Molecular Genetics of Congenital Diaphragmatic Hernia

描述（由申请人提供）：申请人是一名已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医生科学家，专注于研究影响儿童健康的遗传疾病。 该申请将为候选人提供发育生物学，人类遗传学新技术和小鼠遗传学方面的培训。 总体科学目标是确定和表征先天性腹股沟疝（CDH）的基因。 CDH是一种发育异常，其中隔膜区域无法形成，从而允许腹腔内容物侵入通常为发育中的肺保留的空间。 CDH影响大约1/2，500的新生儿。 死亡率为30%至60%，存活的婴儿通常有肺发育不全和肺动脉高压的显著发病率。 本研究的具体目标包括：1）建立一个合作的DNA库研究CDH，2）筛选候选基因的CDH引起的突变，3）确定和定位染色体区域涉及CDH使用新的比较基因组杂交（CGH）技术，和4）表征小鼠模型的CDH。 基于染色体位置和推定功能选择的候选基因将通过直接测序筛选引起CDH的突变。 将通过G带染色体分析和CGH，使用分辨率约为300 kb的超过20，000个重叠克隆的全基因组微阵列，对CHD个体的DNA进行异常筛查。 通过选择性切除COUP-TFII（一种由视黄酸途径调控的转录因子基因），开发了一种模拟人类最常见CDH形式的CDH小鼠模型。 候选人将通过确定COUP-TFII消融对膈肌肌肉组织、神经支配、细胞增殖以及细胞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，并将提供对膈肌发育的分子基础的深入了解。

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.