Chromatin mechanism of molecular motor gene regulation

分子运动基因调控的染色质机制

• 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
• 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

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（由申请人提供）：病理应激诱导心肌转录基因重编程，导致肌病和心力衰竭。然而，人们对病理应激如何触发染色质重组以控制肌病基因表达知之甚少。该研究项目重点关注不同的染色质调节因子如何协同作用，建立抑制性染色质环境，以控制 α-肌球蛋白重链 (¿-MHC) 的表达，α-肌球蛋白重链 (¿-MHC) 是一种分子运动基因，其在应激心脏中的抑制会导致肌病和心力衰竭。将检查四种不同的染色质调节因子（染色质重塑酶、组蛋白甲基转移酶、组蛋白脱乙酰酶和 DNA 甲基转移酶）对 ¿-MHC 的协同抑制。由于化学抑制剂可抑制这些染色质因子，因此有可能开发出细胞膜来增强衰竭心脏中的 ¿- MHC，从而改善心脏功能。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​​染色质调节剂的潜在药物的协同作用/动力学。目标 1：定义 ¿-MHC 抑制和心力衰竭所需的 DNA 甲基转移酶的特定亚型。我们将敲除小鼠心肌中的 DNA 甲基转移酶亚型，并检查其对应激心脏中 ¿-MHC 抑制以及心力衰竭进展的影响。目标 2：确定染色质重塑剂如何整合组蛋白和 DNA 甲基转移酶来沉默 ¿-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）来定义这些对 ¿-MHC 的组蛋白和 DNA 甲基化至关重要的染色质因子之间的相互作用。目标 3：确定组蛋白脱乙酰酶如何与组蛋白和 DNA 甲基转移酶协调来抑制 ¿-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 和免疫共沉淀）来定义这些染色质因子之间的相互作用，这些因子对于组蛋白甲基化和乙酰化以及 ¿-MHC 启动子的 DNA 甲基化至关重要。