Pathologic Mechanisms of Polycystic Kidney Disease

多囊肾的病理机制

基本信息

批准号：
7941657
负责人：
Angela Wandinger-Ness
金额：
$ 8.99万
依托单位：
UNIVERSITY OF NEW MEXICO
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-30 至 2012-09-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7941657
关键词：
Actins Adherens Junction Adhesions Autosomal Dominant Polycystic Kidney Biochemical Biological Assay Biotinylation Caveolins Cell membrane Cell-Cell Adhesion Cells Cholesterol Complex Cytoskeleton DNA Sequence Rearrangement Data Disease Down-Regulation E-Cadherin Endocytosis Epithelial Epithelial Cells Event Exocytosis Family Filopodia Fluorescence Resonance Energy Transfer Fractionation Genes Kinetics LAR tyrosine phosphatase receptor Life Ligation Link Mediating Membrane Membrane Microdomains Membrane Protein Traffic Monitor Monomeric GTP-Binding Proteins Multiprotein Complexes Mutation PKD2 protein Pathologic Pathology Pathway interactions Peptide Signal Sequences Phosphoric Monoester Hydrolases Phosphorylation Phosphotransferases Plasma Polycystic Kidney Diseases Principal Investigator Protein Tyrosine Kinase Protein Tyrosine Phosphatase Proteins Recycling Regulation Regulation of Exocytosis Role Signal Pathway Signal Transduction Signaling Molecule Simulate Site Surface Testing Therapeutic Intervention beta catenin cellular imaging design flotillin insight kinase inhibitor mutant overexpression polycystic kidney disease 1 protein programs receptor research study response sensor src-Family Kinases trafficking

项目摘要

Autosomal Dominant Polycystic Kidney Disease (ADPKD) is caused by mutations in the genes encoding polycystin-1 and/or polycystin-2, but results in epithelial cells with disrupted adherensjunctions and compromised beta-catenin signaling pathways. Our data showthe polycystins in a multiprotein complex with adherens junction components. The complexes are associated with plasma membranemicrodomains containing the structural lipid raft protein flotillin-2, but not similar microdomains containing caveolin. Disruption of the adherens junction complexes is linked to down regulation of LAR family receptortyrosine phosphatases and hyperphosphorylation of the proteins in the complex. Since adherens junctions provide structural stability to the epithelial sheet through connections to the actin cytoskeleton, and such connections are compromised in ADPKD, these alterations are likely to contribute to the disease pathology. We hypothesize that flotillin-2 membrane microdomains represent sites wherethe polycystins are activated and cooperate with signaling molecules to bring about stable cell-cell adhesion. Consequently,when polycystin-1 function is mutant or absent, the signaling to initiate cell adhesionis altered and changes in renalcystogenic potential result. The experiments in this proposalwill elucidate the organization and function of the polycystin-containing multiprotein complexes associated with the flotillin-2 membranemicrodomainsby probing for colocalized tyrosine kinases and phosphatases and by monitoringthe contribution of flotillin-2 rafts to actin remodeling, membranetrafficking and stable cell-cell adhesion. Successfulcompletion of the proposed experiments will provide new, mechanistic information on the temporal sequence of eventsleading from polycystin-1activation to the stabilization of E-cadherin mediated adhesion. These mechanistic insights are expected to be useful for designing therapeutic interventions, particularly those that make use of kinase inhibitors.

常染色体显性多囊肾脏疾病（ADPKD）是由编码基因突变引起的 polycystin-1和/或polycystin-2，但导致上皮细胞具有破坏的粘发和 β-catenin信号通路受损。我们的数据显示了与粘附连接组件。这些复合物与质膜菌群有关含有结构性脂质筏蛋白鞭毛素-2，但不包含小窝蛋白的微域。粘附连接络合物的破坏与降低LAR家族受体的调节有关复合物中蛋白质的磷酸酶和高磷酸化。由于粘附连接提供通过与肌动蛋白细胞骨架的连接到上皮板的结构稳定性，此类连接在ADPKD中受到损害，这些改变可能会导致疾病病理。我们假设氟列蛋白-2膜微区域代表了多囊这位点的位点，并且与信号分子合作以产生稳定的细胞粘附。因此，当polycystin-1时功能是突变体或不存在的，启动细胞粘附的信号传导发生了变化和肾上腺素的变化潜在的结果。该提案中的实验将阐明含多囊菌素的多蛋白质蛋白复合物与氟列蛋白-2膜性罗生物素有关探测共定位的酪氨酸激酶和磷酸酶，并通过监测氟替林-2的贡献氨基肌动蛋白重塑，膜分裂和稳定的细胞粘附。成功完成拟议的实验将提供有关事件的时间顺序的新机械信息从多囊素1激活到e-钙粘着蛋白介导的粘附的稳定。这些机械洞察力预计将对设计治疗干预措施有用，尤其是那些使用激酶的干预措施抑制剂。