In vivo Pathway Discovery in Autosomal Dominant Polycystic Kidney Disease

常染色体显性多囊肾病的体内途径发现

• 批准号: 10434820
• 负责人: Michael J. Caplan
• 金额: $ 128.94万
• 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: 2019-09-09 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10434820
• 关键词: Address; Adult; Affect; Affinity Chromatography; Animal Disease Models; Animal Model; Autosomal Dominant Polycystic Kidney; Biochemical; Bioinformatics; Biological; Biological Models; Biology; Biometry; Catalogs; Cell model; Cells; Characteristics; Cilia; Communities; Complex; Coupled; Cyst; Cystic kidney; Data; Data Set; Defect; Dialysis procedure; Disease; Elements; End stage renal failure; Enterobacteria phage P1 Cre recombinase; Epithelial Cells; Equipment and supply inventories; Evaluation; Fatty Acids; Gene Expression; Genes; Genetic; Genetic Transcription; Genotype; Glucose; Goals; Growth; Individual; Informatics; Inherited; Kidney; Kidney Diseases; Knock-out; Knockout Mice; Life; Liquid substance; Liver Mitochondria; Mendelian disorder; Messenger RNA; Metabolic; Metabolism; Methods; Mitochondria; Modeling; Molecular; Mus; Mutation; National Institute of Diabetes and Digestive and Kidney Diseases; Nature; Normal tissue morphology; Orthologous Gene; PKD2 protein; Pathogenesis; Pathway interactions; Patients; Phase; Physiological; Play; Population; Production; Property; Proteins; Proteomics; Reactive Oxygen Species; Renal function; Renal tubule structure; Research; Research Personnel; Resources; Ribosomes; Role; Signal Pathway; Signal Transduction; Specificity; Structure; System; Time; Tissues; Tracer; Translating; Transplantation; Wild Type Mouse; animal data; base; biological systems; cell type; design; expectation; gene product; human disease; in vivo; in vivo Model; innovation; kidney cortex; metabolic abnormality assessment; metabolic phenotype; metabolomics; mitochondrial metabolism; mouse model; new therapeutic target; novel; polycystic kidney disease 1 protein; programs; protein biomarkers; protein expression; renal epithelium; side effect; solute; targeted treatment; therapeutic development; therapy development; tool; transcriptome; transcriptomics

Abstract Autosomal Dominant Polycystic Kidney Disease (ADPKD) is one of the most common monogenic diseases, affecting >1:1000 individuals worldwide. It is characterized by large fluid-filled renal cysts that remodel, compress and destroy surrounding normal tissue, and that progressively reduce kidney function, leading to end stage renal disease in about 50% of patients by the sixth decade of life. Most ADPKD results from mutations in two genes, PKD1, which encodes the polycystin-1 protein (PC1), and PKD2, which encodes the polycystin-2 protein (PC2). PC1 and PC2 interact with one another and are thought to play a role in cilia signaling. It is generally accepted that the cilium is a central component in the pathways that drive ADPKD pathogenesis. Although their mechanistic connection to the functional PC complex in cilia is unclear, numerous signaling pathways are perturbed in cysts. In the past few years the list of disease-related pathways has grown through new evidence that implicates metabolism as a novel pathway that is profoundly affected in ADPKD and that may both participate in disease pathogenesis and serve as a target for therapeutic development. While it remains to be established whether the newly-identified metabolic derangements that characterize ADPKD are direct drivers of cyst formation, it is clear that the nature and activities of a cell's many and varied intertwined metabolic circuits plays a central role in determining its capacity to invest the energy required in order to participate in the proliferation and active solute and fluid transport that are required for cyst growth. The main goal of this proposal is to provide the research community with novel tools and data sets that will substantially enhance efforts to explore and exploit the metabolic changes that characterize ADPKD. We will produce a uniquely designed and rigorously curated resource based upon novel in vivo models of the cell specific transcriptomic, mitochondrial proteomic and mitochondrial metabolic effects that result from the earliest stages after loss of the PC proteins and that are further informed by the effects of concomitant cilia loss and PC protein reactivation. This program will make use of adult inducible conditional PC knockout mouse models and will employ strategies that will permit conditional isolation of ribosomes (TRAP) and conditional isolation of mitochondria, thus enabling cell-type- specific transcriptomic and mitochondrial proteomic and metabolomic studies. State of the art in vivo metabolic flux studies will be applied to the kidney cortices of these novel genetic mouse models. The results of these analyses will be combined to produce robust biological data sets that will be assembled through application of the requisite informatics mechanisms in order to disseminate these data to the broader research community in near real time. Critically, our in vivo studies are designed to discover the earliest changes that occur after kidney tubules lose polycystin protein expression—at time points well before cysts form. The research team brings together extensive and complementary expertise in ADPKD animal models, PC signaling and biology and in vivo metabolic studies coupled with strong biostatistical and bioinformatics support to produce the proposed resource.

常染色体显性遗传性多囊肾病（ADPKD）是一种常见的单基因遗传性肾病， 疾病，影响世界各地>1：1000的个体。它的特点是巨大的充满液体的肾囊肿重塑， 压缩和破坏周围的正常组织，并逐渐降低肾功能，导致终末期肾病。 约50%的患者在生命的第60个十年时患有阶段性肾病。大多数ADPKD是由以下基因突变引起的： 两个基因，PKD 1，编码多囊蛋白-1蛋白（PC 1），PKD 2，编码多囊蛋白-2 蛋白（PC 2）。PC 1和PC 2相互作用，被认为在纤毛信号传导中发挥作用。是 普遍认为纤毛是驱动ADPKD发病机制的途径中的中心组分。 虽然它们与纤毛中功能性PC复合物的机制尚不清楚，但许多信号转导通路都是通过它们的信号转导途径来实现的。 囊肿内的通路受到干扰。在过去的几年里，与疾病相关的途径的清单已经通过 新的证据表明，代谢是一种新的途径，在ADPKD中受到深刻影响， 两者都参与疾病的发病机理并作为治疗发展的靶点。这个问题仍然没有 确定新发现的ADPKD代谢紊乱是否是直接驱动因素 很明显，细胞的许多不同的相互交织的代谢回路的性质和活动 在决定其投资所需能源的能力方面发挥着核心作用， 增殖和活跃的溶质和流体运输所需的囊肿生长。该提案的主要目标是 是为研究界提供新的工具和数据集，这将大大加强努力， 探索和利用ADPKD的代谢变化特征。我们将创造一个独特的设计， 基于细胞特异性转录组学、线粒体和线粒体的新的体内模型， 蛋白质组和线粒体代谢的影响，从PC蛋白质丢失后的最早阶段产生 并且进一步通过伴随的纤毛损失和PC蛋白再活化的影响来告知。这个程序 将利用成年诱导条件性PC敲除小鼠模型，并将采用允许 核糖体的条件分离（TRAP）和线粒体的条件分离，从而使细胞型- 特异性转录组学和线粒体蛋白质组学和代谢组学研究。体内代谢的最新技术水平 通量研究将应用于这些新的遗传小鼠模型的肾皮质。的结果予以 分析将结合起来，以产生强大的生物数据集，将通过应用 必要的信息学机制，以便将这些数据传播给更广泛的研究界， 接近真实的时间。重要的是，我们的体内研究旨在发现肾脏移植后发生的最早变化。 小管失去多囊蛋白的表达-在囊肿形成之前的时间点。研究团队带来了 在ADPKD动物模型、PC信号传导和生物学以及体内 代谢研究加上强大的生物统计学和生物信息学支持，以产生拟议的资源。