SOX9 regulation of Hexokinase 1 controls neuroinflammatory astrocyte subtypes in neuropathic pain

Neuropathic pain (NeP) results from nerve damage or disease, lacking effective treatments. Astrocytes contribute to long-lasting neuroinflammation in the dorsal horn, driving NeP development. Directly targeting astrocytes is not feasible due to their roles in supporting neuronal homeostasis and pain resolution. Despite this understanding, the heterogeneity of astrocytes and the regulation of deleterious subsets emergence in pain remain less known. Through a comprehensive approach involving metabolomic, single-cell transcriptomic, epigenomic profiling and regional astrocyte-specific perturbation studies, we identified distinct astrocyte clusters under physiological and pathological pain conditions, and elucidated mechanisms by which metabolic regulation of neuroinflammatory astrocyte subsets during pain pathogenesis. We found an astrocyte specifier, Sox9, transcriptionally regulates Hexokinase1 (HK1), a critical enzyme that catalyzes the first step in glucose metabolism irreversibly, contributing to astrocytic glycolysis homeostasis. Initial nerve damage induced abnormal phosphorylation of Sox9, triggering aberrantly activation of HK1 for high-rate glycolysis in astrocytes. Moreover, the excessive lactate production from heightened glycolysis remodeled histones of gene promoters via lactylation, H3K9la, promoting transcriptional modules of genes governing pro-inflammatory and neurotoxic signaling, which induced pathogenic astrocyte properties while reducing beneficial populations, ultimately causing persistent pain state. Importantly, we demonstrate that targeted modulation of the SOX9-HK1-H3K9la axis specifically dampens deleterious astrocyte subsets, promoting long-lasting recovery of NeP. Collectively, our findings unveil a novel immunometabolic mechanism and identify multiple potential targets for effective therapeutic interventions in the treatment of NeP.