YTHDF2-orchestrated mA methylation of BECN1 induces Scoparone-mediated hepatic stellate cell ferroptosis to attenuate liver fibrosis.

Abstract

BACKGROUND: Liver fibrosis represents a dynamically reversible pathological process arising as an adaptive repair response to chronic hepatic insults. Scoparone (SCO), an active constituent of artemisia, has demonstrated therapeutic potential across diverse liver diseases, but its antifibrotic mechanism remains unclear. PURPOSE: This study aims to elucidate the molecular mechanism by which SCO ameliorates liver fibrosis through mA epitranscriptomic regulation of hepatic stellate cell (HSC) ferroptosis. METHODS: Murine liver fibrosis models and human HSC cells were employed to evaluate the therapeutic effects of SCO on liver fibrosis. Single-cell sequencing, spatial transcriptome sequencing, transcriptome sequencing, immunoprecipitation and laser confocal were used to investigate the potential molecular mechanisms. RESULTS: Animal experiments and cellular studies showed that SCO exhibited potent antifibrotic effects, which was attributed to the induction of HSC ferroptosis through mA modification. Integrative transcriptomic and bioinformatic analyses identified BECN1 as a key target for mA methylation regulation of ferroptosis. Mechanistically, SCO may interact with the ASN462 residue of YTHDF2, enhancing its protein stability and expression. Elevated YTHDF2 can facilitate translation of BECN1 mRNA by recognizing mA methylation at the A100 site within the 5'-UTR, leading to SLC7A11 activity inhibition and subsequent ferroptotic cell death in HSCs. Clinically, YTHDF2 and BECN1 expression was downregulated in fibrotic liver tissue specimens, which was associated with a poor prognosis. CONCLUSIONS: These results reveal a novel epitranscriptomic mechanism by which SCO induces HSC ferroptosis to attenuate liver fibrosis by promoting the formation of BECN1-SLC7A11 complex through YTHDF2-mediated mA modification. Thess findings molecular insights and therapeutic rationales for SCO-based antifibrotic therapies.