Chromatin mechanism of molecular motor gene regulation

分子运动基因调控的染色质机制

基本信息

批准号：
8666043
负责人：
CHING-PIN CHANG
金额：
$ 38.22万
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：
2013
资助国家：
美国
起止时间：
2013-06-01 至 2017-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8666043
关键词：
ATP phosphohydrolase Acetylation Animal Model Cardiac Cardiac Myocytes Cardiomyopathies Chemicals Chromatin Co-Immunoprecipitations CpG dinucleotide DNA DNA Methylation DNA Methyltransferase DNA Modification Methylases DNA methyltransferase inhibition Development Drug Targeting Environment Failure Gene Expression Gene Expression Regulation Gene Silencing Genes Genetic Heart Heart Contractilities Heart Hypertrophy Heart failure Histone Deacetylase Histones Human Hypertrophy Knock-out Left Left ventricular structure Lysine Mammals Methods Methylation Molecular Molecular Biology Molecular Motors Morbidity - disease rate Mus Muscle Contraction Myocardium Myopathy Myosin Heavy Chains Operative Surgical Procedures Outcome Patients Performance Pharmaceutical Preparations Process Protein Isoforms Repression Research Resistance Severities Societies Stress Ventricular Cardiac alpha-Myosin Work base chromatin immunoprecipitation chromatin remodeling design histone methyltransferase improved inhibitor/antagonist insight mortality pressure prevent programs promoter public health relevance scaffold small molecule therapeutic target tool

项目摘要

DESCRIPTION (provided by applicant): Pathological stress induces transcriptional gene reprogramming in the heart muscle, leading to myopathy and heart failure. However, little is known about how the pathological stress triggers the restructuring of chromatin to control myopathic gene expression. This research program focuses on how different chromatin-regulating factors act in concert to establish a repressive chromatin environment to control the expression of alpha-myosin heavy chain (¿-MHC), a molecular motor gene whose repression in the stressed hearts contributes to myopathy and heart failure. Four different chromatin regulators-chromatin remodeler, histone methyltransferase, histone deacetylase and DNA methyltransferase-will be examined for their collaborative repression of ¿-MHC. Because chemical inhibitors are available to inhibit these chromatin factors, it is possible to develop drus to boost ¿- MHC in the failing heart to improve cardiac function. Understanding how these chromatin factors (drug targets) interact will help us design new drugs and understand the synergy/dynamics of potential drugs that inhibit different classes of chromatin regulators. Aim 1: Defining specific isoforms of DNA methyltransferase required for ¿-MHC repression and heart failure. We will knock out DNA methyltransferase isoforms in the heart muscle of mice and examine the effects on ¿-MHC repression in the stressed heart and on the progression of heart failure. Aim 2: Determine how chromatin remodeler integrates histone and DNA methyltransferases to silence ¿-MHC. We will use animal models, surgical method, and various molecular biology methods (chromatin immunoprecipitation and quantitative PCR) to define the interactions between these chromatin factors crucial for the histone and DNA methylation of ¿-MHC. Aim 3: Defining how histone deacetylase coordinates with histone and DNA methyltransferases to repress ¿-MHC promoter. We will use animal models, surgical methods, and various molecular biology methods (chromatin immunoprecipitation, quantitative PCR, and co-immunoprecipitation) to define the interactions between these chromatin factors that are essential for histone methylation and acetylation, as well as DNA methylation of ¿-MHC promoter.

描述（由适用提供）：病理压力诱导心肌的转录基因重编程，导致肌病和心力衰竭。然而，关于病理应力如何触发染色质控制肌病基因表达的摘要知之甚少。该研究计划的重点是不同的染色质调节因子如何共同起作用，以建立反射性染色质环境，以控制α-肌球蛋白重链（�-MHC）的表达，这是一种分子运动基因，其在压力心脏中的表达有助于肌病和心脏失败。四个不同的染色质调节剂 - 染色质重塑剂，组蛋白甲基转移酶，组蛋白脱乙酰基酶和DNA甲基转移酶将检查其协作表达�-MHC的协作表达。由于化学抑制剂可用于抑制这些染色质因子，因此有可能发展DRUs以增强心脏失败的MHC以改善心脏功能。了解这些染色质因子（药物靶标）如何相互作用将有助于我们设计新药，并了解抑制不同类别的染色质调节剂的潜在药物的协同/动力学。 AIM 1：定义`-MHC表达和心力衰竭所需的DNA甲基转移酶的特定同工型。我们将在小鼠心脏肌肉中敲出DNA甲基转移酶同工型，并检查对压力心脏和心力衰竭进展的对„ -MHC抑制的影响。 AIM 2：确定染色质重塑剂如何整合组蛋白和DNA甲基转移酶以沉默�-MHC。我们将使用动物模型，手术方法和各种分子生物学方法（染色质免疫沉淀和定量PCR）来定义这些染色质因子对组蛋白和DNA甲基化至关重要的 - MHC的相互作用。 AIM 3：定义Hisstone脱乙酰基酶如何与组蛋白和DNA甲基转移酶坐标以抑制-MHC启动子。我们将使用动物模型，手术方法和各种分子生物学方法（染色质免疫沉淀，定量PCR和共免疫沉淀）来确定这些染色质因子之间对组蛋白甲基化和乙酰化至关重要的相互作用，以及-MHC促进剂的DNA甲基化。