Molecular Genetics of Congenital Diaphragmatic Hernia

描述（申请人提供）：申请人是一名完成临床培训的高级临床遗传学研究员。他的长期目标是成为一名独立的内科科学家，专注于研究影响儿童健康的遗传疾病。该应用程序将为候选人提供发育生物学，人类遗传学和小鼠遗传学新技术方面的培训。总体的科学目标是确定和表征负责先天性膈疝（CDH）的基因。CDH是一种发育异常，其中隔膜区域无法形成，导致腹腔内的内容物侵入通常为发育中的肺保留的空间。大约每2500名新生儿中就有1人感染CDH。死亡率从30%到60%不等，存活下来的婴儿往往有明显的肺发育不全和肺动脉高压的发病率。本研究的具体目标包括：1)建立CDH研究的合作DNA库；2)筛选CDH引起突变的候选基因；3)利用新型比较基因组杂交（CGH）技术鉴定和定位CDH相关的染色体区域；4)表征CDH小鼠模型。根据染色体位置和推测的功能选择候选基因，通过直接测序筛选导致cdh突变的基因。来自冠心病患者的DNA将通过g带染色体分析和CGH筛选异常，使用超过20,000个重叠克隆的全基因组微阵列，分辨率约为300 kb。通过选择性消融COUP-TFII（一种由视黄酸途径调节的转录因子基因），一种模拟人类最常见形式CDH的CDH小鼠模型已经建立。候选人将通过确定COUP-TFII消融对膈肌组织、神经支配、细胞增殖和凋亡的影响来表征这些小鼠。识别和表征导致CDH的基因可能会导致新的预防和治疗策略的发展，并将为隔膜发育的分子基础提供见解。

DESCRIPTION (provided by applicant): The applkicant is a senior clinical genetics fellow who has completed his clinical training. His long-term objective is to become an independent physician scientist focused on studying genetic disorders that impact the health of children. This application will provide the candidate with training in developmental biology, new technologies in human genetics, and mouse genetics. The overall scientific goal is to identify and characterize genes responsible for congenital diaphragmatic hernia (CDH). CDH is a developmental abnormality in which regions of the diaphragm fail to form, allowing the contents of the abdominal cavity to invade the space normally reserved for the developing lung. CDH affects approximately 1 in 2,500 newborns. Mortality ranges from 30 to 60%, and infants who survive often have significant morbidity from pulmonary hypoplasia and pulmonary hypertension. Specific aims for this study include: 1) establishing a collaborative DNA repository for the study of CDH, 2) screening candidate genes for CDH-causing mutations, 3) identifying and mapping chromosomal regions involved in CDH using novel comparative genomic hybridization (CGH) technology, and 4) characterizing a mouse model of CDH. Candidate genes, selected based on chromosome position and putative function, will be screened for CDH-causing mutation by direct sequencing. DNA from individuals with CHD will be screened for abnormalities by G-banded chromosome analysis and CGH using a genome-wide microarray of over 20,000 overlapping clones with a approximately 300 kb resolution. A CDH mouse model that mimics the most common form of CDH in human has been developed by selective ablation of COUP-TFII, a gene for transcription factor regulated by the retinoic acid pathway. The candidate will characterize these mice by determining the effect of COUP-TFII ablation on diaphragm musculature, innervation, cellular proliferation, as well as apoptosis. Identifying and characterizing the genes that cause CDH may lead to the development of new preventative and therapeutic strategies, and will provide insight into the molecular basis of diaphragm development.