Isolation, Identification and Characterization of a Toxin Causing Biliary Atresia

引起胆道闭锁的毒素的分离、鉴定和表征

基本信息

批准号：
8705505
负责人：
MICHAEL A PACK
金额：
$ 50.72万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-30 至 2015-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8705505
关键词：
Affect Animal Disease Models Animal Model Animals Australia Biliary Biliary Atresia Biological Biological Assay Biological Models Biopsy Cattle Cell Culture Techniques Cells Childhood Cholestasis Cirrhosis Clinical Coculture Techniques Complex Coupled Diagnosis Disease Employee Strikes Environment Environmental Exposure Epidemic Etiology Event Extrahepatic Fibrosis Gallbladder Genes Goals Human Inbred BALB C Mice Individual Infection Inflammatory Ingestion Injury International Larva Lead Link Liver Liver diseases Livestock Macaca mulatta Modeling Morphogenesis Mouse Strains Mus Myofibroblast Nature Neonatal Newborn Infant Obstruction Orthologous Gene Pathogenesis Pathway interactions Patients Pattern Phenotype Plants Population Pregnant Women Reporter Rodent Rotavirus Sheep Signal Pathway Specimen Staging Study models Time Tissues Toxin Transgenic Organisms Vertebrates Viral Zebrafish biliary tract cell injury cholangiocyte disease characteristic epidemiologic data genetic risk factor high throughput screening human disease in vitro Model in vivo in vivo Model insight intrahepatic knock-down liver transplantation mature animal molecular marker offspring overexpression precursor cell pregnant research study

项目摘要

DESCRIPTION (provided by applicant): Biliary atresia (BA) is a fibro inflammatory disorder that is the leading cause of neonatal cholestasis and the most common indication for liver transplant in the pediatric population. Although epidemiologic data suggest that BA arises from the interplay of genetic risk factors coupled with environmental exposures, the etiology is unknown. Animal models are limited, and the best-known animal model of the disease, the infection of newborn BALB/c mice with rhesus rotavirus (RRV), has provided important information about immunological aspects of the disease but has been less useful for studying the rapidly progressive fibrosis characteristic of the disease. Insight into the pathogenesis of BA comes from the study of a naturally-occurring animal model of the disease. Over the last 40 years, there have been three epidemics of BA in newborn livestock in Australia associated with ingestion of the plant Dysphania glomulifera by pregnant sheep and cows. Clinical and pathological findings from the affected lambs and calves show striking similarities with human BA, in particular marked fibrosis at the time of diagnosis. We hypothesize that determining the causative toxin will enable the identification of general cholangiocyte damage pathways that lead ultimately to BA and fibrosis in vertebrates. We have thus collected and imported D. glomulifera from the Australian pasture affected by the most recent epidemic and have employed a zebrafish bioassay to successively subfractionate the plant and identify active fractions. We have several fractions with a complexity of 2-20 that, remarkably, cause a BA-like pattern of injury (with atresia of the gallbladder and extrahepatic biliary tree) in zebrafish larvae exposed after biliary morphogenesis has occurred, mimicking human BA. The overall goal of this proposal is therefore to identify the biliary toxins in Dysphania, characterize the cholangiocyte damage pathways they induce, and establish new animal models as a means of understanding the pathogenesis of human BA. This will be achieved through three specific aims: 1. to identify D. glomulifera biliary toxins using the in vivo zebrafish bioassay~ 2. To identify molecular markers and genes that regulate extrahepatic biliary injury in zebrafish larvae treated with D. glomulifera toxins~ and 3. To identify and characterize damage pathways induced by D. glomulifera toxins in mammalian models.

描述（由申请人提供）：胆道闭锁（BA）是一种纤维炎症性疾病，是新生儿胆汁淤积的主要原因，也是儿科人群肝移植的最常见适应症。尽管流行病学数据表明 BA 是由遗传风险因素与环境暴露相互作用引起的，但其病因尚不清楚。动物模型有限，该疾病最著名的动物模型是用恒河猴轮状病毒 (RRV) 感染新生 BALB/c 小鼠，提供了有关该疾病免疫学方面的重要信息，但对于快速研究该疾病的作用不大。该疾病的特征是进行性纤维化。对 BA 发病机制的深入了解来自对该疾病自然发生的动物模型的研究。过去 40 年来，澳大利亚新生牲畜中发生了 3 次 BA 流行，与怀孕的羊和牛摄入 Dysphania glomulifera 植物有关。受影响羔羊和犊牛的临床和病理结果与人类 BA 具有惊人的相似性，特别是在诊断时出现明显的纤维化。我们假设确定致病毒素将能够识别最终导致脊椎动物胆管细胞损伤和纤维化的一般胆管细胞损伤途径。因此，我们从受最近流行病影响的澳大利亚牧场收集并进口了 D. glomulifera，并采用斑马鱼生物测定法对植物进行了连续细分并鉴定了活性组分。我们有几个复杂度为 2-20 的分数，它们在胆道形态发生发生后暴露的斑马鱼幼虫中引起类似 BA 的损伤模式（胆囊和肝外胆管树闭锁），模仿人类 BA。因此，该提案的总体目标是识别言语障碍中的胆道毒素，表征它们诱导的胆管细胞损伤途径，并建立新的动物模型作为了解人类 BA 发病机制的手段。这将通过三个具体目标来实现： 1. 使用体内斑马鱼生物测定来鉴定 D. glomulifera 胆汁毒素~ 2. 鉴定在用 D. glomulifera 毒素处理的斑马鱼幼虫中调节肝外胆道损伤的分子标记和基因~ 和 3 . 鉴定和表征哺乳动物模型中 D. glomulifera 毒素引起的损伤途径。