Genetic analysis of congenital diaphragmatic hernia

描述(申请人提供)：长期目标：我们想要研究人类先天性横隔疝(CDH)的遗传病因。这种常见的出生缺陷与显著的产前死亡率和长期发病率有关。有重要的证据表明基因在CDH的病因中很重要，但适合在这种零星情况下进行基因定位的孟德尔家系很少见。我们目前正在使用阵列比较基因组杂交(阵列CGH)来识别患有CDH和染色体易位的患者的亚显微染色体异常。我们已经确认了一位CDH先证者，他具有从头染色体易位，并且单个细菌人工染色体(BAG)克隆的拷贝数减少，定位于CGH阵列上易位断裂点的区域。我们想要证实CGH的结果，并表明易位断裂点与CDH发病相关的基因缺失或表达改变有关。如果我们能确定这个基因，我们就可以确定该基因在CDH发病机制中的作用程度，在一组非综合征性CDH和CDH及畸形患者中。我们还将使用系统发育/基因组学方法来确定与已鉴定的基因和蛋白质具有相似结构和功能基序的基因和蛋白质同源物。本研究的主要目的是：1.鉴定先天性巨噬细胞增多症患者的染色体断裂点缺失基因(S)或基因表达改变。2.对48例非综合征性CDH患者和24例CDH及其他畸形患者进行测序，以确定该基因在人类CDH发病机制中的参与程度。3.用基因组/系统发育学方法确定与已鉴定的基因和蛋白具有相似结构和功能的基因和蛋白同源物，并在同一CDH患者队列中通过直接测序筛选这些新基因的序列改变。

DESCRIPTION (provided by applicant): Long term objectives: We would like to investigate the genetic etiology of congenital diaphragmatic hernia (CDH) in humans. This common birth defect is associated with a significant prenatal mortality and long-term morbidity. There is significant evidence that genes are important in the etiology of CDH but Mendelian pedigrees suitable for gene mapping in this sporadic condition are rare. We are currently using array comparative genomic hybridization (array CGH) to identify submicroscopic chromosome aberrations in patients who have CDH and chromosome translocations. We have identified a proband with CDH who has a de novo chromosome translocation and reduced copy number for a single bacterial artificial chromosome (BAG) clone localized to the region of a translocation breakpoint on array CGH. We would like to confirm the CGH result and show that the translocation breakpoint is associated with a gene deletion or altered expression of a gene involved in the pathogenesis of CDH. If we can identify this gene, we would determine the degree of involvement of the gene in the pathogenesis of CDH in a cohort of patients with non-syndromic CDH and with CDH and malformations. We would also use a phylogenetic/genomic approach to ascertain gene and protein homologues with similar structural and functional motifs to the identified gene and protein. Our specific aims are 1. To identify a deleted gene(s) or altered expression of a gene at a chromosome breakpoint in a patient with CDH and a de novo chromosome translocation. 2. To determine the degree of involvement of the identified gene in the pathogenesis of CDH in humans by sequencing this gene in a cohort of 48 patients with non-syndromic CDH and in 24 patients CDH and additional malformations. 3. To use a genomic/phylogenetic approach to ascertain gene and protein homologues with similar structure and function to the identified gene and protein and to screen these new genes for sequence alterations by direct sequencing in the same patient cohort with CDH.