Targeting Protein Trafficking in Arrhythmogenic Cardiomyopathy

致心律失常性心肌病中的靶向蛋白质运输

• 批准号: 10301665
• 负责人: Joseph A. Palatinus
• 金额: $ 17.93万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-15 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10301665
• 关键词: Actins; Address; Affect; Age; Animal Model; Arrhythmia; Biology; Birth; Cardiac Myocytes; Cardiovascular Diseases; Cardiovascular system; Cell Culture Techniques; Cell model; Cessation of life; Cicatrix; Co-Immunoprecipitations; Confocal Microscopy; Connexin 43; Connexins; Coronary; Coupling; Data; Defect; Defibrillators; Desmosomes; Devices; Disease; Disease model; Doctor of Philosophy; Echocardiography; Educational workshop; Electron Microscopy; Gap Junctions; Goals; Heart; Heart Diseases; Hela Cells; Institutes; Intercalated disc; International; Ion Channel; Ischemia; Knock-out; Knowledge; Laboratories; Ligation; Link; Location; Mediating; Mentors; Metabolic; Mitochondria; Modeling; Molecular; Monitor; Mus; Mutant Strains Mice; Mutation; Myocardial; Myocardial dysfunction; Names; Normal Statistical Distribution; Outcome; Oxygen; Pathologic; Patients; Phenotype; Physical activity; Physicians; Prevention; Protein Isoforms; Proteins; Recovery; Regulation; Reproducibility; Research; Research Personnel; Research Training; Resolution; Robin bird; Role; Scientist; Signal Transduction; Structure; Sudden Death; Targeted Research; Techniques; Telemetry; Testing; Training Programs; Transfection; Translating; Translational Research; United States; United States National Institutes of Health; Universities; Utah; Virus; Vocational Guidance; Work; arrhythmogenic cardiomyopathy; base; cardioprotection; career development; desmoglein 2; effective therapy; experience; implantation; improved; in vivo; insight; knock-down; medical schools; mutant; new therapeutic target; novel; novel therapeutic intervention; novel therapeutics; preservation; prevent; professor; protein transport; sudden cardiac death; symposium; targeted treatment; trafficking

ABSTRACT This proposal is to support the career development of Joseph Palatinus, MD, PhD, to become an independent physician scientist with a focus on basic/translational research by targeting protein trafficking as a means to prevent and treat arrhythmias and sudden cardiac death (SCD). The PI will have as a primary mentor Professor Robin Shaw, MD PhD, who is world renowned expert in protein trafficking and cardiomyocyte biology. The comprehensive training program is composed of laboratory-based research, coursework, workshops, grantsmanship seminars, scientific conferences (local and international), and career guidance by a mentoring committee. The committee includes the mentors and leverages the combined experience and technical knowledge of 5 leading independent cardiovascular investigators at the Eccles Cardiovascular Research and Training Institute at the University of Utah as well as Dr. Jeffery Saffitz at Harvard Medical School, a world expert on Arrhythmogenic cardiomyopathy. The research proposed will apply the alternatively translated isoform of Connexin 43 (Cx43), dubbed GJA1-20k (and an established actin stabilizing protein), to the Desmoglein 2 (DSG2) mutant model of ACM. Preliminary data has demonstrated disrupted actin organization in both cellular and animal models of a DSG2 mutation which, in the heart is associated with a loss of gap junction trafficking to the intercalated disk and downstream cardiac dysfunction. It is hypothesized that GJA1-20k will lead to recovery of gap junction localization, suppression of arrhythmias and prevention of SCD in the DSG2 mutant model of ACM by directly interacting with and stabilizing cellular actin. Two specific aims are proposed: 1) Is actin dysregulation responsible for the trafficking defects observed in the DSG2 mutant model? And 2) Does GJA1- 20k rescue the cardiomyopathic and arrhythmogenic phenotype of ACM and prevent SCD? High resolution confocal microscopy, proximity ligation immunolabeling, electron microscopy, co- immunoprecipitation, echocardiography, and in vivo telemetry monitoring are some of the advanced techniques which will be used to develop a mechanistic understanding of actin dependent trafficking mediated by GJA1-20k. The results from this study are expected to develop a novel paradigm to treat and prevent arrhythmogenic SCD by targeting protein trafficking.

抽象的 该提案旨在支持 Joseph Palatinus 医学博士、哲学博士的职业发展，使其成为一名 独立医师科学家，专注于基础/转化研究 蛋白质贩运作为预防和治疗心律失常和心源性猝死的手段 （SCD）。 PI 将由医学博士 Robin Shaw 教授作为主要导师，他是世界 蛋白质运输和心肌细胞生物学领域的著名专家。综合培训 项目由基于实验室的研究、课程、研讨会、资助组成 研讨会、科学会议（本地和国际）以及导师的职业指导 委员会。该委员会包括导师，并利用综合经验和 埃克尔斯 5 位领先的独立心血管研究人员的技术知识 犹他大学心血管研究与培训学院以及 Jeffery 博士 哈佛医学院的萨菲茨是致心律失常性心肌病的世界专家。这 拟议的研究将应用 Connexin 43 (Cx43) 的替代翻译亚型， 桥粒芯糖蛋白 2 (DSG2) 被称为 GJA1-20k（以及已确定的肌动蛋白稳定蛋白） ACM突变模型。初步数据表明，两者的肌动蛋白组织均受到破坏。 DSG2 突变的细胞和动物模型，在心脏中，该突变与心脏 间隙连接运输至闰盘和下游心脏功能障碍。这是 假设 GJA1-20k 将导致间隙连接定位的恢复，抑制 通过直接相互作用在 ACM DSG2 突变模型中预防心律失常和 SCD 具有并稳定细胞肌动蛋白。提出了两个具体目标：1) 肌动蛋白失调吗 造成 DSG2 突变体模型中观察到的运输缺陷的原因是什么？ 2) GJA1- 20k 挽救 ACM 的心肌病和致心律失常表型并预防 SCD？高的 分辨率共聚焦显微镜、邻近连接免疫标记、电子显微镜、共 免疫沉淀、超声心动图和体内遥测监测是其中一些 先进技术将用于发展对肌动蛋白的机械理解 由 GJA1-20k 介导的依赖性贩运。这项研究的结果预计 开发一种通过靶向蛋白质来治疗和预防致心律失常性 SCD 的新范例 贩运。