Elucidation of the Development and Function of the Cardiac Conduction System

阐明心脏传导系统的发育和功能

• 批准号: 10039151
• 负责人: William R Goodyer
• 金额: $ 16.65万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10039151
• 关键词: Adult; Advisory Committees; Alleles; Animals; Antibodies; Area; Arrhythmia; Backcrossings; Basic Science; Biological; Biological Assay; Biological Pacemakers; Blood Circulation; CRISPR/Cas technology; Calcium; California; Cardiac; Cardiac Myocytes; Cardiac Output; Cardiac Surgery procedures; Cardiac conduction system; Cardiac development; Cardiology; Cardiovascular system; Categories; Cell Count; Cell Surface Proteins; Cell physiology; Cell surface; Cells; Childhood; Clinical; Communication; Development; Diagnostic; Disease; Dose; Dyes; Echocardiography; Educational workshop; Electrocardiogram; Electrophysiology (science); Exercise; Expression Profiling; Fluorescent Antibody Technique; Foundations; Gene Expression; Gene Expression Profiling; Generations; Genes; Goals; Grant; Harvest; Heart; Heart Rate; Heterogeneity; Human; Human Development; Image; Immunofluorescence Immunologic; In Vitro; International; Intervention; Ion Channel; Knock-out; Knockout Mice; Knowledge; Label; Leadership; Life; Link; Measures; Medicine; Membrane; Mentors; Mentorship; Methods; Microscopy; Molecular; Molecular Analysis; Monitor; Mus; Operative Surgical Procedures; Pediatrics; Phenotype; Physicians; Postdoctoral Fellow; Prevention; Protocols documentation; Publishing; Reporter; Research; Resolution; Role; Safety; Science; Scientist; Signal Transduction; Small Interfering RNA; Stem Cell Research; Structure; System; Systems Development; Techniques; Time; Tissue-Specific Gene Expression; Tissues; Training; Training Activity; Transgenic Organisms; Universities; Variant; Visualization; Work; Writing; base; career; cell type; copine; experience; experimental study; externship; gain of function; genome wide association study; heart cell; heart function; heart rate variability; heart rhythm; heart visualization; iatrogenic injury; imaging modality; in vivo; induced pluripotent stem cell; knock-down; mid-career faculty; near infrared dye; new therapeutic target; nodal myocyte; novel; novel diagnostics; optical imaging; overexpression; prevent; professor; programs; protein transport; single-cell RNA sequencing; skills; stem cells; targeted agent; tool; trafficking; training opportunity; translational medicine; transmission process; voltage

PROJECT SUMMARY/ABSTRACT This five-year proposal will provide a platform for Dr. Goodyer’s successful transition to an independent physician scientist investigating novel mechanisms for the prevention and treatment of cardiac rhythm disorders. Specific mentorship and training opportunities have been tailored to build on the foundation of the applicant. Dr. Goodyer will be mentored by Dr. Sean M. Wu, Associate Professor of Medicine and Pediatrics and Dr. Anne Dubin, Professor of Pediatrics, Section Chief of Pediatric Electrophysiology (EP). Additionally, an outstanding advisory team of internationally-renowned scientists has been selected, each advisor with unique experiences and skillsets in fields ranging from basic science EP to translational medicine. Dr. Goodyer’s training plan lays out a personalized program for developing his proficiency in the following key areas: 1. Use of human induced pluripotent stem cells (hiPSCs) to functionally evaluate novel cardiac conduction system (CCS) genes; 2. Small animal phenotyping skills for in vivo analyses of CCS development and function; 3. Knowledge and techniques in basic science EP; and, 4. Professional development including leadership, grant writing and science communication skills. The training plan includes experiences from Stanford courses on stem cell research (eg. STEMREM 201B: Stem Cells and Human Development), weekly seminars on cardiology and translational medicine, renowned workshops on leadership and communication as well as a tailored externship focused on advanced basic science EP techniques in the lab of advisor Dr. Chiamvimonvat, Professor of Cardiovascular Medicine at University of California Davis. These mentorship and training activities are tailored to enable the candidate to achieve specific research goals aimed at the elucidation of CCS development and function. In Aim 1, Dr. Goodyer will investigate the role of a novel, intracellular, CCS-specific gene Cpne5 (copine 5), uncovered in his recently published work in Circulation Research and associated with human heart rate variation by independent genome wide association studies. The applicant will evaluate the function of Cpne5 in conduction cells by performing in vitro loss- and gain-of-function assays using both isolated mouse and hiPSC-derived CCS cells. In Aim 2, the candidate will further investigate Cpne5 in the context of CCS development and disease in vivo by comprehensive cardiac and electrophysiological phenotyping of CRISPR-Cas9 generated Cpne5 systemic knockout mice. Finally, Aim 3 capitalizes on the applicant’s recent discovery of another previously unknown CCS-specific marker. Specifically, by targeting this cell surface marker the applicant will validate the use of a novel antibody-based optical imaging method for visualizing the CCS in human hearts ex vivo. These studies will provide a proof-of-principle for the in vivo labeling of cardiac substructures and lay the foundation for translational opportunities in the real-time visualization of the CCS during cardiac interventions to prevent accidental intraoperative damage to the CCS.

项目摘要/摘要 这个五年的提案将为戈德尔博士成功过渡到独立的平台 医师科学家研究了预防和治疗心律的新机制 疾病。特定的心态和训练机会是为基础而量身定制的 申请人。 Goodyer博士将由医学和儿科副教授Sean M. Wu博士考虑 儿科教授，小儿电生理学主管（EP）教授Anne Dubin博士。另外， 已经选出了国际知名科学家的杰出顾问团队，每个顾问都有独特的顾问 从基础科学EP到翻译医学等领域的经验和技能。 Goodyer博士 培训计划提出了一个个性化计划，以在以下关键领域发展他的熟练程度：1。 人类诱导的多能干细胞（HIPSC）的功能评估新型心脏传导系统 （CCS）基因； 2。用于CCS开发和功能的体内分析的小动物表型技巧； 3。 基础科学中的知识和技术； 4。专业发展，包括领导，授予 写作和科学沟通技巧。培训计划包括斯坦福课程的经验 干细胞研究（例如StemRem 201B：干细胞和人类发育），每周的半手 心脏病学和转化医学，有关领导力和沟通的著名研讨会以及 量身定制的实习重点是顾问Chiamvimonvat博士实验室的先进基础科学EP技术， 加州大学戴维斯分校心血管医学教授。这些精通和培训活动 量身定制的是为了使候选人能够实现旨在阐明CCS的特定研究目标 发展和功能。在AIM 1中，Goodyer博士将研究一种新颖的细胞内CCS特异性的作用 Gene CPNE5（Copine 5），在他最近发表的循环研究和相关的工作中发现了 与独立基因组广泛关联研究的人类心率变化。申请人将评估 通过使用两者进行体外损失和功能获得测定，CPNE5在传导细胞中的功能 分离的小鼠和hipsc衍生的CCS细胞。在AIM 2中，候选人将进一步调查CPNE5 CCS发育和疾病在体内的背景通过全面的心脏和电生理学 CRISPR-CAS9的表型产生的CPNE5全身敲除小鼠。最后，AIM 3大写 申请人最近发现了另一个以前未知的CCS特定标记。具体而言，通过目标 细胞表面标记申请人将验证使用基于新型抗体的光学成像方法的使用 在体内可视化人类心脏中的CC。这些研究将为体内提供原则证明 心脏子结构的标签并为实时转化机会奠定基础 心脏干预期间CC的可视化，以防止意外术中对CCS的损害。