Quadricistronic system comprising a homing receptor and chimeric antigen receptor for stable genetic modification of cellular immunotherapies
Provided herein are modified NK-92® cells comprising one or more nucleic acids encoding i) a homing receptor, ii) Antigen Binding Protein (ABP) or Chimeric Antigen Recpetor (CAR) that specifically binds to a target antigen, iii) an Fc Receptor such as CD16 or CD16-158V, and/or iv) a cytokine, wherein the nucleic acid sequence is operably linked to a promoter. Further provided herein are modified NK-92® cells comprising one or more nucleic acids encoding i) IL-12 and/or TGF-beta trap, ii) an Antigen Binding Protein (ABP) or Chimeric Antigen Recpetor (CAR) that specifically binds to a target antigen, iii) an Fc Receptor such as CD16 or CD16-158V, and/or iv) a cytokine, wherein the nucleic acid sequence is operably linked to a promoter. Also provided are compositions and kits comprising the modified NK-92® cells, as well as methods of treating cancer using the modified cells.
1. A recombinantly modified natural killer (NK)-92 cell stably transfected with a nucleic acid:
wherein the NK-92 cell has American Type Culture Collection (ATCC) Deposit No. CRL-2407;
wherein the nucleic acid encodes a transforming growth factor (TGF)-beta trap, an anti-programmed cell death 1 ligand 1 (PD-L1) chimeric antigen receptor (CAR), an Fc receptor, and a cytokine;
wherein the TGF-beta trap has the amino acid sequence of SEQ ID NO:66, and wherein the anti-PD-L1 CAR has the amino acid sequence of SEQ ID NO:69; and
wherein the recombinant cell expresses the anti-PD-L1 CAR and the Fc receptor on a cell surface of the recombinant cell.
2. The recombinant NK-92 cell of claim 1 wherein the Fc receptor has the amino acid sequence of SEQ ID NO:12.
3. The recombinant NK-92 cell of claim 2 wherein the amino acid sequence of SEQ ID NO:12 has a F158V mutation.
4. The recombinant NK-92 cell of claim 1 wherein the cytokine has an endoplasmic retention sequence.
5. The recombinant NK-92 cell of claim 1 wherein the cytokine is erIL-2 and has the amino acid sequence of SEQ ID NO:15.
6. The recombinant NK-92 cell of claim 2 wherein the cytokine is erIL-2 and has the amino acid sequence of SEQ ID NO:15.
7. The recombinant NK92 cell of claim 3 wherein the cytokine is erIL-2 and has the amino acid sequence of SEQ ID NO:15.
8. The recombinant NK92 cell of claim 1 wherein the cytokine is erIL-15 and has the nucleic acid sequence of SEQ ID NO:67.
9. The recombinant NK92 cell of claim 1 wherein the nucleic acid that encodes the Fc receptor has the nucleic acid sequence of SEQ ID NO:13.
10. The recombinant NK92 cell of claim 9 wherein the nucleic sequence of SEQ ID NO:13 has a F158V mutation.
11. The recombinant NK92 cell of claim 1 wherein the nucleic acid that encodes the cytokine has the nucleic acid sequence of SEQ ID NO:14.
12. The recombinant NK92 cell of claim 9 wherein the nucleic acid that encodes the cytokine has the nucleic acid sequence of SEQ ID NO:14.
13. The recombinant NK92 cell of claim 10 wherein the nucleic acid that encodes the cytokine has the nucleic acid sequence of SEQ ID NO:14.
14. The recombinant NK92 cell of claim 1 wherein the nucleic acid that encodes the cytokine is a nucleic acid sequence that encodes the amino acid sequence of SEQ ID NO:15.
15. The recombinant NK92 cell of claim 1 wherein TGF-beta trap is a secreted TGF-beta trap.
16. A recombinantly modified NK92 cell stably transfected with a recombinant nucleic acid and expressing from the recombinant nucleic acid an anti programmed cell death 1 ligand 1 (PD-L1) chimeric antigen receptor (CAR) having an amino acid sequence of SEQ ID NO:69, and further expressing from the recombinant nucleic acid a transforming growth factor (TGF)-beta trap having the amino acid sequence of SEQ ID NO:66, and optionally at least one of an Fc receptor, and a cytokine, and wherein the NK-92 cell has ATCC Deposit No. CRL-2407.
17. The recombinant NK-92 cell of claim 16 , wherein the recombinant cell expresses the Fc receptor or the cytokine, and wherein the Fc receptor has the amino acid sequence of SEQ ID NO:12, optionally with a F158V mutation, and wherein the cytokine is erIL-2 and has the amino acid sequence of SEQ ID NO:15.
18. A method of treating a cancer in a subject, comprising administering to the subject a therapeutically effective amount of a composition to the subject, the composition comprising a plurality of recombinant isolated NK-92 cells according to claim 1 , and where the cancer has cancer cells expressing PD-L1.