Genome-wide CRISPR screens identify critical targets to enhance CAR-NK cell antitumor potency

Alexander Biederstädt¹, Rafet Basar², Jeong-Min Park², Nadima Uprety², Rejeena Shrestha², Francia Reyes Silva², Merve Dede³, John Watts², Sunil Acharya², Donghai Xiong², Bin Liu², May Daher², Hind Rafei², Pinaki Banerjee², Ping Li², Sanjida Islam², Huihui Fan⁴, Mayra Shanley², Jingling Jin², Bijender Kumar², Vernikka Woods², Paul Lin², Silvia Tiberti², Ana Karen Nunez Cortes², Xin Ru Jiang⁵, Inci Biederstädt², Patrick Zhang², Ye Li², Seema Rawal², Enli Liu², Luis Muniz-Feliciano², Gary M Deyter², Elizabeth J Shpall⁶, Natalie Wall Fowlkes⁷, Ken Chen³, Katayoun Rezvani⁸

Affiliations

¹ Department of Stem Cell Transplantation and Cellular Therapy, The University of Texas MD Anderson Cancer Center, Houston, TX, USA; Institute for Cell Therapy Discovery and Innovation, The University of Texas MD Anderson Cancer Center, Houston, TX, USA; Department of Medicine III: Hematology & Oncology, School of Medicine, Technical University of Munich, Munich, Germany.
² Department of Stem Cell Transplantation and Cellular Therapy, The University of Texas MD Anderson Cancer Center, Houston, TX, USA; Institute for Cell Therapy Discovery and Innovation, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
³ Department of Bioinformatics and Computational Biology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
⁴ Department of Neurology, the University of Texas McGovern Medical School at Houston, Houston, TX, USA.
⁵ Department of Stem Cell Transplantation and Cellular Therapy, The University of Texas MD Anderson Cancer Center, Houston, TX, USA; Institute for Cell Therapy Discovery and Innovation, The University of Texas MD Anderson Cancer Center, Houston, TX, USA; The University of Texas MD Anderson Cancer Center UTHealth Houston Graduate School of Biomedical Sciences, Houston, TX, USA.
⁶ Department of Stem Cell Transplantation and Cellular Therapy, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
⁷ Department of Stem Cell Transplantation and Cellular Therapy, The University of Texas MD Anderson Cancer Center, Houston, TX, USA; Institute for Cell Therapy Discovery and Innovation, The University of Texas MD Anderson Cancer Center, Houston, TX, USA; Department of Veterinary Medicine & Surgery, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
⁸ Department of Stem Cell Transplantation and Cellular Therapy, The University of Texas MD Anderson Cancer Center, Houston, TX, USA; Institute for Cell Therapy Discovery and Innovation, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. Electronic address: krezvani@mdanderson.org.

PMID: 40845844
PMCID: PMC12396527 (available on 2026-08-18)
DOI: 10.1016/j.ccell.2025.07.021

Genome-wide CRISPR screens identify critical targets to enhance CAR-NK cell antitumor potency

Alexander Biederstädt et al. Cancer Cell. 2025.

. 2025 Aug 18:S1535-6108(25)00327-7.

doi: 10.1016/j.ccell.2025.07.021. Online ahead of print.

Authors

Affiliations

¹ Department of Stem Cell Transplantation and Cellular Therapy, The University of Texas MD Anderson Cancer Center, Houston, TX, USA; Institute for Cell Therapy Discovery and Innovation, The University of Texas MD Anderson Cancer Center, Houston, TX, USA; Department of Medicine III: Hematology & Oncology, School of Medicine, Technical University of Munich, Munich, Germany.
² Department of Stem Cell Transplantation and Cellular Therapy, The University of Texas MD Anderson Cancer Center, Houston, TX, USA; Institute for Cell Therapy Discovery and Innovation, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
³ Department of Bioinformatics and Computational Biology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
⁴ Department of Neurology, the University of Texas McGovern Medical School at Houston, Houston, TX, USA.
⁵ Department of Stem Cell Transplantation and Cellular Therapy, The University of Texas MD Anderson Cancer Center, Houston, TX, USA; Institute for Cell Therapy Discovery and Innovation, The University of Texas MD Anderson Cancer Center, Houston, TX, USA; The University of Texas MD Anderson Cancer Center UTHealth Houston Graduate School of Biomedical Sciences, Houston, TX, USA.
⁶ Department of Stem Cell Transplantation and Cellular Therapy, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
⁷ Department of Stem Cell Transplantation and Cellular Therapy, The University of Texas MD Anderson Cancer Center, Houston, TX, USA; Institute for Cell Therapy Discovery and Innovation, The University of Texas MD Anderson Cancer Center, Houston, TX, USA; Department of Veterinary Medicine & Surgery, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.
⁸ Department of Stem Cell Transplantation and Cellular Therapy, The University of Texas MD Anderson Cancer Center, Houston, TX, USA; Institute for Cell Therapy Discovery and Innovation, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. Electronic address: krezvani@mdanderson.org.

PMID: 40845844
PMCID: PMC12396527 (available on 2026-08-18)
DOI: 10.1016/j.ccell.2025.07.021

Abstract

Adoptive cell therapy using engineered natural killer (NK) cells is a promising approach for cancer treatment, with targeted gene editing offering the potential to further enhance their therapeutic efficacy. However, the spectrum of actionable genetic targets to overcome tumor and microenvironment-mediated immunosuppression remains largely unexplored. We performed multiple genome-wide CRISPR screens in primary human NK cells and identified critical checkpoints regulating resistance to immunosuppressive pressures. Ablation of MED12, ARIH2, and CCNC significantly improved NK cell antitumor activity against multiple treatment-refractory human cancers in vitro and in vivo. CRISPR editing augmented both innate and CAR-mediated NK cell function, associated with enhanced metabolic fitness, increased secretion of proinflammatory cytokines, and expansion of cytotoxic NK cell subsets. Through high-content genome-wide CRISPR screening in NK cells, this study reveals critical regulators of NK cell function and provides a valuable resource for engineering next-generation NK cell therapies with improved efficacy against cancer.

Keywords: CAR-NK cell therapy; adoptive cell therapy; functional perturbomics; genome-wide CRISPR screens; metabolic reprogramming; multiplexed cellular engineering; natural killer cells; precision gene editing; solid tumors; tumor microenvironment.

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Conflict of interest statement

Declaration of interests R.B., N.U., May Daher., H.R., P.B., S.A., M.S., Paul Lin., Y.L., E.L., E.J.S., K.R., and The University of Texas MD Anderson Cancer Center have an institutional financial conflict of interest with Takeda Pharmaceuticals. R.B., S.A., E.L., E.J.S., K.R. and The University of Texas MD Anderson Cancer Center have an institutional financial conflict of interest with Affimed. K.R. participates on the Scientific Advisory Board for Avenge Bio, Virogin Biotech, Navan Technologies, Caribou Biosciences, Bit Bio Limited, Replay Holdings, oNKo Innate, The Alliance for Cancer Gene Therapy ACGT, Innate Pharma and Shinobi Therapeutics. K.R. is the scientific founder of Syena. May Daher participates on the Scientific Advisory Board of Cellsbin. E.J.S. participates on the Scientific Advisory Board for Adaptimmune Limited, Axio Research, Celaid Therapeutics, FibroBiologics, Navan Technologies, New York Blood Center, and Zelluna Immunotherapy. The remaining authors declare no competing interests.

References

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Genome-wide CRISPR screens identify critical targets to enhance CAR-NK cell antitumor potency

Affiliations

Genome-wide CRISPR screens identify critical targets to enhance CAR-NK cell antitumor potency

Authors

Affiliations

Abstract

Conflict of interest statement

References

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