source data of glycogen-G6PD phase separation

Glucose-6-phosphate (G6P) is a key metabolic molecule that regulates reactive oxygen species (ROS) homeostasis by initiating the pentose phosphate pathway (PPP) to generate NADPH that converts H2O2 to water by providing hydrogen. While both glucose phosphorylation and glycogenolysis result in G6P production, here we show that G6P derived from glycogenolysis, rather than glucose phosphorylation, flows to PPP for ROS clearance in CD8+ memory T (Tm) cells and inflammatory macrophages. This is because glycogenolysis-produced G1P allosterically promotes G6P dehydrogenase (G6PD) to bind to glycogen liquid-liquid phase separation (LLPS), thereby recruiting PPP enzymes to form compartmentalization for the rapid cascade reaction. Based on mechanistic elucidation, we demonstrated that G1P can act as an antitumor immunotherapeutic agent by modulating memory fitness and maintenance of tumor-reactive CD8+ T cells. These findings revealed a previously unknown function of glycogen metabolism, which is of paramount importance in the regulation of PPP and redox homeostasis in cells.

葡萄糖-6-磷酸（Glucose-6-phosphate, G6P）是一类关键代谢分子，可通过启动磷酸戊糖途径（pentose phosphate pathway, PPP）生成烟酰胺腺嘌呤二核苷酸磷酸（NADPH），后者通过提供氢将过氧化氢（H₂O₂）还原为水，从而调控活性氧（reactive oxygen species, ROS）稳态。尽管葡萄糖磷酸化与糖原分解均可产生G6P，但本研究证实，在CD8+记忆T（CD8+ memory T, Tm）细胞与炎性巨噬细胞中，源自糖原分解的G6P而非葡萄糖磷酸化生成的G6P，会流向PPP以清除ROS。其核心机制为：糖原分解产生的葡萄糖-1-磷酸（Glucose-1-phosphate, G1P）可别构促进葡萄糖-6-磷酸脱氢酶（G6P dehydrogenase, G6PD）与糖原的液液相分离（liquid-liquid phase separation, LLPS）结合，进而招募PPP通路相关酶类形成功能区室，以支持快速级联反应。基于上述机制阐释，我们证明G1P可通过调控记忆细胞适配性与肿瘤反应性CD8+ T细胞的维持，作为抗肿瘤免疫治疗剂发挥作用。本研究揭示了糖原代谢此前未被报道的全新功能，该功能在调控细胞内PPP通路与氧化还原稳态过程中具有至关重要的意义。