation upon detection of either CD19 or CD20 (or both). It is a single molecule that confers two-input recognition capability upon human T cells engineered to stably express this CAR.
21 . A bispecific chimeric antigen receptor (CAR) comprising, from N to C terminus:
a) a signal sequence;
b) a CD20 scFv comprising a variable light domain followed by a variable heavy domain, wherein the CD20 scFv comprises the variable light domain and the variable heavy domain sequences of SEQ ID NO: 3;
c) a peptide linker;
d) a CD19 scFv comprising a variable heavy domain followed by a variable light domain, wherein the CD19 scFv comprises the variable heavy domain and the variable light domain sequences of SEQ ID NO: 4;
e) a spacer;
f) a transmembrane domain;
g) a co-stimulatory domain; and
h) a cytoplasmic domain.
22 . The bispecific CAR of claim 21 , wherein the signal sequence comprises the sequence of SEQ ID NO: 2.
23 . The bispecific CAR of claim 21 , wherein the peptide linker comprises the sequence (G 4 S) 4 .
24 . The bispecific CAR of claim 21 , wherein the spacer comprises the sequence of SEQ ID NO: 5.
25 . The bispecific CAR of claim 21 , wherein the transmembrane domain comprises the sequence of SEQ ID NO: 6.
26 . The bispecific CAR of claim 21 , wherein the co-stimulatory domain comprises the sequence of SEQ ID NO: 8.
27 . The bispecific CAR of claim 21 , wherein the cytoplasmic domain comprises the sequence of SEQ ID NO: 9.
28 . A method of treating a B-cell leukemia or lymphoma in a patient comprising administering to the patient a population of cells expressing the bispecific CAR of claim 21 .
29 . The method of claim 28 , wherein the population of cells is a population of autologous cells.
30 . The method of claim 28 , wherein the population of cells is a population of T cells, NK cells, or NKT cells.
31 . The method of claim 28 , wherein the B-cell leukemia or lymphoma is B-cell non-Hodgkin lymphoma (NHL).
32 . The method of claim 28 , wherein the B-cell leukemia or lymphoma is B-cell chronic lymphocytic leukemia (CLL).
33 . The method of claim 28 , wherein the population of cells abrogates antigen loss by the B-cell leukemia or lymphoma cells.
34 . The method of claim 28 , further comprising delivering to the patient an additional cancer therapy selected from radiation, hormone therapy, chemotherapy, immunotherapy, and a combination thereof.
35 . The method of claim 28 , wherein the patient is a human patient.
36 . A bispecific chimeric antigen receptor (CAR) comprising, from N to C terminus:
a) a signal sequence comprising SEQ ID NO: 2;
b) a CD20 scFv comprising a variable light domain followed by a variable heavy domain, wherein the CD20 scFv comprises the variable light domain and the variable heavy domain sequences of SEQ ID NO: 3;
c) a peptide linker comprising the sequence (G 4 S) 4 ;
d) a CD19 scFv comprising a variable heavy domain followed by a variable light domain, wherein the CD19 scFv comprises the variable heavy domain and the variable light domain sequences of SEQ ID NO: 4;
e) a spacer comprising the sequence of SEQ ID NO: 5;
f) a transmembrane domain comprising the sequence of SEQ ID NO: 6;
g) a co-stimulatory domain comprising the sequence of SEQ ID NO: 8; and
h) a cytoplasmic domain comprising the sequence of SEQ ID NO: 9.
37 . A method of treating a B-cell leukemia or lymphoma in a patient comprising administering to the patient a population of cells expressing the bispecific CAR of claim 36 .
38 . The method of claim 37 , wherein the population of cells is a population of autologous cells.
39 . The method of claim 37 , wherein the population of cells is a population of T cells, NK cells, or NKT cells.
40 . The method of claim 37 , wherein the B-cell leukemia or lymphoma is B-cell non-Hodgkin lymphoma (NHL).
41 . The method of claim 37 , wherein the B-cell leukemia or lymphoma is B-cell chronic lymphocytic leukemia (CLL).
42 . The method of claim 37 , wherein the population of cells abrogates antigen loss by the B-cell leukemia or lymphoma cells.
43 . The method of claim 37 , further comprising delivering to the patient an additional cancer therapy selected from radiation, hormone therapy, chemotherapy, immunotherapy, and a combination thereof.
44 . The method of claim 37 , wherein the patient is a human patient.