The Genetic Basis of Pediatric Cholestasis

小儿胆汁淤积的遗传基础

基本信息

批准号：
10665248
负责人：
LAURA N BULL
金额：
$ 24.52万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-24 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10665248
关键词：
Actins Address Adult Apical Area Bile fluid Biliary Biological Biological Models Blood CRISPR/Cas technology Candidate Disease Gene Child Childhood Cholestasis Clinical Clinical Data Clinical Trials Collaborations Complement Cytoskeleton DNA Data Data Collection Diabetes Mellitus Diagnosis Diagnostic Digestive System Disorders Disease Disease model Etiology Europe Excretory function Fishes Functional disorder Funding Future Gene Mutation Generations Genes Genetic Genetic Predisposition to Disease Genetic Transcription Genetic study Genome Genomics Goals Health Hepatocyte Hospitals Human Impairment Inherited Injury Institutes Intrahepatic Cholestasis Investigation Kidney Diseases Knock-out Laboratories Life Light Liver Liver diseases London Methylation Mission Modeling Molecular Mutation National Institute of Diabetes and Digestive and Kidney Diseases Nature Neonatal Pathogenesis Pathologic Pathway interactions Patients Pediatric Hospitals Pharmaceutical Preparations Phenotype Physiology Poland Proteins RNA RNA Processing Research Resources Role Sampling Sclerosing Cholangitis Serum Stratification Structure Technology Testing Tight Junctions Tissue Banks Tissues Variant Yin Zebrafish accurate diagnosis advanced disease base bile canaliculus structure bile duct college data resource diagnostic strategy disease phenotype disease prognosis exome gene discovery genetic technology genetic testing genome editing genome sequencing genomic data improved individualized medicine liver function liver transplantation mutant mutation screening next generation sequencing novel protein transport targeted sequencing therapeutic target transcriptome whole genome

项目摘要

PROJECT SUMMARY/ABSTRACT Bile flow is essential for normal liver function. Cholestasis is the term for all phenomena where bile flow is im- paired. Cholestatic liver disease results from persistent cholestasis. Substantial progress has been made in un- derstanding of the mechanisms underlying pediatric cholestatic liver disease, including by the current appli- cants. However, in ~30% of children with isolated (primary) cholestasis (PC) the genetic etiology is not identi- fied, despite routine diagnostic sequencing. A closely related clinical presentation results from bile duct injury, and those presenting early in life are described as having neonatal sclerosing cholangitis (NSC). Accurate diag- nosis improves ability to assess disease prognosis and tailor treatment to the underlying disease etiology. This proposal continues the collaboration between Drs. Bull and Thompson, together with the Northwest Genomics Center. The latter provides access to unparalleled resources for genomic data generation and analysis, which can be applied to the unmatched sample and data resources of the 2 largest pediatric liver centers in Europe (King's College London and Children's Memorial Institute in Warsaw), Dr Bull's laboratory, and the NIDDK- funded Childhood Liver Disease Research Network (ChiLDReN). To complement the genetic findings through assessment of disease mechanisms using zebrafish models, Dr. Yin is joining the project. These resources will address the following challenges. In Aim 1, genetic studies will be performed to identify novel genes underlying PC and NSC, and novel mutations in known cholestasis genes. State-of-the-art genetic technologies will be used, including next-generation sequencing of targeted gene panels, exomes, genomes, and RNA, as well as methylation analysis. Available clinical data and tissue collections will be used to find correlates supportive of the causative nature of identified genetic defects, and to establish features associated with mutation in particu- lar genes. In Aim 2, the pathophysiological mechanisms underlying genetic cholestasis will be investigated in zebrafish models. Null-mutant fish have already been generated for usp53 and lsr; orthologues of 2 genes in which mutations have recently been found in children with cholestasis. We will test the hypothesis that Lsr and Usp53 are critical in maintaining the integrity of bile canaliculi. We will assess whether loss of Lsr or Usp53 impairs bile canalicular structure, blood-bile barrier, and apical and basolateral protein trafficking. We will de- termine the interactions of Lsr and Usp53 with other junctional proteins and actin cytoskeleton in hepatocytes. In conclusion, the proposed studies will result in identification of new disease genes and mutations, illuminate commonalities and distinctions between different genetic forms of cholestasis, and will use model systems to shed light on the biological consequences of loss of these genes, which can be difficult to tease out in human livers with advanced disease. These aims fulfill key goals of ChiLDReN and are central to the mission of the Na- tional Institute of Diabetes and Digestive and Kidney Diseases.

项目概要/摘要胆汁流动对于正常的肝功能至关重要。胆汁淤积是胆汁流动不畅的所有现象的总称。配对。胆汁淤积性肝病是由持续性胆汁淤积引起的。联合国工作取得实质性进展了解小儿胆汁淤积性肝病的潜在机制，包括通过当前的应用行话。然而，约 30% 患有孤立性（原发性）胆汁淤积 (PC) 的儿童的遗传病因尚不明确。尽管进行了常规诊断测序，但仍然存在。与胆管损伤密切相关的临床表现，那些在生命早期出现的患者被描述为患有新生儿硬化性胆管炎（NSC）。准确诊断 nosis 提高了评估疾病预后并根据潜在疾病病因制定治疗方案的能力。这该提案继续了博士之间的合作。布尔和汤普森以及西北基因组公司中心。后者提供了对基因组数据生成和分析的无与伦比的资源的访问，这可应用于欧洲最大的2家儿科肝脏中心无与伦比的样本和数据资源（伦敦国王学院和华沙儿童纪念研究所）、Bull 博士实验室和 NIDDK- 资助儿童肝病研究网络 (ChiLDReN)。通过以下方式补充遗传发现使用斑马鱼模型评估疾病机制，尹博士正在加入该项目。这些资源将应对以下挑战。在目标 1 中，将进行遗传学研究以确定潜在的新基因 PC 和 NSC，以及已知胆汁淤积基因的新突变。最先进的基因技术将使用的，包括目标基因组、外显子组、基因组和 RNA 的下一代测序，以及甲基化分析。现有的临床数据和组织收集将用于寻找支持的相关性确定的遗传缺陷的致病性质，并建立与突变相关的特征，特别是拉尔基因。在目标 2 中，将研究遗传性胆汁淤积的病理生理机制斑马鱼模型。已经为usp53和lsr生成了无效突变鱼； 2个基因的直系同源物最近在患有胆汁淤积的儿童中发现了哪些突变。我们将检验 Lsr 和 Usp53 对于维持胆小管的完整性至关重要。我们将评估是否丢失 Lsr 或 Usp53 损害胆管结构、血胆屏障以及顶端和基底外侧蛋白质运输。我们将去- 终止 Lsr 和 Usp53 与肝细胞中其他连接蛋白和肌动蛋白细胞骨架的相互作用。总之，拟议的研究将导致新疾病基因和突变的鉴定，阐明胆汁淤积的不同遗传形式之间的共性和区别，并将使用模型系统来揭示了这些基因丢失的生物学后果，这在人类中很难弄清楚患有晚期疾病的肝脏。这些目标实现了ChiLDReN的关键目标，并且是Na- 糖尿病、消化和肾脏疾病国家研究所。