DNA N6-methyladenine methylase N6AMT1 controls neuropathic pain through epigenetically modifying Kcnj16 in dorsal horn neurons

Nerve injury-induced aberrant changes in gene expression in spinal dorsal horn neurons are critical for the genesis of neuropathic pain. N6-methyladenine (m(6)A) modification of DNA represents an additional layer of gene regulation. Here, we report that peripheral nerve injury significantly decreased the level of m(6)A-specific DNA methyltransferase 1 (N6amt1) in dorsal horn neurons. This decrease was attributed, at least partly, to a reduction in transcription factor Nr2f6. Rescuing the decrease in N6amt1 reversed the loss of m(6)A at the promoter for inwardly rectifying potassium channel subfamily J member 16 (Kcnj16), mitigating the nerve injury-induced upregulation of Kcnj16 expression in the dorsal horn and alleviating neuropathic pain hypersensitivities. Conversely, mimicking the downregulation of N6amt1 in naive mice erased DNA m(6)A at the Kcnj16 promoter, elevated Kcnj16 expression, and led to neuropathic pain-like behaviors. Therefore, decreased N6amt1 caused by NR2F6 is required for neuropathic pain, likely through its regulation of m(6)A-controlled KCNJ16 in dorsal horn neurons, suggesting that DNA m(6)A modification may be a potential new target for analgesic and treatment strategies.