Background Mechanosensitive ion channels play a key role in heart development, physiology, and disease. However, little is known about the molecular mechanisms of the mechanosensitive nonselective cationic channel Piezo family in cardiac fibrosis. Methods and Results Mice were treated with ISO/Ang-II/TAC to induce cardiac fibrosis. AAV9 carrying POSTN promoter-driven small hairpin RNA targeting YTHDF1, and Piezo2 were administered to ISO mice to investigate their roles in cardiac fibrosis. RNA-seq, single-cell sequencing, and histological and biochemical analyses were performed to determine the mechanism by which YTHDF1 regulates Piezo2 expression in cardiac fibrosis. Piezo2 was reconstituted in YTHDF1-deficient cardiac fibroblasts and mouse hearts to study its effects on cardiac fibroblast autophagy and fibrosis. Piezo2 but not Piezo1 expression increased in experimental cardiac fibrosis and TGF-β1-induced cardiac fibroblasts. Fibroblast-specific Piezo2 deficiency ameliorated fibroblast activation and autophagy and inhibited cardiac fibrosis. Mechanistically, Piezo2 upregulation was associated with elevated m6A mRNA levels. Site-specific m6A modifications at peak_26355 were crucial for regulating the binding of YTHDF1 to Piezo2 mRNA and inducing Piezo2 translation. Notably, Piezo2 epitranscriptomic repression ameliorated experimental cardiac fibrosis. Conclusions We demonstrated a novel epitranscriptomic mechanism through which YTHDF1 recognizes Piezo2 and controls cardiac fibroblast autophagy and fibrosis through m6A-dependent modulation. Our findings provide new insights for the development of preventive measures for cardiac fibrosis.

中文翻译：

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通过 YTHDF1 依赖性 PIEZO2 mRNA m6A 修饰对心脏纤维化的表观转录组学调控


背景机械敏感离子通道在心脏发育、生理学和疾病中起着关键作用。然而，人们对机械敏感非选择性阳离子通道 Piezo 家族在心脏纤维化中的分子机制知之甚少。方法和结果 小鼠用 ISO/ANG-II/TAC 处理以诱导心脏纤维化。将携带 POSTN 启动子驱动的靶向 YTHDF1 的小发夹 RNA 和 Piezo2 的 AAV9 给予 ISO 小鼠，以研究它们在心脏纤维化中的作用。进行 RNA-seq 、 单细胞测序 以及组织学和生化分析，以确定 YTHDF1 调节心脏纤维化中 Piezo2 表达的机制。在 YTHDF1 缺陷的心脏成纤维细胞和小鼠心脏中重构 Piezo2 以研究其对心脏成纤维细胞自噬和纤维化的影响。在实验性心脏纤维化和 TGF-β 1 诱导的心脏成纤维细胞中，Piezo2 而不是 Piezo1 表达增加。成纤维细胞特异性 Piezo2 缺陷改善了成纤维细胞活化和自噬，并抑制了心脏纤维化。从机制上讲，Piezo2 上调与 m6A mRNA 水平升高有关。peak_26355位点特异性 m6A 修饰对于调节 YTHDF1 与 Piezo2 mRNA 的结合和诱导 Piezo2 翻译至关重要。值得注意的是，Piezo2 表观转录组抑制改善了实验性心脏纤维化。结论 我们展示了一种新的表观转录组学机制，YTHDF1 通过该机制识别 Piezo2 并通过 m6A 依赖性调节控制心脏成纤维细胞自噬和纤维化。我们的研究结果为心脏纤维化预防措施的制定提供了新的见解。