Sarcomeric Regulation of Signal Transduction and Cardiomyopathy

信号转导和心肌病的肌节调节

• 批准号: 9976127
• 负责人: Yuxuan Guo
• 金额: $ 11.16万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2020-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9976127
• 关键词: Ablation; Actinin; Actins; Affect; Animals; Binding; CRISPR/Cas technology; Cardiac; Cardiac Myocytes; Cardiomyopathies; Cells; Chromatin; Code; Cytoskeleton; Data; Defect; Dependovirus; Dilatation - action; Dilated Cardiomyopathy; Disease; Disease model; Education; F-Actin; Fibrosis; Gene Expression Profile; Genes; Genetic; Genetic Transcription; Goals; Grant; Heart; Heart Abnormalities; Heart Diseases; Human; Impairment; Individual; Inherited; Knock-out; Knowledge; Lead; Mediating; Mediator of activation protein; Mentorship; Missense Mutation; Modeling; Molecular; Mosaicism; Mus; Mutagenesis; Mutation; Myocardial Contraction; Myocardial dysfunction; Myosin Heavy Chains; Nuclear; Nuclear Translocation; Occupations; Pathogenesis; Pathway interactions; Physiological; Play; Regulation; Research; Research Proposals; Role; Sarcomeres; Scientist; Secondary to; Serum Response Factor; Signal Transduction; Structure; Technology; Testing; Therapeutic; Therapeutic Effect; Training; Transcriptional Regulation; Treatment Efficacy; Ventricular; base; cardiac muscle disease; cardiac tissue engineering; career development; cofactor; crosslink; driving force; familial dilated cardiomyopathy; genome editing; induced pluripotent stem cell; inherited cardiomyopathy; insight; novel; novel therapeutic intervention; polymerization; prevent; targeted treatment; therapeutic development; transcription factor; transcriptomics

PROJECT SUMMARY/ABSTRACT Sarcomeres are specialized cytoskeletal structures in cardiomyocytes (CMs) that generate the force driving the heart contraction. Mutations in genes coding sarcomere components, or “sarcomere mutations”, often cause inherited cardiomyopathies including most cases of inherited dilated cardiomyopathy (DCM). Sarcomere mutations are believed to impair CM contraction, thereby causing the contractile defects in DCM hearts. How- ever, whether other aspects of sarcomere function contribute to DCM pathogenesis has been incompletely studied. This knowledge gap prevents comprehensive understanding of DCM pathogenesis and limits the de- velopment of effective therapeutic strategies. This research proposal tests the overarching hypothesis that sarcomeres regulate signal transduction, which contributes to the pathogenesis of DCM. Specifically, Dr. Guo will dissect the mechanisms by which ac- tinin-2 (ACTN2) mutations perturb serum response factor (SRF) signaling through SRF cofactors MKL1/2, and the contribution of this pathway to DCM. This hypothesis will be tested through three Specific Aims. Aim 1 will study whether ACTN2 activates MKL-SRF signaling by promoting actin polymerization in CMs. Aim 2 will de- termine if then ACTN2-MKL1/2 pathway regulates transcription in CMs by affecting SRF-chromatin binding. Aim 3 will test the hypothesis that ACTN2-MKL-SRF signaling contributes to the pathogenesis of DCM caused by ACTN2 missense mutations. This project will reveal novel molecular insights about DCM pathogenesis. These insights may lead to new therapeutic approaches to treat DCM. This project is built upon state-of-the-art technologies such as adeno-associated virus (AAV)-delivered CRISPR/Cas9-based somatic mutagenesis and cardiac genetic mosaics analysis that were co-developed by Dr. Guo. K99/R00 will further sponsor training in induced pluripotent stem cells (hiPSCs), genome editing, sin- gle-cell transcriptomics and engineered heart tissues, which together with Dr. Guo’s past expertise will estab- lish a powerful and comprehensive technical platform for successful independent research. This grant will also support Dr. Guo’s education from a mentorship team of world-class scientists with relevant expertise and facili- tate Dr. Guo’s career development activities as he transitions toward independence.

项目总结/摘要 肌节是心肌细胞（CM）中的特化细胞骨架结构， 心脏收缩。编码肌节成分的基因突变，或“肌节突变”，通常 引起遗传性心肌病，包括大多数遗传性扩张型心肌病（DCM）。肌节 突变被认为损害CM收缩，从而导致DCM心脏的收缩缺陷。怎么-- 然而，肌节功能的其他方面是否也参与DCM的发病机制尚不完全清楚， 研究了这种知识差距阻碍了对DCM发病机制的全面理解，并限制了对DCM的认识。 制定有效的治疗策略。 这项研究计划测试了肌节调节信号转导的总体假设， 这有助于DCM的发病机制。具体来说，郭博士将剖析的机制，其中ac- 肌动蛋白-2（ACTN 2）突变通过血清反应因子（SRF）辅因子MKL 1/2干扰SRF信号传导， 这一途径对DCM的贡献。这一假设将通过三个具体目标进行检验。目标1将 研究ACTN 2是否通过促进CM中的肌动蛋白聚合来激活MKL-SRF信号传导。目标2将- 最后，如果ACTN 2-MKL 1/2途径通过影响SRF-染色质结合来调节CM中的转录。 目的3将验证ACTN 2-MKL-SRF信号通路参与DCM发病机制的假设， ACTN 2错义突变。该项目将揭示有关DCM发病机制的新分子见解。 这些见解可能会导致新的治疗方法来治疗DCM。 该项目是建立在国家的最先进的技术，如腺相关病毒（AAV）-交付 基于CRISPR/Cas9的体细胞诱变和心脏遗传嵌合体分析， 博士小郭。K99/R 00将进一步赞助诱导多能干细胞（hiPSC）、基因组编辑、单克隆抗体（SINS）、 Gle-cell转录组学和工程心脏组织，加上郭博士过去的专业知识， lish是一个强大而全面的技术平台，用于成功的独立研究。这笔赠款还将 支持郭博士的教育，从一个导师团队的世界一流的科学家与相关的专业知识和设施， 陈述郭博士在向独立过渡期间的职业发展活动。