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

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.