Precision Enhancement of CAR-NK Cells through Non-Viral Engineering and Highly Multiplexed Base Editing.
| العنوان: | Precision Enhancement of CAR-NK Cells through Non-Viral Engineering and Highly Multiplexed Base Editing. |
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| المؤلفون: | Wang M; Department of Pediatrics, University of Minnesota, Minneapolis, MN, USA.; Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA.; Center for Genome Engineering, University of Minnesota, Minneapolis, MN, USA.; Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN, USA., Krueger JB; Department of Pediatrics, University of Minnesota, Minneapolis, MN, USA.; Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA.; Center for Genome Engineering, University of Minnesota, Minneapolis, MN, USA., Gilkey AK; Department of Pediatrics, University of Minnesota, Minneapolis, MN, USA.; Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA.; Center for Genome Engineering, University of Minnesota, Minneapolis, MN, USA., Stelljes EM; Department of Pediatrics, University of Minnesota, Minneapolis, MN, USA.; Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA.; Center for Genome Engineering, University of Minnesota, Minneapolis, MN, USA., Kluesner MG; Department of Pediatrics, University of Minnesota, Minneapolis, MN, USA.; Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA.; Center for Genome Engineering, University of Minnesota, Minneapolis, MN, USA.; Molecular and Cellular Biology Graduate Program, Fred Hutchinson Cancer Center, Seattle, WA, USA., Pomeroy EJ; Department of Pediatrics, University of Minnesota, Minneapolis, MN, USA.; Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA.; Center for Genome Engineering, University of Minnesota, Minneapolis, MN, USA., Skeate JG; Department of Pediatrics, University of Minnesota, Minneapolis, MN, USA.; Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA.; Center for Genome Engineering, University of Minnesota, Minneapolis, MN, USA., Slipek NJ; Department of Pediatrics, University of Minnesota, Minneapolis, MN, USA.; Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA.; Center for Genome Engineering, University of Minnesota, Minneapolis, MN, USA., Lahr WS; Department of Pediatrics, University of Minnesota, Minneapolis, MN, USA.; Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA.; Center for Genome Engineering, University of Minnesota, Minneapolis, MN, USA., Vázquez PNC; Department of Pediatrics, University of Minnesota, Minneapolis, MN, USA.; Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA.; Center for Genome Engineering, University of Minnesota, Minneapolis, MN, USA.; Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN, USA., Zhao Y; Department of Pediatrics, University of Minnesota, Minneapolis, MN, USA.; Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA.; Center for Genome Engineering, University of Minnesota, Minneapolis, MN, USA., Eaton EJ; Department of Pediatrics, University of Minnesota, Minneapolis, MN, USA.; Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA.; Center for Genome Engineering, University of Minnesota, Minneapolis, MN, USA.; Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN, USA., Laoharawee K; Department of Pediatrics, University of Minnesota, Minneapolis, MN, USA.; Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA.; Center for Genome Engineering, University of Minnesota, Minneapolis, MN, USA.; Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN, USA., Webber BR; Department of Pediatrics, University of Minnesota, Minneapolis, MN, USA.; Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA.; Center for Genome Engineering, University of Minnesota, Minneapolis, MN, USA., Moriarity BS; Department of Pediatrics, University of Minnesota, Minneapolis, MN, USA.; Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA.; Center for Genome Engineering, University of Minnesota, Minneapolis, MN, USA. |
| المصدر: | BioRxiv : the preprint server for biology [bioRxiv] 2024 Mar 08. Date of Electronic Publication: 2024 Mar 08. |
| نوع المنشور: | Preprint |
| اللغة: | English |
| بيانات الدورية: | Country of Publication: United States NLM ID: 101680187 Publication Model: Electronic Cited Medium: Internet NLM ISO Abbreviation: bioRxiv Subsets: PubMed not MEDLINE |
| مستخلص: | Natural killer (NK) cells' unique ability to kill transformed cells expressing stress ligands or lacking major histocompatibility complexes (MHC) has prompted their development for immunotherapy. However, NK cells have demonstrated only moderate responses against cancer in clinical trials and likely require advanced genome engineering to reach their full potential as a cancer therapeutic. Multiplex genome editing with CRISPR/Cas9 base editors (BE) has been used to enhance T cell function and has already entered clinical trials but has not been reported in human NK cells. Here, we report the first application of BE in primary NK cells to achieve both loss-of-function and gain-of-function mutations. We observed highly efficient single and multiplex base editing, resulting in significantly enhanced NK cell function. Next, we combined multiplex BE with non-viral TcBuster transposon-based integration to generate IL-15 armored CD19 CAR-NK cells with significantly improved functionality in a highly suppressive model of Burkitt's lymphoma both in vitro and in vivo . The use of concomitant non-viral transposon engineering with multiplex base editing thus represents a highly versatile and efficient platform to generate CAR-NK products for cell-based immunotherapy and affords the flexibility to tailor multiple gene edits to maximize the effectiveness of the therapy for the cancer type being treated. Competing Interests: Conflict of interest statement M.W., E.J.P., M.G.K., B.S.M. and B.R.W. have filed patents covering the methods and approaches outlined in this work. |
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| معلومات مُعتمدة: | R01 AI161017 United States AI NIAID NIH HHS; P30 CA077598 United States CA NCI NIH HHS; U24 OD026641 United States OD NIH HHS; R21 CA237789 United States CA NCI NIH HHS; U54 CA232561 United States CA NCI NIH HHS; R21 AI163731 United States AI NIAID NIH HHS; P01 CA254849 United States CA NCI NIH HHS; R01 AI146009 United States AI NIAID NIH HHS; U54 CA268069 United States CA NCI NIH HHS; P50 CA136393 United States CA NCI NIH HHS |
| تواريخ الأحداث: | Date Created: 20240318 Latest Revision: 20240327 |
| رمز التحديث: | 20240327 |
| مُعرف محوري في PubMed: | PMC10942345 |
| DOI: | 10.1101/2024.03.05.582637 |
| PMID: | 38496503 |
| قاعدة البيانات: | MEDLINE |
| DOI: | 10.1101/2024.03.05.582637 |
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