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: 10686273
• 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/10686273
• 关键词: Accidents; Adult; Advisory Committees; Alleles; Animals; Antibodies; Area; Arrhythmia; Backcrossings; Basic Science; Biological; Biological Assay; Biological Pacemakers; CRISPR/Cas technology; Calcium; California; Cardiac; Cardiac Myocytes; Cardiac Output; Cardiac Surgery procedures; Cardiac conduction system; Cardiology; Cardiovascular system; Categories; Cell Count; Cell Surface Proteins; Cell physiology; Cell surface; Cells; Childhood; Circulation; Clinical; Communication; Development; Disease; Dose; Dyes; Echocardiography; Educational workshop; Electrocardiogram; Electrophysiology (science); Exercise; 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; 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; loss of function; 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 barrier; translational medicine; translational potential; 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 这项为期五年的提案将为古代尔博士成功过渡到独立的 研究心律失常预防和治疗新机制的内科科学家 精神错乱。具体的指导和培训机会已量身定做，以 申请人。古代尔博士将由医学和儿科学副教授肖恩·M·吴博士指导 儿科学教授、儿科电生理学(EP)科长安妮·杜宾博士。此外，一个 评选出了由国际知名科学家组成的优秀顾问团队，每位顾问都拥有独特的 从基础科学EP到转化医学等领域的经验和技能。古代尔医生的 培训计划列出了一个个性化的计划，以发展他在以下关键领域的熟练程度：1.使用 人诱导多能干细胞(HiPSCs)对新型心脏传导系统的功能评价 (CCS)基因；2.用于体内分析CCS发育和功能的小动物表型技术； 基础科学方面的知识和技术；以及，4.专业发展，包括领导力，补助金 写作和科学交流能力。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：干细胞与人类发育)，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博士的实验室里，定制的外部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内科教授。这些辅导和培训活动 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。在目标1中，古代尔博士将研究一种新的、细胞内的、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工作中被发现 通过独立的全基因组关联研究发现人类心率变异。申请者将进行评估 Cpne5在传导细胞中的功能通过使用两种方法进行体外功能丧失和功能获得分析 分离小鼠和HiPSC来源的CCS细胞。在目标2中，候选人将进一步调查Cpne5在 综合心脏和电生理研究CCS的发展和体内疾病 CRISPR-Cas9的表型鉴定产生了Cpne5系统基因敲除小鼠。最后，Aim 3充分利用了 申请人最近发现的另一个先前未知的CCS特异性标记。具体地说，通过瞄准这个 细胞表面标记申请人将验证一种新的基于抗体的光学成像方法的使用 体外人心脏中CCS的可视化研究。这些研究将为体内试验提供原理证明。 标记心脏亚结构，并为实时翻译机会奠定基础 心脏介入治疗期间CCS的可视化，以防止术中CCS的意外损伤。