Nr2f1a promotes atrial maintenance and ventricular growth in the embryonic zebrafish heart

Nr2f1a 促进胚胎斑马鱼心脏的心房维持和心室生长

• 批准号: 10385751
• 负责人: Kendall Martin
• 金额: $ 4.04万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-09 至 2023-04-08
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10385751
• 关键词: Adult; Affect; Aortic coarctation; Arrhythmia; Atrial Heart Septal Defects; Biological Assay; Cardiac; Cardiac Myocytes; Cardiac development; Cells; Chromatin; Congenital Abnormality; Congenital Heart Defects; Data; Defect; Development; Double Outlet Right Ventricle; Embryo; Enhancers; Ensure; Etiology; Family; Generations; Genes; Genetic; Genetic Epistasis; Genetic Predisposition to Disease; Genetic Transcription; Growth; Heart; Heart Atrium; Heart Septal Defects; Hormones; Human; Individual; Maintenance; Methods; Molecular; Molecular Genetics; Morbidity - disease rate; Mus; Mutation; Newborn Infant; Nuclear Family; Nuclear Hormone Receptors; Operative Surgical Procedures; Orphan; Patients; Prevention strategy; Proteins; Reporting; Sinoatrial Node; Structure; Testing; Tissues; Transgenic Organisms; Transposase; Venous; Ventricular; Vertebrates; Work; Zebrafish; apoAI regulatory protein-1; cardiogenesis; experimental study; gene regulatory network; heart cell; infant death; infant morbidity/mortality; member; mortality; mutant; nodal myocyte; novel; novel strategies; protein function; repaired; septal defect; transcription factor; transcriptome sequencing

Project Summary/Abstract Congenital heart defects (CHDs) are the most common type of congenital malformation and the leading cause of birth defect associated infant death. CHDs can affect many different structures within the heart, including the atrial and ventricular septa and outflow tract (OFT). Atrial septal defects (ASDs) are often associated with arrhythmias and conduction defects, which can occur concurrently due to mutations in genes vital for both early development of the cardiac chambers and development of the sinoatrial node (SAN), which houses the pacemaker cells of the heart. While surgical intervention can correct some CHDs, surgery often does not repair associated conduction defects. Furthermore, arrhythmias are the leading cause of morbidity and mortality in adults with CHDs. Mutations in NR2F2, a member of the orphan nuclear hormone receptor transcription factor family, have been associated with multiple types of CHDs, most commonly ASDs but recently ventricular and OFT defects have been reported as well. NR2F2 is specifically expressed in atrial cardiomyocytes (ACs) in both humans and mice, and mouse studies have shown that Nr2f2 is required for atrial development and maintenance; however, the mechanisms by which these proteins function within ACs and how mutations in NR2F2 result in a spectrum of CHDs affecting both the atria and ventricles are not well understood. Recent work from our lab has identified zebrafish Nr2f1a as the functional equivalent of mammalian Nr2f2. Our preliminary data using zebrafish has revealed that in the absence of Nr2f1a there is a progressive ectopic expansion of SAN identity within ACs. Furthermore, integration of RNA-seq and ATAC-seq analysis of isolated ACs suggests that Nr2f1a represses the core SAN gene regulatory network (GRN) by maintaining expression nkx2.5 within ACs. In Aim 1, we will test the hypothesis that Nr2f1a is required to repress SAN identity by directly maintaining expression of Nkx2.5. Additionally, our preliminary data has revealed a novel requirement for Nr2f1a in ventricular development. In Aim 2, we will test the hypothesis that Nr2f1a cell non-autonomously promotes ventricular growth. Ultimately, the proposed studies have the potential to illuminate previously unknown molecular and genetic etiology underlying congenital arrhythmias and CHDs affecting both the atria and ventricles associated with NR2F2 mutations found in humans.

项目概要/摘要 先天性心脏病（CHD）是最常见的先天性畸形类型，也是导致先天性畸形的主要原因 与婴儿死亡相关的出生缺陷。 CHD 会影响心脏内的许多不同结构，包括 心房和心室间隔和流出道 (OFT)。房间隔缺损 (ASD) 通常与 心律失常和传导缺陷，由于对早期心律失常和传导缺陷至关重要的基因突变可能同时发生 心腔的发育和窦房结 (SAN) 的发育，窦房结容纳着 心脏的起搏细胞。虽然手术干预可以纠正一些先心病，但手术通常无法修复 相关的传导缺陷。此外，心律失常是导致患者发病和死亡的主要原因。 患有先心病的成年人。孤儿核激素受体转录因子成员 NR2F2 突变 家族，与多种类型的先心病（CHD）有关，最常见的是自闭症谱系障碍（ASD），但最近出现心室和 OFT 缺陷也有报道。 NR2F2 在心房心肌细胞 (AC) 中特异性表达 人类和小鼠，小鼠研究表明 Nr2f2 是心房发育和维持所必需的； 然而，这些蛋白质在 AC 内发挥作用的机制以及 NR2F2 的突变如何导致 影响心房和心室的冠心病谱尚不清楚。我们实验室最近的工作 确定斑马鱼 Nr2f1a 与哺乳动物 Nr2f2 的功能等效。我们使用斑马鱼的初步数据 研究表明，在 Nr2f1a 缺失的情况下，AC 内 SAN 身份会逐渐异位扩展。 此外，对分离的 AC 进行 RNA-seq 和 ATAC-seq 分析的整合表明 Nr2f1a 抑制 通过维持 AC 内的 nkx2.5 表达来构建核心 SAN 基因调控网络 (GRN)。在目标 1 中，我们将 检验 Nr2f1a 需要通过直接维持 Nkx2.5 的表达来抑制 SAN 身份的假设。 此外，我们的初步数据揭示了 Nr2f1a 在心室发育中的新要求。瞄准 2，我们将检验Nr2f1a细胞非自主地促进心室生长的假设。最终， 拟议的研究有可能阐明以前未知的分子和遗传病因学 发现与 NR2F2 突变相关的影响心房和心室的先天性心律失常和先天性心脏病 在人类中。