The role of the C3 domain of myosin binding protein-C in the pathogenesis of hypertrophic cardiomyopathy

肌球蛋白结合蛋白-C C3结构域在肥厚型心肌病发病机制中的作用

• 批准号: 9232907
• 负责人: Dan F Smelter
• 金额: $ 3.1万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-03-01 至 2019-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9232907
• 关键词: Actins; Adrenergic Agents; Affect; Affinity; Amino Acid Substitution; Amino Acids; Binding; Binding Proteins; Biological Assay; Biomedical Research; C-terminal; C10; Calcium; Cardiac; Cardiac Myocytes; Cardiac Myosins; Cardiovascular Diseases; Charge; Complement 3c; Contractile Proteins; Development; Disease; Disease Progression; Engineering; Environment; Face; Familial Hypertrophic Cardiomyopathy; Gene Transfer; Head; Health; Heart; Heart Diseases; Human; Hypertrophic Cardiomyopathy; Immunoprecipitation; Individual; Inherited; Laboratories; Lead; Ligands; Light; Link; Location; Messenger RNA; Missense Mutation; Molecular; Mus; Mutation; Myosin ATPase; N-terminal; Neonatal; Pathogenesis; Pathogenicity; Pathologic; Peptide Sequence Determination; Phenotype; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphotransferases; Physiological; Primary Myocardial Diseases; Property; Protein Binding Domain; Protein Region; Proteins; Proteomics; Regulation; Reporting; Research; Research Proposals; Role; Sarcomeres; Scientist; Structure; Surface; Surface Plasmon Resonance; Testing; Thick Filament; Tissues; Training Programs; Transcript; Universities; Vertebral column; Wisconsin; adenoviral-mediated; citrate carrier; genetic regulatory protein; heart dimension/size; heart function; insight; interest; mutant; myosin-binding protein C; novel; pre-doctoral; prevent; protein expression; protein protein interaction; public health relevance; restraint; sudden cardiac death; treatment strategy

 DESCRIPTION (provided by applicant): Cardiac myosin binding protein C (cMyBP-C) is a regulatory protein of the cardiac sarcomere that contributes to contractile reserve. Mutations in cMyBP-C are a leading cause of hypertrophic cardiomyopathy (HCM), a disease that affects about 1 in 500 people worldwide. Truncation mutations in cMyBP-C cause HCM primarily through haploinsufficiency; however, the pathogenesis of disease-causing missense mutations is not well understood. The C-terminal domains are essential for incorporation into the sarcomere, while the N-terminal domains modulate contraction through phosphorylation-dependent inhibitory interactions. The role of the central domains in the physiologic function of cMyBP-C is unclear. The R495Q and R502W mutations in the C3 domain of cMyBP-C are among the most prevalent causes of HCM, however exactly how they affect function and contribute to the pathologic phenotype remains unknown. Studies suggest that mutations in this region may disrupt a putative protein binding domain. Given their location in the C3 domain we hypothesize that the R495Q and R502W mutations modulate contractility by modifying an interaction between domain C3 and an as-yet undefined ligand. In this pre-doctoral proposal, we will express mutant R495Q and R502W cMyBP-C in murine three-dimensional cardiac tissue constructs followed by assessment of contractility, calcium handling, calcium sensitivity and responsiveness to beta-adrenergic stimulation. To identify the mechanism linking these mutations to HCM, we will use contemporary proteomics approaches to define the function of the C3 domain and assess whether these mutations alter putative protein-protein interactions. Determining the function of the C3 domain will provide insight into the overall function of cMyBP-C, the pathogenesis of HCM, and may identify novel treatment strategies to prevent or modify disease progression. This pre-doctoral research proposal is part of a comprehensive training program which takes full advantage of the strong research environment at the University of Wisconsin-Madison to enhance the candidate's likely transition to a successful biomedical research scientist.

 描述（由适用提供）：心脏肌球蛋白结合蛋白C（CMYBP-C）是心脏肌膜的调节蛋白，有助于收缩储备。 CMYBP-C中的突变是肥厚性心肌病（HCM）的主要原因，这种疾病影响了全球500人中的大约1人。 CMYBP-C中的截短突变导致HCM原发性通过单倍体不足；但是，尚不清楚引起疾病的错义突变的发病机理。 C末端域对于肌膜的保险至关重要，而N末端结构域通过磷酸化依赖性抑制性相互作用调节收缩。中心领域在CMYBP-C的生理功能中的作用尚不清楚。 CMYBP-C的C3结构域中的R495Q和R502W突变是HCM最普遍的原因之一，但是它们准确地影响功能并有助于病理表型仍然未知。研究表明，该区域的突变可能会破坏推定的蛋白结合结构域。鉴于它们在C3域中的位置，我们假设R495Q和R502W突变通过修改域C3与尚未定义的配体之间的相互作用来调节收缩力。在此博士前建议中，我们将在鼠三维心脏组织构建体中表达突变体R495Q和R502W CMYBP-C，然后评估收缩力，钙处理，钙敏感性和对β-肾上腺素能刺激的反应性。为了确定将这些突变与HCM联系起来的机制，我们将使用当代蛋白质组学方法来定义C3结构域的功能，并评估这些突变是否改变了假定的蛋白质蛋白质相互作用。确定C3结构域的功能将洞悉CMYBP-C，HCM发病机理的整体功能，并可能确定预防或改变疾病进展的新型治疗策略。这项博士前研究建议是一项综合培训计划的一部分，该计划充分利用了威斯康星大学麦迪逊分校的强大研究环境，以增强候选人可能过渡到成功的生物医学研究科学家。