Modeling myosin mechanobiology towards understanding the mechanisms of hypertrophic cardiomyopathy

模拟肌球蛋白力学生物学以了解肥厚型心肌病的机制

基本信息

批准号：
10906499
负责人：
Alison Schroer Vander Roest
金额：
$ 10.36万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-01 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10906499
关键词：
ATP phosphohydrolase Adhesions Advisory Committees Affect Alleles Area Automobile Driving Award Biological Models Biomechanics Biomedical Engineering CRISPR/Cas technology Cardiac Cardiac Myocytes Categories Cell Adhesion Cell-Cell Adhesion Cells Cellular biology Clinical Clustered Regularly Interspaced Short Palindromic Repeats Complex Computer Models Contracts Cues Development Disease Disease Progression Elements Engineering Environment Event Extracellular Matrix Fibroblasts Fibrosis Fluorescence Resonance Energy Transfer Functional disorder Gene Expression Generations Genes Genotype Goals Health Heart Heart Diseases Heart failure Heritability Heterogeneity Human Hypertrophic Cardiomyopathy Hypertrophy ITGB3 gene In Vitro Induced Mutation Integrins Intercellular Junctions Kinetics Knowledge Lead Link MAP Kinase Gene Measurement Measures Mechanics Mediator Mentors Mentorship Modeling Molecular Molecular Biology Mutation Myofibrils Myofibroblast Myosin ATPase Myosin Heavy Chains Organ Pathologic Pathway interactions Patients Pattern Pharmacotherapy Phase Phenotype Point Mutation Positioning Attribute Production Proteins Regulation Relaxation Research Research Support Role Sarcomeres Severity of illness Signal Pathway Signal Transduction System Technical Expertise Techniques Testing Therapeutic Tissues Traction Force Microscopy Training Universities Validation Variant Vinculin beta-Myosin career career development disease phenotype disease-causing mutation experience extracellular improved in silico in vivo individualized medicine induced pluripotent stem cell induced pluripotent stem cell derived cardiomyocytes innovation insight molecular phenotype mutant new therapeutic target novel research and development response sensor single molecule skill acquisition skills species difference tool transcriptome transcriptome sequencing transcriptomics

ATP phosphohydrolase Adhesions Advisory Committees Affect Alleles Area Automobile Driving Award Biological Models Biomechanics Biomedical Engineering CRISPR/Cas technology Cardiac Cardiac Myocytes Categories Cell Adhesion Cell-Cell Adhesion Cells Cellular biology Clinical Clustered Regularly Interspaced Short Palindromic Repeats Complex Computer Models Contracts Cues Development Disease Disease Progression Elements Engineering Environment Event Extracellular Matrix Fibroblasts Fibrosis Fluorescence Resonance Energy Transfer Functional disorder Gene Expression Generations Genes Genotype Goals Health Heart Heart Diseases Heart failure Heritability Heterogeneity Human Hypertrophic Cardiomyopathy Hypertrophy ITGB3 gene In Vitro Induced Mutation Integrins Intercellular Junctions Kinetics Knowledge Lead Link MAP Kinase Gene Measurement Measures Mechanics Mediator Mentors Mentorship Modeling Molecular Molecular Biology Mutation Myofibrils Myofibroblast Myosin ATPase Myosin Heavy Chains Organ Pathologic Pathway interactions Patients Pattern Pharmacotherapy Phase Phenotype Point Mutation Positioning Attribute Production Proteins Regulation Relaxation Research Research Support Role Sarcomeres Severity of illness Signal Pathway Signal Transduction System Technical Expertise Techniques Testing Therapeutic Tissues Traction Force Microscopy Training Universities Validation Variant Vinculin beta-Myosin career career development disease phenotype disease-causing mutation experience extracellular improved in silico in vivo individualized medicine induced pluripotent stem cell induced pluripotent stem cell derived cardiomyocytes innovation insight molecular phenotype mutant new therapeutic target novel research and development response sensor single molecule skill acquisition skills species difference tool transcriptome transcriptome sequencing transcriptomics

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项目摘要

PROJECT SUMMARY: This proposed project focuses on understanding how mutations in beta cardiac myosin with diverse and often opposing effects on myosin molecular function contribute to similar disease phenotypes of hypertrophic cardiomyopathy (HCM): cardiomyocyte (CM) hypertrophy, hypercontractility, and tissue fibrosis. Understanding disease mechanisms and the heterogeneity of phenotypes across multiple scales (molecular, cellular, and clinical) will enable the development of better and more individualized therapies for patients with HCM. The first aim of this proposal will clarify the mechanisms by which altered intracellular forces affect intracellular signaling and CM hypertrophy. The second aim will use computational modeling to examine how alterations to interrelated parameters of myosin function change force production temporally and spatially. The third aim will define how altered extracellular mechanics affect CM and cardiac fibroblast phenotypes. The proposal uses several innovative molecular, cellular, bioengineering, and computational modeling tools to examine and contextualize the role of altered cellular mechanics in driving changes in cardiac cell phenotypes. CRISPR/Cas9 gene editing in human induced pluripotent stem cells provides a model system to investigate the effects of specific mutations in a dynamic cellular context. Micropatterned engineered platforms will be used to improve myofibril alignment and allow direct measurement of intracellular force production by traction force microscopy. Molecular biology and transcriptomic techniques will be used to measure changes in intracellular signaling and gene expression in response to mechanical perturbation. Another innovation is the novel application of a vinculin tension sensor FRET probe to directly measure intracellular forces at sarcomere Z disks and at cellular adhesions. Together these platforms will allow direct validation of temporal and spatial cellular forces predicted using computational modeling (molecular myosin models and cell-specific finite element models). Finally, investigating the effect of altered extracellular mechanics on CM function and cardiac fibroblast activation in engineered environments will give insights into the effects of fibrotic remodeling. The research and career development training plans will enable the transition of Dr. Vander Roest to an independent career. During the mentored (K99 phase) of this proposal, Dr. Vander Roest will develop technical skills in molecular biomechanics, FRET measurements, and transcriptome analysis, and expand her skills in computational modeling in the context of cardiac disease. This training will take place at Stanford University, under the mentorship of Drs. Bernstein and Spudich, as well as an expert trans-disciplinary advisory committee, including 2 experts in computational modeling (Drs. Regnier and Campbell). The development of these skills will support the research plan for the independent phase of this project, combining new skills and experience gained during this award with Dr. Vander Roest’s past experience in myofibroblast mechanobiology to facilitate a successful transition to independence.

项目摘要：该拟议项目的重点是了解β心脏肌球蛋白中的突变如何潜水员和对肌球蛋白分子功能的经常影响有助于类似的疾病表型肥厚性心肌病（HCM）：心肌细胞（CM）肥大，超收缩性和组织纤维化。了解多个尺度的疾病机制和表型的异质性（分子，细胞和临床）将能够发展出更好，更个性化的疗法适用于HCM的患者。该提案的第一个目的将阐明改变细胞内的机制力影响细胞内信号传导和CM肥大。第二个目标将使用计算建模到检查如何暂时改变肌球蛋白功能的相互关联参数的变化。空间。第三个目标将定义细胞外力学改变如何影响CM和心脏成纤维细胞表型。该提案使用几种创新的分子，细胞，生物工程和计算建模工具来检查和背景化改变的细胞力学在推动心脏变化中的作用细胞表型。人类诱导多能干细胞中的CRISPR/CAS9基因编辑提供了模型系统研究在动态细胞环境中特定突变的影响。微图案工程平台将用于改善肌原纤维排列，并允许通过牵引力显微镜。分子生物学和转录组技术将用于测量变化响应机械扰动的细胞内信号传导和基因表达。另一个创新是杂种蛋白张力传感器FRET探针的新颖应用直接测量肌膜的细胞内力 Z磁盘和细胞粘附。这些平台将允许直接验证临时和空间使用计算建模预测的细胞力（分子肌球蛋白模型和细胞特异性有限元模型）。最后，研究改变细胞外力学对CM功能和心脏成纤维细胞的影响工程环境中的激活将使您深入了解纤维化重塑的影响。研究和职业发展培训计划将使Vander Roest博士过渡到独立职业。在该提案的指导（K99阶段）期间，范德·罗斯特（Vander Roest）博士将发展技术技能分子生物力学，FRET测量和转录组分析，并扩展她的技能心脏病背景下的计算建模。该培训将在斯坦福大学举行在博士的心态下。伯恩斯坦（Bernstein）和斯普迪奇（Spudich）以及专家跨学科咨询委员会，包括2位计算建模专家（Regnier和Campbell博士）。这些技能的发展将支持该项目独立阶段的研究计划，结合了新技能和获得的经验在这个奖项期间，范德·罗斯特（Vander Roest 成功过渡到独立。