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 本五年计划书中描述的综合培训计划旨在帮助施陶德博士做好准备， 过渡到职业生涯作为一个富有成效的独立调查员，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，施陶德博士将扩大 他在诱导多能干细胞衍生的心肌细胞表型在单细胞水平的技能，以及 获得致病性肌球蛋白突变的生物物理分析和高通量检测设计的新技能， 分析.此外，他将获得大量的经验和培训，指导，科学演示， 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 一个令人印象深刻的团队，由世界知名的心血管生物学专家组成，指导他，包括他的共同导师， Drs. Mark Mercola和Euan阿什利，以及他的导师委员会成员Joseph Wu博士， 玛琳·拉宾诺维奇和唐·贝斯他们的支持将为施陶德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的机制， 几乎一半的心力衰竭病例都是由心室肌引起的。具体而言，该提案侧重于 儿童限制性心肌病（RCM），一种严重的遗传性疾病，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 了解这种疾病的分子机制。这项提案利用了新的干细胞 RCM模型结合舒张功能的高通量测量来探讨这些机制。 在目标1中，将表征和比较RCM的多个细胞模型，以确定是否不同 不同种类的RCM突变通过相似或不同的机制起作用。AIM2使用功能基因组学方法 以确定不同的突变是否引起不同的致病机制，这些机制集中在一个类似的 临床表现目的3重点关注一个独特的线，从患者的严重，儿科发作的RCM引起的 心肌肌球蛋白的一种新生突变。在这一目标中，一种新的多尺度方法将生物物理效应 肌球蛋白分子上的这种RCM突变与整个细胞的生理变化有关，并将其与 类似的，以前表征的肌球蛋白突变，导致肥厚性心肌病，一个更 常见但一般不太严重的疾病。