Signaling Node at TSC2 S1365 Potently Regulates T-Cell Differentiation and Improves Adoptive Cellular Cancer Therapy

Abstract

AbstractMTORC1 integrates signaling from the immune microenvironment to regulate T-cell activation, differentiation, and function. TSC2 in the tuberous sclerosis complex potently regulates mTORC1 activation. CD8+ T-cells lacking TSC2 have constitutively enhanced mTORC1 activity and generate potent effector T cells; however sustained mTORC1 activation prevents generation of long-lived memory CD8+ T-cells. Here we show manipulating TSC2 at Ser1365 potently regulates activated but not basal mTORC1 signaling in T cells. Unlike non-stimulated TSC2 knockout cells, CD8+ T-cells expressing mutant TSC2-S1365A (SA) have normal basal mTORC1 activity. PKC and T-cell Receptor (TCR) stimulation induces TSC2 S1365 phosphorylation and preventing this with the SA mutation markedly increases mTORC1 activation and T-cell effector function. Consequently, CD8+ SA T-cells display greater effector responses while retaining their capacity to become long-lived memory T-cells. CD8+ SA T-cells also display enhanced effector function under hypoxic and acidic conditions. In murine and human solid-tumor models, CD8+ SA T-cells used as adoptive cell therapy have greater anti-tumor immunity than WT CD8+ T-cells. These findings reveal an upstream mechanism to regulate mTORC1 activity in T-cells. The TSC2-SA mutation enhances both T-cell effector function and long-term persistence/memory formation, supporting a novel approach to engineer better CAR-T cells to treat cancer.