Characterization of the urofacial syndrome gene

描述（由申请人提供）：Urofacial（Ochoa）综合征（UFS，OMIM #236730）是一种常染色体隐性遗传疾病，其特征为先天性阻塞性尿路病和面部表情异常。患者有遗尿，尿路感染，肾积水和排尿功能障碍的结果，神经源性膀胱。先前的全基因组扫描将疾病基因定位于染色体10 q23-q24。目前，我们已经分析了33个多态性SNP，并将疾病基因缩小到标记D10 S2499和D10 S603之间的1.8Mb基因组DNA。本课题的目的是利用高通量突变筛选技术，通过定位克隆的方法来鉴定缺陷基因。在过去的几十年里，定位克隆已经被用于识别许多人类疾病基因，当它们的基因组位置被缩小到小区域时，例如，几百kb的DNA由于大多数孟德尔性状是罕见和零星的，很难获得信息丰富的系谱，以缩小疾病间隔到这样的小区域。这一现实严重阻碍了疾病表型缺陷基因的表征。目前的UFS区间包含1.8Mb的基因组DNA，这对于传统的定位克隆来说太大了。因此，我们提出了一种新的克隆策略。我们将使用高通量SpectruMeetings系统进行突变筛查，并根据人类基因组资源对UFS间隔内的所有候选基因进行表征。这种高通量突变筛选系统和人类基因组资源的结合将使我们能够从这1.8Mb的基因组间隔中鉴定UFS基因。UFS为研究神经源性膀胱、隐匿性神经源性膀胱和其他排尿功能障碍提供了一个很好的模型。这种毁灭性的疾病影响了大量的美国人和世界上其他人，并且可能受到神经起源的相同先天性疾病的影响。UFS的表征将使我们能够创建动物模型来研究神经源性膀胱的分子机制。本研究所采用的策略可能对人类疾病基因的定位克隆具有重要意义。一旦我们证实了这种方法在UFS中的可行性，它就可以应用于克隆具有大基因组间隔的人类疾病基因座，例如，这些疾病位点具有几Mb甚至10 Mb的基因组DNA。

DESCRIPTION (provided by applicant): The Urofacial (Ochoa) syndrome (UFS, OMIM #236730) is an autosomal recessive disease characterized by congenital obstructive uropathy and abnormal facial expression. The patients have enuresis, urinary tract infection, hydronephrosis and voiding dysfunction as a result of a neurogenic bladder. Previous genome-wide scan localized the disease gene to chromosome 10q23-q24. Currently, we have analyzed 33 polymorphic SNPs and narrowed the disease gene to a 1.8Mb of genomic DNA between markers D10S2499 and D10S603. The goal of this propossal is to characterize the defective gene by positional cloning with the adavnced high throughput mutation screening techniques. For the past several decades, positional cloning has been used to identify many human disease genes when their genomic locations are narrowed to small regions, e.g., several hundred-kb of DNA. As most of the mendelian traits are rare and sporadic, it is difficult to obtain informative pedigrees to narrow the disease interval to such small regions. This reality significantly hampered the characterization of the defective genes for the disease phenotypes. The current UFS interval contains 1.8Mb of genomic DNA which is too big for conventional positional cloning. As a result, we propose a novel strategy for the cloning. We will use the high throughput SpectruMedix system for mutation screening, and all of the candidate genes within the UFS interval will be characterized based on the human genome resources. This combination of high throughput mutation screening system and human genome resources would allow us to identify the UFS gene from this 1.8Mb of genomic interval. UFS provides an excellent model for the studies of neurogenic bladders, occult neurogenic bladders and other voiding dysfunctions. Such devastating disorders affect a large number of Americans and other people in the world, and are probably affected by the same congenital disorder of neurological origin. Characterization of UFS would allow us to create animal models to study the molecular mechanism underlining neurogenic bladders. The strategy applied in this proposal may have significant implications for positional cloning of the human disease genes. Once we have confirmed the feasibility of this approach in UFS, it can be applied for cloning of human disease loci with large genomic intervals, e.g., those disease loci with several Mb to even 10Mb of genomic DNA.