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

Murad R Mamedov1,2, Shane Vedova3,4, Jacob W Freimer3,4,5

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Gamma delta (γδ) T cells target cancer by recognizing stress signals. Activating AMP-activated protein kinase (AMPK) boosts cancer cell stress signals, enhancing γδ T cell killing.

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Area of Science:

  • Immunology
  • Cancer Biology
  • Cellular Metabolism

Background:

  • γδ T cells are potent anticancer effectors recognizing conserved stress signals on transformed cells.
  • Vγ9Vδ2 T cells, a major human γδ T cell subset, recognize the BTN2A1-BTN3A1 complex on target cells.
  • Mechanisms linking cancer cell stress to γδ T cell targeting are not fully understood.

Purpose of the Study:

  • To identify pathways regulating γδ T cell killing and BTN3A expression in cancer cells.
  • To elucidate the role of cellular metabolism in γδ T cell-mediated cytotoxicity.
  • To explore therapeutic strategies for enhancing γδ T cell anticancer activity.

Main Methods:

  • Genome-wide CRISPR screens in target cancer cells.
  • Analysis of gene expression, protein modifications, and membrane trafficking.
  • Investigation of metabolic pathway disruptions and AMP-activated protein kinase (AMPK) signaling.
  • Assessment of Vγ9Vδ2 T cell killing in cell line and patient-derived tumor organoid models.

Main Results:

  • CRISPR screens revealed multilayered regulation of BTN3A cell surface expression and γδ T cell activation.
  • Disruption of cancer cell metabolic pathways, especially ATP production, altered BTN3A levels.
  • AMPK activation by metabolic stress induces BTN2A1 and BTN3A expression.
  • AMPK activation in cancer models increased BTN2A1-BTN3A complex expression and Vγ9Vδ2 T cell killing.

Conclusions:

  • Metabolic stress in cancer cells, mediated by AMPK, upregulates the BTN2A1-BTN3A complex.
  • This AMPK-dependent mechanism enhances Vγ9Vδ2 T cell recognition and killing of cancer cells.
  • Targeting metabolic pathways and activating AMPK offers a novel strategy to boost γδ T cell immunotherapy.