Identification and Characterization of Genes in Congenital Diaphragmatic Hernia

先天性膈疝基因的鉴定和表征

• 批准号: 8280344
• 负责人: Margaret J Wat
• 金额: $ 4.26万
• 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: 2010-07-01 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8280344
• 关键词: 8p23.1; Abdomen; Affect; Anterior; Biological Assay; Candidate Disease Gene; Caring; Cells; Chest; Child; Chromosomal Duplication; Chromosome Deletion; Chromosomes; Clinical; Congenital Abnormality; Congenital diaphragmatic hernia; Custom; Cytogenetic Analysis; DNA; Defect; Development; Disease; Evaluation; Exons; Family; Genes; Genetic; Genetic Counseling; Genome; Genotype; Goals; Heart; Hernia; Human; Hybridization Array; In Vitro; Incidence; Individual; Knowledge; Lead; Life; Live Birth; Liver; Lung; Maps; Medical; Methods; Molecular; Mus; Mutation; Organ; Pathway interactions; Patients; Peritoneal; Phenotype; Physicians; Play; Pulmonary Hypertension; Resources; Respiratory Diaphragm; Role; Scientist; Screening procedure; Series; Tendon structure; Tretinoin; Up-Regulation; base; cohort; comparative genomic hybridization; design; gene discovery; human embryonic stem cell; in vivo; innovation; mortality; mouse model; novel; novel strategies; prevent; research study

DESCRIPTION (provided by applicant): The goal of this proposal is to identify and characterize novel genes for congenital diaphragmatic hernia (CDH). CDH occurs in ~1/3000 live births and has a mortality rate of 30-60% caused mostly by pulmonary hypoplasia and pulmonary hypertension. 30-40% of CDH cases are associated with other severe birth defects including heart anomalies. Although genetic factors clearly play an important role in diaphragm development only a few CDH genes have been identified and little is known about the mechanisms by which they cause CDH. The Specific Aims for this study are: 1) map and identify genes that contribute to human CDH, 2) determine the histopathologic mechanisms that lead to development of anterior CDH in the Sox7 mouse model, and 3) determine if Sox7 interacts with Gata4 in development of anterior CDH. Array comparative genomic hybridization (aCGH) has been shown to be an effective non-biased method for identifying genes in sporadic disorders like CDH. DNA from a cohort of 150 CDH patients will be screened for small CDH-related chromosomal deletions/duplications using a custom designed exon-focused aCGH array. Candidate genes will be selected from these regions and screened for additional mutations in our CDH cohort. Genotype-phenotype correlations will then be made based on clinical information from individual patients. aCGH studies have revealed that one of the most common chromosomal regions deleted in CDH is a portion of 8p23.1 which contains the S0X7 gene. A newly-created Sox7 mouse model of CDH will be used to explore the histopathologic mechanisms that underlie the formation of anterior CDH. Studies will look for changes that could compromise the structural integrity of the diaphragm and evidence of abnormal expansion of the peritoneal mesothelial folds that separate the diaphragm from the liver. It is likely that these mechanisms will be similar to those which cause anterior CDH in humans. Sox7 is required for upregulation of Gata4, another CDH-related gene located in the CDH critical region on 8p23.1. In vivo mouse studies will be used to determine if Sox7 and Gata4 interact genetically in anterior diaphragm development followed by in vitro studies to determine the molecular basis of this interaction. These studies will help identify genetic changes that cause some children to develop congenital diaphragmatic hernia (CDH), a life threatening birth defect. The knowledge gained will help doctors provide better medical care and genetic counseling to affected families and may lead to new ways to prevent or treat these hernias. Since 30-40% of all children with CDH have other severe birth defects, these studies may also help physicians and scientists understand what causes these defects and how to treat them.

描述（由申请人提供）：该提案的目标是识别和表征先天性膈疝（CDH）的新基因。 CDH 发生于约 1/3000 的活产中，死亡率为 30-60%，主要由肺发育不全和肺动脉高压引起。 30-40% 的 CDH 病例与其他严重出生缺陷有关，包括心脏异常。尽管遗传因素显然在膈肌发育中发挥着重要作用，但仅鉴定了少数 CDH 基因，并且对其引起 CDH 的机制知之甚少。这项研究的具体目标是：1) 绘制和鉴定有助于人类 CDH 的基因，2) 确定导致 Sox7 小鼠模型中前部 CDH 发育的组织病理学机制，以及 3) 确定 Sox7 是否在前部 CDH 发育过程中与 Gata4 相互作用。阵列比较基因组杂交 (aCGH) 已被证明是一种有效、无偏见的方法，可用于识别 CDH 等散发性疾病中的基因。将使用定制设计的外显子聚焦 aCGH 阵列，对来自 150 名 CDH 患者的 DNA 进行筛查，以发现与 CDH 相关的小型染色体缺失/重复。将从这些区域中选择候选基因，并在我们的 CDH 队列中筛选其他突变。然后将根据个体患者的临床信息建立基因型-表型相关性。 aCGH 研究表明，CDH 中最常见的染色体缺失区域之一是 8p23.1 的一部分，其中包含 S0X7 基因。新创建的 Sox7 小鼠 CDH 模型将用于探索前部 CDH 形成的组织病理学机制。研究将寻找可能损害膈肌结构完整性的变化，以及将膈肌与肝脏分开的腹膜间皮褶皱异常扩张的证据。这些机制很可能与导致人类前部 CDH 的机制相似。 Sox7 是 Gata4 上调所必需的，Gata4 是位于 8p23.1 上 CDH 关键区域的另一个 CDH 相关基因。小鼠体内研究将用于确定 Sox7 和 Gata4 在前膈发育中是否存在遗传相互作用，随后进行体外研究以确定这种相互作用的分子基础。 这些研究将有助于识别导致一些儿童患上先天性膈疝（CDH）的基因变化，这是一种危及生命的出生缺陷。获得的知识将帮助医生为受影响的家庭提供更好的医疗护理和遗传咨询，并可能带来预防或治疗这些疝气的新方法。由于 30-40% 的 CDH 儿童有其他严重的出生缺陷，这些研究也可能帮助医生和科学家了解导致这些缺陷的原因以及如何治疗它们。