Proteomics of Biliary Atresia

描述（申请人提供）：胆道闭锁（Biliary Atresia， 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难以诊断和治疗。此外，BA的病理生理和病因尚不清楚。由于一些BA患儿可通过Kasai门肠造口术立即手术治疗，因此需要更快速、更敏感和特异性地识别BA患儿的诊断方法。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着进展。我们建议采用荧光二维凝胶电泳、计算机辅助图像分析、自动蛋白质斑点提取和蛋白质水解消化结合测序质谱技术对BA和新生儿肝炎（NH）婴儿血清样本进行分析。然后，我们将使用基于计算机的统计算法进行数据分析，以确定蛋白质模式，预测哪些婴儿患有BA，哪些婴儿患有NH。在这些研究的同时，我们建立了一个小鼠模型系统来研究胆管结扎对血清蛋白丰度的影响。该模型系统将提供关于胆道闭锁婴儿血清蛋白变化主要由胆道阻塞引起的见解。我们的人类婴儿血清的研究极大地促进了我们参与最近创建的胆道闭锁研究联盟（BARC）。这项由9个主要儿科医学中心开展的合作研究将收集患者临床数据和生物样本，并由许多BARC相关研究人员进行分析。虽然样本将被编码以保护患者身份，但不同的研究小组将对来自同一患者的样本和临床数据进行许多不同类型的研究，这一事实将大大增加我们蛋白质组学分析的价值。

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.