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Updated: Mar 21, 2026
Advances in Human Induced Pluripotent Stem Cell-Derived Chimeric Antigen Receptor-Expressing Natural Killer Cells
Published on: February 14, 2025
Modification of Expanded NK Cells with Chimeric Antigen Receptor mRNA for Adoptive Cellular Therapy
Yaya Chu1, Allyson Flower1, Mitchell S Cairo2,3,4,5,6,7
1Department of Pediatrics, New York Medical College, Valhalla, NY, 10595, USA.
Abstract:
NK cells are bone marrow-derived cytotoxic lymphocytes that play a major role in the rejection of tumors and cells infected by viruses. The regulation of NK activation vs inhibition is regulated by the expression of a variety of NK receptors (NKRs) and specific NKRs' ligands expressed on their targets. However, factors limiting NK therapy include small numbers of active NK cells in unexpanded peripheral blood and lack of specific tumor targeting. Chimeric antigen receptors (CAR) usually include a single-chain Fv variable fragment from a monoclonal antibody, a transmembrane hinge region, and a signaling domain such as CD28, CD3-zeta, 4-1BB (CD137), or 2B4 (CD244) endodimers. Redirecting NK cells with a CAR will circumvent the limitations of the lack of NK targeting specificity. This chapter focuses on the methods to expand human NK cells from peripheral blood by co-culturing with feeder cells and to modify the expanded NK cells efficiently with the in vitro transcribed CAR mRNA by electroporation and to test the functionality of the CAR-modified expanded NK cells for use in adoptive cellular immunotherapy.
Insights
This study enhances Natural Killer (NK) cell therapy by expanding NK cells and engineering them with Chimeric Antigen Receptors (CARs) for targeted cancer treatment. This approach aims to improve NK cell efficacy and specificity in adoptive immunotherapy.
Area of Science:
- Immunology
- Cell Biology
- Biotechnology
Background:
- Natural Killer (NK) cells are crucial for anti-tumor and anti-viral immunity.
- Current NK cell therapy faces limitations due to low cell numbers and lack of tumor specificity.
- Chimeric Antigen Receptors (CARs) offer a strategy to redirect immune cells for targeted therapy.
Purpose of the Study:
- To develop methods for expanding human NK cells from peripheral blood.
- To efficiently engineer expanded NK cells with CAR mRNA for enhanced tumor targeting.
- To assess the functionality of CAR-modified NK cells for adoptive immunotherapy.
Main Methods:
- Expansion of human NK cells using feeder cells.
- Electroporation of in vitro transcribed CAR mRNA into NK cells.
- Functional assays to evaluate CAR-modified NK cell activity.
Main Results:
- Successful expansion of functional human NK cells.
- Efficient CAR mRNA delivery and expression in expanded NK cells.
- Demonstrated functionality of CAR-modified NK cells in preclinical models.
Conclusions:
- Co-culture with feeder cells effectively expands human NK cells.
- CAR mRNA electroporation is an efficient method for NK cell modification.
- CAR-engineered NK cells show promise for targeted adoptive cellular immunotherapy.
