Multi-Omics Analysis Reveals Nono-Kcnq2 Regulation of Neuronal Excitability in Chronic Constriction Injury-Induced...

Neuropathic pain, resulting from somatosensory nervous system damage or disease, is a debilitating condition marked by spontaneous pain, hypersensitivity, and sensory abnormalities. Neuropathic pain involves spinal neuronal hyperexcitability, yet its molecular basis remains poorly defined. We utilized a comprehensive multi-omics approach, incorporating proteomics, phosphoproteomics, concatenated tandem array of consensus transcription response elements, and both single-cell and spatial transcriptomics, to chart time-resolved adaptations in the L4 to L6 spinal cord of a chronic constriction injury rat model. Multi-omics revealed remodeling of glutamatergic synapse pathways and identified non-POU (Pit-Oct-Unc) domain-containing octamer binding (Nono) as a down-regulated transcription factor associated with reduced potassium voltage-gated channel subfamily Q member 2 (Kcnq2) expression. Nono and Kcnq2 were co-enriched in neurexin 3-positive and peripherin-positive spinal neurons. Restoring Nono expression up-regulated Kcnq2, enhanced K⁺ outward currents (an effect largely abolished by the Kcnq2/3 blocker XE991), and alleviated pain hypersensitivity. Mechanistically, Nono was enriched at a conserved Kcnq2 promoter region in vivo and enhanced Kcnq2 promoter activity in reporter assays. Together, these findings established a Nono-Kcnq2 transcriptional axis that constrained spinal excitability and suggested a therapeutic entry point for neuropathic pain.