Endothelin Signaling and Actions in Renal Mesangium

肾系膜中的内皮素信号传导和作用

基本信息

批准号：
9143751
负责人：
ANDREY SOROKIN
金额：
$ 33.28万
依托单位：
MEDICAL COLLEGE OF WISCONSIN
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-20 至 2019-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9143751
关键词：
Adaptor Signaling Protein Adult Apoptotic Applications Grants Area Biomedical Research Calcium Capillary Endothelial Cell Cell Cycle Regulation Cell Proliferation Cell physiology Cells Complex Coupled DBL Oncoprotein Diabetic Nephropathy Disease Endothelial Cells Endothelin Endothelin Receptor Endothelin-1 Engineering Epithelial Cells Excision Extracellular Matrix Extracellular Matrix Proteins FOXO3A gene Family Fibrosis Filtration GTP-Binding Proteins Gene Targeting Generations Genes Glomerular Capillary Glomerular Mesangial Cell Goals Health Human Hypertension Inbred Strains Rats Injury Kidney Kidney Diseases Kidney Failure Knock-in Knock-out Longevity Mediating Modification Molecular Mutate Mutation Nephritis Oxidative Stress Parietal Pathogenesis Pathology Patients Peptides Phagocytosis Phosphorylation Site Play Production Protein Isoforms Proteins Rat Strains Rattus Regulation Renal function Renal glomerular disease Research Personnel Role Signal Pathway Signal Transduction Structure of glomerular mesangium Testing Urine age related blood filtration cell type glomerular function glomerulosclerosis in vivo innovation member mesangial cell mutant novel podocyte protein protein interaction rat genome transcription factor zinc finger nuclease

项目摘要

DESCRIPTION (provided by applicant): In renal mesangium endothelin-1 (ET-1) exerts excessive contraction, proliferation and extracellular matrix accumulation leading to glomerulosclerosis and kidney failure. The molecular mechanisms of ET-1 actions in renal mesangium are insufficiently studied. In the current grant application we aim to prove that novel ET-1 mediated signaling pathways, discovered by us in cultured glomerular mesangial cells (GMC), play principal role in glomerular diseases in vivo when ET-1 production is increased and renal mesangium is dysfunctional. To achieve these goals we have generated unique rat strains in which we precisely modified rat genome using engineered Zinc Finger Nucleases (ZFNs) in combination with innovative in vivo knock-in strategy. Until recently the precise modification of rat genome was not possible, but the generation of targeted gene changes using ZFNs in inbred rat strains has become one of the major breakthroughs in the field dramatically increasing opportunities of investigators in utilizing rats for biomedical research. In our preliminary studie we have discovered novel signaling pathway stimulated by ET-1 in GMC which involves the formation of multiunit signaling complex including adaptor protein p66 Shc. We hypothesize that ET-1 signaling via adaptor protein p66 Shc in renal mesangium in vivo is contributing to kidney pathologies associated with abnormal function of renal mesangial cells. In specific aim 1 we will test whether ET-1-mediated signaling via p66 Shc contributes to renal injury in glomerular diseases associated with enhanced ET-1 production and abnormal glomerular function. We will induce anti-Thy-1.1 nephritis and hypertension-induced nephropathy in rats which either lack p66 Shc protein or express endogenous p66 Shc with introduced mutations. The extent of renal injury will be assessed. In specific aim 2 we will use primary GMC derived from wild type and genetically modified rat strains to uncover the molecular mechanism of p66 Shc signaling in renal mesangium. We will test the hypothesis that p66 Shc promotes GMC proliferation via inactivation of transcription factor FOXO3a and restricts GMC contractility through regulation of calcium influx. These studies are important because abnormal GMC function is detected in the majority of patients with hypertension induced nephropathy and glomerulosclerosis. The elucidation of mechanisms of ET-1-induced renal pathologies will result in understanding of the mechanisms underlying proliferation-associated and oxidative stress related renal glomerular diseases.

描述（由申请人提供）：在肾脏中心素-1（ET-1）中，施加过度收缩，增殖和细胞外基质积累，导致肾小球硬化和肾衰竭。肾小肌中ET-1作用的分子机制不足。在当前的赠款应用中，我们旨在证明我们在培养的肾小球肾小球细胞（GMC）中发现的新型ET-1介导的信号通路，当ET-1产生增加并且肾小管中性症是功能障碍时，在体内肾小球疾病中起主要作用。为了实现这些目标，我们已经产生了独特的大鼠菌株，在这些大鼠菌株中，我们使用工程锌指核酸酶（ZFN）与创新的体内敲入策略结合使用了工程锌指核酸酶（ZFN）。直到最近，不可能进行大鼠基因组的精确修饰，但是在近交大鼠菌株中使用ZFN的靶向基因变化的产生已成为该领域的主要突破之一，这显着增加了研究人员利用大鼠进行生物医学研究的机会。在我们的初步研究中，我们发现了GMC中ET-1刺激的新型信号传导途径，该途径涉及形成多振子信号传导复合物，包括衔接蛋白p66 SHC。我们假设体内肾小球膜中通过衔接蛋白p66 SHC的ET-1信号传导有助于与肾脏中膜细胞功能异常功能相关的肾脏病理。在特定目标1中，我们将测试ET-1介导的信号通过P66 SHC是否有助于与增强ET-1产生和异常肾小球功能相关的肾小球疾病的肾脏损伤。我们将诱导抗Thy-1.1肾炎和高血压诱导的大鼠肾病，而大鼠缺乏p66 SHC蛋白或带有引入突变的表达内源性p66 SHC。将评估肾脏损伤的程度。在特定目标2中，我们将使用源自野生型和转基因大鼠菌株的主要GMC来揭示肾小肌中p66 SHC信号传导的分子机制。我们将检验以下假设：p66 SHC通过转录因子FOXO3A促进GMC增殖，并通过调节钙涌入来限制GMC收缩性。这些研究很重要，因为在大多数高血压诱导肾病和肾小球硬化症患者中检测到异常的GMC功能。 ET-1诱导的肾脏病理机制的阐明将导致了解与增殖相关和氧化应激相关肾脏肾小球疾病的基础机制。