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

• 批准号: 10597658
• 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
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-06-01 至 2024-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10597658
• 关键词: Address; Affect; Alleles; Animal Model; Autosomal Dominant Polycystic Kidney; Bile Duct Epithelium; Biogenesis; Bioinformatics; Biological; Biological Assay; Board Certification; 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; Heterozygote; 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; Mucins; 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; autosome; 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

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/摘要惠特尼·贝斯博士是一位获得董事会认证的肾病学术专家，长期 对遗传学的研究兴趣，目标是成为一名内科科学家并做出重大贡献 对遗传性肾脏疾病的研究和治疗。最常见的遗传性肾病，常染色体 显性多囊肾病(ADPKD)，通常由PKD1或PKD2基因突变引起， 编码多囊蛋白-1(PC1)和多囊蛋白-2。Besse博士试图通过以下方式更好地了解PC1的功能 识别和描述对这一功能至关重要的基因的作用。她已经确定了基因的突变 内质网(ER)中的编码蛋白也会导致难以区分的肾/肝囊肿，但在 无PKD1或PKD2突变。在这些ER的小鼠模型中定义的导师实验室 肾/肝囊肿因PC1功能剂量不足而形成的基因。这项提议将推进这些 通过扩展遗传分析方法/模式和扩大遗传未解决的队列进行的研究 显性遗传性多囊肾和肝病(PKD/PLD)定义新的候选疾病4例 基因。这项研究已经以这种方式确定了五个与雌激素受体相关的候选基因和第六个基因 已经在初步研究中得到了实验验证。五位新候选人将有系统地 通过评估CRISPR/Cas9产生的细胞系中PC1的生物发生、运输和纤毛表达来进行评估。 具有与临床数据相关联的外显子组序列的大型精确医学数据库也将使用 评估临床变异和表型外显性的候选基因。关于这方面的初步研究 一项提案发现，编码ER Hsp40伴侣的DNAJB11突变导致PKD/PLD，原因是 PC1功能受损。有报道称，携带DNAJB11基因突变的患者也会发展为肾小管间质 肾脏疾病。DNAJB11是引起PKD/PLD的几种蛋白质之一，也与ER蛋白有关 易位孔(易位子)。这项提案将研究这些细胞之间的蛋白质相互作用关系。 几种易位基因相关的多囊疾病基因产物。我们将探索DNAJB11的影响 特别是关于PC1的功能特性，关于UPR的激活，以及它对致病菌贩运的影响 PC1的错义变体。我们还将研究肾小管间质肾病的动物和细胞模型。 基于多囊细胞和UPR基因的联合失活，以更好地解决我们对 人类这些基因的突变可能导致PKD/PLD和肾小管间质慢性肾脏疾病。Dr。 Besse的培训将包括正式学习、由专家导师/顾问指导的教育、参与 研讨会/会议，以及使用拟议的新方法方法来创造科学贡献。 这一培训，加上耶鲁大学广泛的专业知识和资源，如外显子组测序，研究 注册、鼠标模型等将为Besse博士提供开发 成功的独立研究遗传性肾脏疾病的机制。