Endothelial METTL3 (Methyltransferase-Like 3) Inhibits Fibrinolysis by Promoting PAI-1 (Plasminogen Activator Inhibitor-1) Expression Through Enhancing Jun Proto-Oncogene N6-Methyladenosine Modification.
Endothelial METTL3 (Methyltransferase-Like 3) Inhibits Fibrinolysis by Promoting PAI-1 (Plasminogen Activator Inhibitor-1) Expression Through Enhancing Jun Proto-Oncogene N6-Methyladenosine Modification.
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内皮细胞METTL3（甲基转移酶样3）通过增强Jun原癌基因n6 -甲基腺苷修饰促进PAI-1（纤溶酶原激活物抑制剂-1）表达抑制纤维蛋白溶解。
DOI：
10.1161/atvbaha.121.316414
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发表时间：
2021-12
期刊：
影响因子：
--
通讯作者：
Wen A
中科院分区：
文献类型：
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作者：
Bai Q;Lu Y;Chen Y;Zhang H;Zhang W;Wu H;Wen A
文本中提供了补充数字内容。 METTL3（甲基转移酶样蛋白3）介导的N6-甲基腺苷修饰是真核生物mRNA上最丰富的RNA修饰，在多种生理和病理过程中发挥着至关重要的作用。然而，N6-甲基腺苷修饰是否在血栓形成中发挥作用尚不清楚。本研究旨在确定 METTL3 在内皮细胞介导的血栓形成中的作用。 RNA测序和实时定量PCR显示，在METTL3敲低的人脐静脉内皮细胞中，PAI-1（纤溶酶原激活剂抑制剂-1）的表达下调。体外实验表明 METTL3 抑制纤维蛋白溶解。机械方面，RNA甲基化测序和meRIP实时定量PCR表明METTL3催化JUN mRNA 3'UTR上的N6-甲基腺苷修饰。 Western blotting分析显示METTL3促进JUN蛋白表达。染色质免疫沉淀分析表明，JUN 与人脐静脉内皮细胞中的 PAI-1 启动子结合。此外，用脂多糖攻击的小鼠导致血管中 METTL3 表达更高。内皮特异性敲低 Mettl3 可降低血浆中活性 PAI-1 的表达，并减弱内毒素血症期间肝脏和肺部的纤维蛋白沉积。我们的研究表明，METTL3 介导的 N6-甲基腺苷修饰在纤维蛋白溶解中发挥着至关重要的作用，并且是治疗血栓性疾病的潜在靶点。
Supplemental Digital Content is available in the text. METTL3 (methyltransferase-like protein 3)-mediated N6-methyladenosine modification is the most abundant RNA modification on eukaryote mRNAs and plays a crucial role in diverse physiological and pathological processes. However, whether N6-methyladenosine modification has function in thrombosis is unknown. This study aims to determine the role of METTL3 in the endothelial cells-mediated thrombosis. RNA-sequencing and real-time quantitative PCR revealed that the expression of PAI-1 (plasminogen activator inhibitor-1) was downregulated in METTL3 knockdown human umbilical vein endothelial cells. In vitro experiments showed that METTL3 suppressed fibrinolysis. Mechanically, RNA methylation sequencing and meRIP-quantitative real-time PCR showed that METTL3 catalyzed N6-methyladenosine modification on 3′ UTR of JUN mRNA. Western blotting analysis showed that METTL3 promoted JUN protein expression. Chromatin immunoprecipitation analysis demonstrated that JUN bound to the PAI-1 promoter in human umbilical vein endothelial cells. Furthermore, mice challenged with lipopolysaccharide resulted in higher METTL3 expression in vessels. Endothelial-specific knockdown of Mettl3 decreased expression of active PAI-1 in plasma and attenuated fibrin deposition in livers and lungs during endotoxemia. Our study reveals that METTL3-mediated N6-methyladenosine modification plays a crucial role in fibrinolysis and is an underlying target for the therapy of thrombotic disorders.