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Expression of chimeric antigen receptors in natural killer cells with a regulatory-compliant non-viral method

Cancer Gene Therapy volume 17, pages 147–154 (2010)Cite this article

Abstract

Natural killer (NK) cells hold promise for cancer therapy. NK cytotoxicity can be enhanced by expression of chimeric antigen receptors that re-direct specificity toward target cells by engaging cell surface molecules expressed on target cells. We developed a regulatory-compliant, scalable non-viral approach to engineer NK cells to be target-specific based on transfection of mRNA encoding chimeric receptors. Transfection of eGFP mRNA into ex vivo expanded NK cells (N=5) or purified unstimulated NK cells from peripheral blood (N=4) resulted in good cell viability with eGFP expression in 85±6% and 86±4%, 24 h after transfection, respectively. An mRNA encoding a receptor directed against CD19 (anti-CD19-BB-z) was also transfected into NK cells efficiently. Ex vivo expanded and purified unstimulated NK cells expressing anti-CD19-BB-z exhibited enhanced cytotoxicity against CD19+ target cells resulting in ⩾80% lysis of acute lymphoblastic leukemia and B-lineage chronic lymphocytic leukemia cells at effector target ratios lower than 10:1. The target-specific cytotoxicity for anti-CD19-BB-z mRNA-transfected NK cells was observed as early as 3 h after transfection and persisted for up to 3 days. The method described here should facilitate the clinical development of NK-based antigen-targeted immunotherapy for cancer.

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Acknowledgements

We express their gratitude to Dr Gary Moroff and Ms Amy Neuschaferand at American Red Cross Holland Lab (15601 Crabbs Branch Way, Rockville, MD) for training and usage of a Gammacell 1000 irradiator.

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Authors and Affiliations

  1. MaxCyte Inc, Gaithersburg, MD, USA

    L Li, L N Liu, S Feller, C Allen, R Shivakumar, J Fratantoni, L A Wolfraim, N Chopas, S Dzekunov & M Peshwa

  2. Departments of Oncology and Pathology, St Jude Children's Research Hospital, Memphis, TN, USA

    H Fujisaki & D Campana

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  1. L Li
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Correspondence to L Li.

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Li, L., Liu, L., Feller, S. et al. Expression of chimeric antigen receptors in natural killer cells with a regulatory-compliant non-viral method. Cancer Gene Ther 17, 147–154 (2010). https://doi.org/10.1038/cgt.2009.61

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  • DOI: https://doi.org/10.1038/cgt.2009.61

Keywords

  • NK cells
  • chimeric antigen receptor
  • electroporation
  • non-viral
  • transfection
  • mRNA

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