Silencing of METTL3 attenuates cardiac fibrosis induced by myocardial infarction via inhibiting the activation of cardiac fibroblasts
Silencing of METTL3 attenuates cardiac fibrosis induced by myocardial infarction via inhibiting the activation of cardiac fibroblasts
复制标题
METTL3沉默通过抑制心肌成纤维细胞的活化减轻心肌梗死引起的心肌纤维化
DOI：
10.1096/fj.201903169r
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发表时间：
2020-11-05
期刊：
影响因子：
4.8
通讯作者：
Pan, Zhenwei
中科院分区：
文献类型：
--
作者：
Li, Tingting;Zhuang, Yuting;Pan, Zhenwei
心脏纤维化的特征是心脏成纤维细胞的活化和细胞外基质的积聚。胃L3是甲基转移酶复合物的一个组成部分，参与与哺乳动物发育和疾病进展相关的多种生物学过程。然而，胃L3在心脏纤维化中的作用仍然未知。我们在体外用TGF-β 1（20 ng/mL）进行成纤维细胞活化，并用慢病毒建立体内小鼠模型，以评估胃L3对心脏成纤维细胞增殖和胶原形成的影响。甲基化RNA免疫沉淀（MeRIP）用于确定潜在的纤维化调节基因。在慢性心肌梗死小鼠的心脏纤维化组织和经TGF-β 1处理的培养的心脏成纤维细胞（CFs）中，胃L3的表达水平增加。胃L3的增强表达促进增殖和成纤维细胞向肌成纤维细胞的转化以及胶原积累，而胃L3的沉默则相反。通过携带pIL 3 siRNA的慢病毒沉默pIL 3可显着减轻MI小鼠的心脏纤维化。转录组和N6-甲基腺苷（m（6）A）分析显示，沉默胃L3后，胶原相关基因的表达和m（6）A水平发生了变化。胃L3介导的m（6）A修饰对心脏纤维化的发展至关重要，为操纵纤维化和相关心脏疾病提供了分子靶点。
Cardiac fibrosis is characterized by the activation of cardiac fibroblasts and accumulation of extracellular matrix. METTL3, a component of methyltransferase complex, participates in multiple biological processes associated with mammalian development and disease progression. However, the role of METTL3 in cardiac fibrosis is still unknown. We performed fibroblasts activation with TGF-beta 1 (20 ng/mL) in vitro and established in vivo mouse models with lentivirus to assess the effects of METTL3 on cardiac fibroblasts proliferation and collagen formation. Methylated RNA immunoprecipitation (MeRIP) was used to define the potential fibrosis-regulated gene. The expression level of METTL3 was increased in cardiac fibrotic tissue of mice with chronic myocardial infarction and cultured cardiac fibroblats (CFs) treated with TGF-beta 1. Enforced expression of METTL3 promoted proliferation and fibroblast-to-myofibroblast transition and collagens accumulation, while silence of METTL3 did the opposite. Silence of METTL3 by lentivirus carrying METTL3 siRNA markedly alleviated cardiac fibrosis in MI mice. Transcriptome and N6-methyladenosine (m(6)A) profiling analyses revealed that the expression and m(6)A level of collagen-related genes were altered after silence of METTL3. METTL3-mediated m(6)A modification is critical for the development of cardiac fibrosis, providing a molecular target for manipulating fibrosis and the associated cardiac diseases.