sox9b function in cardiomyocyte and great vessel development

sox9b 在心肌细胞和大血管发育中的功能

基本信息

批准号：
9979907
负责人：
Jessica Susan Plavicki
金额：
$ 24.12万
依托单位：
OCEAN STATE RESEARCH INSTITUTE, INC.
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/9979907
关键词：
5&apos Untranslated Regions Address Adult Affect Air Pollution Animal Model Aortic Valve Stenosis Biology Blood Vessels Candidate Disease Gene Cardiac Cardiac Myocytes Cardiac development Cardiopulmonary Cardiovascular system Centers of Research Excellence Cessation of life Clinical Data Code Congenital Abnormality Congenital Heart Defects Data Defect Development Diffusion Dominant-Negative Mutation Ebstein&apos s Anomaly Echocardiography Embryo Endothelial Cells Endothelium Environmental Pollution Etiology Exposure to Gene Expression Genes Genetic Genetic Diseases Genetic Epistasis Genetic Transcription Genetic study Goals Health Heart Heart Atrium Heart Diseases Heart Valves Heart failure Human Immunohistochemistry Incidence Individual Infant Mortality Laboratories Lead Lithium Mammals Mediating Mediator of activation protein Modeling Molecular Molecular Target Morphogenesis Mus Mutation Oxygen Patent Foramen Ovale Patients Pesticides Phenocopy Phenotype Plastics Play Quantitative Reverse Transcriptase PCR Reporting Research Risk Factors Role Tetralogy of Fallot Toxic Environmental Substances Transgenic Organisms United States Vascular Endothelial Cell Ventricular Septal Defects WNT Signaling Pathway Zebrafish campomelic dysplasia cardiogenesis cell type congenital heart disorder epidemiologic data gene function heart function in vivo infant morbidity inhibitor/antagonist innovation knock-down loss of function malformation optogenetics pleiotropism prevent promoter therapeutic target tool transcription factor transcriptome sequencing wasting

项目摘要

PROJECT SUMMARY/ABSTRACT Congenital abnormalities of the heart and great vessels are the leading cause of infant mortality and morbidity in the United States yet the etiology of most congenital defects remains unknown. Approximately 90% of congenital heart and vessel defect cases are multifactorial. Exposure to environmental contaminants is thought to significantly contribute to the incidence of defects as well adult heart disease. Our long-term goal is to prevent and treat congenital defects and adult heart disease by understanding, in detail, the functions of genes that are critical for cardiovascular development and also disrupted by exposure to environmental toxicants. We have identified a candidate gene, SOX9, that is expressed in multiple cell types in the developing and mature heart and is downregulated in animal models following exposure to environmental toxicants present in air pollution, e-waste, plastics, and pesticides. Human mutations in the SOX9 coding region result in Campomelic Dysplasia (CD), a severe genetic disorder that usually results in death. A number of different congenital heart and great vessel defects have been reported in patients with CD as well as in individuals with mutations affecting SOX9 function. Together, the human loss of function data suggests that SOX9 plays several critical roles in cardiac and great vessel development. However, Sox9b/Sox9 functions in cardiomyocyte and great vessel development have not been investigated using animal models. Our preliminary data indicate that zebrafish Sox9b is essential for cardiomyocyte development and function as well as great vessel formation. Our immediate goal is to identify the molecular mechanisms that mediate the cardiac and great vessel phenotypes observed following loss of Sox9b function in zebrafish and to determine if Sox9b functions are conserved by mammalian Sox9. We are using an innovative multi-species approach and a combination of genetic tools to manipulate zebrafish Soxb and murine Sox9 during cardiovascular development. In Aim 1.1, we will determine if Sox9b is an inhibitor of canonical WNT signaling in the developing zebrafish heart. In Aim 1.2, we will inhibit Sox9b function in embryonic cardiomyocytes and use RNAseq to identify additional molecular targets of Sox9b during zebrafish heart development. In Aim 2, we use the cell-type specific manipulations and optogenetics to determine how loss of Sox9b function leads to the great vessel phenotypes and to determine the relationship between Sox9b, Nkx2.5, and Stat4, two known mediators of great vessel development. In Aim 3, we will extend our understanding of Sox9b function in cardiac development by examining how loss of Sox9 in mouse cardiomyocytes and endothelial cells affects cardiac and great vessel development as well as how loss of Sox9 in the murine heart impacts canonical WNT signaling. Together, these studies will help us understand the endogenous functions of Sox9b/Sox9 and how genetic and/or environmental disruptions in SOX9 function can contribute to congenital heart and great vessel defects in humans.

项目概要/摘要心脏和大血管的先天性异常是婴儿死亡和发病的主要原因在美国，大多数先天性缺陷的病因仍然未知。大约90% 先天性心脏和血管缺陷病例是多因素的。人们认为暴露于环境污染物显着增加缺陷以及成人心脏病的发生率。我们的长期目标是通过详细了解基因的功能来预防和治疗先天性缺陷和成人心脏病这对心血管发育至关重要，但也会因接触环境毒物而受到干扰。我们已鉴定出一个候选基因 SOX9，该基因在发育和成熟的多种细胞类型中表达暴露于空气中环境毒物后的动物模型中心脏的表达下调污染、电子垃圾、塑料和农药。 SOX9 编码区的人类突变导致 Campomelic 发育不良（CD），一种严重的遗传性疾病，通常会导致死亡。多个不同的先天性心脏据报道，CD 患者以及携带突变的个体存在大血管缺陷影响 SOX9 功能。总之，人类功能丧失数据表明 SOX9 发挥着几个关键作用在心脏和大血管发育中发挥作用。然而，Sox9b/Sox9 在心肌细胞中发挥作用，并且具有重要作用。尚未使用动物模型研究血管发育。我们的初步数据表明斑马鱼 Sox9b 对于心肌细胞的发育和功能以及大血管的形成至关重要。我们的近期目标是确定介导心脏和大血管的分子机制斑马鱼 Sox9b 功能丧失后观察到的表型，并确定 Sox9b 功能是否由哺乳动物 Sox9 保守。我们正在使用创新的多物种方法和组合在心血管发育过程中操纵斑马鱼 Soxb 和鼠 Sox9 的遗传工具。在目标 1.1 中，我们将确定 Sox9b 是否是发育中的斑马鱼心脏中典型 WNT 信号传导的抑制剂。瞄准 1.2，我们将抑制胚胎心肌细胞中的Sox9b功能，并使用RNAseq来鉴定额外的斑马鱼心脏发育过程中 Sox9b 的分子靶标。在目标 2 中，我们使用细胞类型特异性操作和光遗传学以确定 Sox9b 功能的丧失如何导致大血管表型并确定 Sox9b、Nkx2.5 和 Stat4（两个已知的大血管调节因子）之间的关系发展。在目标 3 中，我们将通过以下方式扩展对 Sox9b 在心脏发育中功能的理解：检查小鼠心肌细胞和内皮细胞中 Sox9 的缺失如何影响心脏和大血管发育以及小鼠心脏中 Sox9 的缺失如何影响典型的 WNT 信号传导。一起，这些研究将帮助我们了解 Sox9b/Sox9 的内源性功能以及遗传和/或 SOX9 功能的环境破坏可能导致先天性心脏和大血管缺陷人类。