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: 8738648
• 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/8738648
• 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

DESCRIPTION (provided by applicant): 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 o 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.

描述（由申请人提供）：人类多囊肾病和纤毛之间的功能联系部分基于其突变导致肾囊肿形成的基因与其蛋白质产物在纤毛-基体复合体中表达的基因之间的重叠。在这两类基因中的任何一类发生突变后，常见表型肾囊肿的出现导致了受多囊蛋白-1 (PC1) 或多囊蛋白-2 (PC2) 损失影响的细胞途径与结构完整纤毛损失后受影响的细胞途径的概念方程。该提议基于我们的新观察，即如果结构完整的纤毛同时被消融，则 Pkd1 或 Pkd2 失活后囊肿的形成会显着减慢。这 这种效应存在于 Pkd1 和 Pkd2 的成年发病和早期发育小鼠模型中，并且与纤毛消融的遗传机制无关。这些发现为 PC1/PC2 复合物抑制的途径提供了遗传证据，该途径需要完整的纤毛在缺乏 PC1/PC2-纤毛依赖性囊肿激活 (CDCA) 途径的情况下产生最大的囊肿促进信号。该提案的目的是确定 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的具体决定因素，其在多囊蛋白失活后的活性受到完整纤毛的存在或不存在的调节。我们将使用一系列高度相关的体内小鼠模型来研究已知和新颖的候选CDCA途径，这些模型包含CDCA的所有阶段。我们将通过转录组学和蛋白质组学发现方法来补充这些定向研究。我们已将整合素信号转导确定为候选CDCA活性。我们将探索多囊蛋白在纤毛整合素信号传导中的作用，并确定该途径的哪些成分在纤毛中活跃。我们将使用细胞中的基因敲除和小鼠条件性敲除模型来确定整合素信号传导是否是CDCA的一个组成部分，如果是，我们将对其进行治疗靶向以确定ADPKD的临床前疗效。整个项目提供了关于 ADPKD 发病机制的两项新发现，每一项都具有巨大的转化潜力。