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Updated: Jan 4, 2026
Advances in Human Induced Pluripotent Stem Cell-Derived Chimeric Antigen Receptor-Expressing Natural Killer Cells
Published on: February 14, 2025
A Genetically Engineered Primary Human Natural Killer Cell Platform for Cancer Immunotherapy
Emily J Pomeroy1, John T Hunzeker1, Mitchell G Kluesner1
1Department of Pediatrics, University of Minnesota, Minneapolis, MN 55455, USA; Masonic Cancer Center, University of Minnesota, Minneapolis, MN 55455, USA; Center for Genome Engineering, University of Minnesota, Minneapolis, MN 55455, USA.
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.
Insights
This study presents a new CRISPR/Cas9 method to engineer natural killer (NK) cells for cancer immunotherapy. Gene-edited NK cells show enhanced activity and cancer cell killing, offering a promising therapeutic approach.
Area of Science:
- Immunology
- Genetics
- Biotechnology
Background:
- Natural killer (NK) cells are crucial for cancer immunotherapy.
- Inhibitory signaling can limit NK cell effectiveness.
- Engineering NK cells can enhance their anti-cancer activity.
Purpose of the Study:
- To develop an efficient CRISPR/Cas9 method for editing human NK cells.
- To enhance NK cell cytotoxicity by knocking out inhibitory genes.
- To establish a platform for generating engineered NK cells for cancer therapy.
Main Methods:
- Utilized CRISPR/Cas9 gene editing in primary human peripheral blood NK cells.
- Efficiently knocked out inhibitory genes ADAM17 and PDCD1.
- Demonstrated gene knockin capability using recombinant adeno-associated virus serotype 6 (rAAV6).
Main Results:
- Achieved up to 90% editing efficiency in primary NK cells.
- Gene-edited NK cells exhibited significantly improved activity, cytokine production, and cancer cell cytotoxicity.
- Expanded gene-edited NK cells to clinically relevant numbers without activity loss.
Conclusions:
- The developed CRISPR/Cas9 platform enables efficient engineering of primary NK cells.
- Engineered NK cells show enhanced anti-cancer properties.
- This method provides a feasible approach for developing universal NK cell-based cancer immunotherapies.
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