Downregulation of ciRNA‐ Kat6b in dorsal spinal horn is required for neuropathic pain by regulating Kcnk1 in miRNA‐26a‐dependent manner

AIMS: Nerve injury‐induced maladaptive changes in gene expression in the spinal neurons are essential for neuropathic pain genesis. Circular RNAs (ciRNA) are emerging as key regulators of gene expression. Here, we identified a nervous‐system‐tissues‐specific ciRNA‐Kat6 with conservation in humans and mice. We aimed to investigate whether and how spinal dorsal horn ciRNA‐Kat6b participates in neuropathic pain. METHODS: Unilateral sciatic nerve chronic constrictive injury (CCI) surgery was used to prepare the neuropathic pain model. The differentially expressed ciRNAs were obtained by RNA‐Sequencing. The identification of nervous‐system‐tissues specificity of ciRNA‐Kat6b and the measurement of ciRNA‐Kat6b and microRNA‐26a (miRNA‐26a) expression level were carried out by quantitative RT‐PCR. The ciRNA‐Kat6b that targets miRNA‐26a and miRNA‐26a that targets Kcnk1 were predicted by bioinformatics analysis and verified by in vitro luciferase reports test and in vivo experiments including Western‐blot, immunofluorescence, and RNA–RNA immunoprecipitation. The correlation between neuropathic pain and ciRNA‐Kat6b, miRNA‐26a, or Kcnk1 was examined by the hypersensitivity response to heat and mechanical stimulus. RESULTS: Peripheral nerve injury downregulated ciRNA‐Kat6b in the dorsal spinal horn of male mice. Rescuing this downregulation blocked nerve injury‐induced increase of miRNA‐26a, reversed the miRNA‐26a‐triggered decrease of potassium channel Kcnk1, a key neuropathic pain player, in the dorsal horn, and alleviates CCI‐induced pain hypersensitivities. On the contrary, mimicking this downregulation increased the miRNA‐26a level and decreased Kcnk1 in the spinal cord, resulting in neuropathic pain‐like syndrome in naïve mice. Mechanistically, the downregulation of ciRNA‐Kat6b reduced the accounts of miRNA‐26a binding to ciRNA‐Kat6b, and elevated the binding accounts of miRNA‐26a to the 3′ untranslated region of Kcnk1 mRNA and degeneration of Kcnk1 mRNA, triggering in the reduction of KCNK1 protein in the dorsal horn of neuropathic pain mice. CONCLUSION: The ciRNA‐Kat6b/miRNA‐26a/Kcnk1 pathway in dorsal horn neurons regulates the development and maintenance of neuropathic pain, ciRNA‐Kat6b may be a potential new target for analgesic and treatment strategies.