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: 8440700
• 负责人: Daryl Armstrong Scott
• 金额: $ 29.02万
• 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/8440700
• 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; 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.

描述（由申请人提供）：我们的长期目标是了解先天性隔膜疝（CDH）的分子基础 - 一种常见的生命威胁性先天缺陷。然而，我们鉴定CDH发育中涉及的关键分子相互作用的能力受到了已知CDH相关基因的普遍匮乏的阻碍，这是对CDH发育构成的组织病理学变化的不完全理解，并且缺乏基于细胞/培养的系统，该系统无法研究diaphragm的发育。可以通过研究与CDH相关的基因（例如心脏）的发展，可以最大程度地限制这些局限性，这些基因更容易研究。由于多达40％的CDH患者也患有心血管畸形（CVM），因此可能在这些器官之间共享某些关键的开发程序/途径。在此提案中，我们将通过研究CDH/CVM最小删除区域中的SOX7和GATA4-TWO类视网膜类似的转录因子来识别这些共同点，这些转录因子是8p23.1染色体上的最小删除区域，使用新型小鼠模型，基于细胞的模型，基于细胞的系统和独特的人力资源。 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导致或易感CDH发育的类视视视网想相关基因。我们已经表明，一部分Sox7小鼠会形成前CDH，类似于GATA4小鼠中的CDH。我们将首先确定该模型中CDH易感性的组织病理学变化，然后通过使用NKX3-2 CRE有条件地破坏其表达方式来确定Sox7和Gata4在后膜片中的作用。为了定义SOX7在心脏发育中的作用，我们将在体外区分p19细胞中改变SOX7的表达，并分析Sox7 - / - 胚胎和小鼠的体内心脏表型，其中Sox7表达在其中使用NKX2-5 CRE在第一和第二心脏场中破坏了Sox7的表达。然后，将使用标准分子技术鉴定这些表型的分子变化。我们还将确定SOX7与GATA4启动子的直接结合是否负责SOX7依赖性GATA4的调节。通过比较单一和双重杂合小鼠中CDH和CVM的发病率和严重程度，我们还将确定Sox7是否与体内GATA4相互作用。为了确认其在人类CDH中的各个角色，并发现新的基因型/表型相关性，我们将在一组CDH患者中确定Sox7，GATA4和其他两个与类视黄素相关的转录因子（coup-TFII和FOG2）中的有害变化。在短期内，这些研究将有助于我们了解隔膜和心脏中由Sox7和Gata4调节的关键分子和生物途径，它们的破坏会导致CDH和CVM的发展。反过来，这可能导致为这些出生缺陷的新型治疗或预防策略的发展。