Genetic Studies of Sarcomere-based Cardiac Diseases

基于肌节的心脏病的遗传学研究

• 批准号: 7837512
• 负责人: Xiaolei Xu
• 金额: $ 24.19万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-15 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7837512
• 关键词: Address; Adult; Alleles; Antisense Technology; Binding; Biological Models; C-terminal; Cardiovascular Diseases; Data; Development; Dilated Cardiomyopathy; Disease; Embryo; Embryonic Development; Exhibits; Exons; Filament; Fishes; Genes; Genetic; Genomics; Goals; Heart Diseases; Homologous Gene; Human; Human Genetics; Hypertrophic Cardiomyopathy; Individual; Investigation; Knock-out; Length; Link; Location; Logic; Mammals; Molecular; Muscle Contraction; Muscular Dystrophies; Mutagenesis; Mutation; Myofibrillogenesis; Myosin ATPase; N-terminal; Pathogenesis; Phenotype; Phosphotransferases; Protein Isoforms; Proteins; RNA Splicing; Research Personnel; Sarcomeres; Series; Signaling Molecule; Striated Muscles; Technology; Testing; Udd-Markesbery Muscular Dystrophy; Zebrafish; base; connectin; genetic analysis; in vivo; knock-down; mutant; novel; programs; research study; titin kinase

DESCRIPTION (provided by applicant): The sarcomere is the basic contractile unit in striated muscle. While mutations in sarcomeric proteins have been linked to various human cardiovascular diseases, it is unclear why mutations in different sarcomeric genes or even different mutations in the same gene result in distinct diseases. The goal of this proposal is to use titin as a paradigm to investigate molecular mechanisms of sarcomere assembly and sarcomeric diseases. We have identified six titin mutants in zebrafish from mutagenesis screens and demonstrated that phenotypes from one mutant allele resemble human dilated cardiomyopathy. We recently identified the full-length genomic sequence for zebrafish titin (tinl) and cloned the other titin homologue (tin2) in zebrafish. Our preliminary data support the central hypothesis of this proposal, which predicts that different domains of Titin participate in distinct steps of myofibrillogenesis disruption of which results in different sarcomeric diseases. In our Specific Aim 1, we will investigate the functional divergence of tinl and tin2. By leveraging morpholino technology, we will test the hypothesis that different isoforms of titin have different functions in myofibrillogenesis. In our Specific Aim 2, we will test the hypothesis that different domains of titin have different functions in myofibrillogenesis. We will generate a series of truncations of both tinl and tin2 in vivo to reveal the functions of the C-terminal domains, including the kinase domain. We will also investigate the function of a Titin N-terminal domain by knocking out a titin interacting protein, Tcap. In our Specific Aim3, we will determine the pathological consequences of different titin mutations in zebrafish. We will first identify the location of mutations in all six titin mutants and then characterize the pathological phenotypes in both embryonic and adult fish. By generating and characterizing a series of titin mutations, these experiments will deepen our understanding of why mutations in different titin exons result in distinct structural changes of the sarcomere and different phenotypes of sarcomeric diseases, including dilated cardiomyopathy, hypertrophic cardiomyopathy, and muscular dystrophy.

描述（由申请人提供）：肌节是横纹肌的基本收缩单位。虽然肌节蛋白的突变与各种人类心血管疾病有关，但尚不清楚为什么不同肌节基因的突变或甚至同一基因的不同突变会导致不同的疾病。本研究的目的是以肌联蛋白为研究范式，探讨肌节组装和肌节疾病的分子机制。我们已经确定了6个肌联蛋白突变体在斑马鱼的诱变筛选，并证明从一个突变等位基因的表型类似于人类扩张型心肌病。我们最近鉴定了斑马鱼肌联蛋白（tin 1）的全长基因组序列，并克隆了斑马鱼的另一个肌联蛋白同源物（tin 2）。我们的初步数据支持这一提议的中心假设，该假设预测肌联蛋白的不同结构域参与肌原纤维生成的不同步骤，从而导致不同的肌节疾病。在我们的具体目标1中，我们将研究tin 1和tin 2的功能分歧。通过利用吗啉技术，我们将测试的假设，不同亚型的肌联蛋白有不同的功能，在肌原纤维形成。在我们的具体目标2中，我们将测试肌联蛋白的不同结构域在肌原纤维发生中具有不同功能的假设。我们将在体内产生一系列tin 1和tin 2的截短，以揭示C-末端结构域的功能，包括激酶结构域。我们还将通过敲除肌联蛋白相互作用蛋白Tcap来研究肌联蛋白N-末端结构域的功能。在我们的特定目标3中，我们将确定斑马鱼中不同肌联蛋白突变的病理后果。我们将首先确定所有六个肌联蛋白突变体的突变位置，然后在胚胎和成鱼的病理表型的特点。通过产生和表征一系列肌联蛋白突变，这些实验将加深我们对为什么不同肌联蛋白外显子中的突变导致肌节的不同结构变化和肌节疾病（包括扩张型心肌病、肥厚型心肌病和肌营养不良症）的不同表型的理解。