IGF2BP2 stabilizes SLC39A14 mRNA via m6A modification to promote ferroptosis in cerebral ischemia-reperfusion injury.

Abstract

Cerebral ischemia-reperfusion injury (CIRI) is a leading cause of neurological disability with few effective therapeutic strategies, and ferroptosis has been proven to participate in CIRI progression while its upstream regulatory mechanisms remain poorly defined. Herein, we performed transcriptomic analysis on GSE162072 and GSE58720 datasets, and constructed mouse tMCAO/R and neuronal OGD/R models to recapitulate CIRI. Multiple assays including Longa scoring, TTC staining, H&E staining and Evans blue assay were applied to evaluate neurological function, cerebral infarction, histopathology and blood-brain barrier (BBB) integrity. We also quantified ferroptosis-related markers and detected m6A modification, RNA-protein interaction and mRNA stability via commercial kits, dot blot, MeRIP-qPCR, RIP-qPCR and actinomycin D assays. The results revealed that SLC39A14 was markedly upregulated in CIRI, accompanied by neuronal ferroptosis activation. Knockdown of SLC39A14 alleviated iron overload, lipid peroxidation and ferroptotic neuronal death both in vitro and in vivo. Mechanistically, the m6A reader IGF2BP2 was elevated in CIRI, which directly bound SLC39A14 mRNA to improve its stability and sustain its expression. In vivo AAV-mediated SLC39A14 silencing reduced infarct volume, maintained BBB integrity and relieved neurological deficits. Collectively, IGF2BP2 facilitates CIRI development by driving neuronal ferroptosis through m6A-dependent regulation of SLC39A14 mRNA stability, which may serve as a promising therapeutic target for CIRI.