A Genetically Engineered Primary Human Natural Killer Cell Platform for Cancer Immunotherapy
Enhancing natural killer (NK) cell cytotoxicity by blocking inhibitory signaling could lead to improved NK-based cancer immunotherapy. Thus, we have developed a highly efficient method for editing the genome of human NK cells using CRISPR/Cas9 to knock out inhibitory signaling molecules. Our method efficiently edits up to 90% of primary peripheral blood NK cells. As a proof-of-principle we demonstrate highly efficient knockout of ADAM17 and PDCD1, genes that have a functional impact on NK cells, and demonstrate that these gene-edited NK cells have significantly improved activity, cytokine production, and cancer cell cytotoxicity. Furthermore, we were able to expand cells to clinically relevant numbers, without loss of activity. We also demonstrate that our CRISPR/Cas9 method can be used for efficient knockin of genes by delivering homologous recombination template DNA using recombinant adeno-associated virus serotype 6 (rAAV6). Our platform represents a feasible method for generating engineered primary NK cells as a universal therapeutic for cancer immunotherapy.
Journal
Molecular TherapyAuthors
Funding
Genetically Modified Natural Killer Cells for Cancer ImmunotherapyExpression of recombinant Fc receptors by engineered NK cells to enhance cancer cell killingNovel Immunomodulation and Facilitation of “Suppression Proof” CAR NK cell against Ewing sarcomaImmunology Training ProgramCancer Center Support GrantNational Cancer Institute FundingUS Department of Defense FundingChildren's Cancer Research Fund FundingProstate Cancer Foundation FundingRandy Shaver Cancer Research and Community Fund Funding