STRUCTURE AND FUNCTION OF CD2AP IN THE KIDNEY

CD2AP 在肾脏中的结构和功能

基本信息

批准号：
9055158
负责人：
JEFFREY H MINER
金额：
$ 46.99万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2000
资助国家：
美国
起止时间：
2000-08-15 至 2020-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9055158
关键词：
Actins Acute Adult Appointment Attention Attenuated Biological Biological Assay Biopsy Cell Adhesion Cells Chronic Kidney Failure Coupled Data Disease Filtration Foot Process Founder Effect Functional disorder Funding G Protein-Coupled Receptor Genes GTPase-Activating Proteins Genes Goals Grant Guanosine Triphosphate Phosphohydrolases IL4 gene In Vitro Injury Interleukin-13 Interleukin-4 Kidney Kidney Diseases Knock-in Mouse Knowledge Ligands Location Membrane Modeling Monomeric GTP-Binding Proteins Mus Pathogenesis Pathway interactions Patients Phenotype Physiological Play Population Process Proteinuria Publications Publishing Renal glomerular disease Reporting Research Role STAT6 gene Signal Transduction Stress Fibers Structure Surveys Technology Testing Therapeutic Transgenes Transgenic Mice Transgenic Organisms Work base cytokine embryonic stem cell in vivo mouse model novel novel therapeutics podocyte public health relevance rho rho GTP-Binding Proteins treatment strategy

Actins Acute Adult Appointment Attention Attenuated Biological Biological Assay Biopsy Cell Adhesion Cells Chronic Kidney Failure Coupled Data Disease Filtration Foot Process Founder Effect Functional disorder Funding G Protein-Coupled Receptor Genes GTPase-Activating Proteins Genes Goals Grant Guanosine Triphosphate Phosphohydrolases IL4 gene In Vitro Injury Interleukin-13 Interleukin-4 Kidney Kidney Diseases Knock-in Mouse Knowledge Ligands Location Membrane Modeling Monomeric GTP-Binding Proteins Mus Pathogenesis Pathway interactions Patients Phenotype Physiological Play Population Process Proteinuria Publications Publishing Renal glomerular disease Reporting Research Role STAT6 gene Signal Transduction Stress Fibers Structure Surveys Technology Testing Therapeutic Transgenes Transgenic Mice Transgenic Organisms Work base cytokine embryonic stem cell in vivo mouse model novel novel therapeutics podocyte public health relevance rho rho GTP-Binding Proteins treatment strategy

展开

项目摘要

DESCRIPTION (provided by applicant): This is the third renewal application for this project to understand the pathophysiology of proteinuric kidney diseases. Progress over the last 20 years has focused attention on the podocyte, at least in part because of our pioneering work on CD2AP. In the last funding period, we hypothesized that foot process effacement, a universal feature of proteinuria, was the physiological manifestation of Rac activation. Previous in vitro studies showed that Rac controlled actin cytoskeletal structures like the lamellopodia and membrane ruffling. We postulated that podocytes maintain high levels of Rho activity, which functions to enhance cell adhesion by controlling the formation of actin cables known as stress fibers. To prove our hypothesis, we showed that the acute activation of Rac induced foot process effacement and proteinuria. However, others in the field have argued that it is Rho activation that is the cause of effacement and proteinuria. Because all of this work is based on transgenics, which are difficult to compare because of the number of copies of the transgene and because of random integration, we propose to generate mice using ES cell technology, where a single copy of the transgene is targeted to a specific chromosomal locus. This will allow different transgenics to be compared to each other. In specific aim #1, we will compare and contrast three transgenic mice that we generated that allow for inducible expression of constitutively active Cdc42, RhoA and Rac1. In specific aim #2, we will test whether the activation of RhoA can attenuate the activation of Rac1 and proteinuria in several different models of podocyte injury. Since GPCRs are a major regulator of Rho in cells, and as we believe that Rho activation to be a potential strategy for the treatment of proteinuria, we propose to first characterize all of the GPCRs expressed in podocytes. We will then test whether ligands for this GPCRs can activate Rho using both cultured podocytes and a novel minced kidney assay that we developed in the last funding period. We believe that a better understanding of the mechanistic cell biological causes of foot process effacement and proteinuria will allow for novel therapeutics to be developed that will allow this process to be reversed.

描述（由应用程序提供）：这是该项目了解蛋白尿肾脏疾病的病理生理学的第三个续订应用。在过去的20年中，进步将注意力集中在足细胞上，至少部分是因为我们在CD2AP上的开创性工作。在最后一个资金期间，我们假设脚步能量是蛋白尿的普遍特征，是RAC激活的物理表现。先前的体外研究表明，RAC控制肌动蛋白细胞骨架结构，例如层状膜和膜褶皱。我们假设足细胞维持高水平的RHO活性，从而通过控制称为应激纤维的肌动蛋白电缆的形成来增强细胞粘附。为了证明我们的假设，我们证明了RAC的急性激活诱导的脚部过程能量和蛋白尿。然而，该领域的其他人认为，RHO激活是能量和蛋白尿的原因。由于所有这些工作都是基于转换，因此由于转换的副本数量以及随机整合而难以比较，因此我们建议使用ES细胞技术生成小鼠，其中转换的单个副本针对特定的染色体基因座。这将允许将不同的转换相互比较。在特定的目标＃1中，我们将比较和对比三只转基因小鼠，这些小鼠允许诱导表达组成型活性Cdc42，RhoA和Rac1。在特定的目标＃2中，我们将测试RhoA的激活是否可以在几种不同模型的足细胞损伤模型中减弱Rac1和protinuria的激活。由于GPCR是细胞中RHO的主要调节剂，并且由于我们认为Rho激活是治疗蛋白尿的潜在策略，因此我们提出首先表征在足细胞中表达的所有GPCR。然后，我们将测试该GPCR的配体是否可以使用培养的足细胞和我们在上一个资金期间开发的新型肾脏测定法也可以激活Rho。我们认为，更好地理解足部工艺能量和蛋白尿的机械细胞生物学原因，可以开发出新的治疗，从而可以逆转此过程。