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.
1. A polypeptide comprising a CD19-OR-CD20 chimeric antigen receptor (CAR), comprising in order: anti CD20 scFv comprising a light chain variable region and heavy chain variable region from SEQ ID NO:3; (G4S)4 linker; anti-CD19 scFv comprising a heavy chain variable region and light chain variable region from SEQ ID NO:4; spacer domain of SEQ ID NO:5; transmembrane domain of SEQ ID NO:6; 4-1 BB cytoplasmic signaling domain of SEQ ID NO:8; CD3 zeta signaling domain of SEQ ID NO:9.
2. The polypeptide of claim 1 , wherein the polypeptide further comprises a sortable tag.
3. A nucleic acid encoding a polypeptide according to claim 1 .
4. A cell comprising the polypeptide of claim 1 .
5. The cell of claim 4 , wherein the cell is a T cell.
6. The polypeptide of claim 2 , wherein the polypeptide further comprises a T2A cleavage site between the CAR and sortable tag.
7. The polypeptide of claim 6 , wherein the T2A cleavage site comprises SEQ ID NO:10.
8. The polypeptide of claim 1 , wherein the CAR consists of, in order: anti CD20 scFv having the light chain variable region and heavy chain variable region from SEQ ID NO:3; (G4S)4linker; anti-CD19 scFv having the heavy chain variable region and light chain variable region from SEQ ID NO:4; the spacer domain of SEQ ID NO:5; the transmembrane domain of SEQ ID NO:6; the 4-1 BB cytoplasmic signaling domain of SEQ ID NO:8; and the CD3 zeta signaling domain of SEQ ID NO:9.
9. The polypeptide of claim 1 , wherein the polypeptide further comprises a signal sequence of SEQ ID NO:2.
10. A cell comprising the nucleic acid of claim 3 .