Chimeric antigen receptor-modified NK-92 cells
Provided are NK-92 cells expressing a chimeric antigen receptor (CAR). The CAR can comprise an intracellular domain of FcεRIγ. Also described are methods for treating a patient having or suspected of having a disease that is treatable with NK-92 cells, such as cancer or a viral infection, comprising administering to the patient NK-92-CAR cells.
1. A genetically modified NK cell carrying a non-viral vector encoding a membrane bound recombinant chimeric antigen receptor (CAR),
wherein the CAR comprises in a single polypeptide chain:
an extracellular scFv domain according to SEQ ID NO:51,
a hinge domain,
a transmembrane domain, and
a FcεRIγ signaling domain according to SEQ ID NO:1; and
wherein the NK cell is an NK-92 cell.
2. The genetically modified NK cell of claim 1 , wherein the hinge domain comprises a CD8 hinge domain according to SEQ ID NO:6 or SEQ ID NO:45, and/or wherein the transmembrane domain comprises a CD28 transmembrane domain according to SEQ ID NO:7.
3. The genetically modified NK cell of claim 1 , further carrying a membrane bound recombinant CD16.
4. The genetically modified NK cell of claim 1 , further comprising a recombinant cytokine with an endoplasmic retention sequence.
5. The genetically modified NK cell of claim 1 , wherein the CAR comprises in a single polypeptide chain: an extracellular scFv domain according to SEQ ID NO:51, a hinge domain according to SEQ ID NO:6 or SEQ ID NO:45, a transmembrane domain according to SEQ ID NO:7, and a FcεRIγ signaling domain according to SEQ ID NO:1.
6. A genetically modified NK cell, comprising:
a recombinant nucleic acid encoding a chimeric antigen receptor (CAR), wherein the recombinant nucleic acid is a transfected plasmid;
wherein the NK cell is an NK-92 cell;
wherein the CAR comprises in a single polypeptide chain an extracellular binding domain according to SEQ ID NO:51, a hinge domain, a transmembrane domain, and a FcεRIγ signaling domain according to SEQ ID NO:1.
7. The genetically modified NK cell of claim 6 , wherein the recombinant nucleic acid is an RNA.
8. The genetically modified NK cell of claim 7 , wherein the RNA is a polycistronic RNA that further encodes a CD16 and/or a cytokine with an endoplasmic retention sequence.
9. The genetically modified NK cell of claim 6 , wherein the extracellular binding domain comprises a scFv.
10. The genetically modified NK cell of claim 6 , wherein the extracellular binding domain specifically binds to a tumor-specific antigen, a tumor associated antigen, or a patient- and tumor-specific antigen.
11. The genetically modified NK cell of claim 10 , wherein the tumor-specific antigen is PD-L1.
12. The genetically modified NK cell of claim 6 , wherein the hinge domain and/or the transmembrane domain comprise a CD8 hinge domain and/or a CD28 transmembrane domain.
13. A method of treating cancer in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of the genetically modified NK cells of claim 1 , thereby treating the cancer.
14. The method of claim 13 , further comprising a step of administering at least one additional therapeutic entity selected from the group consisting of a viral cancer vaccine, a bacterial cancer vaccine, a yeast cancer vaccine, N-803, an antibody, a stem cell transplant, and a tumor targeted cytokine.
15. The method of claim 13 , wherein the cancer is selected from leukemia, acute lymphocytic leukemia, acute myelocytic leukemia, chronic leukemias, chronic myelocytic (granulocytic) leukemia, chronic lymphocytic leukemia, polycythemia vera, lymphomas, Hodgkin's disease, non-Hodgkin's disease, multiple myeloma, Waldenstrom's macroglobulinemia, heavy chain disease, solid tumors including, but not limited to, sarcomas and carcinomas such as fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyo sarcoma, colon carcinoma, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinomas, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilm's tumor, cervical cancer, testicular tumor, lung carcinoma, small cell lung carcinoma, bladder carcinoma, epithelial carcinoma, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, menangioma, melanoma, neuroblastoma and retinoblastoma.
16. A method of treating a viral infection in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of the genetically modified NK cells of claim 1 , thereby treating the viral infection.
17. The method of claim 16 , further comprising administering an antiviral drug.