Multiscale investigation of cardiomyopathy-associated mutations in metavinculin

美维库林心肌病相关突变的多尺度研究

基本信息

批准号：
10558703
负责人：
Samantha Kirstin Barrick
金额：
$ 7.86万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-02-01 至 2024-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10558703
关键词：
Actins Address Affect Amino Acids Area Attention Binding Biological Assay Biological Process Cardiac Cardiac Myocytes Cardiomyopathies Cell-Matrix Junction Cells Coupling Dependence Development Dilated Cardiomyopathy Disease Education Educational process of instructing Extracellular Matrix Fellowship Generations Heart Heart Diseases Human Hydrogels Impairment Individual Induced Mutation Intercalated disc Intercellular Junctions Investigation Lead Ligands Measurement Measures Mediating Microfilaments Mission Molecular Muscle Muscle Cells Mutation Organizational Change Pathogenicity Pathologic Patients Physiological Play Process Protein Isoforms Proteins RNA Splicing Research Role Sarcomeres Site Stress Tail Testing Traction Traction Force Microscopy Training United States National Institutes of Health Universities VCL gene Ventricular Remodeling Vinculin Washington alpha helix biophysical techniques disease phenotype disease-causing mutation experimental study induced pluripotent stem cell derived cardiomyocytes inherited cardiomyopathy live cell imaging live cell microscopy mechanical force mechanotransduction medical schools mouse model mutant novel therapeutics optic trap optical traps programs research and development single molecule skills stem cells transmission process

项目摘要

Project Summary/Abstract Familial cardiomyopathies are genetic heart diseases that involve ventricular remodeling and altered cardiac contractility. These diseases are often caused by mutations in proteins within the sarcomere, the fundamental contractile unit of cardiomyocytes. Mutations in non-sarcomeric proteins involved in mechanotransduction, the process by which cells sense and respond to mechanical force, have also been implicated in cardiomyopathy but have received considerably less attention. For instance, studies of human patients have identified cardiomyopathy-associated mutations in metavinculin, the muscle-specific isoform of the ubiquitous mechanotransducer vinculin, but how these mutations lead to the disease phenotype is not well-understood. Structural studies have shown that the 68-amino acid insert that differentiates metavinculin from vinculin replaces the first alpha-helix in the actin-binding vinculin tail domain. Although this alpha-helix does not directly bind actin, the metavinculin insert results in drastic changes of the organization of actin filaments by metavinculin compared to vinculin, suggesting an allosteric effect on actin binding. The proposed research will test the hypothesis that the pathogenic mechanism of cardiomyopathy caused by mutations in metavinculin involves disruption of cardiac mechanotransduction through impairment of the force-dependent binding of metavinculin to actin. Single- molecule force measurements of metavinculin binding to actin will directly address whether force stabilizes binding of metavinculin to actin, as has been previously demonstrated for vinculin, as well as the effect of pathogenic mutations on this force-dependent binding. The cellular consequences of altered force dependence of metavinculin-actin binding will be investigated in stem cell-derived cardiomyocytes that carry the disease- causing mutations using live-cell imaging of sarcomerogenesis (the establishment of new sarcomeres) and traction force microscopy. These experiments will also be carried out on metavinculin-null cardiomyocytes to elucidate the role of metavinculin in sarcomerogenesis and cellular contractility. The training provided under this fellowship will take place at the Washington University School of Medicine, a world leader in biomedical education and research. The proposed research aligns with the strategic objectives of the NIH by addressing the normal biological function of metavinculin and the pathobiological mechanism underlying the onset and progression of cardiomyopathy caused by mutations in metavinculin. In addition, the proposed training plan will contribute to the strategic objective of developing a scientific workforce capable of accomplishing the NIH’s mission by supporting the development of research, teaching, and professional skills required for the PI to establish a successful independent research program in the field of cardiac mechanobiology.