GSK-3 regulates CD4-CD8 cooperation needed to generate super-armed CD8+ cytolytic T cells against tumors

Moes, B.; Janna, J.; Kazanova, A.; LIU, C.; Gao, Y.; Ponnoor, N. A.; Castoun-Puckett, L.; Oroya, A.; Nguyen, H. L.; Cabald, A. L.; Tu, T. H.; Rudd, C. E.

doi:10.1101/2025.03.08.642085

bioRxiv · 10.1101/2025.03.08.642085

GSK-3 regulates CD4-CD8 cooperation needed to generate super-armed CD8+ cytolytic T cells against tumors

Moes, B.·Janna, J.·Kazanova, A.·LIU, C.·Gao, Y.·Ponnoor, N. A.·Castoun-Puckett, L.·Oroya, A.·Nguyen, H. L.·Cabald, A. L.·Tu, T. H.·Rudd, C. E.

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Abstract

While immune checkpoint blockade (ICB) has revolutionized cancer treatment, the key T-cell signaling pathways responsible for its potency remain unclear. GSK-3 is an inhibitory kinase that is most active in resting T-cells. In this study, we demonstrate that GSK-3 facilitates PD-1 blockade, an effect seen by modulating CD4 T-cell help for CD8+ CTL responses against ICB resistant tumors. We show that GSK-3 controls metabolic reprogramming towards glycolysis and synergizes with PD-1 to induce a transcriptional program that reduces suppressive CD4+ Treg numbers while generating super-armed effector-memory CD8+ CTLs that express an unprecedented 7/9 granzymes from the genome. Crucially, we found that GSK-3 cooperates with PD-1 blockade to determine the dependency of CD8+ CTLs on help from CD4+ T-cells. Our study unravels a novel cooperative PD-1 blockade-dependent signaling pathway that potentiates CTL responses against tumors, offering a new strategy to overcome immunotherapy resistance by modulating CD4+ helper and CD8+ cytotoxic functions. SignificanceThis study demonstrates for the first time that GSK-3 controls the crosstalk between CD4+ and CD8+ T cells, synergizing with anti-PD-1 therapy to overcome resistance to checkpoint blockade and to generate super-armed CD8+ effector cells in cancer immunotherapy. This newly uncovered GSK-3-dependent CD4-CD8 T-cell crosstalk mechanism presents a new approach to enhance anti-PD-1 immunotherapy.

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Moes, B., Janna, J., Kazanova, A., LIU, C., Gao, Y., Ponnoor, N. A., Castoun-Puckett, L., Oroya, A., Nguyen, H. L., Cabald, A. L., Tu, T. H., Rudd, C. E.. 2025-03-12. GSK-3 regulates CD4-CD8 cooperation needed to generate super-armed CD8+ cytolytic T cells against tumors. https://doi.org/10.1101/2025.03.08.642085

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