Bispecific or-gate chimeric antigen receptor responsive to cd19 and cd20

Abstract

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.