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：
8518316
负责人：
MICHAEL A PACK
金额：
$ 48.94万
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/8518316
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

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

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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，也是小儿种群中肝移植的最常见迹象。尽管流行病学数据表明BA是由遗传危险因素与环境暴露的相互作用引起的，但病因尚不清楚。动物模型是有限的，这种疾病的最著名动物模型是恒河猴轮状病毒（RRV）的新生儿BALB/c小鼠的感染，它提供了有关该疾病免疫学方面的重要信息，但对于研究该疾病的快速进行性纤维化特征的有用程度较小。对BA发病机理的洞察来自研究该疾病的天然动物模型。在过去的40年中，澳大利亚新生儿牲畜的三种流行病与通过怀孕的绵羊和奶牛摄入植物吞咽困难的植物吞噬植物有关。受影响的羔羊和犊牛的临床和病理发现表现出与人类BA的惊人相似性，特别是在诊断时明显的纤维化。我们假设确定致病毒素将使一般胆管细胞损伤途径鉴定，这最终导致脊椎动物的BA和纤维化。因此，我们已经从受到最新流行病影响的澳大利亚牧场中收集并进口了D. glomulifera，并采用了斑马鱼生物测定法，将植物延伸并确定活跃的分数。我们有几个分数，复杂性为2-20，显着地导致斑马鱼幼虫发生胆道形态发生后发生的斑马鱼幼虫的损伤模式（胆囊闭锁和肝外胆道上）发生了模仿。因此，该提案的总体目标是确定烦躁的胆汁毒素，以它们诱导的胆管细胞损伤途径来表征，并建立新的动物模型作为理解人类BA发病机理的一种手段。这将通过三个特定的目的来实现：1。使用体内斑马鱼生物测定〜2。确定D. glomulifera胆汁毒素〜2。确定分子标记物和基因，这些标志物和基因在斑马鱼幼虫中调节肝脏外liar损伤，并用D. glomulifera d. glomulifera toxins and和3。以识别和表征D. d。n. glomulifera tode。