Biological Basis of Phenotypes and Clinical Outcomes in Biliary Atresia

项目摘要/摘要 这是一个竞争性的更新应用，研究临床表型和结果的生物学基础。 胆道闭锁，新生儿胆汁淤积最常见的原因。这种疾病是由纤维炎症引起的。 肝外胆管梗阻，出现于婴儿期早期。尽管接近尾声的进程几乎一致- 肝硬化期，手术/药物治疗的不同反应和疾病的进展率提示 存在导致疾病不同表型或阶段的未被认识的生物过程。 在该奖项之前的任期内，我们发现了通过IL-8、肿瘤坏死因子和成分增加的信号转导的证据 炎性小体在胆管损伤发病机制中的作用及分子回路的同时激活 依赖IL-33诱导组织修复。我们还确定了基质金属蛋白酶-7在胆管上皮损伤中的关键作用。 作为胆道闭锁的高度敏感和特异的生物标志物。在为此应用程序做准备时，我们 应用计算机建模和高级分析挖掘肝脏转录组，发现了一个14基因 预测天然肝脏存活2年的签名，并将谷胱甘肽代谢确定为新的 治疗的目标是抑制纤维化。利用血清蛋白质组学，我们还发现了血清蛋白质 与门静脉高压症确定为晚期纤维化的儿童隔离。这些数据构成了 在三个相互关联的目标中提出的新研究的基础：1)发现 胆道闭锁的结果和发病机制，2)确定门静脉高压症的生物标志物 肝脏疾病的进展，以及3)确定胆道闭锁组织损伤的发病机制。 AIM 1的实验将使用来自大型队列的RNAseq数据来挖掘基因组和分子路径 预测临床结果，随后对胆道闭锁小鼠模型和 通过靶向肝脏代谢回路抑制纤维化的临床前试验中的新生儿纤维化。实验 Aim 2将使用血清蛋白质组学数据来研究SEMA6B、sFRP3、COMMD7、VCAM1和BMX如何 单独或联合作为门脉高压的生物标志物。目标3中的实验将 从胆道闭锁受试者的肝脏中提取胆汁类器官并检验与胆道闭锁缺陷相关的假说 细胞成熟和胆管细胞纤维化的激活如何是胆汁的重要机制 导管损伤。通过应用高度互补的方法来研究来自足够大小的队列的组织 ，我们的实验将提供对疾病的新生物标记物的洞察，他们的 在发病机制中的作用，以及如何根据生物终点对新的临床试验进行个性化。

PROJECT SUMMARY/ABSTRACT This is a competing renewal application studying the biological basis of clinical phenotype and outcome of biliary atresia, the most common cause of neonatal cholestasis. The disease results from a fibro-inflammatory obstruction of extrahepatic bile ducts and present in early infancy. Despite nearly uniform progression to end- stage cirrhosis, the variable response to surgical/medical treatment and rate of progression of disease suggest the existence of unrecognized biological processes that are driving different phenotypes or stages of disease. In the previous tenure of the award, we found evidence of increased signaling via IL-8, TNF, and components of the inflammasome in pathogenesis of bile duct injury, and the simultaneous activation of molecular circuits dependent on IL-33 to induce tissue repair. We also identified a key role for MMP-7 in bile duct epithelial injury and as a highly sensitive and specific biomarker for biliary atresia. In preparation for this application, we applied computer modeling and high analytics to mine the hepatic transcriptome and found a 14-gene signature that predicts 2-year survival with the native liver and identifies glutathione metabolism as a new therapeutic target to suppress fibrosis. Using serum proteomics, we also uncovered serum proteins that segregate with children with advanced fibrosis as determined by portal hypertension. These data form the foundation for the new studies proposed in three inter-related aims: 1) To discover molecular determinants of outcome and pathogenesis of biliary atresia, 2) To identify biomarkers of portal hypertension during progression of liver disease, and 3) To define pathogenic mechanisms of tissue injury in biliary atresia. Experiments for Aim 1 will use RNAseq data from a large cohort to mine gene groups and molecular pathways that predict clinical outcome, followed by complementary studies in mouse models of biliary atresia and neonatal fibrosis in pre-clinical trials to suppress fibrosis by targeting metabolic circuits in the liver. Experiments for Aim 2 will use data from serum proteomics to investigate how SEMA6B, sFRP3, COMMD7, VCAM1, and BMX perform as biomarkers of portal hypertension individually or in combination. And experiments in Aim 3 will derive biliary organoids from the liver of subjects with biliary atresia and test hypothesis related to defects of cell maturation and to how the activation of fibrogenesis in cholangiocytes is an important mechanisms of bile duct injury. By applying highly complementary approaches to study tissues from adequately sized cohorts that have been phenotyped prospectively, our experiments will provide insight into new biomarkers of disease, their role in pathogenesis, and how new clinical trials can be personalized based on biological end-point.