Elucidation of key lactylation- and palmitoylation-related genes and their related mechanisms in the development of neuropathic pain.

Neuropathic pain (NP) is a debilitating disorder with heterogeneous etiologies and limited reliable biomarkers for diagnosis and therapeutic targeting. This study aimed to identify key lactylation- and palmitoylation-related genes involved in NP and to elucidate their potential regulatory mechanisms. NP-related transcriptomic datasets (GSE124272, GSE150408, and GSE95849) were retrieved from the Gene Expression Omnibus (GEO). Differentially expressed genes associated with lactylation and palmitoylation were identified and analyzed using functional enrichment, correlation analysis, and protein-protein interaction networks. Feature genes were selected using support vector machine-recursive feature elimination and XGBoost algorithms. Diagnostic performance was assessed using receiver operating characteristic (ROC) curve analysis, and a predictive nomogram was constructed. Immune infiltration was evaluated by single-sample gene set enrichment analysis, while gene set enrichment analysis, miRNA-mRNA, and transcription factor-mRNA network analyses were performed to explore regulatory mechanisms. Independent clinical validation was conducted by quantitative real-time PCR using peripheral blood samples from NP patients (n = 17) and matched healthy controls (n = 17). Hematopoietic cell kinase (HCK; lactylation-related) and WASP actin nucleation-promoting factor (WAS; palmitoylation-related) were identified as NP feature genes. Both genes were significantly upregulated in NP samples across training and validation GEO datasets and were independently confirmed in the clinical cohort. HCK and WAS demonstrated diagnostic utility (area under the curve > 0.7 in GEO analyses), and a nomogram based on these genes showed favorable calibration and decision-curve benefit. Expression of both genes was positively correlated with infiltration of neutrophils, monocytes, and macrophages and was enriched in the NOD-like receptor signaling pathway. Regulatory network analysis revealed interactions between HCK and miRNAs including hsa-miR-21-5p and hsa-miR-146a-5p, and between WAS and miRNAs including hsa-miR-34a-5p and hsa-miR-16-5p. Both genes were predicted to be regulated by the transcription factor SPI1. WAS and HCK represent promising peripheral blood biomarkers for NP and may contribute to NP pathogenesis through immune cell-mediated neuroinflammation and NOD-like receptor signaling, highlighting potential targets for future diagnostic and therapeutic development.