1. A Genetically Engineered Primary Human Natural Killer Cell Platform for Cancer Immunotherapy.
- Author
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Pomeroy EJ, Hunzeker JT, Kluesner MG, Lahr WS, Smeester BA, Crosby MR, Lonetree CL, Yamamoto K, Bendzick L, Miller JS, Geller MA, Walcheck B, Felices M, Webber BR, Starr TK, and Moriarity BS
- Subjects
- ADAM17 Protein genetics, Animals, CRISPR-Cas Systems, Cytotoxicity, Immunologic genetics, Dependovirus, Female, Gene Knockout Techniques, Healthy Volunteers, Humans, K562 Cells, Mice, Mice, Inbred NOD, Mice, SCID, Ovarian Neoplasms pathology, Parvovirinae genetics, Programmed Cell Death 1 Receptor genetics, Treatment Outcome, Xenograft Model Antitumor Assays, Adoptive Transfer methods, Cell Engineering methods, Genetic Engineering methods, Killer Cells, Natural immunology, Ovarian Neoplasms therapy
- Abstract
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., (Copyright © 2019 The American Society of Gene and Cell Therapy. Published by Elsevier Inc. All rights reserved.)
- Published
- 2020
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