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: 10200801
• 负责人: Michael J. Caplan
• 金额: $ 131.27万
• 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/10200801
• 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.

摘要 常染色体显性多囊肾病（ADPKD）是最常见的单基因遗传病之一。 疾病，影响全球超过 1:1000 的个体，其特征是巨大的充满液体的肾囊肿，可重塑， 压缩并破坏周围的正常组织，并逐渐降低肾功能，导致最终 大约 50% 的患者在 60 岁时患有阶段性肾病，大多数 ADPKD 是由突变引起的。 两个基因，PKD1，编码多囊蛋白 1 蛋白 (PC1)，PKD2，编码多囊蛋白 2 PC1 和 PC2 相互作用，并被认为在纤毛信号传导中发挥作用。 人们普遍认为纤毛是驱动 ADPKD 发病机制的核心组成部分。 尽管它们与纤毛中功能性 PC 复合体的机制联系尚不清楚，但许多信号传导 在过去的几年里，与疾病相关的途径不断增多。 新证据表明代谢是 ADPKD 中深受影响的一种新途径，并且可能 两者都参与疾病发病机制并作为治疗开发的靶点。 确定新发现的 ADPKD 特征代谢紊乱是否是直接驱动因素 囊肿形成的过程中，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 在确定其投资参与参与所需能源的能力方面发挥着核心作用 包囊生长所需的增殖以及活性溶质和液体运输是该提案的主要目标。 是为研究界提供新颖的工具和数据集，这将大大加强努力 探索和利用 ADPKD 的代谢变化，我们将生产出独特设计和的产品。 基于细胞特异性转录组、线粒体的新颖体内模型的严格策划的资源 PC 蛋白丢失后最早阶段产生的蛋白质组和线粒体代谢效应 并且通过伴随的纤毛损失和 PC 蛋白重新激活的影响进一步了解该程序。 将利用成人诱导条件 PC 敲除小鼠模型，并采用允许的策略 核糖体的条件分离（TRAP）和线粒体的条件分离，从而使细胞类型 特定的转录组学和线粒体蛋白质组学和代谢组学研究。 通量研究将应用于这些新型遗传小鼠模型的肾皮质。 分析将被结合起来产生强大的生物数据集，这些数据集将通过应用进行组装 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 至关重要的是，我们的体内研究旨在发现肾脏发生后最早的变化。 研究小组发现，在囊肿形成之前的时间点，肾小管就失去了多囊蛋白的表达。 汇集了 ADPKD 动物模型、PC 信号传导和生物学以及体内方面广泛且互补的专业知识 代谢研究加上强大的生物统计学和生物信息学支持来产生拟议的资源。