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.

项目概要 此五年提案中描述的全面培训计划旨在帮助 Staudt 博士做好准备 转型为一名富有成效的独立研究者，专注于破译分子 舒张性心脏病的潜在机制。在所描述的研究过程中，Staudt 博士将扩展 他在单细胞水平上诱导多能干细胞衍生的心肌细胞表型分析方面的技能，以及 获得致病性肌球蛋白突变的生物物理分析和高通量测定设计的新技能 分析。此外，他还将在指导、科学演示和培训方面获得丰富的经验和培训。 授予写作。这些技能对于他成功过渡到独立至关重要。该提案汇集了 一个由世界知名心血管生物学专家组成的令人印象深刻的团队为他提供指导，包括他的共同导师， 博士。马克·默科拉 (Mark Mercola) 和尤安·阿什利 (Euan Ashley) 以及他的指导委员会成员 Drs.吴约瑟夫, 马琳·拉宾诺维奇和唐·贝尔斯。他们的支持将为 Staudt 博士提供资源和指导 需要在 K08 奖项结束时在独立的学术生涯中取得成功。 该项目的科学目的是揭示导致舒张功能障碍（一种异常僵硬）的机制 心室肌的损伤导致了近一半的心力衰竭病例。具体而言，该提案重点关注 小儿限制性心肌病（RCM）是一种严重的遗传性疾病，其特征是孤立的、严重的 舒张功能障碍。由于相对缺乏治疗方法，患有这种疾病的患者几乎没有治疗选择 了解这种疾病的分子机制。该提案利用了新型干细胞 RCM 模型与舒张功能的高通量测量相结合来探索这些机制。 在目标 1 中，将对 RCM 的多种细胞模型进行表征和比较，以确定是否存在不同的 各类 RCM 突变通过相似或不同的机制发挥作用。目标 2 使用功能基因组学方法 确定不同的突变是否会引发不同的致病机制，而这些机制会集中在相似的机制上 临床表现。目标 3 重点关注由以下原因引起的严重儿科发病 RCM 患者的独特系列： 心肌肌球蛋白的从头突变。为了实现这一目标，一种新颖的多尺度方法将生物物理效应联系起来 肌球蛋白分子上的 RCM 突变对整个细胞的生理变化的影响，并将其与 类似的、先前描述的肌球蛋白突变会导致肥厚性心肌病，这是一种更常见的疾病 常见但通常不太严重的疾病。