Translational Control of Cardiac Fibrosis

心脏纤维化的转化控制

• 批准号: 10220123
• 负责人: Peng Yao
• 金额: $ 38.5万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-11 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10220123
• 关键词: Alleles; Amino Acids; Amino Acyl-tRNA Synthetases; Apoptosis; Biological Process; Cardiac; Cardiac Myocytes; Cardiomyopathies; Cardiovascular Diseases; Cardiovascular Manifestation; Chemicals; Chinese Herbs; Codon Nucleotides; Collagen; Coupled; Cyclic AMP-Dependent Protein Kinases; DNA; DNA Sequence Alteration; Data; Development; Dilated Cardiomyopathy; Dose; Down-Regulation; Elongation Factor; Enzymes; Event; FURIN gene; Fibrinogen; Fibroblasts; Gene Expression; Gene Targeting; Genes; Genetic; Genetic Transcription; Genetic Translation; Glutamic Acid; Halofuginone; Heart; Heart Diseases; Heart Hypertrophy; Heart failure; Herbal Medicine; Housekeeping; Human; Hypertrophy; Infusion procedures; Interleukin-11; Isoproterenol; Knock-out; Knockout Mice; Ligation; Mammals; Mediating; Messenger RNA; Modeling; Modification; Molecular Biology; Molecular Target; Morbidity - disease rate; Mus; Myocardial Infarction; Myocardial Ischemia; Pathologic; Pathology; Pathway interactions; Patients; Pharmacology; Phenotype; Play; Post-Transcriptional Regulation; Process; Proline; Proline-Specific tRNA; Protein Biosynthesis; Proteins; Regulatory Pathway; Role; Signal Transduction; Stimulus; Stress; Testing; Therapeutic Effect; Therapeutic Intervention; Transfer RNA; Transforming Growth Factor beta; Translational Activation; Translational Repression; Translations; United States; Up-Regulation; activating transcription factor; antifibrotic treatment; base; conditional knockout; constriction; coronary fibrosis; druggable target; genome wide association study; glutamyl-prolyl-tRNA synthetase; improved; inhibitor/antagonist; insight; mRNA Decay; mortality; mouse genome; mouse model; new therapeutic target; novel; novel therapeutic intervention; polysome profiling; prevent; preventive intervention; proline-tRNA; screening; targeted treatment; transcriptome sequencing; transcriptomics; translation factor

Project Summary: Cardiovascular disease (CVD) is the leading cause of morbidity and mortality worldwide. Heart failure (HF), a major form of CVD, results from cardiomyocyte (CM) hypertrophy and apoptosis, combined with cardiac fibroblast (CF) activation and proliferation. Although TGF-b-IL-11 axis induced protein synthesis of pro-fibrotic genes have been observed, the mechanisms that promote pro-fibrotic mRNA translation in CF have not been identified. Therefore, filling this gap and identifying the pharmacologically targetable translational control determinants of cardiac fibrosis is of paramount importance. Aminoacyl-tRNA synthetases (ARSs) catalyze the ligation of amino acids to cognate tRNAs. In mammals, glutamyl-prolyl-tRNA synthetase (EPRS) catalyzes the attachment of glutamic acid (E) and proline (P), to their cognate tRNAs for protein synthesis. By screening of ARSs induced in TGF-b- activated human CFs, human ARSs with SNPs in CHD, mouse ARSs associated with ISO-induced cardiomyopathy by GWAS, and ISO-induced ARSs in mouse failing hearts, we have identified EPRS as the key ARS involved in various cardiac pathologies. In this project, we uncovered EPRS as an integrated node downstream of multiple pro-fibrotic stimuli. We found that activating transcription factor 5 acts as an upstream transcriptional regulator of EPRS. Halofuginone (Halo), a Chinese herbal medicine-derived chemical compound, is a prolyl-tRNA synthetase-specific inhibitor. Low-dose Halo, as well as genetic knockout of one allele of EPRS in the mouse genome, reduces cardiac hypertrophy and fibrosis in multiple HF mouse models. We employed RNA-Seq and polysome profiling-Seq in Halo- treated fibroblasts to define gene expression changes at the transcriptomic and translatomic level upon EPRS inhibition. We identified novel proline codon rich (PRR) genes in addition to collagens, which are upregulated by EPRS at the posttranscriptional level and may play critical roles in cardiac fibrosis. Inactivation of EPRS promotes translational repression of PRR mRNA coupled with enhanced mRNA decay. This effect requires inactivation of eIF5A, an elongation factor for decoding Pro-rich codons. Our central hypothesis is: MI stress-induced EPRS promote cardiac fibrosis via increased Pro-tRNAPro pool, and enhanced stabilization and translational activation of pro-fibrotic PRR mRNAs in CFs. We will test this hypothesis by pursuing 3 aims. Aim 1. Determine if downregulation, globally and in CFs, and pharmacological inhibition of EPRS prevent or reverse cardiac fibrosis in HF models. Aim 2. Determine novel preferential downstream PRR pro-fibrotic mRNA targets of EPRS and upstream regulatory pathway of EPRS. Aim 3. Determine mechanisms of eIF5A-mediated translational control and mRNA decay of PRR genes. This project will promote the development of novel therapeutic approaches by inhibiting translation factors, and identification of novel pharmacological targets.

项目摘要： 心血管疾病（CVD）是全球发病率和死亡率的主要原因。心脏衰竭 （HF）是CVD的一种主要形式，是由心肌细胞（CM）肥大和凋亡产生的 心脏成纤维细胞（CF）激活和增殖。尽管TGF-B-IL-11轴诱导蛋白质合成 已经观察到了促纤维化基因，这些机制促进了促纤维化mRNA翻译 在CF中尚未确定。因此，填补这一空白并在药理上识别 心脏纤维化的目标转化控制决定因素至关重要。 氨基酰基-TRNA合成酶（ARSS）催化氨基酸与同源性TRNA的连接。在 哺乳动物，谷氨酸 - 丙酰基-TRNA合成酶（EPRS）催化谷氨酸（E）的附着和 脯氨酸（p），用于蛋白质合成的同源trNA。通过筛选在TGF-B-中诱导的ARSS 激活的人CF，具有CHD的SNP的人类ARS，与ISO诱导相关的小鼠ARSS GWAS的心肌病，以及小鼠心脏失败的ISO诱导的ARS，我们确定EPR是 涉及各种心脏病理的关键AR。在这个项目中，我们发现了EPRS 多个促纤维化刺激的下游集成节点。我们发现激活转录因子 5充当EPRS的上游转录调节器。 halofuginone（Halo），一种中草药 药物衍生的化合物是一种脯氨酰tRNA合成酶特异性抑制剂。低剂量光环，AS 以及小鼠基因组中一个EPR等位基因的遗传敲除，减少心脏肥大和 多种HF小鼠模型中的纤维化。我们在光环中使用了RNA-Seq和多质体分析序列 - 处理的成纤维细胞在转录组和翻译水平上定义基因表达变化 EPR抑制。除了胶原蛋白外，我们还确定了新型脯氨酸密码子（PRR）基因 在转录后水平上被EPRS上调，并可能在心脏纤维化中起关键作用。 EPRS的灭活促进了PRR mRNA的翻译抑制与增强的mRNA 衰变。这种效果需要使EIF5A失活，EIF5A是解码富富密码子的伸长因子。 我们的中心假设是：MI应力诱导的EPRS通过增加的促进性促进心脏纤维化 池，增强了CFS促纤维化PRR mRNA的稳定和翻译激活。我们将 通过追求3个目标来检验这一假设。目标1。确定在全球和CFS上的下调以及 HF模型中EPR的药理抑制预防或反向心脏纤维化。目标2。确定 新型EPRS和上游调节性的新型优先下游PRR促纤维化mRNA靶标 EPRS的途径。目标3。确定EIF5A介导的翻译控制和mRNA的机制 PRR基因的衰变。该项目将通过 抑制翻译因子和新型药理靶标的鉴定。