2. Academic Validation
  3. CRISPR screens decode cancer cell pathways that trigger γδ T cell detection

CRISPR screens decode cancer cell pathways that trigger γδ T cell detection

  • Nature. 2023 Aug 30. doi: 10.1038/s41586-023-06482-x.
Murad R Mamedov 1 2,  Shane Vedova # 3 4,  Jacob W Freimer # 3 4 5,  Avinash Das Sahu 6 7 8,  Amrita Ramesh 9,  Maya M Arce 3 4,  Angelo D Meringa 10,  Mineto Ota 3 4 5,  Peixin Amy Chen 3 4,  Kristina Hanspers 11,  Vinh Q Nguyen 3 12 13 14,  Kirsten A Takeshima 3,  Anne C Rios 15 16,  Jonathan K Pritchard 5 17,  Jürgen Kuball 10 18,  Zsolt Sebestyen 10,  Erin J Adams 9 19,  Alexander Marson 20 21 22 23 24 25 26 27
Affiliations

Affiliations

  • 1 Gladstone-UCSF Institute of Genomic Immunology, San Francisco, CA, USA. murad.mamedov@ucsf.edu.
  • 2 Department of Medicine, University of California, San Francisco, San Francisco, CA, USA. murad.mamedov@ucsf.edu.
  • 3 Gladstone-UCSF Institute of Genomic Immunology, San Francisco, CA, USA.
  • 4 Department of Medicine, University of California, San Francisco, San Francisco, CA, USA.
  • 5 Department of Genetics, Stanford University, Stanford, CA, USA.
  • 6 Department of Data Science, Dana-Farber Cancer Institute, Boston, MA, USA.
  • 7 Department of Biostatistics, Harvard T.H. Chan School of Public Health, Boston, MA, USA.
  • 8 UNM Comprehensive Cancer Center, University of New Mexico, Albuquerque, NM, USA.
  • 9 Department of Biochemistry and Molecular Biology, University of Chicago, Chicago, IL, USA.
  • 10 Center for Translational Immunology, University Medical Center Utrecht, Utrecht, the Netherlands.
  • 11 Institute of Data Science and Biotechnology, Gladstone Institutes, San Francisco, CA, USA.
  • 12 Department of Surgery, University of California, San Francisco, San Francisco, CA, USA.
  • 13 Diabetes Center, University of California, San Francisco, San Francisco, CA, USA.
  • 14 UCSF CoLabs, University of California, San Francisco, San Francisco, CA, USA.
  • 15 Princess Máxima Center for Pediatric Oncology, Utrecht, the Netherlands.
  • 16 Oncode Institute, Utrecht, the Netherlands.
  • 17 Department of Biology, Stanford University, Stanford, CA, USA.
  • 18 Department of Hematology, University Medical Center Utrecht, Utrecht, the Netherlands.
  • 19 Committee on Immunology, University of Chicago, Chicago, IL, USA.
  • 20 Gladstone-UCSF Institute of Genomic Immunology, San Francisco, CA, USA. alexander.marson@ucsf.edu.
  • 21 Department of Medicine, University of California, San Francisco, San Francisco, CA, USA. alexander.marson@ucsf.edu.
  • 22 Diabetes Center, University of California, San Francisco, San Francisco, CA, USA. alexander.marson@ucsf.edu.
  • 23 Department of Microbiology and Immunology, University of California, San Francisco, San Francisco, CA, USA. alexander.marson@ucsf.edu.
  • 24 Innovative Genomics Institute, University of California-Berkeley, Berkeley, CA, USA. alexander.marson@ucsf.edu.
  • 25 UCSF Helen Diller Family Comprehensive Cancer Center, University of California, San Francisco, San Francisco, CA, USA. alexander.marson@ucsf.edu.
  • 26 Parker Institute for Cancer Immunotherapy, University of California, San Francisco, San Francisco, CA, USA. alexander.marson@ucsf.edu.
  • 27 Institute for Human Genetics, University of California, San Francisco, San Francisco, CA, USA. alexander.marson@ucsf.edu.
  • # Contributed equally.
PMID: 37648854 DOI: 10.1038/s41586-023-06482-x
Abstract

γδ T cells are potent Anticancer effectors with the potential to target tumours broadly, independent of patient-specific neoantigens or human leukocyte antigen background1-5. γδ T cells can sense conserved cell stress signals prevalent in transformed cells2,3, although the mechanisms behind the targeting of stressed target cells remain poorly characterized. Vγ9Vδ2 T cells-the most abundant subset of human γδ T cells4-recognize a protein complex containing butyrophilin 2A1 (BTN2A1) and BTN3A1 (refs. 6-8), a widely expressed cell surface protein that is activated by phosphoantigens abundantly produced by tumour cells. Here we combined genome-wide CRISPR screens in target Cancer cells to identify pathways that regulate γδ T cell killing and BTN3A cell surface expression. The screens showed previously unappreciated multilayered regulation of BTN3A abundance on the cell surface and triggering of γδ T cells through transcription, post-translational modifications and membrane trafficking. In addition, diverse genetic perturbations and inhibitors disrupting metabolic pathways in the Cancer cells, particularly ATP-producing processes, were found to alter BTN3A levels. This induction of both BTN3A and BTN2A1 during metabolic crises is dependent on AMP-activated protein kinase (AMPK). Finally, small-molecule activation of AMPK in a cell line model and in patient-derived tumour organoids led to increased expression of the BTN2A1-BTN3A complex and increased Vγ9Vδ2 T cell receptor-mediated killing. This AMPK-dependent mechanism of metabolic stress-induced ligand upregulation deepens our understanding of γδ T cell stress surveillance and suggests new avenues available to enhance γδ T cell Anticancer activity.

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