Molecular Genetics of Congenital Diaphragmatic Hernia

申请人描述(申请人提供)：申请者为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内科科学家，专注于研究影响儿童健康的遗传疾病。这项申请将为应聘者提供发育生物学、人类遗传学和老鼠遗传学方面的新技术方面的培训。总的科学目标是识别和表征导致先天性横隔膜疝气(CDH)的基因。CDH是一种发育异常，即横隔区不能形成，使腹腔内容物侵入正常为发育中的肺保留的空间。大约每2500名新生儿中就有一名患有CDH。死亡率从30%到60%不等，存活的婴儿通常有明显的肺发育不良和肺动脉高压的发病率。本研究的具体目的包括：1)建立研究CDH的DNA协作库；2)筛选CDH突变的候选基因；3)利用新的比较基因组杂交(CGH)技术鉴定和定位与CDH相关的染色体区域；4)鉴定CDH小鼠模型。根据染色体位置和可能的功能选择的候选基因将通过直接测序来筛选导致CDH的突变。来自冠心病患者的DNA将通过G-显带染色体分析和CGH进行筛查，方法是使用一个由大约300 kb分辨率的20,000多个重叠克隆组成的全基因组微阵列。通过选择性地切除COUP-TFII基因，建立了一种模仿人类最常见形式的CDH的CDH小鼠模型。COUP-TFII是一种受维甲酸途径调节的转录因子基因。候选人将通过确定COUP-TFII消融对横隔肌肌肉组织、神经支配、细胞增殖和细胞凋亡的影响来表征这些小鼠。识别和鉴定导致CDH的基因可能会导致新的预防和治疗策略的发展，并将提供对横隔膜发育的分子基础的洞察。

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.