Defining the molecular regulators of valvular delamination via multi-omic dissection of Ebstein’s Anomaly

通过 Ebstein 异常的多组学解剖定义瓣膜分层的分子调节因子

基本信息

批准号：
10606574
负责人：
ALEXANDER Flaherty MERRIMAN
金额：
$ 5.27万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-01 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10606574
关键词：
ATAC-seq Adult Anatomy Apical Automobile Driving Bar Codes Biological Biological Assay Biological Models Biology Blood flow Cardiac Cell Communication Cell Differentiation process Cell Maturation Cell physiology Cells Childhood Chromatin Clinical Collaborations Complex Congenital Heart Defects Data Defect Development Disease Dissection Doctor of Philosophy Dyes Ebstein&apos s Anomaly Electrophysiology (science)Elements Endothelium Event Failure Fellowship Fostering Functional disorder Future Gene Expression Profile Genomics Goals Heart Heart Atrium Heart Transplantation Heart Valves Human Genetics Individual Lateral Location Machine Learning Maintenance Maps Mediating Mentorship Mesenchymal Mesenchyme Molecular Morbidity - disease rate Morphogenesis Morphology Mus Muscle Myocardial Myocardium National Research Service Awards Operative Surgical Procedures Pathogenesis Patients Phenotype Population Preceptorship Process Prosthesis RNA Regenerative Medicine Regulation Research Resolution Right ventricular structure Scientist Secondary to Secure Signal Pathway Signal Transduction Slide Source Surgeon System Techniques Testing Therapeutic Time Tissue Sample Training Tricuspid valve structure Variant Ventricular cardiac tissue engineering career clinical training epigenomics epithelial to mesenchymal transition gene regulatory network genetic analysis high throughput screening human disease human tissue in vivo Model information gathering innovation interstitial cell mRNA Expression malformation mortality mouse model multiple omics novel novel therapeutics patient oriented periostin pharmacologic programs prospective single cell sequencing single-cell RNA sequencing spatiotemporal stem cell biology tool transcriptomics

项目摘要

PROJECT SUMMARY/ABSTRACT Cardiac valves are critical to the maintenance of unidirectional blood flow in the heart. Congenital and acquired valvulopathies are a major source of morbidity and mortality in both the pediatric and adult populations. Current therapeutic strategies are limited, often requiring surgical replacement with suboptimal prostheses. Valvulogenesis begins with formation of the endocardial cushions via epithelial-to-mesenchymal transition (EMT). Many of the signaling pathways regulating this EMT event have been defined; however, little is known about the molecular regulators of post-EMT valvulogenesis. Ebstein’s Anomaly, a rare congenital heart defect, is characterized by variably dysplastic, muscularized tricuspid valve leaflets that are often tethered to the underlying myocardium, resulting in apical displacement of the annulus and atrialization of the right ventricle. This defect is often attributed to a failure of valvular delamination – a critical morphogenetic step of post-EMT valvulogenesis during which the primordial valve leaflets separate from the underlying myocardium. Previous studies suggest that altered differentiation dynamics of valvular interstitial cells (VICs) may contribute to the pathogenesis of this valvular defect. I propose to define the molecular regulators of valvular delamination via a multi-omic dissection of Ebstein’s Anomaly, leveraging an Ebstein’s Anomaly murine model system, primary human tissue, and human genetics data. In my first aim, I will use spatiotemporal single cell RNA sequencing and human genetics analyses to identify the atrioventricular cushion signaling interactions regulating valvular delamination. In my second aim, I will use an integrated single cell RNA/ATAC sequencing approach to define the gene regulatory networks driving VIC fate determination. My primary sponsor, Dr. Deepak Srivastava, has extensive expertise in developmental cardiac biology, human genetics, and stem cell biology. My co- sponsor, Dr. Chun (Jimmie) Ye, has an expertise in experimental and computational single cell genomics. Their collective expertise will assure that I receive the necessary training and mentorship to complete the proposed research. To obtain the rare tissue samples required for my project, I have formed a multi-center collaboration with two high volume Ebstein’s Anomaly patient centers. Additionally, I have secured expertise in human genetics and machine learning from Dr. Jingjing Li, Ph.D., in support of the human genetics component of my project. Concurrently, I am engaging in a longitudinal clinical preceptorship in pediatric cardiothoracic surgery with my clinical training co-sponsor, Dr. Peter Kouretas, Surgical Director of the UCSF Pediatric Heart Transplantation Program. Overall, the proposed research will elucidate previously uncharacterized mechanisms of late atrioventricular valvular morphogenesis and inform future efforts toward the development of novel regenerative-medicine based therapeutics for congenital and acquired valvulopathies. F30 NRSA fellowship support would foster my expertise in developmental cardiac biology and computational genomics, furthering me toward my ultimate career goal of becoming an academic pediatric cardiothoracic surgeon-scientist.

项目摘要/摘要心脏瓣膜对于维持心脏的单向血流至关重要。先天性和被收购瓣膜病是小儿和成人人群中发病率和死亡率的主要来源。当前的治疗策略是有限的，通常需要用次优的假体进行手术替代。瓣膜发生始于通过上皮到间质转变形成心内膜垫（EMT）。已经定义了调节此EMT事件的许多信号通路；但是，鲜为人知关于EMT后瓣膜发生的分子调节剂。 Ebstein的异常，一种罕见的先天性心脏缺陷，其特征是多样性发育不良，肌肉发达的三尖瓣小叶，通常被束缚在基础心肌，导致环的顶端位移和右心室的心房化。该缺陷通常归因于瓣膜分层的失败 - emt后的关键形态发生步骤原始瓣膜小叶与基础心肌分离的瓣膜发生。以前的研究表明，瓣膜间质细胞（VIC）的分化动力学改变可能有助于该瓣膜缺陷的发病机理。我建议定义瓣膜分层的分子调节剂通过对Ebstein异常的多摩变解剖，利用Ebstein的异常鼠模型系统，原代人组织和人类遗传学数据。在我的第一个目标中，我将使用时空单细胞RNA 测序和人类遗传学分析，以识别调节调节的房屋缓冲信号传导相互作用瓣膜分层。在第二个目标中，我将使用集成的单细胞RNA/ATAC测序方法定义驱动VIC脂肪测定的基因调节网络。我的主要赞助商Deepak Srivastava博士，在发育性心脏生物学，人遗传学和干细胞生物学方面具有广泛的专业知识。我的共同赞助商Chun（Jimmie）Ye博士拥有实验和计算单细胞基因组学专家。他们的集体专业知识将假设我会收到必要的培训和心态来完成拟议的研究。为了获得我项目所需的稀有组织样本，我组成了多中心协作 Ebstein的两个大容量的异常患者中心。此外，我还确保了人类的专业知识遗传学和机器从博士的Jingjing Li博士学习，以支持我的人类遗传学成分项目。同时，我正在小儿心胸外科手术中进行纵向临床诊所伴随着我的临床培训共同赞助者，UCSF儿科心脏外科主任彼得·库雷塔斯（Peter Kouretas）博士移植程序。总体而言，拟议的研究将阐明先前未表征的机制晚期心室瓣膜形态发生，并为发展新颖的努力提供了努力基于再生医学疗法的先天性和获得的瓣膜病。 F30 NRSA奖学金支持将促进我在发育性心脏生物学和计算基因组学方面的专业知识，我进一步朝着我成为学术儿科心胸外科医生科学家的最终职业目标。