In vitro Expression of Hormone Regulated Genes: COUP-TFII in CDH

描述（由申请人提供）：先天性膈疝（CDH）是一种危及生命的出生缺陷，是儿童死亡和行动不便的主要原因。病理异常的特点是腹部内容物（可能包括胃、肝、肠和脾）不适当地突出，通过不正确形成的隔膜进入胸腔，损害肺的生长发育。除了腹部内容物突出引起的继发性缺陷外，肺发育本身也可能受到影响，并导致这些患者的CDH表型。尽管这种疾病在350多年前就被描述出来了，但对CDH的病因却知之甚少。使用在肠系膜中缺失COUP-TFII的条件零突变体，我们发现COUP-TFII的组织特异性零突变体表现出Bochdalek CDH，这是最常见的CDH形式。COUP-TFII是孤儿核受体的一员，在胚胎发育期间膈肌和肺形成的关键区域表达。消融前肠间质，包括肝后间质板（PHMP）的COUP-TFII，导致膈肌畸形和PHMP与体壁的适当附着失败。最近，通过阵列CGH和Fish分析，15q26.1-26.2内的一个关键区域在CDH患者中被删除。COUP-TFII是位于该关键区域的四个已知基因之一。结合我们的COUP-TFII条件突变体的结果，遗传分析表明COUP-TFII是最常见类型Bochdalek CDH的最有可能的候选基因。为了描述COUP-TFII如何调节膈肌和肺的发育，以及膈肌正常形成的扰动如何导致CDH，提出了三个具体目标：1)。条件阴性小鼠横膈膜发育过程中缺陷的分析2）. 分析COUP-TFII在肺发育中的作用及其与CDH的关系3)。筛选和鉴定CDH患者的COUP-TFII突变。通过这项研究，我们希望对膈肌和肺的形态发生以及这些器官的畸形如何导致这种毁灭性的先天性缺陷提供重要的见解。最终，通过筛查CDH患者的COUP-TFII突变，肯定会提高建立诊断试剂盒的翻译前景，并为一些CDH患者设计干预措施。

DESCRIPTION (provided by applicant): Congenital diaphragmatic hernia (CDH), a life-threatening birth defect, is a major cause of pediatric mortality and mobility. The pathological anomalies are characterized by the inappropriate protrusion of the abdominal contents, possibly including the stomach, liver, intestines and spleen, through an improperly formed diaphragm into the thoracic cavity which impairs lung growth and development. In addition to the secondary defect resulting from protrusion of abdominal contents, lung development itself may be also effected and contribute to CDH phenotypes in these patients. Although the disease was described more than 350 years ago, the etiology of CDH is poorly understood. Using a conditional null mutant in which COUP-TFII is deleted in the mesentery, we showed that tissue specific null mutants of COUP-TFII exhibit Bochdalek CDH, the most common form of CDH. COUP-TFII, a member of orphan nuclear receptors, is expressed in regions critical for the formation of the diaphragm and lung during embryonic development. Ablation of COUP-TFII in the foregut mesenchyme, including the post-hepatic mesenchymal plate (PHMP), results in the malformation of the diaphragm and the failure of appropriate attachment of the PHMP to the body wall. Recently a critical region within 15q26.1-26.2 has been mapped by array CGH and Fish analysis to be deleted in CDH patients. COUP-TFII is one of the four known genes resided within this critical region. Together with results of our COUP-TFII conditional mutants, the genetic analysis implicates COUP-TFII as most likely candidate gene for the most common type of Bochdalek CDH. To delineate how COUP-TFII regulates diaphragm and lung development and how perturbation of proper diaphragm formation might result in CDH, three specific aims are proposed: 1). Analysis of the defects exhibited by the conditional null mice during diaphragm development; 2). Analyze the role of COUP-TFII in lung development and its relationship to CDH; 3). Screen and characterize COUP-TFII mutations in CDH patients. With the proposed study, we hope to provide important insights on morphogenesis of the diaphragm and lung and how malformation of these organs leads to such devastating congenital defects. And eventually, by screening CDH patients for the mutations in COUP-TFII, it will certainly improve the translational prospect in setting up a diagnostic kit and devise an intervention for some CDH patients.