The m6A methyltransferase METTL3 promotes hypoxic pulmonary arterial hypertension
The m6A methyltransferase METTL3 promotes hypoxic pulmonary arterial hypertension
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DOI：
10.1016/j.lfs.2021.119366
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发表时间：
2021-03-19
期刊：
影响因子：
6.1
通讯作者：
Yan, Gaoliang
中科院分区：
文献类型：
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作者：
Qin, Yuhan;Qiao, Yong;Yan, Gaoliang
目的：N-6-甲基腺苷（N-6-methyladenosine，m（6）A）是哺乳动物mRNA中最常见的RNA内部化学修饰。越来越多的证据表明，在心血管疾病，包括心脏肥大，心力衰竭，缺血性心脏病，血管钙化，再狭窄和主动脉瘤的关键作用的m（6）A。然而，m（6）A是否参与了缺氧性肺动脉高压（HPH）的发生和发展仍不清楚。本研究旨在探讨胃关键转移酶L3在HPH发生发展中的作用。材料与方法：采用肺动脉平滑肌细胞（PASMCs）和低氧大鼠模型，研究胃关键转移酶L3介导的m（6）A在HPH发生发展中的作用。采用EdU、transwell和TUNEL法检测细胞增殖、迁移和凋亡率。m（6）A RNA甲基化定量试剂盒和m（6）A-qPCR检测总m（6）A水平和m（6）A水平的PTEN mRNA。RNA免疫沉淀法检测胃L3与PTEN mRNA的相互作用。关键发现：胃L3 mRNA和蛋白在体内和体外均异常上调。此外，胃L3的下调可减弱PASMCs的增殖和迁移。此外，低氧条件下PASMCs中m（6）A结合蛋白YTHDF 2表达明显增加。YTHDF 2可识别m（6）A介导的PTEN mRNA，促进PTEN的降解。结论：缺氧诱导的PASMCs增殖中，PTEN表达下调，可能通过激活PI 3 K/Akt信号通路而导致PASMCs过度增殖。意义：胃L3/YTHDF 2/PTEN轴在缺氧诱导的PASMCs增殖中发挥重要作用，为HPH的治疗提供了新靶点。
Aims: N-6-methyladenosine (m(6)A) is the most prevalent internal chemical RNA modification in mammal mRNAs. Accumulating evidence has shown the critical role of m(6)A in cardiovascular diseases including cardiac hypertrophy, heart failure, ischemic heart disease, vascular calcification, restenosis, and aortic aneurysm. However, whether m(6)A participates in the occurrence and development of hypoxic pulmonary hypertension (HPH) remains largely unknown. The present study aims to explore the role of key transferase METTL3, in the development of HPH.Materials and methods: Pulmonary artery smooth muscle cells (PASMCs) and hypoxic rat models were used to research the METTL3-mediated m(6)A in HPH. EdU, transwell and TUNEL were performed to evaluate the proliferation, migration and apoptosis rates. m(6)A RNA Methylation Quantification Kit and m(6)A-qPCR were utilized to measure the total m(6)A level and m(6)A level of PTEN mRNA. RNA immunoprecipitation was used to detect the interaction between METTL3 and PTEN mRNA.Key findings: Both METTL3 mRNA and protein were found abnormally upregulated in vivo and in vitro. Furthermore, downregulation of METTL3 attenuated PASMCs proliferation and migration. In addition, m(6)A binding protein YTHDF2 was found significantly increased in PASMCs under hypoxia. YTHDF2 recognized METTL3 mediated m(6)A modified PTEN mRNA and promoted the degradation of PTEN. Decreased PTEN led to over-proliferation of PASMCs through activation of PI3K/Akt signaling pathway.Significance: METTL3/YTHDF2/PTEN axis exerts a significant role in hypoxia induced PASMCs proliferation, providing a novel therapeutic target for HPH.