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Publications

Journal Article Epublish

PubMed: 40248694 DOI: 10.3389/fimmu.2025.1557766

Nucleofection-based screening of chimeric antigen receptor candidates in human natural killer cells.

Front Immunol 16:1557766

Authors: Chelsia Qiuxia Wang, Leonard Leong, Lei Yang, Shengli Xu, Kong-Peng Lam, Andre Boon-Hwa Choo, Andy Hee-Meng Tan

Affiliations

  • Immune Cell Manufacturing, Bioprocessing Technology Institute (BTI), Agency for Science, Technology and Research (ASTAR), Singapore, Singapore.
  • Biotherapeutics Development, Bioprocessing Technology Institute (BTI), Agency for Science, Technology and Research (ASTAR), Singapore, Singapore.
  • Singapore Immunology Network, Agency for Science, Technology and Research (ASTAR), 8A Biomedical Grove, Immunos, Singapore, Singapore.
  • Departments of Physiology, Yong Loo Lin School of Medicine, National University of Singapore (NUS), Singapore, Singapore.
  • Departments of Microbiology and Immunology, Yong Loo Lin School of Medicine, National University of Singapore (NUS), Singapore, Singapore.
  • Departments of Pediatrics, Yong Loo Lin School of Medicine, National University of Singapore (NUS), Singapore, Singapore.
  • School of Biological Sciences, Nanyang Technological University (NTU), Singapore, Singapore.
  • Department of Biomedical Engineering, Faculty of Engineering, National University of Singapore (NUS), Singapore, Singapore.
  • Food, Chemical and Biotechnology Cluster, Singapore Institute of Technology (SIT), Singapore, Singapore.

Abstract

Chimeric antigen receptor (CAR)-modified cell therapy products approved for clinical treatment of hematological malignancies have hitherto been based on T cells. NK cells represent a promising immune cell type that can be considered for CAR engineering due to their potential to be generated as off-the-shelf allogeneic cellular therapy. Viral transduction of NK cells with CARs has been fraught with challenges of long process time and poor CAR transduction efficiency. Here, we describe the development of an optimized protocol for electroporation-based delivery of CAR mRNA into NK cells expanded from human peripheral blood mononuclear cells in the presence of co-stimulating feeder cells. This enabled rapid assessment of the functional capacity of NK cells transiently expressing various CARs to kill liquid and solid tumor cells . Ultimately, we anticipate that such an approach will enable selection of CAR candidates for their subsequent clinical applicability and manufacturability.

MeSH

Cells, Killer Cells, Natural, Immune System Diseases, Receptors, Antigen, Receptors, Chimeric Antigen, Therapeutics