2. Academic Validation
  3. m6A-mediated silencing of RNF41 by METTL3/YTHDC1 disrupts autophagy to drive intervertebral disc degeneration

m6A-mediated silencing of RNF41 by METTL3/YTHDC1 disrupts autophagy to drive intervertebral disc degeneration

  • Cell Biol Toxicol. 2025 Nov 3;41(1):147. doi: 10.1007/s10565-025-10093-9.
Bo Yao 1,  Li Wan 1,  Jia Deng 1,  Zhengyu Chen 1,  Lei Zhao 1,  Weiguo Wang 2,  Ziwei Han 3
Affiliations

Affiliations

  • 1 Department of Spine Surgery, Want Want Hospital, Changsha, 410016, Hunan, China.
  • 2 Department of Spine Surgery, The Third Xiangya Hospital of Central South University, Changsha, 410013, Hunan, China. [email protected].
  • 3 Department of Spine Surgery, Want Want Hospital, Changsha, 410016, Hunan, China. [email protected].
PMID: 41177830 DOI: 10.1007/s10565-025-10093-9
Abstract

Background: Intervertebral disc (IVD) degeneration (IDD) represents a predominant origin of low back pain and disability, yet current therapeutic interventions remain suboptimal. Emerging evidence highlights Autophagy activation as a therapeutic strategy against IDD. This study investigates the mechanistic interplay between N6-methyladenosine (m6A) modifications and Autophagy dysregulation in IDD pathogenesis.

Methods: Bioinformatics analysis identified ring finger protein 41 (RNF41) as a key autophagy-IDD intersection gene. Functional validation utilized tert-butyl hydroperoxide (TBHP)-treated human nucleus pulposus (NP) cells to assess RNF41's effects on senescence (CDKN2A), Autophagy (LC3-II/p62), Apoptosis (TUNEL), inflammation (IL-18/IL-1β), and extracellular matrix (ECM) homeostasis (aggrecan/MMP). Key m6A regulators modulating Autophagy were screened via correlation analysis. In vivo validation employed adeno-associated virus (AAV)-mediated methyltransferase-like 3 (METTL3)/RNF41 delivery in puncture-induced IDD rat models.

Results: RNF41 expression was downregulated in human IVD tissues. Overexpression of RNF41 mitigated TBHP-induced senescence, Apoptosis, activated AMPK/mTOR-mediated Autophagy, suppressed inflammation, and restored ECM balance. The Autophagy inhibitor chloroquine (CQ) abolished the protective effects of RNF41 overexpression on degenerative NP cells. Mechanistically, METTL3/YTHDC1 co-regulation in degenerative NP cells mediated m6A hypermethylation of RNF41 mRNA, shortening its half-life via YTHDC1-dependent decay. Intradiscal METTL3-silencing AAV attenuated puncture-induced disc loss and histopathological degeneration, whereas RNF41-silencing AVV exacerbated ECM disruption and annular disorganization.

Conclusion: METTL3/YTHDC1-mediated m6A modification drives IDD progression by silencing RNF41, thereby impairing Autophagy and ECM integrity. Targeting this axis offers a clinically actionable strategy to delay disc degeneration, particularly in patients with early-stage IDD. This evidence establishes RNF41's role as a theragnostic biomarker and therapeutic targe, enabling precision-guided interventional approaches.

Keywords

AMPK/mTOR; Autophagy; Intervertebral disc degeneration; N6-methyladenosine; RNF41.

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