The Genetic Basis of Pediatric Cholestasis

小儿胆汁淤积的遗传基础

• 批准号: 8700964
• 负责人: LAURA N BULL
• 金额: $ 55.92万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8700964
• 关键词: Adult; Alleles; Benign recurrent intrahepatic cholestasis; Bile fluid; Biological Assay; Birth; Candidate Disease Gene; Cell Line; Child; Childhood; Cholestasis; Code; Collection; Complex; Country; Custom; DNA; Data; Development; Diabetes Mellitus; Diagnosis; Diagnostic; Diagnostic Procedure; Diagnostic tests; Digestive System Disorders; Disease; Disease Management; Education; Europe; Evaluation; Exons; Failure; Functional RNA; Funding; Gene Mutation; Genes; Genetic; Genetic Predisposition to Disease; Genetic Techniques; Goals; Health; Hereditary Disease; Hospitals; Impairment; Inherited; Institutes; Intestines; Investigation; Kidney Diseases; Knowledge; Laboratories; Lead; Life; Light; Liver; Liver diseases; London; Methodology; Mission; Mothers; Mutate; Mutation; National Institute of Diabetes and Digestive and Kidney Diseases; Nature; Normal tissue morphology; Nucleic Acid Regulatory Sequences; Pathway interactions; Patients; Performance; Phenotype; Physiology; Placenta; Poland; Progressive intrahepatic cholestasis; Proteins; RNA Splicing; Regulatory Element; Research; Research Design; Resources; Role; Sampling; Sorting - Cell Movement; Structure; Techniques; Testing; Tissues; Transcript; Transplantation; United States National Institutes of Health; Variant; base; cohort; college; cost effective; data mining; design; exome sequencing; experience; gene discovery; genetic technology; genetic variant; improved; in vitro Assay; liver transplantation; next generation sequencing; novel; public health relevance; screening; tool

DESCRIPTION (provided by applicant): Before birth many functions required of the liver can be replaced by the placenta/mother. It is therefore in the first few months of life that the majoriy of congenital or genetic liver disease will present. The great majority of these diseases are cholestatic diseases, in which there is failure to make, or drain, bile from the liver into the intestine. A clinically defined group of patients was identified some years ago, and their disorder termed progressive familial intrahepatic cholestasis (PFIC). Significant advances in the understanding of the genetic basis of PFIC and related disorders have been made by the current applicants. However, we know that between one third and one half of these patients remain without a precise genetic diagnosis. This is partly due to limitations in the currently employed diagnostic pathways, which are expensive and lengthy; however, it is due to a greater degree to our not having yet discovered all the causative genes. In this proposal, we bring together sample resources from the NIH-funded Childhood Liver Disease Research and Education Network (ChiLDREN), Dr. Bull's laboratory, and the 2 largest pediatric liver centers in Europe. By combining these sample resources and our experience in the field, with recent dramatic advances in genetic technology, we are poised to achieve the goals outlined here. Aim 1 of the current study will implement cost-effective, custom- designed next-generation sequencing assays, and supporting techniques, to screen patients from our large sample resource for mutation in key known cholestasis genes (i.e. genes mutated in forms of cholestasis). In so doing, we will define a cohort of patients without detected mutation, who are therefore suitable for inclusion in studies aimed at new gene discovery. Aim 2A will identify novel cholestasis genes and mutations using a powerful recently developed technique, termed 'whole-exome' sequencing. This technique, in which coding sequence of all known genes is sequenced, is remarkably efficient and cost-effective. Exome sequence of a subset of patients will be generated and analyzed to identify mutations and candidate disease genes, and results confirmed in a larger patient set. The data gained from the gene discovery components of the study will be used to refine a diagnostic testing pathway in an iterative fashion. Aim 2B will investigate some newly identified genes and variants to assess their functional roles and consequences. Transcript structure and levels, protein levels and localization, as well as ultrastructural features, may be assessed. Mutations in regulatory elements may be assessed using in vitro assays. Patient liver tissue and appropriate cell lines will be used. The result of ur proposed studies will be identification of new genes and mutations involved in these diseases, which will inevitably shed light on basic physiology, and lead to development of robust, rapid and affordable diagnostic testing. These goals are central to both the strategy of the Childhood Liver Disease Research and Education Network (ChiLDREN) and the mission of the National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK), as stated in its Action Plan for Liver Disease Research.

描述（由申请人提供）：在出生之前，肝脏/母亲需要许多功能。因此，在生命的头几个月中，先天性或遗传性肝病的多元将出现。这些疾病中的绝大多数是胆汁疾病，其中未能使肝脏胆汁散落到肠道中。几年前，确定了一组临床定义的患者，他们的疾病 称为进行性家庭肝内胆汁淤积（PFIC）。当前申请人对PFIC和相关疾病的遗传基础的理解和相关疾病的理解取得了重大进展。但是，我们知道，这些患者中有三分之一至一半保持没有精确的遗传诊断。这部分是由于当前使用的诊断途径的限制，这些诊断途径昂贵且冗长。但是，这是由于我们尚未发现所有因果基因的程度。在此提案中，我们将NIH资助的儿童肝病研究和教育网络（儿童），Bull博士实验室和欧洲2个最大的儿科肝脏中心的样本资源汇总在一起。通过将这些样本资源和我们在该领域的经验结合在一起，以及遗传技术的最新进步，我们准备实现此处概述的目标。当前研究的目标1将实施具有成本效益，定制设计的下一代测序测定法和支持技术，以筛选患者与我们的大型样本资源中的大型样本资源中的突变中的突变（即以胆汁淤积形式突变的基因）。这样一来，我们将定义一组未检测到的突变的患者，因此适合纳入针对新基因发现的研究。 AIM 2A将使用一种强大的最近开发的技术鉴定新的胆汁淤积基因和突变，称为“全异常”测序。该技术在所有已知基因的编码序列中进行了测序，具有非常有效且具有成本效益的技术。将生成并分析一部分患者的外显子组序列，以鉴定突变和候选疾病基因，并在较大的患者组中确认结果。从研究的基因发现成分中获得的数据将用于以迭代方式完善诊断测试途径。 AIM 2B将研究一些新鉴定的基因和变体，以评估其功能作用和后果。可以评估转录本结构和水平，蛋白质水平和定位以及超微结构特征。可以使用体外测定法评估调节元件中的突变。将使用患者肝组织和适当的细胞系。 UR提出的研究的结果将是鉴定这些疾病中涉及的新基因和突变，这将不可避免地阐明基本生理学，并导致稳健，快速和负担得起的诊断测试的发展。这些目标既是儿童肝病研究和教育网络（儿童）的战略，以及国家糖尿病和消化研究所和肾脏疾病（NIDDK）的使命，正如其肝病行动计划中所指出的那样。