COMPOSITIONS AND METHODS FOR IMPROVED NK CELL THERAPIES
The present disclosure relates to the preparation and use of CAR-NK cells which are modified by a nucleic acid targeting compound.
1 . A CAR natural killer (NK) cell-derived effector cell population comprising:
a population of NK cells expressing a chimeric antigen receptor (CAR), the CAR comprising an extracellular domain which specifically binds a predetermined antigen, a transmembrane domain, and a cytoplasmic co-stimulatory signaling domain, wherein the nucleic acid of the NK cells have been modified by reaction with a nucleic acid targeting compound that reacts directly with the nucleic acid, wherein the NK cells are present in the population in a therapeutically effective amount for treatment of a malignancy that expresses the predetermined antigen.
2 . The CAR-NK cell-derived effector cell population of claim 1 , wherein the population of NK cells expressing a chimeric antigen receptor comprises a population of activated NK cells expressing a chimeric antigen receptor.
3 . The CAR-NK cell-derived effector cell population of claim 1 , wherein the reaction of the nucleic acid of the NK cells with the nucleic acid targeting compound results in interstrand cross-links and/or adducts in the nucleic acid.
4 . The CAR-NK cell-derived effector cell population of claim 1 , wherein the nucleic acid of the NK cells have been modified by reaction with the nucleic acid targeting compound so that the NK cells are attenuated for proliferation.
5 . The CAR-NK cell-derived effector cell population of claim 1 , wherein the nucleic acid targeting compound is a nucleic acid alkylator.
6 . The CAR-NK cell-derived effector cell population of claim 5 , wherein the nucleic acid alkylator is a FRALE such as β-alanine, N-(acridin-9-yl), 2-[bis(2-chloroethyl)amino]ethyl ester.
7 . The CAR-NK cell-derived effector cell population of claim 1 , wherein the nucleic acid targeting compound is activated by illumination.
8 . The CAR-NK cell-derived effector cell population of claim 7 , wherein the nucleic acid targeting compound is a psoralen compound activated by UVA illumination.
9 . The CAR-NK cell-derived effector cell population of claim 8 , wherein the NK cells comprise psoralen-induced interstrand crosslinks introduced between the strands of the genomic DNA.
10 . The CAR-NK cell-derived effector cell population of claim 9 , wherein the interstrand crosslinks inhibit replication of the NK cells.
11 . The CAR-NK cell-derived effector cell population of claim 1 , wherein the psoralen is 4′-(4-amino-2-oxa)butyl-4,5′,8-trimethylpsoralen, 4′aminomethyl 4,5′,8trimethylpsoralen (AMT), 5-methoxy psoralen, trioxalen 4, 5′8-trimethylpsoralen, or 8-methoxy psoralen.
12 . The CAR-NK cell-derived effector cell population of any claim 1 , wherein at least a portion of the NK cells produce one or more cytokines.
13 . The CAR-NK cell-derived effector cell population of claim 12 , wherein at least a portion of the NK cells produce one or more cytokines selected from the group consisting of IFN-γ, GM-CSF, TNF and IL-10.
14 . The CAR-NK cell-derived effector cell population of claim 1 , wherein at least a portion of the NK cells express one or more surface markers selected from the group consisting of CD56, CD16, CD27 and NKp46.
15 . The CAR-NK cell-derived effector cell population of claim 1 , wherein greater than 90% of the NK cells in the population are non-proliferating.
16 . The CAR-NK cell-derived effector cell population of claim 1 , wherein greater than 90% of the activated NK cells in the population are non-proliferating.
17 . The CAR-NK cell-derived effector cell population of claim 1 , wherein the predetermined antigen is a cancer antigen.
18 . The CAR-NK cell-derived effector cell population of claim 17 , wherein the predetermined antigen is selected from the antigens listed in Table 1.
19 . The CAR-NK cell-derived effector cell population of claim 17 , wherein the cancer is selected from the group consisting of lung cancer, melanoma, breast cancer, prostate cancer, colon cancer, renal cell carcinoma, ovarian cancer, neuroblastoma, rhabdomyosarcoma, leukemia and lymphoma.
20 . A method of inducing an immune response to at least one predetermined antigen in a subject, comprising administering to the subject a CAR-NK cell-derived effector cell population of claim 1 in an amount sufficient to induce an anti-tumor response to a cancer in the subject, wherein the cancer expresses the predetermined antigen.
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