Engineered natural killer (NK) cells and compositions and methods thereof
The present invention provides engineered Natural Killer (NK) cells and methods of producing engineered NK cells. The engineered NK cells and compositions containing the engineered NK cells are useful for treating diseases such as cancer.
1. An engineered NK cell that is derived from a primary cell obtained from a human subject or is derived from a clonal cell line, in which a gene encoding FcRγ chain is genetically disrupted to knockout FcRγ in the engineered cell, wherein the genetic disruption of the gene encoding FcRγ increases antibody dependent cellular cytotoxicity (ADCC) via CD16 expressed on the surface of the engineered cell compared to a NK cell that is not genetically engineered with the genetic disruption.
2. The engineered NK cell of claim 1 , wherein the expression of FcRγ chain is reduced by greater than or greater than about 50% as compared to the expression of the protein in the NK cell that is not genetically engineered with the genetic disruption.
3. The engineered NK cell of claim 1 , wherein the expression of FcRγ chain expressed in the cell is undetectable in an immunoblot assay.
4. The engineered NK cell of claim 1 , that is derived from a primary cell obtained from a human subject.
5. The engineered NK cell of claim 1 , wherein the CD16 is a heterologous CD16 that comprises a CD16-activating mutation.
6. A method of producing an engineered NK cell, that is derived from a primary cell obtained from a human subject or is derived from a clonal cell line, in which a gene encoding FcRγ chain is genetically disrupted to knockout FcRγ in the engineered cell, wherein the genetic disruption of the gene encoding FcRγ increases antibody dependent cellular toxicity (ADCC) via CD16 expressed on the surface of the engineered cell compared to a NK cell that is not genetically engineered with the genetic disruption.
7. A composition comprising an effective amount of the engineered NK cell of claim 1 .
8. A kit comprising the engineered cell of claim 1 and an additional agent for treatment of a disease, wherein the additional agent is an antibody or an Fc-fusion protein.
9. The engineered NK cell of claim 1 , wherein the CD16 is a heterologous CD16 that comprises a mutation that results in a higher affinity of CD16 to IgG1.
10. The engineered NK cell of claim 1 , wherein the CD16 is a heterologous CD16 that comprises a 158V mutation.
11. The method of claim 6 , wherein the NK cell is derived from a primary cell obtained from a human subject.
12. The method of claim 6 , wherein the method results in reduced expression of FcRγ chain that is reduced by greater than or greater than about 50% as compared to the expression in the NK cell in that is not genetically engineered.
13. The method of claim 6 , wherein the method results in FcRγ chain expression level that is not detectable by an immunoblot assay.
14. The method of claim 6 , wherein the genetic disruption is by an endonuclease.
15. The method of claim 14 , wherein the endonuclease is a TAL nucleases, meganucleases, zinc-finger nucleases, an Argonaute nuclease or a CRISPR enzyme in combination with a guide RNA.
16. The method of claim 6 , wherein the CD16 is a heterologous CD16 and the method comprises introducing a nucleic acid encoding the heterologous CD16 into the NK cell.
17. The method of claim 16 , wherein the heterologous CD16 comprises a mutation that results in a higher affinity of CD16 to IgG1.
18. The method of claim 17 , wherein the heterologous CD16 comprises a 158V mutation.