Hypermethylation of miR-181b in monocytes is associated with coronary artery disease and promotes M1 polarized phenotype via PIAS1-KLF4 axis
Hypermethylation of miR-181b in monocytes is associated with coronary artery disease and promotes M1 polarized phenotype via PIAS1-KLF4 axis
复制标题
DOI：
10.21037/cdt-20-407
复制
发表时间：
2020-08-01
影响因子：
2.4
通讯作者：
Sun, Yanming
中科院分区：
文献类型：
--
作者：
Wang, Zhonghua;Li, Chunlei;Sun, Yanming
背景：microRNAs表达异常参与巨噬细胞极化和动脉粥样硬化的发生发展。除了microRNA，DNA甲基化的改变被认为是最常见的表观遗传学变化之一。本研究旨在探讨冠心病患者外周血单核细胞中miR-181b甲基化状态的改变及其机制。方法：检测冠心病患者和健康对照外周血单核细胞中miR-181b甲基化状态。然后，我们用miR-181b模拟物转染单核细胞，并确定miR-181b对巨噬细胞表型转换和炎症反应的作用。通过MethyLight PCR和焦磷酸测序确定的rniR-181b启动子处的DNA甲基化水平在CAD患者中显著增加。基于TargetScan数据库，我们确定MASI为miR-181b的靶基因，并通过Dual-Luciferase分析、定量PCR和免疫印迹等方法研究了miR-181b与PIAS1的相互作用。结果：miR-181b基因启动子区甲基化直接导致miR-181b活性和表达的降低。通过定量PCR确定miR-181b的过表达减少M1极化并促进M2极化。虽然PIAS 1的敲低诱导单核细胞中的KLF 4降解和SUMO化，但miR-181 b模拟物通过抑制PIAS 1逆转KLF 4 SUMO化。结论：miR-181b高甲基化可通过激活巨噬细胞PIAS1和KLF4的SUMO化，诱导M1极化，促进动脉粥样硬化的发生。
Background: Dysregulated microRNAs are involved in the macrophage polarization and atherosclerotic development. Apart from microRNAs, alteration in DNA methylation is considered as one of the most frequent epigenetic changes. The purpose of the research is to investigate the altered methylation status of miR-181b in the circulating monocytes from patients with coronary artery disease (CAD) and explore the underlying mechanisms.Methods: We examined the methylation status of miR-181b in purified circulating monocytes from patients with CAD and healthy controls. We then transfected monocytes with miR-181b mimics and determined the role of miR-181b on the phenotypic switch of macrophages and inflammatory response. DNA methylation levels determined by MethyLight PCR and pyrosequencing at the promoter of rniR-181b significantly increased in CAD patients. Based on TargetScan database, we identified MASI as the target gene of miR-181b and explored the interaction of miR-181b and PIAS1 by Dual-Luciferase assay, quantitative PCR and immunoblots. We also investigated the role of miR-181b and PIAS1 on macrophage polarization and inflammation.Results: Hypermethylation at the promoter of miR-181b directly contributed to the decrease of miR-181b activity and expression. Overexpression of miR-181b reduced M1 polarization and facilitated M2 polarization determined by quantitative PCR. While knockdown of PIAS1 induced KLF4 degradation and SUMOylation in monocytes, miR-181b mimics reverse the KLF4 SUMOylation via suppression of PIAS1. Moreover, KLF4 SUMOylation by PIAS1 reversed M1 polarization induced by depletion of miR-181b in monocytes.Conclusions: Hypermethylation of miR-181b induces M1 polarization and promotes atherosclerosis through activation of PIAS1 and KLF4 SUMOylation in macrophages.