Deciphering Myofilament Modifications in Ischemic Cardiomyopathy

破译缺血性心肌病中的肌丝修饰

• 批准号: 8605545
• 负责人: Ying Ge
• 金额: $ 36.87万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-03-01 至 2017-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8605545
• 关键词: Address; Biological Markers; Cardiac; Cardiac Muscle Contraction; Cardiomyopathies; Chronic; Clinical; Complement; Congestive Heart Failure; Contractile Proteins; Country; Detection; Development; Diagnostic; Disease; Disease Progression; Early Diagnosis; Early treatment; Epidemic; Etiology; Failure; Family suidae; Functional disorder; Heart; Heart Diseases; Heart failure; Human; Indium; Isometric Exercise; Knowledge; Left; Left Ventricular Remodeling; Link; Mass Spectrum Analysis; Measures; Mechanics; Mediating; Microfilaments; Modeling; Modification; Molecular; Monitor; Morbidity - disease rate; Muscle Cells; Muscle relaxation phase; Myocardial; Myocardial Infarction; Myocardial Ischemia; Myocardium; Outcome; Patients; Phosphorylation; Play; Post-Translational Protein Processing; Preparation; Prognostic Marker; Property; Protein Isoforms; Proteins; Proteomics; RNA Splicing; Recombinants; Regulation; Relaxation; Reproducibility; Research; Role; Sampling; Sarcomeres; Signal Transduction; Site; Skin; Staging; Technology; United States; Variant; Ventricular; acute coronary syndrome; base; clinically relevant; improved; in vivo; innovation; insight; mortality; mutant; new therapeutic target; novel; outcome forecast; protein expression; public health relevance; research study; stoichiometry; success; tandem mass spectrometry; tool

DESCRIPTION (provided by applicant): Ischemic cardiomyopathy (ICM) is the most prevalent type of chronic heart diseases in the United States and a common underlying etiology of congestive heart failure. Nonetheless the underlying molecular mechanism of ICM remains unclear. Moreover, the prognosis of chronic ICM is very poor if left untreated. Therefore it is highly important to gain a better understanding of the disease mechanism and to identify specific biomarkers for early detection and treatment of ICM. Myofilament proteins in the sarcomeres not only play essential roles in cardiac contractility but also are critical elements in signal reception and transduction during the onset and progression to heart failure. The hypothesis is that post-myocardial infarction (MI) maladaptive cardiac remodeling can result in altered protein modifications in myofilaments that are associated with cardiac dysfunction and offer potential diagnostic and prognostic biomarkers of ICM. Although multiple myofilament post-translational modifications (PTMs) are believed to act in concert in regulating cardiac function, a comprehensive analysis of myofilament proteins simultaneously and assess all synergistic PTM changes related to cardiac function is lacking. Herein, we will employ an innovative integrated "top-down" protein mass spectrometry (MS)-based disease proteomics platform to simultaneously examine myofilament proteins extracted from healthy and diseased swine and human myocardium and identify all disease-related changes in myofilament modifications. Aim 1 will identify altered protein modifications in myofilaments from diseased swine myocardium of post-MI ICM model and determine their functional consequences. Aim 2 will determine myofilament alterations in end-stage failing human myocardium from ICM patients and their functional consequences. Aim 3 will assess the functional effects of novel myofilament modifications in Ca2+-mediated cardiac muscle contraction and relaxation. Mechanical properties (the isometric force and the Ca2+-sensitivity of the force) will be measured in skinned myocardial preparations in parallel to proteomics experiments. Furthermore, we will assess regional systolic and diastolic function to establish mechanistic links between specific proteomic changes and in vivo and ex vivo cardiac function. With the completion of this project, we expect to identify multiple functionally significant alterations in myofilaments and understand how these alterations act in concert to modulate maladaptive signaling during post-MI left ventricular remodeling to failure. The success of our research will provide new insights into the mechanisms underlying ICM and will identify new candidate biomarkers for early detection of the presence and progression of ICM.

描述(由申请人提供)：缺血性心肌病(ICM)是美国最常见的慢性心脏病类型，也是充血性心力衰竭的常见潜在病因。然而，ICM的潜在分子机制仍不清楚。此外，如果不进行治疗，慢性ICM的预后非常差。因此，更好地了解ICM的发病机制，识别特异性的生物标志物，对ICM的早期发现和治疗具有重要意义。肌节中的肌丝蛋白不仅在心脏收缩能力中起重要作用，而且也是 心力衰竭发病和进展过程中的信号接收和转导。该假说认为，心肌梗死(MI)后适应性不良的心脏重塑可导致与心功能障碍相关的肌丝蛋白修饰改变，并为ICM提供潜在的诊断和预后生物标志物。尽管多个肌丝翻译后修饰(PTM)被认为在调节心脏功能方面起协同作用，但 同时对肌丝蛋白进行全面分析，并评估与心功能有关的所有PTM变化的协同性，这一点尚缺乏。在此，我们将采用一个创新的基于“自上而下”的蛋白质质谱仪(MS)的疾病蛋白质组学平台，同时检测从健康和患病的猪和人类心肌中提取的肌丝蛋白，并识别肌丝修饰中所有与疾病相关的变化。目的1鉴定心肌梗死后ICM模型中病变猪心肌肌丝的蛋白修饰，并确定其功能后果。目的2将确定来自ICM患者的终末期衰竭人类心肌的肌丝变化及其功能后果。目的3将评估新的肌丝修饰在钙离子介导的心肌收缩和松弛中的功能效应。机械特性(等长力和力的钙敏感性)将在蛋白质组学实验的同时在带皮心肌标本中进行测量。此外，我们将评估局部收缩和舒缩功能，以建立特定蛋白质组变化与体内和体外心脏功能之间的机制联系。随着这个项目的完成，我们希望确定肌丝的多种功能显著变化，并了解这些变化如何协同作用，在心肌梗死后左室重构失败期间调节不良适应信号。我们研究的成功将为ICM的机制提供新的见解，并将为早期发现ICM的存在和进展寻找新的候选生物标志物。