Redirecting glucose flux during in vitro expansion generates epigenetically and metabolically superior anti-tumor T cells for adoptive cell therapies (CUT&Tag)

Cellular therapies are uniquely living drugs that must survive and persist within the body to achieve anti-tumor efficacy. In vitro culture of therapeutic T cells often relies on hypermetabolic conditions to maximize expansion. We show that this non-physiologic environment generates metabolically and functionally impaired T cells that more readily rely on aerobic glycolysis than T cells found in vivo. To target this shift in glycolytic metabolism, we employ dichloroacetate (DCA) during in vitro expansion resulting in elevated mitochondrial capacity and stemness properties pre-infusion - ultimately improving anti-tumor efficacy. DCA-conditioned T cells surprisingly show no elevated intratumoral function, but have an immediate and long-term survival benefit. DCA conditioning decreases reliance on extracellular glucose by promoting the usage of more physiologic carbon sources. This metabolic shift increases histone acetylation at key longevity genes dependent on citrate export from mitochondria. Blockade of mitochondrial citrate export – and thus formation of cytosolic acetyl-coA – reverses the beneficial effects of DCA. Our data suggest a simple means of correcting in vitro expansion of adoptive cell therapies for enhanced anti-tumor efficacy.