Proteomics of Biliary Atresia

描述(申请人提供)：胆道闭锁(BA)是儿童最严重的肝脏疾病，也是儿童肝移植最常见的适应症。然而，这种进行性硬化性胆管病仍然很难诊断和处理。此外，BA的病理生理学和病因学仍知之甚少。由于一些患有BA的婴儿受益于立即进行Kasai门肠吻合术的外科治疗，因此需要更快速、更敏感和更具特异性的诊断方法来识别BA婴儿。随着新的蛋白质组学技术和新的基于计算机的图像分析和预测统计算法的出现，最近在使用血清蛋白质谱分析根据疾病状态或预后分离患者的能力方面取得了显著进展。我们建议使用荧光双向凝胶电泳法、计算机辅助图像分析、自动蛋白质斑点提取和蛋白质分解消化相结合的方法分析来自BA和新生儿肝炎(NH)的人血清样本，然后使用测序质谱仪进行蛋白质鉴定。然后，我们将使用基于计算机的统计算法进行数据分析，以确定预测哪些婴儿患有BA和哪些婴儿患有NH的蛋白质模式。在这些研究的同时，我们建立了一个小鼠模型系统来检测胆管结扎对血清蛋白丰度的影响。该模型系统将深入了解胆道闭锁婴儿的血清蛋白变化主要是由胆道梗阻引起的。我们参与了最近成立的胆道闭锁研究联盟(BARC)，极大地促进了我们对婴儿血清的研究。这项由9个主要儿科医学中心进行的合作研究将收集患者的临床数据和生物样本，这些数据和生物样本将由许多BARC相关研究人员分析。尽管样本将被编码以保护患者身份，但事实上，来自同一患者的样本和临床数据将由不同的小组进行许多不同类型的研究，这将极大地增加我们蛋白质组分析的价值。

DESCRIPTION (provided by applicant): Biliary Atresia (BA) is the most serious liver disease of childhood and the most common indication for pediatric liver transplantation. This progressive sclerosing cholangiopathy, however, remains difficult to diagnose and to manage. In addition, the pathophysiology and etiology of BA remains poorly understood. Because some infants with BA benefit from immediate surgical treatment with a Kasai portoenterostomy, diagnostic methods to identify infants with BA more rapidly and with greater sensitivity and specificity are needed. With the advent of new proteomic technology and new computer based image analysis and predictive statistical algorithms, recent dramatic advances have been made in the ability to use serum protein profile analysis to segregate patients by disease state or prognosis. We propose to analyze human serum samples from infants with BA and neonatal hepatitis (NH) using a combination of fluorescent two dimensional gel electrophoresis, computer assisted image analysis, automated protein spot picking and proteolytic digestion followed by protein identification using sequencing mass spectroscopy. We will then use computer based statistical algorithms for data analysis to identify protein pattems that predict which infants have BA and which infants have NH. In parallel with these studies, we have established a mouse model system to examine the effect of bile duct ligation on the abundance of serum proteins. This model system will provide insight about which serum protein changes in infants with biliary atresia result primarily from biliary tract obstruction. Our studies of human infant serum are greatly facilitated by our participation in the recently created Biliary Atresia Research Consortium (BARC). This collaborative study by 9 major pediatric medical centers will collect patient clinical data and biological samples that will be analyzed by many BARC associated investigators. Although the samples will be coded to protect patient identity, the fact that many different types of research will be performed by different groups on samples and clinical data from the same patients will greatly add to the value of our proteomic analyses.