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: 10703399
• 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/10703399
• 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; 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; Persons; Phenotype; Physical activity; Physicians; Prevention; Protein Isoforms; Proteins; Recovery; Regulation; Reproducibility; Research; Research Personnel; Resolution; Robin bird; Role; Scientist; Signal Transduction; Sudden Death; Targeted Research; Techniques; Telemetry; Testing; Training; Training Programs; Transfection; Translating; Translational Research; United States; United States National Institutes of Health; Universities; Utah; Virus; Vocational Guidance; Work; arrhythmogenic cardiomyopathy; 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.

摘要 这项提议是为了支持约瑟夫·帕拉蒂纳斯的职业发展，医学博士，博士，成为一名 独立的内科科学家，专注于基础/翻译研究 蛋白质转运作为预防和治疗心律失常和心脏性猝死的手段 (SCD)。国际象棋将由世界知名的医学博士罗宾·肖教授担任主要导师 著名的蛋白质运输和心肌细胞生物学专家。综合训练 课程包括基于实验室的研究、课程、工作坊、资质 研讨会、科学会议(本地和国际)，以及由导师提供职业指导 委员会审议阶段。该委员会包括导师，并利用综合经验和 ECCLES的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和培训研究所以及杰弗里博士 哈佛医学院的萨菲兹说，他是致心律失常心肌病的世界专家。这个 建议的研究将应用连接蛋白43(Cx43)的替代翻译异构体， 被命名为GJA1-20k(和一个已建立的肌动蛋白稳定蛋白)，与桥粒蛋白2(DSG2) ACM的突变模型。初步数据显示，在这两种情况下，肌动蛋白的组织都受到了破坏 DSG2突变的细胞和动物模型，该突变在心脏中与 缝隙连接转运至间盘和下游心功能不全。它是 假设GJA1-20k将导致缝隙连接局部化的恢复，抑制 DSG2基因突变ACM模型的心律失常及SCD的预防 并稳定细胞肌动蛋白。提出了两个具体的目标：1)肌动蛋白是否失调 对在DSG2突变模型中观察到的贩运缺陷负责？2)GJA1是否- 20K挽救ACM的心肌病和致心律失常表型，预防SCD？高 分辨率共聚焦显微镜，邻近连接免疫标记，电子显微镜，共聚焦显微镜 免疫沉淀、超声心动图和活体遥测监测是其中的一部分 先进的技术将被用来发展对肌动蛋白的机械性理解 由GJA1-20K调解的依赖贩运。这项研究的结果预计将 开发靶向蛋白质治疗和预防致心律失常的SCD的新范式 贩卖人口。