Mechanisms of Polycystin and Cilia Function in ADPKD

多囊蛋白和纤毛在 ADPKD 中的功能机制

• 批准号: 8615251
• 负责人: STEFAN SOMLO
• 金额: $ 36.21万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-20 至 2018-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8615251
• 关键词: Ablation; Adult; Affect; Animal Model; Antibodies; Autosomal Dominant Polycystic Kidney; Binding; Biological Assay; Cell Line; Cell model; Cells; Cilia; Complement; Complex; Cyst; Cystic kidney; Data; Development; Disease; Duct (organ) structure; Epithelial Cells; Equation; Fibronectin Receptors; Fibronectins; Gene Dosage; Gene Expression Profile; Gene Silencing; Genes; Genetic; Genetic Models; Genotype; Human; In Vitro; Individual; Integrins; Kidney; Kidney Failure; Knock-out; Left; Ligands; Membrane; Modeling; Molecular; Molecular Profiling; Mus; Mutation; Nephrons; PKD2 protein; Pathogenesis; Pathway interactions; Phenotype; Polycystic Kidney Diseases; Principal Investigator; Property; Proteins; Proteomics; Relative (related person); Role; Series; Severities; Signal Pathway; Signal Transduction; Staging; Structure; Time; Transgenic Organisms; Translations; base; bile duct; early onset; expectation; in vivo; inhibitor/antagonist; kinetosome; mouse model; mutant; novel; polycystic kidney disease 1 protein; preclinical efficacy; programs; public health relevance; recombinase; reconstitution; transcriptomics

Program Director/Principal Investigator (Last, First, Middle): Somlo, Stefan Project Summary/Abstract The functional connection between human polycystic kidney diseases and cilia is based in part on the overlap between genes whose mutation results in kidney cyst formation and genes whose protein products are expressed in the cilia-basal body complex. The occurrence of a common phenotype, kidney cysts, following mutation in either of these classes of genes has led to a conceptual equation of cellular pathways affected by loss of polycystin-1 (PC1) or polycystin-2 (PC2) with those affected following loss of structurally intact cilia. This proposal is based on our novel observation that cyst formation following inactivation of either Pkd1 or Pkd2 is markedly slowed if structurally intact cilia are concomitantly ablated. This effect is present in adult onset and early developmental mouse models of both Pkd1 and Pkd2 and is independent of the genetic mechanism of cilia ablation. The findings provide genetic evidence for a pathway that is inhibited by the PC1/PC2 complex and that requires intact cilia to produce maximal cyst promoting signals in the absence of PC1/PC2-a Cilia Dependent Cyst Activating (CDCA) pathway. The objective of this proposal are to identify the components of the CDCA pathway. To achieve this objective, we will define the specific determinants of cyst progression whose activity following inactivation of polycystins is modulated by the presence or absence of intact cilia. We will use a highly correlated series of in vivo mouse models that subsume all stages of CDCA to investigate known and novel candidate CDCA pathways. We will complement these directed studies with transcriptomic and proteomic discovery approaches. We have identified integrin signaling as a candidate CDCA activity. We will explore the role of polycystins in integrin signaling in cilia and determine which components of this pathway are active in cilia. We will use gene knockdown in cells and conditional knockout models in mice to determine whether integrin signaling is a component of CDCA and if so, we will target it therapeutically to determine preclinical efficacy in ADPKD. The overall program offers two novel discoveries regarding the pathogenesis of ADPKD that each have substantial potential for translation. PHS 398/2590 (Rev. 06/09) Page 59 Continuation Format Page

计划总监/首席研究员（最后，第一，中间）：索姆洛，斯特凡 项目摘要/摘要 人多囊性肾脏疾病和纤毛之间的功能连接部分基于重叠 在突变导致肾脏囊肿形成的基因与蛋白质产物的基因之间 在纤毛基体体复合体中表达。发生的常见表型，肾囊肿，遵循 这两类基因中的任何一个突变都导致了受细胞途径的概念方程 在结构完整的纤毛后丧失后，多囊素-1（PC1）或Polycystin-2（PC2）的丧失。 该提议是基于我们的新发现，即PKD1或 如果结构完整的纤毛同时消融，则PKD2明显减慢。这种效果存在于成人中 PKD1和PKD2的发作和早期发育小鼠模型，与遗传无关 纤毛消融机制。这些发现为抑制的途径提供了遗传证据 PC1/PC2复合物，这需要完整的纤毛在没有的情况下产生最大的囊肿来促进信号 PC1/PC2-A纤毛依赖性囊肿激活（CDCA）途径。该提议的目的是确定 CDCA途径的组成部分。为了实现这一目标，我们将定义 囊肿进展，其活性在多囊这症后的活性是由存在或不存在调节的 完整的纤毛。我们将使用一系列高度相关的体内鼠标模型，该模型包含CDCA的所有阶段 研究已知和新颖的候选CDCA途径。我们将与这些定向研究补充 转录组和蛋白质组学发现方法。我们已经确定整联蛋白信号传导是候选者 CDCA活性。我们将探讨多囊这在整合素信号传导中的作用，并确定哪个 该途径的组成部分在纤毛中是活跃的。我们将在细胞中使用基因敲低和条件敲除 小鼠中的模型以确定整联蛋白信号传导是否是CDCA的组成部分，如果是，我们将针对它 在治疗上确定ADPKD中的临床前功效。整个计划提供了两个新颖的发现 关于ADPKD的发病机理，每种发病机理都有巨大的翻译潜力。 PHS 398/2590（修订版06/09）第59页延续格式页面