Characterization of the urofacial syndrome gene

描述(申请人提供)：尿面部(Ochoa)综合征(UFS，OMIM#236730)是一种常染色体隐性遗传病，特征是先天性梗阻性尿路病变和面部表情异常。由于神经源性膀胱，患者有遗尿、尿路感染、肾积水和排尿功能障碍。之前的全基因组扫描将疾病基因定位在染色体10q23-q24上。目前，我们已经分析了33个多态SNPs，并将疾病基因缩小到D10S2499和D10S603标记之间的1.8Mb基因组DNA。本研究的目的是利用已有的高通量突变筛选技术，通过定位克隆的方法对缺陷基因进行鉴定。在过去的几十年里，位置克隆被用来识别许多人类疾病基因，当它们的基因组位置被缩小到很小的区域时，例如几百kb的DNA。由于孟德尔的大多数特征是罕见的和零星的，很难获得信息丰富的家系，以将疾病间隔缩小到如此小的地区。这一现实严重阻碍了对疾病表型缺陷基因的表征。目前的UFS间隔包含1.8Mb的基因组DNA，对于传统的位置克隆来说太大了。因此，我们提出了一种新的克隆策略。我们将使用高通量的spectruMedex系统进行突变筛选，并将根据人类基因组资源对UFS区间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的基因组间隔中识别UFS基因。UFS为神经源性膀胱、隐匿性神经源性膀胱和其他排尿功能障碍的研究提供了一个极好的模型。这种破坏性的疾病影响着大量的美国人和世界上的其他人，而且可能受到同样的神经起源的先天性疾病的影响。UFS的特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。这一建议中应用的策略可能对人类疾病基因的定位克隆具有重要意义。一旦我们在UFS中确认了这种方法的可行性，它就可以用于克隆具有大基因组间隔的人类疾病基因座，例如那些基因组DNA长达几个Mb甚至10Mb的疾病基因座。

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.