Molecular Analysis of Ureteric Reflux

描述（由申请人提供）： 反流性肾病（RN）是一种与原发性膀胱输尿管反流（VUR）相关的肾脏瘢痕形成综合征。已知原发性VUR表现为常染色体显性遗传。然而，原发性VUR的遗传缺陷及其分子发病机制尚不清楚。目前VUR和RN的诊断涉及侵入性和昂贵的放射学检查。因此，在兄弟姐妹和后代中进行VUR筛查以及在原发性VUR患者中进行RN筛查不是常规做法。早期发现VUR对预防RN有重要意义，抗生素预防可有效降低瘢痕形成率。该提案的长期目标是开发用于VUR和RN的早期诊断和监测的侵入性较小的测试，以及基于对分子发病机制的更好理解来开发治疗策略。假设原发性VUR和RN患者的输尿管组织和尿液中存在特征性基因和蛋白表达模式。定制点样cDNA微阵列将用于研究差异gone表达。表面增强激光解吸/电离飞行时间质谱与串联质谱联用将用于研究差异蛋白质表达。然而，后一种方法限于小于70 kD的分子。对于较大的分子，将采用二维凝胶电泳。将原发性VUR患者的输尿管组织与继发性VUR患者和肾移植供体的正常对照进行比较。将原发性和继发性VUR伴或不伴RN患者的尿样与年龄和性别匹配的正常儿童进行比较。数据分析将采用最先进的生物信息学方法。将考虑混杂变量，包括年龄、性别和是否存在肾小球蛋白尿。鉴定VUR和RN患者的特异性尿生物标志物可能会导致进一步了解分子发病机制，以及原发性VUR和RN的微创筛查试验的发展。将建立原发性VUR和RN的临床数据库，以建立生物标志物和表型之间的相关性，并测试这些标志物检测原发性VUR和RN的灵敏度和特异性。

DESCRIPTION (provided by applicant): Reflux nephropathy (RN) is a syndrome of renal scarring associated with primary vesico-ureteral reflux (VUR). Primary VUR is known to exhibit autosomal dominant inheritance. However, the genetic defect of primary VUR and its molecular pathogenesis are not well understood. The current diagnosis of VUR and RN involves radiologic tests which are invasive and costly. Therefore VUR screening in siblings and offspring and RN screening in patients with primary VUR are not routinely practised. Early detection of VUR is valuable for the prevention of RN since the incidence of scarring can be reduced effectively by antiobiotic prophylaxis. The long-term goals of this proposal are to develop less-invasive tests for early diagnosis and monitoring of VUR and RN as well as to develop therapeutic strategies based on a better understanding of the molecular pathogenesis. The hypothesis is that characteristic gene and protein expression patterns exist in ureteric tissue as well as in urine of patients with primary VUR and RN. Custom-spotted cDNA microarrays will be used to study differential gone expression. Surface-enhanced laser desorption/ionization time-of-flight mass spectroscopy coupled with tandem mass spectroscopy will be used to study differential protein expression. This latter method is however limited to molecules smaller than 70 kD. For larger molecules, two-dimensional gel electrophoresis will be employed. Ureteric tissue from patients with primary VUR will be compared with those from patients with secondary VUR and normal controls from renal transplant donors. Urine samples from patients with primary and secondary VUR with and without RN will be compared with age- and gender-matched normal children. State-of-the-art bioinformatics methodologies will be used for data analysis. Confounding variables, including age, gender and presence of glomerular proteinuria will be considered. Identification of specific urinary biomarkers in patients with VUR and RN may lead to further understanding of the molecular pathogenesis, as well as the development of less-invasive screening tests for primary VUR and RN. A clinical database of primary VUR and RN will be set up to establish correlations between biomarkers and phenotype and to test the sensitivity and specificity of these markers for detecting primary VUR and RN.