Molecular genetic bases of polycystic and tubulointerstitial kidney diseases

多囊肾和肾小管间质性肾病的分子遗传学基础

基本信息

批准号：
10132310
负责人：
Whitney Elise Besse
金额：
$ 16.79万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-06-01 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10132310
关键词：
Address Affect Alleles Animal Model Autosomal Dominant Polycystic Kidney Bile Duct Epithelium Biogenesis Bioinformatics Biological Biological Assay CRISPR/Cas technology Candidate Disease Gene Cell Culture Techniques Cell Line Cell model Cell physiology Cells Chronic Kidney Failure Cilia Client Clinical Clinical Data Co-Immunoprecipitations Cohort Studies Complex Core Protein Coupled Cyst Databases Dependence Disease Ductal Epithelial Cell Education Endoplasmic Reticulum Enrollment Family Fibrosis Gene Proteins Genes Genetic Genetic Heterogeneity Goals Health Hepatic Cyst Human Impairment Individual Inherited Investigation Kidney Kidney Diseases Kidney Failure Knock-out Laboratories Link Liver Liver diseases Mentors Methodology Methods Modality Modeling Modernization Molecular Molecular Chaperones Molecular Genetics Mus Mutation PKD1 gene PKD2 gene PKD2 protein PRKCSH protein Pathogenesis Pathogenicity Pathway interactions Patients Penetrance Phenotype Physicians Polycystic Kidney Diseases Property Protein translocation Proteins Publishing Radiology Specialty Renal tubule structure Renin Reporting Research Resources Role Scientist Therapeutic Training Translating UMOD gene Up-Regulation Validation Variant XBP1 gene base career cohort dosage exome exome sequencing experience gene discovery gene function gene product genetic analysis human disease improved insight interest interstitial loss of function man meetings mouse model mutant novel overexpression polycystic kidney disease 1 protein precision medicine protein complex proteostasis recruit response trafficking

项目摘要

Project Summary/Abstract Dr. Whitney Besse is a board-certified academic nephrologist with long-standing research interest in genetics whose goal is to become a physician scientist and make significant contributions to the study and treatment of genetic kidney diseases. The most common genetic kidney disease, autosomal dominant polycystic kidney disease (ADPKD), is typically caused by mutations in the PKD1 or PKD2 genes, encoding polycystin-1 (PC1) and polycystin-2. Dr. Besse seeks to better understand the function of PC1 by identifying and describing the role of genes essential to this function. She has identified mutations in genes encoding proteins in the endoplasmic reticulum (ER) that also cause indistinguishable kidney/liver cysts but in the absence of PKD1 or PKD2 mutations. The mentor's laboratory defined in mouse models for these ER genes that kidney/liver cysts form due to insufficient PC1 functional dosage. This proposal will advance these studies by expanding genetic analysis methods/modalities and expanding a cohort of genetically unresolved cases of dominantly inherited polycystic kidney and liver disease (PKD/PLD) to define novel candidate disease genes. The study has identified five promising ER-associated candidate genes in this manner and a sixth gene has been experimentally validated in preliminary studies. The five new candidates will be systematically assessed by evaluating PC1 biogenesis, trafficking and cilia expression in CRISPR/Cas9-generated cell lines. Large precision medicine databases with exome sequences linked to clinical data will also be queried with the candidate genes to evaluate clinical variation and penetrance of phenotypes. Preliminary studies in this proposal identified mutations in DNAJB11, encoding an ER HSP40 chaperone as causing PKD/PLD due to impaired PC1 function. Patients with DNAJB11 mutations have also been reported to develop tubulointerstitial kidney disease. DNAJB11 is one of several proteins causing PKD/PLD that also associate with the ER protein translocation pore (translocon). This proposal will examine the protein interacting relationships among these several translocon associated polycystic disease gene products. We will explore the effects of DNAJB11 specifically on PC1 functional properties, on activation of the UPR, and its effect on trafficking of pathogenic missense variants of PC1. We will also investigate animal and cell models of tubulointerstitial kidney disease based on combined inactivation of polycystic and UPR genes to better resolve our understanding of how human mutations in these genes can result in both PKD/PLD and tubulointerstitial chronic kidney disease. Dr. Besse's training will include formal study, directed education by expert mentor/advisors, participation in seminars/meetings, and use of the proposed new methodologic approaches to create scientific contributions. This training, coupled with the extensive expertise and resources at Yale such as exome sequencing, study enrollment, mouse models, etc. will provide Dr. Besse with the expertise and experience needed to develop a successful independent career studying the mechanisms of genetic kidney disease.

项目摘要/摘要Whitney Besse博士是一名经过董事会认证的学术肾病学家对遗传学的研究兴趣，其目标是成为医师科学家并做出重大贡献研究和治疗遗传肾脏疾病。最常见的遗传肾脏疾病，常染色体主要的多囊性肾脏疾病（ADPKD）通常是由PKD1或PKD2基因突变引起的，编码polycystin-1（PC1）和polycystin-2。 Besse博士试图更好地了解PC1的功能识别和描述基因对此功能必不可少的作用。她已经确定了基因的突变在内质网中编码蛋白质（ER），该蛋白质也引起无法区分的肾脏/肝囊肿，但在没有PKD1或PKD2突变。导师的实验室在鼠标模型中为这些ER定义由于PC1功能剂量不足，肾脏/肝囊肿形成的基因。该提议将推进这些通过扩大遗传分析方法/模态并扩大遗传上未解决的队列的研究主要遗传多囊肾脏和肝病（PKD/PLD）的病例定义了新型候选病基因。该研究以这种方式鉴定了五个有希望的ER相关候选基因和第六个基因在初步研究中已通过实验验证。五个新候选人将系统地通过评估CRISPR/CAS9生成的细胞系中的PC1生物发生，运输和纤毛表达评估。具有与临床数据相关的外显子序列的大型精确医学数据库也将与候选基因评估表型的临床变异和渗透率。初步研究提案确定了DNAJB11中的突变，编码ER HSP40伴侣是由于引起PKD/PLD的原因 PC1功能受损。据报道，患有DNAJB11突变的患者会发展为肾小管间隙肾脏疾病。 DNAJB11是引起PKD/PLD的几种蛋白质之一，也与ER蛋白相关易位孔（转运）。该提案将研究其中的蛋白质相互作用关系几种易位的多囊病基因产物。我们将探索DNAJB11的影响专门针对PC1功能性能，激活UPR及其对致病性运输的影响 PC1的错义变体。我们还将研究微管间质肾脏疾病的动物和细胞模型基于多囊和UPR基因的综合失活，以更好地解决我们对如何的理解这些基因中的人类突变会导致PKD/PLD和Tubulointerstitial慢性肾脏疾病。博士 Besse的培训将包括正规研究，专家导师/顾问的指导教育，参与研讨会/会议，并使用拟议的新方法学方法来创造科学贡献。这种培训，再加上耶鲁大学的广泛专业知识和资源，例如外部测序，研究注册，鼠标模型等将为Besse博士提供开发的专业知识和经验成功研究遗传肾脏疾病机制的成功独立职业。