Targeting Protein Trafficking in Arrhythmogenic Cardiomyopathy

摘要 这项建议是为了支持约瑟夫Palatinus，医学博士，博士的职业发展，成为一个 独立的医生科学家，专注于基础/转化研究， 蛋白质运输作为预防和治疗心律失常和心源性猝死的手段 （SCD）。PI将有一个主要的导师教授罗宾肖，医学博士，谁是世界 著名的蛋白质运输和心肌细胞生物学专家。综合训练 计划是由实验室为基础的研究，课程，研讨会，granitarian 研讨会、科学会议（当地和国际），以及由导师提供的职业指导 以马克思委员会包括导师，并利用他们的综合经验， Eccles 5位领先的独立心血管研究者的技术知识 犹他州大学心血管研究和训练所的Jeffery博士 Saffitz是哈佛医学院的一位心律失常性心肌病专家。的 提出的研究将应用连接蛋白43（Cx43）的替代翻译同种型， 被称为GJA 1 - 20 k（和一种已建立的肌动蛋白稳定蛋白）的桥粒芯糖蛋白2（DSG 2） ACM的突变模型。初步数据表明，在这两种细胞中， DSG 2突变的细胞和动物模型，在心脏中与心脏功能丧失相关， 间隙连接运输到闰盘和下游心脏功能障碍。是 假设GJA 1 - 20 k将导致间隙连接定位的恢复， 在ACM的DSG 2突变模型中通过直接相互作用预防心律失常和SCD 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 负责DSG 2突变模型中观察到的运输缺陷？和2）GJA 1- 20 k拯救ACM的心肌病和致炎表型并预防SCD？高 分辨率共聚焦显微镜，邻位连接免疫标记，电子显微镜，共- 免疫沉淀、超声心动图和体内遥测监测是一些 先进的技术，将用于发展肌动蛋白的机械理解 由GJA 1 - 20 k介导的依赖性贩运。这项研究的结果预计将 通过靶向蛋白质开发治疗和预防致瘤性SCD的新模式 贩卖人口

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.