Mapping Disease Pathways for Biliary Atresia

绘制胆道闭锁的疾病途径

• 批准号: 9904315
• 负责人: RAKESH K. SINDHI
• 金额: $ 50.42万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9904315
• 关键词: Affect; Alpha-mannosidase; Animal Model; Bile fluid; Biliary; Biliary Atresia; Bioinformatics; Biological; Birth; California; Candidate Disease Gene; Cardiac; Caucasians; Cell Mobility; Child; Childhood; Cicatrix; Cilia; Collaborations; Congenital Abnormality; Congenital atresia of extrahepatic bile duct; Data Analyses; Development; Developmental Gene; Disease Pathway; Embryo; Epidermal Growth Factor; Erinaceidae; Etiology; Excretory function; Experimental Models; Extrahepatic; Gene Expression Regulation; Gene Proteins; Genes; Genetic Predisposition to Disease; Genomics; Genotype; Growth Factor Gene; Heart; Hepatic; Hepatology; Hospitals; Human; Impairment; Inflammation; Inflammatory; Inflammatory Response; International; Knockout Mice; Knowledge; Lead; Left; Liver; Liver Failure; London; Lung; Maps; Modeling; Morphogenesis; Mus; Organ; Pathogenesis; Pathology; Pathway interactions; Pattern; Pediatric Hospitals; Phenotype; Philadelphia; Predisposition; Public Health; Refractory; Resources; SNP array; Sampling; Signal Pathway; Signal Transduction; Single Nucleotide Polymorphism; Situs Inversus; Small Interfering RNA; Stress; Stress Response Signaling; Susceptibility Gene; Systems Analysis; Systems Biology; Targeted Resequencing; Techniques; Testing; Tissues; Transforming Growth Factors; Transplant-Related Disorder; Transplantation; Transplantation Surgery; University resources; Variant; Work; Zebrafish; airway epithelium; base; bile duct; bioinformatics resource; biological adaptation to stress; causal variant; cell motility; cilium biogenesis; cohort; college; gene function; genome wide association study; knock-down; liver transplantation; member; novel; prevent; protein protein interaction; response; targeted sequencing; transcriptome; transcriptome sequencing; transplant centers; whole genome

This project will map disease pathways for biliary atresia (BA), which has failed all therapy and requires liver transplantation (LTx), using genetic susceptibility as a common basis. BA causes liver failure at birth and accounts for up to half of all pediatric LTx, worldwide, but has an uncertain etiology. BA leads to scarred and atretic bile ducts, which fail to drain the liver. Some children also have anomalous left-right patterning of extrahepatic organs. Preliminary genome-wide association study (GWAS) of single nucleotide polymorphisms (SNPs) in BA cases, and other evidence from human BA and experimental models lead us to propose the following specific aims: Aim 1. We will confirm whether BA associates with SNPs in manosidase-1-α-2 (MAN1A2), and in genes which signal via hedgehog, epidermal and transforming growth factors and genes for ciliogenesis. These pathways are abnormal in liver from BA cases, zebrafish with knockdown of man1a2, and Man1a2 -/- (knockout) mice. We will identify novel potentially causal variants with targeted sequencing of significant SNP loci. Aim 2. We will confirm whether knockdown of man1a2 and other candidates induces biliary dysgenesis and cardiac and hepatic heterotaxy in zebrafish, impaired ciliogenesis in the ex vivo mouse airway epithelia model of ciliogenesis, and dysregulation of the abovementioned developmental pathways in the liver transcriptome. Aim 3. We will construct putative pathways for BA with those significant SNP loci and dysregulated genes which show interactions in an integrated systems analysis of data from Aims 1 and 2. These loci will also implicate aberrant inflammatory and stress responses. We will corroborate pathways with combined perturbations of a developmental gene with an inflammatory and a stress response gene in zebrafish and ex vivo ciliogenesis models. We expect that aberrant developmental, inflammatory and stress response signaling contributes to hepatic and extrahepatic BA. We will genotype the largest homogeneous cohort of 800 Caucasian BA cases with LTx from three of the world's largest pediatric LTx centers: the Children's Hospital of Pittsburgh, King's College Hospital, London, UK, Birmingham Children's Hospital, UK. We will perform whole genome transcriptome (RNA) sequencing of liver samples from 80 of these cases. We will use experimental and bioinformatics resources of the Universities of Pittsburgh and California, San Diego, and the Center for Applied Genomics, Philadelphia. At the end of our project, we will have expanded our knowledge about pathogenesis of BA and related birth defects and identified candidate strategies to prevent or overcome delayed biliary morphogenesis. The international collaboration of leaders in hepatology (Kelly, Dhawan, Squires), transplant surgery (Sharif, Sindhi), genomics (Weeks, Hakonarson, Higgs), pathology (Ranganathan) and systems biology/bioinformatics (Subramanian, Higgs) is well suited to this task.

该项目将绘制胆道闭锁（BA）的疾病路径，BA已失败，需要肝脏 移植（LTx），使用遗传易感性作为共同的基础。BA在出生时导致肝功能衰竭， 占全球所有儿科LTx的一半，但病因不确定。BA导致疤痕和 阻塞的胆管无法排出肝脏有些孩子也有异常的左右模式， 肝外器官单核苷酸多态性的全基因组关联研究 （SNP）在BA病例中的作用，以及来自人类BA和实验模型的其他证据，使我们提出了 具体目标：目标1。我们将确认BA是否与β-淀粉酶-1-α-2的SNPs相关 （MAN 1A 2），以及通过刺猬、表皮和转化生长因子发出信号的基因，以及 纤毛发生这些途径在来自BA病例的肝脏、敲低man 1a 2的斑马鱼和 Man 1a 2-/-（敲除）小鼠。我们将通过靶向测序确定新的潜在致病变异， 显著SNP位点。目标2.我们将确认敲除man 1a 2和其他候选基因是否会诱导胆汁性肝硬化。 斑马鱼中的发育不全和心脏和肝脏异位，离体小鼠气道中的纤毛发生受损 纤毛发生的上皮细胞模型，以及上述肝脏发育途径的失调 转录组目标3.我们将用这些显著的SNP位点构建BA的推定途径， 在来自目标1和2的数据的整合系统分析中显示相互作用的失调基因。这些 基因座也将涉及异常的炎症和应激反应。我们会确认 在斑马鱼中，发育基因与炎症和应激反应基因的干扰， 体内纤毛发生模型。我们认为异常的发育、炎症和应激反应信号 有助于肝和肝外BA。我们将对800名白人中最大的同质队列进行基因分型， 来自世界上三个最大的儿科LTx中心的LTx BA病例：匹兹堡儿童医院， 英国伦敦国王学院医院、英国伯明翰儿童医院。我们将进行全基因组 转录组（RNA）测序来自这些病例中的80个的肝脏样品。我们将使用实验和 匹兹堡大学、加州大学、圣地亚哥大学和应用生物信息学中心的生物信息学资源 费城基因组在我们的项目结束时，我们将扩大我们对发病机制的认识， BA和相关的出生缺陷，并确定了预防或克服迟发性胆道疾病的候选策略 形态发生肝病学（Kelly，Dhawan，Squires），移植 外科（谢里夫，信德），基因组学（威克斯，Hakonarson，希格斯），病理学（Ranganathan）和系统 生物学/生物信息学（Subramanian，Higgs）非常适合这项任务。