Generating Mouse Mutants with Diabetic Nephropathy

描述(由申请人提供)： AMDCC的目标是开发糖尿病并发症的动物模型，以忠实地复制在人类中观察到的糖尿病并发症。这项建议将提供一种糖尿病肾病(DN)小鼠模型，重点关注两个关键的内皮保护通路：eNOS和前列环素合成酶(PGIs)。这些途径不仅共同定位于内皮细胞内，而且它们的活性也通过细胞水平的过氧硝酸酯相互关联，这两种途径都与人类糖尿病肾病、神经病变、视网膜病变和大血管疾病有关。在之前的资金周期中，Vanderbilt AMDCC网站调查了小鼠糖尿病肾病(DN)的遗传基础。这些研究发现，系统性eNOS缺失是将C57BL/6小鼠从耐药品系转变为糖尿病肾病易感品系的关键遗传修饰因素。内皮型一氧化氮合酶(eNOS或NOSIII)的基因破坏，而不是ApoE或LDLR的基因破坏与C57BL/6糖尿病肾病的显著加速有关，其特征不仅表现为强烈的蛋白尿，而且伴随着肾功能(GFR)的显著系膜溶解和扩张。ENOS作为人类糖尿病肾病风险的临床相关修饰物的参与得到了临床研究的支持，临床研究表明，eNOS Glu298Asp多态的糖尿病患者不仅表现出eNOS活性降低，而且肾功能衰竭的风险增加[Noiri，2002#6975；Shin Shin，2004#8934}。越来越多的证据表明内皮功能障碍与糖尿病并发症的发病机制有关，尤其是肾病和大血管疾病(Schalkwijk，2005#9302)。前列环素合成酶(PGIs)也有类似的多态性，尽管它们在糖尿病肾病进展中的具体作用尚未确定。 目前的建议有两个特定的目标：目的1将确定内皮eNOS在糖尿病肾病进展中的作用；同时确定内皮前列环素合成酶在糖尿病肾病进展中的作用。为了实现这一点，我们将产生有条件的靶向(花)eNOS和PGIS等位基因，并将这些小鼠与Tie2mERCre小鼠杂交，允许从内皮细胞中对这些等位基因进行时间控制的删除。这些研究应该能够剖析这些生化途径在小鼠糖尿病肾病进展中的作用。

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.