SIRPalpha-SILENCED NATURAL KILLER (NK) CELLS

An isolated population of modified human Natural Killer (NK) cells, comprising a reduced Signal Regulatory Protein Alpha (SIRPα) function when compared to an NK cell population having an unmodified SIRPα function, wherein said modified human NK cells effectively kill a population of cancer cells that express CD47 in an in vitro NK assay.

The isolated population of modified human NK cells of claim 1, wherein said modified human NK cells lack an HLA-I expressed on a cell surface.

The isolated population of modified human NK cells of claim 1, wherein said modified human NK cells comprise a knockout of both alleles of a B2M gene.

The isolated population of modified human NK cells of claim 2, wherein said modified human NK cells lack an HLA-II expressed on said cell surface.

The isolated population of modified human NK cells of claim 2, wherein said modified human NK cells comprise a knockout of both alleles of a CIITa gene.

The isolated population of modified human NK cells of claim 3, wherein said modified human NK cells lack an HLA-II expressed on said cell surface.

The isolated population of modified human NK cells of claim 3, wherein said modified human NK cells comprise a knockout of both alleles of the CIITa gene, said cells further comprising a CD47 transgene.

The population of modified human NK cell population of claim 1, wherein said cancer cell killing occurs faster than with said NK cell population having an unmodified SIRPα function.

The population of modified human NK cells claim 1, wherein said modified human NK cells are primary NK cells.

The population of modified human NK cells of claim 1, wherein said reduced SIRPα function results from a genetic modification to said population of modified human NK cells.

The population of modified human NK cells of claim 1, wherein said reduced SIRPα function results from an interfering nucleic acid molecule.

The population of modified human NK cells of claim 1, wherein said reduced SIRPα function results from a molecule that binds to said SIRPα on the surface of said modified human NK cells.

The population of modified human NK cells of claim 1, wherein said cancer is selected from the group consisting of acute myeloid leukemia, non-small cell lung cancer, urinary bladder neoplasms, hepatocellular carcinoma, melanoma, Merkel Cell carcinoma, triple negative breast cancer, ovarian cancer, renal cell carcinoma, colorectal cancer, and a sarcoma.

The population of modified human NK cells of claim 1, wherein said NK cells are derived from an induced pluripotent stem cell (IPSC).

The population of modified human NK cells of claim 1, wherein said NK cells are derived from an embryonic stem cell (ESC).

The population of modified human NK cells of claim 1, wherein said NK cells comprise a chimeric antigen receptor (CAR-NK).

An isolated population of modified human Natural Killer (NK) cells, comprising a reduced Signal Regulatory Protein Alpha (SIRPα) function when compared to a NK cell having an unmodified SIRPα function, wherein said modified human NK cells effectively kill a population of hypoimmune cells that express CD47 in an in vitro NK assay.

A method of treating cancer, comprising administering the population of modified human NK cells of claim 1 to a subject.

A method of making the isolated population of modified human NK cells of claim 1, comprising modifying SIRPα+ NK cells to become SIRPα− using a transcription activator-like effector nuclease, clustered regularly interspaced short palindromic repeats)/Cas9(CRISPR-Cas9), or Zinc Finger nuclease technology.

A method of making the population of modified human NK cells of claim 1, comprising downregulating a SIRPα expression in said population of modified human NK cells using small interfering RNA (siRNA), antisense oligonucleotides (ASO), locked nucleic acids (LNA), splice switching oligonucleotides (SSO), or sno-derived RNA (sdRNA).