Silencing of METTL14 attenuates experimental intracerebral hemorrhage by suppressing TFRC-mediated ferroptosis

Ferroptosis is emerging as a pathological mechanism of intracerebral hemorrhage (ICH), and inhibiting Ferroptosis contributes to improving prognosis. N6-methyladenosine (m6A) methylation is a common RNA modification that is involved in disease progression. This study aimed to explore the effect of METTL14, a m6A transmethylase, on Ferroptosis and the molecular mechanism, and identify its role in ICH progression. PC12 cells were stimulated with hemin to induce injury, and mice were injected with type IV collagenase to generate the ICH model. Ferroptosis was evaluated by detecting Fe2+, Reactive Oxygen Species, and reduced glutathione levels. M6A methylation was assessed by methylated-RNA immunoprecipitation (Me-RIP), RIP, dual-luciferase reporter assay, and actinomycin D treatment. The results showed that hemin induced Ferroptosis of PC12 cells. METTL14 was highly expressed in hemin-treated PC12 cells and ICH mice, and knockdown of METTL14 reversed the promotion of Ferroptosis induced by hemin. Mechanically, METTL14 knockdown inhibited m6A modification of TFRC and reduced its stability at RNA level, which was recognized by IGF2BP2. Overexpression of TFRC reversed the inhibition of Ferroptosis caused by METTL14 silence both in vitro and in vivo. In conclusion, silencing of METTL14 ameliorates ICH by inhibiting Ferroptosis through suppressing m6A methylation of TFRC. These findings suggest that targeting m6A methylation and Ferroptosis may be a promising strategy for ICH therapy.

Ferroptosis; Intracerebral hemorrhage; METTL14; TFRC; m6A methylation.