LncRNA mechanism of heart failure

LncRNA心力衰竭机制

基本信息

批准号：
10021030
负责人：
CHING-PIN CHANG
金额：
$ 54.92万
依托单位：
ALBERT EINSTEIN COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-06-15 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10021030
关键词：
3-Dimensional Adult Binding Biological Assay Biology Cardiac Cardiac Myocytes Chemicals Chromatin Chromatin Loop Clustered Regularly Interspaced Short Palindromic Repeats Code DNA Development Disease Outcome Echocardiography Electrophoretic Mobility Shift Assay Elements Enhancers Failure Fetal Heart Foundations Fractionation Functional disorder Gene Expression Gene Expression Regulation Genes Genetic Models Genetic Transcription Genome Heart Heart Hypertrophy Heart failure Histopathology Human Hybrids Hypertrophy Immunoprecipitation In Vitro Knock-in Knock-out Knockout Mice Mediating Methods Modeling Modification Molecular Molecular Conformation Morbidity - disease rate Mus Muscle Development Myocardium Myopathy Myosin Heavy Chains Names Pathogenesis Pathologic Pharmacotherapy Protein Isoforms Proteins RNA Research Role SMARCA4 gene Signal Transduction Societies Stress Structure Technology Therapeutic Tissue-Specific Gene Expression Tissues Transcript Transcriptional Regulation Transgenic Organisms Translations Treatment Failure Untranslated RNA Work base biological adaptation to stress cardioprotection chromatin remodeling chromosome conformation capture constriction design fetal frontier helicase human disease in vivo induced pluripotent stem cell insight molecular marker mortality mouse genetics muscle regeneration novel pressure programs promoter response ribosome profiling structural genomics success

项目摘要

Project Summary The genome contains numerous non-coding DNA and RNA elements that far outnumber protein- coding genes, presenting a drastically new perspective of the transcriptional circuits. Among noncoding DNA elements, enhancers are the DNA fragments that activate target gene promoter by acting on gene promoters, independent of their distance or orientation to target genes. Many enhancers are recently found to encode noncoding RNAs, giving rise to an emerging class of enhancer RNAs (eRNAs). The eRNA mechanisms of heart failure, particularly at its interface with eDNA and chromatin, are largely unknown. We recently discovered a cardiac-specific eRNA capable of protecting the heart against pathological hypertrophy and failure. This program will focus on new mouse genetic models to define the molecular function of this newly identified eRNA in controlling enhancer function, 3-dimensional chromatin looping, cardiac gene expression, and pathological hypertrophy. Given that RNAs can be chemically modified and delivered as a drug for therapy, the success of this program will lay down a foundation for designing new mechanism-based therapy. Aim 1: Determining the in vivo sufficiency of an eRNA in cardioprotection. We will use transgenic and knock in/out technology of mouse genetics to define the cardioprotective role of this eRNA. Methods include CRISPR-enabled inducible and cardiomyocyte-specific eRNA modifications, transaortic constriction, histopathology, echocardiography, pressure-volume loop, and molecular marker studies. Aim 2: Defining how an eRNA interacts with its cognate eDNA in 3-dimensional chromatin looping and gene regulation. We will determine molecular interactions between an eRNA and its cognate eDNA in forming eRNA-eDNA hybrid duplex and in chromatin looping to target genes for transcription regulation under different pathophysiological conditions. Methods include mouse genetics, chromatin conformation capture, quantitative PCR, helicase assay, electric mobility shift assays, and DNA-RNA duplex immunoprecipitation. Aim 3: Defining eRNA-eDNA interactions in normal and diseased human hearts. We will use human heart tissues and iPS-derived cardiomyocytes to define the evolutionary conservation of eRNA-eDNA interactions and eDNA three-dimensional chromatin looping to target genes. Methods include ribosome profiling, in vitro transcription and translation, qPCR, chromatin fractionation and conformation capture, DNA-RNA duplex immunoprecipitation, and iPS-based technology.

项目摘要基因组包含许多非编码DNA和RNA元素，这些元素远远超过了蛋白质编码基因，对转录电路的新观点产生了巨大的新观点。之中非编码DNA元件，增强子是通过通过作用于基因启动子，与目标基因的距离或方向无关。许多最近发现增强剂编码非编码RNA，从而产生新兴类别增强剂RNA（ERNAS）。心力衰竭的ERNA机制，尤其是在与埃德娜（Edna）和染色质在很大程度上未知。我们最近发现了心脏特异性的Erna 保护心脏免受病理肥大和衰竭的影响。该计划将重点放在新小鼠遗传模型来定义该新鉴定的ERNA的分子功能增强子功能，三维染色质循环，心脏基因表达和病理肥大。鉴于RNA可以化学修饰并作为治疗药物递送该计划的成功将为设计新的基于机制的治疗而奠定基础。 AIM 1：确定心脏保护中ERNA的体内充足性。我们将使用转基因和敲入/淘汰小鼠遗传学的技术来定义ERNA的心脏保护作用。方法包括启用CRISPR的诱导型和心肌细胞特异性ERNA修饰收缩，组织病理学，超声心动图，压力量环和分子标记研究。 AIM 2：定义ERNA如何与3维染色质循环中的同源EDNA相互作用和基因调节。我们将确定Erna及其同源之间的分子相互作用 EDNA形成ERNA-EDNA杂交双链体和染色质循环至靶基因进行转录在不同的病理生理条件下进行调节。方法包括小鼠遗传学，染色质构象捕获，定量PCR，解旋酶测定，电动迁移率分析和DNA-RNA 双链免疫沉淀。目标3：定义正常和患病的人类心脏中的Erna-Edna相互作用。我们将使用人类心脏组织和IPS衍生的心肌细胞定义ERNA-EDNA的进化保守相互作用和EDNA三维染色质与靶基因循环。方法包括核糖体分析，体外转录和翻译，qPCR，染色质分馏和构象捕获，DNA-RNA双链免疫沉淀和基于IPS的技术。