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

项目总监/首席研究员（最后、第一、中间）：Somlo、Stefan 项目概要/摘要 人类多囊肾病和纤毛之间的功能联系部分基于重叠 突变导致肾囊肿形成的基因和蛋白质产物为 在纤毛-基底体复合体中表达。常见表型肾囊肿的发生如下 这两类基因中的任何一类的突变都导致了受以下因素影响的细胞途径的概念方程： 多囊蛋白-1 (PC1) 或多囊蛋白-2 (PC2) 丢失，以及结构完整的纤毛丢失后受影响的人。 该提议基于我们的新观察，即 Pkd1 或 Pkd1 失活后囊肿形成 如果结构完整的纤毛同时被消融，Pkd2 会明显减慢。这种效应存在于成人中 Pkd1 和 Pkd2 的发病和早期发育小鼠模型，并且与遗传无关 纤毛消融机制。这些发现为受抑制的途径提供了遗传证据 PC1/PC2 复合体需要完整的纤毛才能在缺乏 PC1/PC2 复合物的情况下产生最大的囊肿促进信号 PC1/PC2-纤毛依赖性囊肿激活 (CDCA) 途径。该提案的目标是确定 CDCA 途径的组成部分。为了实现这一目标，我们将定义具体的决定因素 囊肿进展，其活性在多囊蛋白失活后受到存在或不存在的调节 完整的纤毛。我们将使用一系列高度相关的体内小鼠模型，其中包含 CDCA 的所有阶段 研究已知和新颖的候选CDCA途径。我们将补充这些定向研究 转录组和蛋白质组发现方法。我们已确定整合素信号传导作为候选者 CDCA 活动。我们将探索多囊蛋白在纤毛整合素信号传导中的作用，并确定哪些 该途径的成分在纤毛中活跃。我们将在细胞中使用基因敲低和条件敲除 小鼠模型以确定整合素信号传导是否是CDCA的一个组成部分，如果是，我们将针对它 治疗以确定 ADPKD 的临床前疗效。整个计划提供了两个新发现 关于 ADPKD 的发病机制，每一个都具有巨大的转化潜力。 PHS 398/2590 (Rev. 06/09) 第 59 页 继续格式页