Related Experiment Video
Updated: Apr 11, 2026
Advances in Human Induced Pluripotent Stem Cell-Derived Chimeric Antigen Receptor-Expressing Natural Killer Cells
Published on: February 14, 2025
Gain-of-function enhancers optimize CAR-NK cell-based anti-cancer immunotherapy
Emma Wong1, Fernando Souza-Fonseca-Guimaraes1
1Frazer Institute, Faculty of Health, Medicine and Behavioural Sciences, The University of Queensland, Woolloongabba, QLD, Australia.
Abstract:
Schematic overview of the two-stage screening approach used to identify NK cell fitness genes. (A) CRISPRa mechanism, showing dCas9-VP64-mediated upregulation of target genes. (B) Whole-genome CRISPRa screening in HER2-CAR-NK92 cells transduced with a CRISPR sgRNA library and transferred into mice bearing HT29 tumours, followed by tumour collection and next-generation sequencing (NGS). (C) Barcoded ORF mini-screen in primary peripheral blood NK (PBNK) cells transduced with HER2-CAR and an ORF library, transferred into HT29 tumour-bearing mice, with subsequent tumour collection and NGS analysis.
Insights
This study identifies natural killer (NK) cell fitness genes using a two-stage CRISPR screening approach. These findings enhance understanding of NK cell function for potential cancer immunotherapy applications.
Area of Science:
- Immunology
- Molecular Biology
- Cancer Research
Background:
- Natural killer (NK) cells are crucial for innate immunity and cancer surveillance.
- Enhancing NK cell fitness is a key strategy for improving adoptive cell therapies.
- CRISPR activation (CRISPRa) offers a powerful tool for functional genomics screens.
Purpose of the Study:
- To identify genes that regulate the fitness and function of chimeric antigen receptor (CAR)-engineered NK cells.
- To develop and validate a two-stage screening approach for NK cell fitness genes.
Main Methods:
- A whole-genome CRISPRa screen was performed in HER2-CAR-NK92 cells in a mouse xenograft model.
- A barcoded open reading frame (ORF) mini-screen was conducted in primary peripheral blood NK (PBNK) cells engineered with HER2-CAR.
- Both screens utilized next-generation sequencing (NGS) to identify genes modulating NK cell fitness in the tumor microenvironment.
Main Results:
- The study successfully implemented a two-stage CRISPRa screening strategy to identify NK cell fitness genes.
- The methodology allowed for the assessment of gene function in both engineered cell lines and primary NK cells within a tumor-bearing mouse model.
Conclusions:
- The developed screening approach is effective for discovering genes that impact NK cell fitness and anti-tumor activity.
- This research provides a foundation for future studies aimed at optimizing NK cell-based immunotherapies.
