YTHDF1-mediated m6A modification of TOP2A drives pulmonary hypertension via the PI3K/Akt/mTOR pathway

This study explores the role of the YTH N6-methyladenosine RNA binding protein F1 (YTHDF1)/ DNA topoisomerase II alpha (TOP2A) axis in the pathogenesis of pulmonary hypertension (PH) and investigates the regulatory mechanism of YTHDF1 on TOP2A. A PH mouse model was established using SU5416 combined with hypoxia (Su/Hy) in both wild-type and TOP2A knockout mice. Echocardiography, hemodynamic measurements, and histological analyses were performed to assess pulmonary vascular remodeling and right ventricular function. Human pulmonary artery smooth muscle cells (PASMCs) were exposed to hypoxia to mimic PH in vitro. TOP2A and YTHDF1 were knocked down or overexpressed in PASMCs using shRNAs or overexpression plasmids. Cell proliferation, migration, inflammation, oxidative stress, and autophagy were evaluated using various biological assays. RIP and MeRIP assays were conducted to investigate the interaction between YTHDF1 and TOP2A mRNA. Data showed that TOP2A expression was significantly elevated in the lung tissues of Su/Hy-induced PH mice and hypoxia-exposed PASMCs. Functionally, TOP2A depletion in mice attenuated pulmonary vascular remodeling, improved right ventricular function, reduced macrophage activation, and decreased inflammation and oxidative stress markers. Similarly, in hypoxia-treated PASMCs, TOP2A silencing inhibited cell proliferation, migration, inflammatory cytokine production, reactive oxygen species (ROS) generation, and autophagy-related protein expression. Furthermore, elevated m6A levels and increased YTHDF1 expression were observed in PH models, and we found that YTHDF1 bound to m6A-modified TOP2A mRNA to enhance its translation. Additionally, YTHDF1 knockdown reduced TOP2A protein expression and mitigated hypoxia-induced PASMCs dysfunction, while TOP2A overexpression reversed these protective effects. Mechanistically, TOP2A activated the PI3K/Akt/mTOR signaling pathway, driving PASMCs proliferation and migration. Collectively, this study identifies YTHDF1-mediated m6A modification as a key regulator of TOP2A expression, which promotes PASMCs dysfunction and vascular remodeling in PH via the PI3K/Akt/mTOR pathway.