Probing the Molecular Mechanisms of Diastolic Dysfunction Using Patient-Specific Stem Cells

利用患者特异性干细胞探讨舒张功能障碍的分子机制

• 批准号: 10739782
• 负责人: David Wells Staudt
• 金额: $ 16.6万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-11 至 2028-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10739782
• 关键词: Age Months; Award; Biochemical; Biology; Biophysics; CRISPR interference; CRISPR/Cas technology; Calcium; Cardiac Myocytes; Cardiac Myosins; Cardiomyopathies; Cardiovascular system; Cell Line; Cell model; Cells; Childhood; Clinical; Collaborations; Complex; DNA Sequence Alteration; Data; Data Set; Defect; Diagnosis; Disease; Disease model; Functional disorder; Funding; Genetic; Genetic Diseases; Genomic approach; Goals; Grant; Heart; Heart Diseases; Heart Transplantation; Heart failure; Human; Hyperactivity; Hypertrophic Cardiomyopathy; Impairment; Individual; Laboratories; Leadership; Link; Measurement; Measures; Mediating; Medical; Mentors; Mentorship; MicroRNAs; Molecular; Molecular Probes; Morbidity - disease rate; Motor; Muscle; Mutation; Myocardium; Myosin ATPase; Outcome; Pathogenicity; Pathway interactions; Patients; Pharmaceutical Preparations; Phenotype; Physicians; Physiological; Physiology; Productivity; Program Description; Proteins; Rare Diseases; Relaxation; Research Personnel; Resources; Rest; Restrictive Cardiomyopathy; Scientist; Symptoms; Testing; Therapeutic; Thick; Training; Training Programs; Translating; Troponin; Troponin T; Ventricular; Work; Writing; biophysical analysis; career; de novo mutation; design; disease-causing mutation; experience; functional genomics; high throughput analysis; high throughput screening; improved; in vitro Model; individualized medicine; induced pluripotent stem cell; induced pluripotent stem cell derived cardiomyocytes; insight; knock-down; member; mortality; mutant; novel; novel therapeutics; pediatric patients; skills; stem; stem cell model; stem cells; therapeutic target; therapy development; treatment strategy; ventricular hypertrophy

PROJECT SUMMARY This comprehensive training program described in this five-year proposal is designed to prepare Dr. Staudt to transition to a career as a productive independent investigator focused on deciphering the molecular mechanisms underlying diastolic heart disease. Over the course of the described studies, Dr. Staudt will expand his skills in induced pluripotent stem cell-derived cardiomyocyte phenotyping at the single-cell level, as well as acquire new skills in biophysical analysis of pathogenic myosin mutations and high-throughput assay design and analysis. Further, he will obtain substantial experience and training in mentorship, scientific presentation, and grant writing. These skills will be crucial for his successful transition to independence. This proposal assembles an impressive team of world-renowned experts in cardiovascular biology to guide him, including his co-mentors, Drs. Mark Mercola and Euan Ashley, as well as the members of his mentorship committee, Drs. Joseph Wu, Marlene Rabinovich, and Don Bers. Their support will provide the resources and mentorship that Dr. Staudt needs to succeed in an independent academic career at the conclusion of the K08 award. The scientific aim of this project is to uncover mechanisms leading to diastolic dysfunction, an abnormal stiffness of the ventricular muscle that contributes to nearly half of heart failure cases. Specifically, this proposal focuses on Pediatric Restrictive Cardiomyopathy (RCM), a severe genetic disorder characterized by isolated, profound diastolic dysfunction. Patients with this disease have few treatment options, stemming from a relative lack of understanding of the molecular mechanisms underlying this disease. This proposal leverages novel stem cell models of RCM combined with high-throughput measurement of diastolic function to probe these mechanisms. In Aim 1, multiple cellular models of RCM will be characterized and compared to determine whether different classes of RCM mutations act via similar or divergent mechanisms. Aim 2 uses a functional genomics approach to determine whether different mutations evoke distinct pathogenic mechanisms that converge on a similar clinical presentation. Aim 3 focuses on a unique line from a patient with severe, pediatric onset RCM caused by a de novo mutation in cardiac myosin. In this aim, a novel multi-scale approach links the biophysical effects of this RCM mutation on myosin molecules to the physiologic changes in whole cells, and compares this to a comparable, previously characterized myosin mutation that causes Hypertrophic Cardiomyopathy, a more common but generally less severe disease.

项目摘要 该五年提案中描述的全面培训计划旨在为Staudt博士做好准备 过渡到职业为生产性独立研究者，专注于破译分子 舒张性心脏病的基础机制。在描述的研究过程中，Staudt博士将扩大 他在单细胞水平的诱导多能干细胞衍生的心肌细胞表型中的技能以及 获得致病性肌球蛋白突变和高通量测定设计的生物物理分析的新技能 分析。此外，他将获得指导，科学演示和培训的丰富经验和培训 授予写作。这些技能对于他成功过渡到独立至关重要。该建议组装 一支令人印象深刻的世界知名专家心血管生物学专家的团队，包括他的联合官员，包括他的联合官员 博士。 Mark Mercola和Euan Ashley以及他的指导委员会成员Drs。约瑟夫·吴， 玛琳·拉比诺维奇（Marlene Rabinovich）和唐·贝尔斯（Don Bers）。他们的支持将为Staudt博士提供资源和指导 需要在K08奖的结论中取得独立学术职业的成功。 该项目的科学目的是发现导致舒张功能障碍的机制，这是异常刚度 心室肌肉的近一半是心力衰竭病例。具体而言，该提议重点 关于儿科限制性心肌病（RCM），一种严重的遗传疾病，其特征是孤立的，深刻的 舒张功能障碍。患有这种疾病的患者几乎没有治疗选择，这是由于相对缺乏 了解该疾病的分子机制。该建议利用新型干细胞 RCM的模型与舒张功能的高通量测量相结合，以探测这些机制。 在AIM 1中，将对RCM的多个蜂窝模型进行表征，并比较确定是否不同 RCM突变类别通过类似或不同的机制作用。 AIM 2使用功能基因组学方法 确定不同的突变是否引起在类似的融合的不同的致病机制 临床表现。 AIM 3专注于由严重，儿科发病RCM的患者的独特线条 心脏肌球蛋白中的从头突变。在此目标中，一种新颖的多尺度方法将 肌球蛋白分子对整个细胞的生理变化的RCM突变，并将其与A进行比较 可比较的，先前描述的肌球蛋白突变会导致肥厚性心肌病，更多 常见但通常不那么严重的疾病。