Retinoid-Related Genes in Diaphragm and Cardiac Development

膈肌和心脏发育中的类维生素A相关基因

• 批准号: 7860177
• 负责人: Daryl Armstrong Scott
• 金额: $ 31.85万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-15 至 2015-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7860177
• 关键词: 8p23.1; Anterior; Binding; Binding Sites; Biological; Biological Assay; Cardiac; Cardiovascular system; Cells; Chromosomes; Congenital Abnormality; Congenital Heart Defects; Congenital diaphragmatic hernia; Defect; Development; Diagnosis; Embryo; Genes; Genetic; Genotype; Goals; Heart; Heterozygote; Human; Human Resources; In Vitro; Incidence; Individual; Knowledge; Lead; Life; Luciferases; Modeling; Molecular; Mus; Organ; Pathway interactions; Patients; Peritoneal; Phenotype; Play; Pleural; Reporter; Respiratory Diaphragm; Retinoids; Role; Severities; Signal Pathway; System; Techniques; Tretinoin; Up-Regulation; Xenopus; apoAI regulatory protein-1; base; cardiogenesis; chromatin immunoprecipitation; cohort; human data; in vivo; malformation; mouse model; novel; novel therapeutics; prevent; programs; promoter; public health relevance; transcription factor

DESCRIPTION (provided by applicant): Our long-term goal is to understand the molecular basis of congenital diaphragmatic hernia (CDH)-a common life threatening birth defect. However, our ability to identify key molecular interactions involved in CDH development is hindered by a general paucity of known CDH-related genes, an incomplete understanding of the histopathologic changes that underlie CDH development and the lack of a cell based/culture based system in which to study diaphragm development. These limitations could be minimized by studying CDH-related genes that are also involved in the development of related organs, such as the heart, that are more amenable to study. Since up to 40% of individuals with CDH also have cardiovascular malformations (CVMs), it is likely that some key developmental programs/pathways are shared between these organs. In this proposal we will identify these commonalities by studying SOX7 and GATA4-two retinoid-responsive transcription factors located together in the CDH/CVM minimal deleted region on chromosome 8p23.1-using novel mouse models, cell based systems, and unique human resources. Specific aims for this proposal include: 1) identify the histopathologic changes associated with disruption of Sox7 and Gata4 in the diaphragm, 2) define the role of Sox7 in cardiac development, 3) identify the molecular basis of SOX7-dependent up regulation of Gata4 and determine if Sox7 interacts genetically with Gata4 in diaphragm and cardiac development and 4) identify deleterious changes in retinoid-related genes that cause or predispose to the development of CDH. We have shown that a portion of Sox7 mice develop anterior CDH similar to that seen in Gata4 mice. We will first determine the histopathologic changes that predispose to CDH in this model and then determine the role of Sox7 and Gata4 in the posterior diaphragm by conditionally disrupting their expression using an Nkx3-2 Cre. To define Sox7's role in heart development we will alter Sox7 expression in differentiating P19 cells in vitro and analyze the in vivo cardiac phenotype of Sox7-/- embryos and mice in which Sox7 expression has been disrupted in the first and second heart fields using an Nkx2-5 Cre. The molecular changes that underlie these phenotypes will then be identified using standard molecular techniques. We will also determine if direct binding of SOX7 to the Gata4 promoter is responsible for SOX7-dependent up regulation of Gata4. By comparing the incidence and severity of CDH and CVMs in single and double heterozygous mice, we will also determine if Sox7 interacts genetically with Gata4 in vivo. To confirm their individual roles in human CDH, and to discover novel genotype/phenotype correlations, we will identify deleterious changes in SOX7, GATA4 and two other retinoid-related transcription factors, COUP-TFII and FOG2, in a cohort of CDH patients. In the short term, these studies will help us to understand the key molecular and biological pathway regulated by SOX7 and GATA4 in both the diaphragm and heart whose disruption leads to development of CDH and CVMs. This, in turn, may lead to the development of novel therapeutic or preventative strategies for these birth defects. PUBLIC HEALTH RELEVANCE: SOX7 and GATA4 are two genes that are important for both diaphragm and heart development. These studies will help us understand how changes in SOX7 and GATA4 cause these life- threatening birth defects and what other genes may be involved. This knowledge may lead to the development of new ways to diagnose, prevent or treat these birth defects.

描述(由申请者提供)：我们的长期目标是了解先天性横隔疝(CDH)的分子基础--一种常见的危及生命的出生缺陷。然而，我们识别CDH发育中的关键分子相互作用的能力受到以下因素的阻碍：已知CDH相关基因的普遍缺乏，对CDH发育背后的组织病理学变化的不完全了解，以及缺乏基于细胞/培养的系统来研究横隔膜的发育。通过研究CDH相关基因，可以最大限度地减少这些限制，这些基因也参与相关器官的发育，如心脏，这些器官更容易研究。由于高达40%的CDH患者还患有心血管畸形(CVM)，很可能这些器官之间共享一些关键的发育程序/途径。在这项建议中，我们将通过研究SOX7和GATA4-两个一起位于染色体8p23.1上CDH/CVM最小缺失区域的维甲酸反应转录因子-来识别这些共同点，使用新的小鼠模型、基于细胞的系统和独特的人力资源。这一建议的具体目的包括：1)确定与横隔膜中Sox7和Gata4基因中断相关的组织病理学变化，2)确定Sox7在心脏发育中的作用，3)确定SOX7依赖的Gata4上调的分子基础，并确定Sox7是否在横隔膜和心脏发育中与Gata4发生遗传相互作用，以及4)确定导致或倾向于CDH发生的维甲酸相关基因的有害变化。我们已经证明，SOX7小鼠的一部分出现了与Gata4小鼠相似的前部CDH。我们将首先确定在该模型中易患CDH的组织病理学变化，然后通过使用Nkx3-2 Cre有条件地干扰Sox7和Gata4的表达来确定Sox7和Gata4在后隔膜中的作用。为了确定Sox7的S在心脏发育中的作用，我们将在体外改变Sox7在P19细胞分化中的表达，并使用Nkx2-5 CRE分析Sox7-/-胚胎和第一和第二心脏区域Sox7表达中断的小鼠的体内心脏表型。这些表型背后的分子变化将使用标准分子技术进行鉴定。我们还将确定SOX7与Gata4启动子的直接结合是否导致SOX7依赖的Gata4上调。通过比较CDH和CMS在单杂合子和双杂合子小鼠中的发生率和严重程度，我们还将确定Sox7是否在体内与Gata4存在遗传相互作用。为了确认它们在人类CDH中的个体作用，并发现新的基因型/表型相关性，我们将在CDH患者队列中鉴定SOX7、GATA4和另外两个维甲类相关转录因子COUP-TFII和FOG2的有害变化。在短期内，这些研究将有助于我们了解SOX7和GATA4在横隔膜和心脏中调控的关键分子和生物学途径，这些途径的破坏导致CDH和CMS的发生。这反过来可能导致针对这些出生缺陷的新的治疗或预防策略的开发。 与公共卫生相关：SOX7和GATA4是两个对横隔膜和心脏发育都很重要的基因。这些研究将帮助我们了解SOX7和GATA4的变化如何导致这些危及生命的出生缺陷，以及哪些其他基因可能参与其中。这一知识可能会导致开发新的方法来诊断、预防或治疗这些出生缺陷。