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: 8233924
• 负责人: Daryl Armstrong Scott
• 金额: $ 30.58万
• 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/8233924
• 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（由申请人提供）：我们的长期目标是了解先天性腹股沟疝（CDH）的分子基础-一种常见的危及生命的出生缺陷。然而，我们的能力，以确定关键的分子相互作用参与CDH的发展是阻碍了一般缺乏已知的CDH相关基因，不完全了解的组织病理学变化，CDH的发展和缺乏一个基于细胞/培养为基础的系统，在其中研究隔膜的发展。这些限制可以通过研究与CDH相关的基因来最小化，这些基因也参与了相关器官的发育，如心脏，这些器官更适合研究。由于高达40%的CDH患者也患有心血管畸形（CVM），因此这些器官之间可能共享一些关键的发育程序/途径。在这项提案中，我们将通过研究SOX 7和GATA 4-两个类维生素A反应性转录因子一起位于染色体8p23.1上的CDH/CVM最小缺失区-使用新的小鼠模型，基于细胞的系统和独特的人力资源来确定这些共性。这项建议的具体目标包括：1）鉴定与隔膜中Sox 7和Gata 4破坏相关的组织病理学变化，2）确定Sox 7在心脏发育中的作用，3）鉴定Gata 4的SOX 7依赖性上调的分子基础，并确定Sox 7是否在膈肌和心脏发育中与Gata 4遗传相互作用，以及4）鉴定类维生素A的有害变化。导致或易患CDH的相关基因。我们已经表明，Sox 7小鼠的一部分发展前部CDH类似于在Gata 4小鼠中看到的。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Nkx 3 -2 Cre有条件地破坏其表达来确定Sox 7和Gata 4在后膈中的作用。为了定义Sox 7在心脏发育中的作用，我们将改变Sox 7在体外分化的P19细胞中的表达，并使用Nkx 2 -5 Cre分析Sox 7-/-胚胎和小鼠的体内心脏表型，其中Sox 7在第一和第二心脏区域的表达已被破坏。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SOX 7与Gata 4启动子的直接结合是否负责SOX 7依赖的Gata 4上调。通过比较单杂合子和双杂合子小鼠中CDH和CVM的发病率和严重程度，我们还将确定Sox 7是否与Gata 4在体内发生遗传相互作用。为了证实它们在人类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SOX 7、GATA 4和另外两种维甲酸相关转录因子COUP-TFII和FOG 2的有害变化。在短期内，这些研究将帮助我们了解在膈肌和心脏中由SOX 7和GATA 4调节的关键分子和生物学途径，其破坏导致CDH和CVM的发展。反过来，这可能会导致这些出生缺陷的新的治疗或预防策略的发展。 公共卫生相关性：SOX 7和GATA 4是两个对横膈膜和心脏发育都很重要的基因。这些研究将帮助我们了解SOX 7和GATA 4的变化如何导致这些危及生命的出生缺陷，以及可能涉及哪些其他基因。这些知识可能会导致诊断，预防或治疗这些出生缺陷的新方法的发展。