Molecular genetic bases of polycystic and tubulointerstitial kidney diseases

项目摘要/摘要Whitney Besse博士是一位经过委员会认证的学术肾病学家， 对遗传学有研究兴趣，目标是成为一名医生科学家并做出重大贡献 遗传性肾病的研究和治疗。最常见的遗传性肾病，常染色体 显性多囊肾病（ADPKD）通常由PKD 1或PKD 2基因突变引起， 编码多囊蛋白-1（PC 1）和多囊蛋白-2。Besse博士试图通过以下方式更好地了解PC 1的功能： 识别和描述对该功能至关重要的基因的作用。她发现了基因突变 编码内质网（ER）中的蛋白质，也可引起难以区分的肾/肝囊肿，但在 不存在PKD 1或PKD 2突变。导师的实验室在小鼠模型中定义了这些ER 由于PC 1功能剂量不足而形成肾/肝囊肿的基因。这项提案将推动这些 通过扩大遗传分析方法/模式和扩大一组遗传学未解决的 显性遗传性多囊肾和肝病（PKD/PLD）病例，以定义新的候选疾病 基因.这项研究已经以这种方式确定了五个有希望的ER相关候选基因， 已经在初步研究中得到了实验验证。五位新候选人将系统地 通过评估CRISPR/Cas9产生的细胞系中的PC 1生物发生、运输和纤毛表达来评估。 具有与临床数据相关联的外显子组序列的大型精密医学数据库也将使用 候选基因，以评估临床变异和表型的变异率。对此的初步研究 一项提案确定了DNAJB 11的突变，编码ER HSP 40伴侣蛋白，导致PKD/PLD， PC 1功能受损。据报道，具有DNAJB 11突变的患者也会出现肾小管间质性疾病 肾病DNAJB 11是引起PKD/PLD的几种蛋白质之一，也与ER蛋白相关 易位孔（translocon）。该建议将研究这些蛋白质之间的相互作用关系 几种易位子相关的多囊病基因产物。我们将探索DNAJB 11的作用， 特别是对PC 1的功能特性，对UPR的激活，及其对病原体运输的影响， PC 1的错义变体。我们还将研究肾小管间质性肾病的动物和细胞模型 基于多囊和UPR基因的联合失活，以更好地解决我们对如何 这些基因中的人类突变可导致PKD/PLD和肾小管间质慢性肾病。博士 Besse的培训将包括正式的学习，由专家导师/顾问指导的教育， 研讨会/会议，以及使用拟议的新方法来创造科学贡献。 这种培训，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 招募，小鼠模型等将为Besse博士提供开发一种 研究遗传性肾病机制的成功独立职业生涯。

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.