Glutamyl-Prolyl-tRNA Synthetase Regulates Proline-Rich Pro-Fibrotic Protein Synthesis During Cardiac Fibrosis
Glutamyl-Prolyl-tRNA Synthetase Regulates Proline-Rich Pro-Fibrotic Protein Synthesis During Cardiac Fibrosis
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DOI：
10.1161/circresaha.119.315999
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发表时间：
2020-08-28
影响因子：
20.1
通讯作者：
Yao, Peng
中科院分区：
文献类型：
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作者：
Wu, Jiangbin;Subbaiah, Kadiam C. Venkata;Yao, Peng
原理：促纤维化基因的蛋白质合成增加是心脏纤维化和心力衰竭的共同特征。尽管如此，心脏纤维化过程中促纤维化基因翻译控制的关键因素和分子机制仍不清楚。目的：目的探讨双功能氨酰-tRNA合成酶（ARS）、谷氨酰-脯氨酰-tRNA合成酶（EPRS）在心肌纤维化翻译调控中的作用。方法和结果：对人类和小鼠心力衰竭的多个公开数据集进行再分析的结果表明，EPRS在各种心脏致病过程中充当了ARS之间的综合节点。我们使用我们的人类和小鼠心脏样本队列证实，与非衰竭心脏相比，在衰竭心脏中EPRS在mRNA和蛋白水平上被诱导（大约增加1.5-2.5倍）。使用CRISPR-Cas9技术或以Postn-Cre-依赖性方式（Ehrs（flox/+）;Postn（MCM/+））基因敲除Eprs的一个等位基因在异丙肾上腺素、横主动脉缩窄和心肌梗死（MI）诱导的心力衰竭小鼠模型中强烈减少心脏纤维化（减少约50%）。使用PRS（脯氨酰-tRNA合成酶）特异性抑制剂卤夫酮抑制EPRS可显著降低心脏成纤维细胞以及TGF-β（转化生长因子-β）激活的肌成纤维细胞中富含脯氨酸的胶原蛋白的翻译效率（TE）。在TGF-β刺激下，EPRS的过表达增加了原代心脏成纤维细胞中胶原蛋白的表达。在常山酮处理的成纤维细胞中使用转录组范围的RNA-Seq和多核糖体分析-Seq，我们确定了除胶原蛋白外的多种新的富含Pro的基因，如Ltbp 2（潜伏性TGF-β结合蛋白2）和Sulf 1（硫酸酯酶1），这些基因由EPRS间接调控。SULF 1在人类和小鼠肌成纤维细胞中高度富集。在原代心脏成纤维细胞培养系统中，siRNA介导的SULF 1敲低减弱了心脏肌成纤维细胞活化和胶原沉积。SULF 1的过表达促进TGF-β诱导的肌成纤维细胞活化并部分拮抗常山酮治疗的抗纤维化作用结论：我们的研究结果表明，EPRS优先控制翻译激活脯氨酸密码子丰富的促纤维化基因在心脏成纤维细胞和增强病理性心脏重塑。
Rationale: Increased protein synthesis of profibrotic genes is a common feature in cardiac fibrosis and heart failure. Despite this observation, critical factors and molecular mechanisms for translational control of profibrotic genes during cardiac fibrosis remain unclear. Objective: To investigate the role of a bifunctional ARS (aminoacyl-tRNA synthetase), EPRS (glutamyl-prolyl-tRNA synthetase) in translational control of cardiac fibrosis. Methods and Results: Results from reanalyses of multiple publicly available data sets of human and mouse heart failure, demonstrated that EPRS acted as an integrated node among the ARSs in various cardiac pathogenic processes. We confirmed that EPRS was induced at mRNA and protein levels (approximate to 1.5-2.5-fold increase) in failing hearts compared with nonfailing hearts using our cohort of human and mouse heart samples. Genetic knockout of one allele ofEprsglobally (Eprs(+/-)) using CRISPR-Cas9 technology or in a Postn-Cre-dependent manner (Eprs(flox/+);Postn(MCM/+)) strongly reduces cardiac fibrosis (approximate to 50% reduction) in isoproterenol-, transverse aortic constriction-, and myocardial infarction (MI)-induced heart failure mouse models. Inhibition of EPRS using a PRS (prolyl-tRNA synthetase)-specific inhibitor, halofuginone, significantly decreases translation efficiency (TE) of proline-rich collagens in cardiac fibroblasts as well as TGF-beta (transforming growth factor-beta)-activated myofibroblasts. Overexpression of EPRS increases collagen protein expression in primary cardiac fibroblasts under TGF-beta stimulation. Using transcriptome-wide RNA-Seq and polysome profiling-Seq in halofuginone-treated fibroblasts, we identified multiple novel Pro-rich genes in addition to collagens, such as Ltbp2 (latent TGF-beta-binding protein 2) and Sulf1 (sulfatase 1), which are translationally regulated by EPRS. SULF1 is highly enriched in human and mouse myofibroblasts. In the primary cardiac fibroblast culture system, siRNA-mediated knockdown of SULF1 attenuates cardiac myofibroblast activation and collagen deposition. Overexpression of SULF1 promotes TGF-beta-induced myofibroblast activation and partially antagonizes anti-fibrotic effects of halofuginone treatment. Conclusions: Our results indicate that EPRS preferentially controls translational activation of proline codon rich profibrotic genes in cardiac fibroblasts and augments pathological cardiac remodeling.