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: 10473531
• 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/10473531
• 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; Disease; Dose; Dyes; Echocardiography; Educational workshop; Electrocardiogram; Electrophysiology (science); Exercise; 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; diagnostic tool; 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; 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.

项目概要/摘要 这项为期五年的提案将为 Goodyer 博士成功转型为独立专家提供一个平台。 医师科学家研究预防和治疗心律的新机制 在此基础上定制了特定的指导和培训机会。 Goodyer 博士将由医学和儿科副教授 Sean M. Wu 博士指导。 Anne Dubin 博士，儿科教授，儿科电生理学 (EP) 科长。 甄选国际知名科学家组成的优秀顾问团队，每位顾问都有独特的见解 Goodyer 博士在基础科学 EP 到转化医学等领域拥有丰富的经验和技能。 培训计划制定了个性化计划，以提高他在以下关键领域的熟练程度： 1. 使用 人类诱导多能干细胞（hiPSC）对新型心脏传导系统进行功能评估 (CCS) 基因；2. CCS 发育和功能体内分析的小动物表型分析技能； 基础科学 EP 的知识和技术；以及 4. 专业发展，包括领导力、资助 培训计划包括斯坦福大学课程的经验。 干细胞研究（例如 STEMREM 201B：干细胞和人类发展），每周研讨会 心脏病学和转化医学、著名的领导力和沟通研讨会以及 在顾问 Chiamvimonvat 博士的实验室中，量身定制的实习项目专注于先进的基础科学 EP 技术， 加州大学戴维斯分校心血管医学教授这些指导和培训活动。 专为使候选人能够实现旨在阐明 CCS 的特定研究目标而量身定制 在目标 1 中，Goodyer 博士将研究一种新型细胞内 CCS 特异性的作用。 基因 Cpne5（copine 5），在他最近发表的《循环研究》和相关研究中发现 申请人将通过独立的全基因组关联研究评估人类心率变化。 通过使用两者进行体外功能丧失和获得检测来确定 Cpne5 在传导细胞中的功能 在目标 2 中，候选人将进一步研究分离的小鼠和 hiPSC 衍生的 CCS 细胞。 通过综合心脏和电生理学了解 CCS 发展和体内疾病的背景 最后，Aim 3 利用了 CRISPR-Cas9 生成的 Cpne5 系统性敲除小鼠的表型。 申请人最近发现了另一种以前未知的 CCS 特异性标记，特别是通过针对此标记。 细胞表面标记申请人将验证基于抗体的新型光学成像方法的使用 这些研究将为体内 CCS 提供原理验证。 标记心脏亚结构并为实时转化机会奠定基础 在心脏介入治疗期间对 CCS 进行可视化，以防止术中意外损坏 CCS。