The Role of Protein Degradation Systems for Glomerular Protein Homeostasis

蛋白质降解系统对肾小球蛋白质稳态的作用

• 批准号: 429327206
• 负责人: Professorin Dr. Catherine Meyer-Schwesinger
• 金额: --
• 依托单位: Institut für Zelluläre und Integrative Physiologie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2019
• 资助国家: 德国
• 起止时间: 2018-12-31 至 2023-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/429327206?language=en
• 关键词: Role; Protein; Degradation; Systems; Glomerular

Three types of glomerular cells, which form a functional syncytium, achieve renal filtration. Of these, podocytes and endothelial cells are separated by the glomerular basement membrane and ultimately compose the three-layered glomerular filtration barrier (GFB), which is thought to impart both size-selective and charge-selective properties. Mesangial cells occupy the space inbetween the GFB to provide structural support and to indirectly participate in filtration by reducing the glomerular surface area by contraction. Typical for glomerular injury is the deposition of proteins originating from the plasma in form of subepithelial, subendothelial and mesangial deposits. Under physiologic conditions glomerular deposition of protein is neglectable even though permeability of the GFB to plasma protein is only partial, suggesting that preventive mechanisms must exist. Impairment of these mechanisms could result in pathologic glomerular protein deposition, which in turn would affect glomerular function. The basis of glomerular protein homeostasis is unknown partly due to the complexity of protein degradation systems and due to the complexity of the glomerular syncytium. In this proposal we hypothesize, that protein uptake and intracellular degradation through the two major degradative systems, namely the ubiquitin-proteasomal system (UPS) and the autophagosomal lysosomal system (ALS) interplay in glomerular cells and contribute to maintain the integrity of the glomerular filter in a cell-specific manner. Preliminary investigations demonstrate that intra- and extracellular protein homeostasis of mesangial and endothelial cells predominantly depends on an intact ALS, whereas podocytes in first line depend on the UPS. Thereby, inhibition of the UPS promotes subepithelial IgG deposition and proteinuria whereas ALS inhibition results in mesangial IgG deposition. To dissect the unknown physiologic and pathophysiologic importance of the UPS and ALS for the intra- and extracellular protein homeostasis of glomerular cells we defined three aims which will be addressed with novel techniques of glomerular cell-specific analyses. (1) Define the differential cell-specific basal activity of the UPS and the ALS in glomerular cells. (2) Comparative analyses of the consequence of clinically used proteasomal inhibitors on the protein homeostasis of glomerular cells. (3) Comparative analyses of the consequence of genetic cell-specific proteasomal and lysosomal impairment for the unchallenged glomerular syncytium and after IgG exposition. The results will provide the missing basis for i.e. understanding the mechanisms of glomerular protein deposition, for understanding conflicting data on adverse drug effects of proteasomal inhibitors, and of the different glomerular phenotypes of mutations involving UPS and ALS proteins.

三种类型的肾小球细胞形成一个功能合体，实现肾滤过。其中，足细胞和内皮细胞被肾小球基底膜隔开，最终组成三层肾小球滤过屏障(GFB)，它被认为具有大小选择和电荷选择特性。肾小球系膜细胞占据GFB之间的空间，提供结构支持，并通过收缩缩小肾小球表面积，间接参与滤过。典型的肾小球损伤是源自血浆的蛋白质以上皮下、内皮下和系膜沉积的形式沉积。在生理条件下，肾小球蛋白沉积是可以忽略的，即使GFB对血浆蛋白的通透性只是部分的，提示必须存在预防机制。这些机制的破坏可能导致病理性肾小球蛋白沉积，进而影响肾小球功能。肾小球蛋白质动态平衡的基础尚不清楚，部分原因是蛋白质降解系统的复杂性以及肾小球合体的复杂性。在这个方案中，我们假设，蛋白质摄取和细胞内降解通过两个主要的降解系统，即泛素-蛋白酶体系统(UPS)和自噬溶酶体系统(ALS)在肾小球细胞中相互作用，并以细胞特异性的方式有助于维持肾小球滤器的完整性。初步研究表明，肾小球系膜细胞和血管内皮细胞的细胞内和细胞外蛋白质平衡主要依赖于完整的ALS，而一线足细胞则依赖于UPS。因此，抑制UPS可促进上皮下免疫球蛋白沉积和蛋白尿，而抑制ALS则导致系膜免疫球蛋白沉积。为了剖析UPS和ALS对肾小球细胞内和细胞外蛋白质稳态的未知生理学和病理生理学意义，我们定义了三个目标，这些目标将通过新的肾小球细胞特异性分析技术来解决。(1)确定肾小球细胞中UPS和ALS的细胞特异性基础活性的差异。(2)临床应用蛋白酶体抑制剂对肾小球细胞蛋白质稳态影响的比较分析。(3)遗传细胞特异性蛋白酶体和溶酶体损伤对未激发的肾小球合体和暴露后的肾小球的影响的比较分析。这些结果将为理解肾小球蛋白沉积的机制，理解蛋白酶体抑制剂不良药物效应的相互矛盾的数据，以及涉及UPS和ALS蛋白的不同肾小球突变表型提供缺失的基础。