Inhibition of the METTL3/m6A/miR-34a-5p axis suppresses trigeminovascular activation in nitroglycerin-induced migraine via the Wnt/β-catenin pathway

J Headache Pain. 2025 Oct 2;26(1):197. doi: 10.1186/s10194-025-02144-7.

Abstract

Background: Neuroinflammation is a key driver of migraine pathogenesis, particularly by promoting neuronal sensitization. METTL3-mediated m6A methylation has emerged as a critical epigenetic regulator in neuroinflammatory responses. This study aims to investigate the role of METTL3 in migraine, focusing on its m6A-dependent regulatory mechanisms.

Methods: A rat migraine model was induced via chronic intermittent nitroglycerin administration. Behavioral tests assessed migraine-like symptoms. Protein and RNA levels of METTL3, miR-34a-5p, and downstream targets were analyzed using Western blot, qPCR, ELISA, and immunofluorescence. The interaction among METTL3, miR-34a-5p, and Wnt1 was validated through Co-IP, RIP, and luciferase reporter assays.

Results: METTL3 expression was significantly upregulated in the trigeminal ganglia of migraine rats. Knockdown of METTL3 reduced trigeminovascular system (TGVS) activation and alleviated migraine symptoms. Mechanistically, METTL3 enhanced miR-34a-5p expression via m6A modification, leading to suppression of the Wnt1/β-catenin pathway. Overexpression of miR-34a-5p further aggravated migraine-related responses by inhibiting Wnt1 signaling.

Conclusion: METTL3 contributes to migraine pathogenesis through m6A-dependent upregulation of miR-34a-5p, which suppresses the Wnt1/β-catenin axis and promotes TGVS activation. Targeting this pathway may offer new therapeutic avenues for migraine.

Keywords: METTL3; MiR-34a-5p; Migraine; Wnt1/β-catenin pathway.

MeSH terms

  • Adenosine* / analogs & derivatives
  • Adenosine* / metabolism
  • Animals
  • Disease Models, Animal
  • Male
  • Methyltransferases* / genetics
  • Methyltransferases* / metabolism
  • MicroRNAs* / metabolism
  • Migraine Disorders* / chemically induced
  • Migraine Disorders* / metabolism
  • Nitroglycerin
  • Rats
  • Rats, Sprague-Dawley
  • Trigeminal Ganglion* / metabolism
  • Wnt Signaling Pathway* / drug effects
  • Wnt Signaling Pathway* / physiology
  • Wnt1 Protein* / metabolism

Substances

  • MicroRNAs
  • Nitroglycerin
  • Methyltransferases
  • MIRN34 microRNA, rat
  • Wnt1 Protein
  • Wnt1 protein, rat
  • Adenosine