LncRNA mechanism of heart failure

LncRNA心力衰竭机制

基本信息

批准号：
9037706
负责人：
CHING-PIN CHANG
金额：
$ 40.15万
依托单位：
INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-04-01 至 2018-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9037706
关键词：
Adult Animals Antisense RNA Biochemical Biological Assay Cardiac Cardiac Myocytes Cardiomyopathies Cell Culture Techniques Chemicals Chromatin Chromatin Remodeling Factor Code Complex Data Development Electrophoretic Mobility Shift Assay Episome FDA approved Failure Feedback Fetal Heart Fibrosis Foundations Gene Expression Gene Expression Regulation Gene Targeting Genes Genetic Genetic Models Health Heart Heart Hypertrophy Heart failure Homeostasis Human Hypertrophy Immunoprecipitation Knock-out Methods Modeling Molecular Molecular Genetics Morbidity - disease rate Mus Myocardium Myopathy Myosin Heavy Chains Neonatal Nucleosomes Nucleotides Outcome Pathogenesis Pharmacotherapy Proteins RNA RNA Splicing Regulation Reporter Repression Role SMARCA4 gene Societies Stress Switch Genes Technology Tissues Transcript Transgenic Organisms Untranslated RNA Western Blotting Work Yeasts arm base chromatin immunoprecipitation chromatin remodeling design epigenetic regulation heart cell heart function human disease in vivo induced pluripotent stem cell insight mortality pressure prevent programs promoter protein complex restoration success

项目摘要

DESCRIPTION (provided by applicant): Pathological stress induces transcriptional gene reprogramming in the heart muscle, leading to cardiac myopathy and heart failure. Such stress-induced gene reprogramming requires regulation at the chromatin level by chromatin-regulating factors. The activity of chromatin regulators can be modulated by an emerging class of RNAs-the long noncoding RNAs (lncRNAs), which are long RNA transcripts with low/no protein-coding potential. The role of lncRNAs in hypertrophy and heart failure, however, is essentially unknown. Most studies of lncRNAs are conducted in cell cultures or in yeast, and mouse genetic models are lacking. This program will focus on a new mouse genetic model to define the molecular function of a newly identified lncRNA in controlling cardiac gene expression, hypertrophy, and failure. Because 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 for heart failure. Aim 1: Defining the in vivo regulation of an lncRNA in the heart. We will use transgenic and knockout technology of mouse genetics to define the genetic and molecular mechanisms by which this lncRNA expression is regulated in the heart. Methods also include immunostaining, western blot, quantitative PCR, chromatin immunoprecipitation (ChIP), and RNA immunoprecipitation (RNA-IP) analyses. Aim 2: Determining how this lncRNA interacts with chromatin-regulating factor in the heart. We will use molecular, cellular, and biochemical methods to determine the molecular mechanism by which the lncRNA controls the molecular functions of chromatin regulators in the hearts. The methods include RNA-IP, ChIP, quantitative PCR, electric mobility shift assays, episome-based reporter assays, and nucleosome assembly. Aim 3: Defining the function of human lncRNA-protein complex. We will use human heart tissues and iPS-derived heart cells to demonstrate the presence of lncRNA complex. Also, we will use molecular, cellular, and biochemical methods to define the function of human lncRNA complex. The methods include RNA-IP, ChIP analysis of small amount of tissues, quantitative PCR, electric mobility shift assays, episome-based reporter assays, and iPS-based technology.

描述（由申请人提供）：病理压力诱导心肌的转录基因重编程，导致心脏肌病和心力衰竭。这种应激诱导的基因重编程需要通过染色质调节因子在染色质水平上调节。染色质调节剂的活性可以通过新兴类别的RNA类别的非编码RNA（LNCRNA）来调节，后者是具有低/无蛋白质编码势的长RNA转录本。但是，LNCRNA在肥大和心力衰竭中的作用本质上是未知的。 LNCRNA的大多数研究都是在细胞培养物或酵母中进行的，并且缺乏小鼠遗传模型。该程序将集中在新的小鼠遗传模型上，以定义新鉴定的LNCRNA在控制心脏基因表达，肥大和衰竭中的分子功能。由于RNA可以化学修饰并作为治疗药物进行化学修饰，因此该计划的成功将为设计基于机制的心力衰竭治疗而奠定基础。目标1：定义心脏中lncRNA的体内调节。我们将使用小鼠遗传学的转基因和基因敲除技术来定义该LNCRNA表达在心脏中调节的遗传和分子机制。方法还包括免疫染色，蛋白质印迹，定量PCR，染色质免疫沉淀（CHIP）和RNA免疫沉淀（RNA-IP）分析。目标2：确定该LNCRNA如何与心脏中染色质调节因子相互作用。我们将使用分子，细胞和生化方法来确定lncRNA控制心脏中染色质蛋白蛋白调节剂的分子功能的分子机制。这些方法包括RNA-IP，芯片，定量PCR，电动迁移率分析，基于沿激体的报告基因分析和核小体组装。目标3：定义人lncRNA-蛋白质复合物的功能。我们将使用人类心脏组织和IPS衍生的心脏细胞来证明LNCRNA复合物的存在。此外，我们将使用分子，细胞和生化方法来定义人lncRNA复合物的功能。这些方法包括RNA-IP，少量组织的芯片分析，定量PCR，电动迁移率分析，基于ISCONOME的记者测定和基于IPS的技术。