OKHSC COBRE: THE ROLE OF THE UBIQUITIN SYSTEM IN RETINAL DEGENERATION

OKHSC COBRE：泛素系统在视网膜变性中的作用

基本信息

批准号：
8360280
负责人：
Scott M Plafker
金额：
$ 16.48万
依托单位：
UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-07-01 至 2012-09-09
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8360280
关键词：
Angiotensin Type 1a Receptor Animal Model Animals Binding Biological Models Cell Cycle Cell model Centers of Research Excellence Complement Complex Complications of Diabetes Mellitus Diabetes Mellitus Diabetic Nephropathy Disease End stage renal failure Event Funding Genetic Glucose Grant Histologic Hyperglycemia Hypertrophy Incidence Kidney Kidney Failure Knockout Mice Learning Maps Mentors Methods National Center for Research Resources Oklahoma Physiological Principal Investigator Rattus Research Research Infrastructure Resources Retinal Degeneration Rodent Model Role S Phase Signal Transduction Source Streptozocin System Tubular formation Ubiquitin Ubiquitin-Conjugating Enzymes United States National Institutes of Health cost design diabetic hemodynamics inhibitor/antagonist mesangial cell response tissue/cell culture tool

项目摘要

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Diabetic nephropathy is a major complication of diabetes and the leading cause of end-stage renal disease. The earliest morphological changes accompanying diabetic nephropathy are hemodynamic and structural changes in the renal glomerular and tubular compartments. At the cellular level, hypertrophy of the mesangial cells is a hallmark of these early changes. Multiple lines of experimental evidence have revealed a prominent role for the interplay of the angiotensin II receptor type 1A (AT1 A) and p27Kip1 in diabetic mesangial cell hypertrophy. p27Kip1 is a cell cycle inhibitor that normally must be degraded to enable progression from G1 to S phase. During hyperglycemia, p27Kip1 is inappropriately stabilized through signaling events from the AT1 A. An effector of the AT1A called RCBTB1 is required for the induction of cellular hypertrophy, but the function of RCBTB1 and how it contributes to hypertrophy following hyperglycemia are unknown. We have discovered that RCBTB1 is a component of the ubiquitin system and that the same domain of RCBTB1 that binds to the AT1A also interacts with UBE2E3, a ubiquitin conjugating enzyme. Our hypothesis is that UBE2E3 and RCBTB1 modulate the AT1A in a ubiquitin-dependent fashion to contribute to p27Kip1-dependent, mesangial cell hypertrophy. Our model system is rat mesangial cells, a well-characterized glomerular cell model that recapitulates the hypertrophy observed in diabetic nephropathy. Our specific aims are: (1) to biochemically map the complexes formed between the AT1 A, RCBTB1, and UBE2E3, and (2) to determine if disrupting the expression/function of RCBTB1 and UBE2E3 alters p27Kip stabilization and cellular hypertrophy in response to high glucose. These studies will be done in tissue culture cells and complemented by studies in an STZ-rodent model of diabetic nephropathy. Kidneys from these animals will be analyzed histologically, immunohistologically, and biochemically for hypertrophy and p27Kip1 accumulation. To complement these aims, we will generate mice null for UBE2E3 or RCBTB1. These animals will provide us with powerful tools to study UBE2E3 and RCBTB1 by genetic and physiological methods in validated animal models of diabetic disease. Collectively, the information learned from these studies may contribute to the rational design of new treatment options for diabetic nephropathy. Such therapies are desperately needed as the incidence of diabetes and renal failure escalate

该副本是利用资源的众多研究子项目之一由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持而且，副投影的主要研究员可能是其他来源提供的包括其他NIH来源。列出的总费用可能代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。糖尿病性肾病是糖尿病的主要并发症，也是终末期肾脏疾病的主要原因。伴随糖尿病性肾病的最早形态变化是肾肾小球和管状区室中的血液动力学和结构变化。在细胞水平，肥大肾小球细胞是这些早期变化的标志。多种实验证据揭示了血管紧张素II受体1A型（AT1 A）和P27KIP1在糖尿病性膜细胞肥大中的重要作用。 P27KIP1是一种细胞周期抑制剂，通常必须降解以启用从G1到S相的进展。在高血糖期间，P27KIP1通过来自AT1 A的信号事件不适当地稳定。诱导细胞肥大的AT1A的效应子是AT1A的效应子，但RCBTB1的功能以及在高血糖过度血糖后对肥大的贡献是未知的。我们发现RCBTB1是泛素系统的一个组成部分，并且与AT1A结合的RCBTB1的相同域也与ube2e3相互作用，ube2e3是一种泛素结合酶。我们的假设是，UBE2E3和RCBTB1以泛素依赖性的方式调节AT1A，以促进P27KIP1依赖性的中键细胞肥大。我们的模型系统是大鼠肾小球细胞，这是一种良好的肾小球细胞模型，可概括糖尿病中观察到的肥大肾病。我们的具体目的是：（1）生化绘制AT1 A，RCBTB1和UBE2E3之间形成的复合物，以及（2）确定是否破坏RCBTB1和UBE2E3的表达/功能是否改变了RCBTB1和UBE2E3的表达/功能，改变了P27KIP稳定化和对高glucose响应的细胞肥大。这些研究将在组织培养细胞中进行，并在糖尿病肾病模型中的研究中得到补充。这些动物的肾脏将在组织学，免疫组织学和生化上进行肥大和P27KIP1积累的分析。为了补充这些目标，我们将为ube2e3生成无效的老鼠或RCBTB1。这些动物将通过遗传和生理方法在经过验证的糖尿病疾病模型中通过遗传和生理方法为研究UBE2E3和RCBTB1提供强大的工具。从这些研究中学到的信息总的来说，可能有助于糖尿病性肾病的新治疗选择的合理设计。迫切需要这种疗法，因为糖尿病和肾衰竭的发生率不断升级