Generating Mouse Mutants with Diabetic Nephropathy

产生患有糖尿病肾病的小鼠突变体

基本信息

批准号：
7896027
负责人：
RAYMOND C. HARRIS
金额：
$ 9.82万
依托单位：
VANDERBILT UNIVERSITY
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：
2009-09-10 至 2011-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7896027
关键词：
Acceleration Acetylcholine Address Albuminuria Alleles Animal Model Apolipoprotein E Attention Biochemical Pathway C57BL/6 Mouse Cardiovascular Diseases Cardiovascular system Cessation of life Clinical Research Complications of Diabetes Mellitus Development Diabetic Nephropathy Dissection Endothelial Cells Endothelium Enzymes Event Excretory function Exhibits Functional disorder Funding Genes Genetic Genetic Polymorphism Genotype Goals Guidelines Human Hyperglycemia Inbred Strain Injury Kidney Diseases Kidney Failure Link Maintenance Microalbuminuria Modeling Mus Mutant Strains Mice Neuropathy Oxidative Stress PTGS2 gene Pathogenesis Pathway interactions Peroxonitrite Play Prostacyclin synthase Prostaglandins I Renal function Research Personnel Resistance Retinal Diseases Risk Role Site Source Structure Testing Vasodilation clinically relevant diabetic enzyme activity human NOS3 protein human disease macrovascular disease man mortality mouse model novel therapeutics prevent programs resistant strain type I diabetic urinary

项目摘要

DESCRIPTION (provided by applicant): The goal of the AMDCC is to develop animal models of diabetic complications that faithfully reproduce diabetic complications observed in humans. This proposal will provide a model of mouse model of diabetic nephropathy (DN) focusing on two key protective endothelial pathways: eNOS and prostacyclin synthase (PGIS). These pathways are not only co-localized within the endothelial cells but their activity is also biochemically interrelated through cellular levels of peroxynitrate, and both that have been implicated in human diabetic nephropathy, neuropathy, retinopathy and macrovascular disease. In the previous funding cycle, the Vanderbilt AMDCC site investigated the genetic underpinnings of diabetic nephropathy (DN) of mice. Those studies identified systemic eNOS deletion as a critical genetic modifier that converts C57BL/6 mice from a resistant strain to one that is susceptible to DN. Genetic disruption of endothelial nitric oxide synthase (eNOS or NOSIII), but not ApoE or LDLR was associated with a marked acceleration of DN in C57BL/6, not only characterized by a robust albuminuria, but also by dramatic mesangiolysis and expansion with decrease renal function (GFR). The involvement of eNOS as a clinically relevant modifier for risk of human diabetic nephropathy is bolstered by clinical studies showing that diabetics with an eNOS Glu298Asp polymorphism not only exhibit decreased eNOS activity but also an accelerated risk of renal failure {Noiri, 2002 #6975; Shin Shin, 2004 #8934}. Accumulating evidence implicates endothelial dysfunction in the pathogenesis of diabetic complications, particularly nephropathy and macrovascular disease {Schalkwijk, 2005 #9302}. Similarly polymorphisms have been identified in prostacyclin synthase (PGIS), although their specific role in the progression of diabetic nephropathy has not been established, The present proposal has two specific aims: Aim 1 will determine the role of endothelial eNOS in the progression of diabetic nephropathy; while To determine the role of Endothelial prostacyclin synthase in the progression of diabetic nephropathy. To achieve this we will generate conditionally targeted (floxed) eNOS and PGIS alleles, and cross these mice with a Tie2mERCre mouse, allowing temporally controlled deletion of these alleles specifically from the endothelium. These studies should allow the dissection of the role of these biochemical pathways in the progression of diabetic nephropathy in mice.

描述（由申请人提供）： AMDCC的目的是开发糖尿病并发症的动物模型，这些模型忠实地再现了人类观察到的糖尿病并发症。该提案将提供糖尿病性肾病（DN）的小鼠模型，该模型重点介绍了两种关键的保护性内皮途径：eNOS和前列环素合酶（PGIS）。这些途径不仅在内皮细胞中共定位，而且它们的活性也通过过氧亚硝酸盐的细胞水平在生物化学上相互关联，这两者都与人类糖尿病性肾病，神经病，视感病变和大型疾病有关。在上一个融资周期中，范德比尔特AMDCC站点研究了小鼠糖尿病性肾病（DN）的遗传基础。这些研究将全身性eNOS缺失确定为一种关键的遗传修饰符，将C57BL/6小鼠从抗性菌株转换为易受DN敏感的菌株。内皮一氧化氮合酶（eNOS或NOSIII）的遗传破坏，而不是ApoE或LDLR与C57BL/6中DN的显着加速有关，不仅具有强大的白蛋白尿素，而且具有巨大的蛋白酶，而且还具有戏剧性的Mesangicalsysiss and with with with with n downalise insange and。（GFR）。临床研究表明，eNOS作为人类糖尿病肾病风险的临床相关修饰符的参与表明，具有eNOS GLU298ASP多态性的糖尿病患者不仅表现出eNOS活性的降低，而且还表现出肾衰竭的加速风险{Noiri，2002＃6975; Shin Shin，2004＃8934}。积累的证据暗示了糖尿病并发症的发病机理，尤其是肾病和大血管疾病{Schalkwijk，2005＃9302}。同样，在前列环蛋白合酶（PGI）中已经确定了多态性，尽管尚未确定它们在糖尿病肾病进展中的特定作用本提案具有两个具体的目的：AIM 1将决定内皮eNOS在糖尿病肾病进展中的作用；同时确定内皮前列环素合酶在糖尿病肾病进展中的作用。为了实现这一目标，我们将生成有条件的靶向（Floxed）eNOS和PGIS等位基因，并用Tie2mercre小鼠越过这些小鼠，从而使这些等位基因专门从内皮中进行了时间控制的缺失。这些研究应允许解剖这些生化途径在小鼠糖尿病肾病进展中的作用。