Neuronal ACVR1-mediated H3K18 lactylation drives NLRP3 pyroptosis to sustain neuropathic pain via metabolic-epigenetic coupling.

OBJECTIVE

We investigated whether neuronal activin A receptor type 1 (ACVR1) drives glycolytic lactate production to fuel histone H3K18 lactylation (H3K18la), thereby activating NLRP3-dependent neuronal pyroptosis and sustaining pain, while assessing therapeutic reversal.

METHODS

We employed the spared nerve injury (SNI) model in mice to study spinal ACVR1 signaling. Chronic pain was quantified via mechanical withdrawal thresholds (PWTs) and cold hypersensitivity. We integrated biochemical assays, flow-cytometry-like immunofluorescence, and multi-omics, including RNA sequencing (RNA-seq) and H3K18la chromatin immunoprecipitation sequencing (ChIP-seq) to map the ACVR1-dependent landscape. Pharmacological interventions utilized the ACVR1 inhibitor LDN-193189 and the lactylation inhibitor oxamate administered from post-operative day (POD) 8 to 14.

RESULTS

SNI induced mechanical allodynia and cold hypersensitivity, with sustained neuronal ACVR1 upregulation, glycolytic reprogramming, and spinal lactate elevation. H3K18la accumulated in NeuN+ neurons concurrent with NLRP3 inflammasome activation and gasdermin D-mediated pyroptosis. ACVR1 overexpression in HT22 cells recapitulated lactate production, H3K18la, and pyroptotic signaling. Exogenous lactate to naïve mice induced pain-like behavior with lactylation-dependent pyroptosis. Critically, delayed ACVR1 inhibition or oxamate after pain establishment reversed hypersensitivity while suppressing spinal lactate, H3K18la, and pyroptotic markers. Integrated RNA-seq and ChIP-seq identified Slc9a3 as a high-confidence ACVR1 target.

CONCLUSIONS

ACVR1-driven glycolysis promotes lactate-dependent H3K18la that activates neuronal NLRP3 pyroptosis to sustain neuropathic pain. Pharmacological blockade of ACVR1 or lactylation reverses established pain, positioning this immunometabolic axis as a tractable therapeutic target.