Molecular Signaling in Cardiac Sarcomeres

心脏肌节的分子信号传导

• 批准号: 7919144
• 负责人: R John Solaro
• 金额: $ 37.61万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-06-01 至 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7919144
• 关键词: 5' -AMP-activated protein kinase; AMP-activated protein kinase kinase; Admission activity; Aerobic; Affect; Binding; Cardiac; Ceramides; Complement; Complex; Consumption; Coupling; Data; Development; Diagnostic Procedure; Dilated Cardiomyopathy; Disease; Embryo; Energy Supply; Evaluation; Failure; Familial Hypertrophic Cardiomyopathy; Fiber; Financial compensation; Funding; Glycolysis; Heart; Heart Diseases; Heart Hypertrophy; Heart failure; Homeostasis; Hospitals; Hypertrophy; In Situ; Investigation; Link; Metabolic; Metabolic Control; Metabolic Pathway; Metabolism; Microfilaments; Modeling; Modification; Molecular; Mus; Muscle Cells; Mutation; Neonatal; Pathway interactions; Peptides; Phenotype; Phosphorylation; Phosphorylation Site; Phosphotransferases; Post-Translational Protein Processing; Protein Isoforms; Proteins; Proteome; Pump; Reporting; Research; Role; Sarcomeres; Signal Pathway; Signal Transduction; Site; Skin; Sphingolipids; Stress; Testing; Therapeutic; Thin Filament; Troponin; Troponin I; adenylate kinase; base; hemodynamics; link protein; mutant; novel; novel diagnostics; oxidation; p21 activated kinase; programs; protein kinase D; research study; skeletal; sphingosine 1-phosphate; stressor

Our long term objective Project 1 is to understand how signals at the level of the cardiac sarcomere serve to coordinate energy supply and energy consumption. Our aims test the hypothesis that modifications at the level of the sarcomeric proteins serve as significant sites of signal convergence in the progression to heart failure. Our preliminary data indicate a complex coupling, which involves promotion of signaling pathways induced by altered sarcomeric function and that coordinately control energy supply and energy consumption through reciprocol post-translational modifications of sarcomeric proteins. The experiments include investigation of novel findings including: i) differential activation of AMP activated kinase (AMPK) in aerobic conditions in hearts expressing mutant troponin I (cTnl) linked to familial hypertrophic cardiomyopathy (FHC), ii) phosphorylation of cTnl by AMPK; iii) identification of novel cTnl sites of phosphorylation associated with PKCe activation and dilated cardiomyopathy, iii) data predicting metabolically driven sphingolipid signaling to the sarcomeres, and iv) evidence for functionally significant cTnl intra-molecular interactions The specific aims are: Aim #1. To compare the role of AMPK (AMP activated protein kinase) as a signaling mechanism coordinating energy supply and energy consumption in normal hearts and hearts stressed by expression of sarcomeric proteins inducing increases in Ca-sensitivity and FHC. Aim #2. To determine the temporal association of the cardiac phenotype of mice expressing PKCe and demonstrating dilated cardiomyopathy with sarcomeric phosphorylation and whether the phenotype is altered by expression of a non-phosphorylatable mutant Tnl lacking the unique N-terminus. Aim #3. To determine the functional significance of interactions of regions of cTnl with itself and with other thin filament protein sites, potentially significant in coordinating energy demand and supply and modified by AMP kinase (AMPK), protein kinase D (PKD), and PKCe. Approaches to the aims includes studies at the level of the in situ beating heart, isolated myocytes, and skinned fibers with focus on dynamics and evaluation of myofilament Ca-sensitivity and the sarcomere sub-proteome. This project interacts closely with and complements the aims of the other three projects. All three cores strongly support this project. Data generated by the experiments proposed will open a new avenue of research linking metabolic signaling with reciprocal signaling to the sarcomeres, and provide molecular mechanisms of significance in the development of novel diagnostic and therapeutic strategies important in heart failure.

我们的长期目标项目1是了解心脏肉瘤级别的信号如何用于协调能源供应和能源消耗。我们的目的检验了以下假设：在肉瘤蛋白水平上的修饰是信号收敛的重要位点，在进展到心力衰竭中。我们的初步数据表明了一个复杂的耦合，涉及促进由肉瘤功能改变引起的信号通路，并通过对肉瘤蛋白的翻译后修饰进行协调控制能量供应和能量消耗。实验包括对新发现的研究，包括：i）在表达与家族性肥大性心肌病（FHC）相关的心脏表达突变型肌钙蛋白I（CTNL）中，AMP激活激酶（AMPK）的差异激活与CTNL与家族性肥厚性心肌病（FHC）相关的差异； iii）鉴定与PKCE激活和扩张心肌病有关的磷酸化的新型CTNL位点，iii）数据预测肉瘤的代谢驱动的鞘脂信号传导，iv）证据，以及功能上有效的CTNL CTNL ctnl内部内部相互作用的证据。为了将AMPK（AMP激活蛋白激酶）的作用比较作为一种信号传导机制，该机制在正常心脏和心脏中协调能量供应和能量消耗，这是由于肌肉蛋白表达诱导CA-敏感性和FHC的增加而压力的。目标＃2。为了确定表达PKCE的小鼠的心脏表型的时间缔合，并证明了与肉瘤磷酸化的扩张性心肌病，以及是否通过缺乏独特的N-末端的非磷酸化突变体TNL的表达来改变表型。目标＃3。确定功能 CTNL区域与自身以及与其他薄丝蛋白位点的相互作用的重要性，在协调能量需求和供应方面具有显着意义，并由AMP激酶（AMPK）（AMPK），蛋白激酶D（PKD）和PKCE修饰。目的的方法包括对原位跳动心脏，孤立的心肌细胞和皮肤纤维的研究，重点是动力学和肌膜CA-敏感性和肌膜亚蛋白质体的评估。该项目与其他三个项目的目标紧密互动并补充。这三个核心都强烈支持这个项目。提出的实验生成的数据将 开辟了一条新的研究途径，将代谢信号与互惠信号传导与肉瘤联系起来，并为在心力衰竭中重要的新型诊断和治疗策略的发展提供了显着性的分子机制。