Compositions and methods for targeting CD13 and TIM-3 with CAR T cells to treat acute myeloid leukemia
The present invention includes compositions and methods for treating AML utilizing bispecific CARs. In certain aspects, the invention includes a bispecific split CAR which binds CD13 and TIM-3 on AML cells.
1. A bispecific chimeric antigen receptor (CAR) comprising a first antigen binding domain capable of binding CD13, a first intracellular domain, a second antigen binding domain capable of binding TIM-3, a transmembrane domain, and a second intracellular domain, wherein the first antigen binding domain comprises the amino acid sequence set forth in SEQ ID NO: 1.
2. The bispecific CAR of claim 1 , wherein the first and/or second antigen binding domain is selected from the group consisting of an antibody, a nanobody, a Fab, and an scFv.
3. The bispecific CAR of claim 1 , wherein the second antigen binding domain comprises the amino acid sequence set forth in SEQ ID NO: 6.
4. The bispecific CAR of claim 1 , wherein the second antigen binding domain comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 20, 22, 24, 26, 28, 30, and 32.
5. The bispecific CAR of claim 1 , wherein the second antigen binding domain is encoded by a nucleotide sequence selected from the group consisting of SEQ ID NO: 19, 21, 23, 25, 27, 29, and 31.
6. The bispecific CAR of claim 1 , wherein the transmembrane domain comprises CD28.
7. The bispecific CAR of claim 1 , wherein the first intracellular domain is selected from the group consisting of 4-1BB, CD28, and CD3 zeta.
8. The bispecific CAR of claim 1 , wherein the second intracellular domain is selected from the group consisting of 4-1BB, CD28, and CD3 zeta.
9. The bispecific CAR of claim 1 , wherein the bispecific CAR further comprises a hinge domain selected from the group consisting of a CD8 hinge, an IgG3s hinge, and an IgG4m hinge.
10. The bispecific CAR of claim 1 , wherein the bispecific CAR is encoded by a nucleotide sequence selected from the group consisting of SEQ ID NOs: 33-39.
11. A modified T cell or precursor thereof, comprising the bispecific CAR of claim 1 .
12. The cell of claim 11 , wherein the T cell is autologous.
13. A nucleic acid encoding the bispecific CAR of claim 1 .
14. A method for treating cancer in a subject in need thereof, the method comprising administering to the subject a modified T cell or precursor thereof comprising the bispecific CAR of claim 1 .
15. The method of claim 14 , wherein the cancer is acute myeloid leukemia (AML).