A Genetically Engineered Primary Human Natural Killer Cell Platform for Cancer Immunotherapy

Pomeroy, E. J.; Hunzeker, J. T.; Kluesner, M. T.; Crosby, M. R.; Lahr, W. S.; Bendzick, L.; Miller, J. S.; Webber, B. R.; Geller, M. A.; Walcheck, B.; Felices, M.; Starr, T. K.; Moriarity, B. S.

doi:10.1101/430553

bioRxiv · 10.1101/430553

A Genetically Engineered Primary Human Natural Killer Cell Platform for Cancer Immunotherapy

Pomeroy, E. J.·Hunzeker, J. T.·Kluesner, M. T.·Crosby, M. R.·Lahr, W. S.·Bendzick, L.·Miller, J. S.·Webber, B. R.·Geller, M. A.·Walcheck, B.·Felices, M.·Starr, T. K.·Moriarity, B. S.

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Abstract

Tumors can evade natural killer (NK) cells by activating inhibitory pathways. We therefore have developed a highly efficient CRISPR/Cas9-based method for editing the genome of peripheral blood human NK cells (PB-NKs) to knock out ADAM17 and PD1 or knock-in genes using recombinant AAV6. Our method allows editing of PB-NKs at efficiencies reaching 90%, equivalent to methods reported for primary human T cells. Moreover, we demonstrate that ADAM17 and PD1 KO PB-NKs have significantly improved activity, cytokine production, and cancer cell cytotoxicity. Our platform represents a feasible method for generating engineered NK cells as a universal therapeutic for cancer immunotherapy.

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Pomeroy, E. J., Hunzeker, J. T., Kluesner, M. T., Crosby, M. R., Lahr, W. S., Bendzick, L., Miller, J. S., Webber, B. R., Geller, M. A., Walcheck, B., Felices, M., Starr, T. K., Moriarity, B. S.. 2018-09-29. A Genetically Engineered Primary Human Natural Killer Cell Platform for Cancer Immunotherapy. https://doi.org/10.1101/430553

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