The role of CD2AP-interacting proteins in the pathogenesis of focal segmental glomerulosclerosis - an in vivo screening approach

局灶节段性肾小球硬化(FSGS)是成人最常见的肾小球疾病，也是终末期肾病的主要原因之一，但其发病机制尚不清楚。在FSGS的发展中起主要作用的蛋白质之一是与狭缝隔膜相关的CD2AP。本项目的目的是研究CD2AP在FSGS发病机制中的22个相互作用因子的功能特征。为此，针对先前确定的候选基因的shRNAs将在CD2AP和突触素杂合子的小鼠中以诱导和足细胞特异性的方式表达。这些小鼠已被证明对FSGS的发展具有更高的敏感性。一项革命性的技术使直接从ES细胞产生100%转基因小鼠成为可能。将对这些动物进行蛋白尿筛查，以确定在疾病发展中发挥作用的候选动物。鉴定出的蛋白质将通过尿液分析和包括电子显微镜在内的组织学方法在体内进行表征。分离的足细胞将通过不同的分子和细胞生物学技术进行分析，包括与缝隙隔膜蛋白和肌动蛋白细胞骨架成分的相互作用研究，以确定新的治疗靶点。更具体和个性化的治疗策略是我们患者预防终末期肾脏疾病和透析治疗的关键。

Although the focal segmental glomerulosclerosis (FSGS) is the most frequent glomerulopathy of the adult and one of the major reasons for end-stage renal disease, its pathomechanisms are poorly understood. One of the proteins that plays a major role in the development of FSGS is the slit diaphragm associated CD2AP. The aim of this project is the functional characterization of 22 interactors of CD2AP in the pathogenesis of FSGS. To this end, shRNAs against the previously identified candidate genes will be expressed in an inducible and podocyte-specific way in mice that are heterozygous for CD2AP and synaptopodin. These mice have been shown to exhibit an increased susceptibility for the development of FSGS. A revolutionary technique makes it possible to generate 100% transgenic mice directly from ES cells. These animals will be screened for proteinuria, to identify candidates that play a role in the development of the disease. The identified proteins will be characterized in vivo by urine analysis and histology including electron microscopy. Isolated podocytes will be analyzed by different molecular and cell biological techniques including interaction studies with slit diaphragm proteins and components of the actin cytoskeleton to identify new therapeutic targets. More specific and individualized therapeutic strategies are the key to prevent end stage renal disease and dialysis treatment for our patients.