Glomerular Capillary Wall: Development and Disease

描述：肾小球毛细血管壁由三部分组成：(1)内皮细胞层；(2)肾小球基底膜；(3)外上皮细胞层(足细胞)。这种独特的结构构成了肾小球滤过屏障，这一点已经得到了很好的证实，但这种结构是如何在肾脏器官发生过程中发展起来的，如何在正常成熟中保持，以及在疾病中受到损害的，还不完全清楚。我们申请的目的是建立一个重点和互补的研究项目组，定义GBM的发展和组装以及选定的肾小球疾病的分子发病机制。具体地说，项目1将检测某些层粘连蛋白和IV型胶原突变小鼠的肾小球异常表型，并通过后肾移植和可诱导的、细胞选择性的转基因表达策略挽救这些表型。项目2将结合经典的分子和细胞方法，使用质谱学、X射线结晶学和表面等离子体共振技术研究IV型胶原网络的形成和与肾小球细胞整合素的相互作用。项目3将绘制致病的Goodppure和Alport同种异体抗原在IV型胶原上的分子位置，检查表达人源化IV型胶原的小鼠和Alport小鼠模型中抗GBM疾病的发展。项目4将使用反义敲击和实时成像技术等方法，研究多头蛇和斑马鱼胚胎作为简化模型生物体内细胞外基质组装和细胞-基质相互作用的分子调节。该方案旨在促进各项目之间的广泛交流和资源共享，这些项目还得到一个行政核心(A)和一个分子识别核心(B)的支持。我们期待这个项目将从根本上揭示有关肾小球结构在健康和疾病方面的新信息。

Description: The kidney glomerular capillary wall is comprised of three elements: (1) an inner endothelial cell layer; (2) the glomerular basement membrane (GBM); and (3) an outer epithelial cell layer (podocytes). That this unique structure constitutes the glomerular filtration barrier has been well established, but how this structure develops during kidney organogenesis, is maintained in maturation normally, and becomes damaged in disease, is not fully understood. The purpose of our application is to establish a focused and complementary group of research projects that will define the development and assembly of the GBM and the molecular pathogenesis underlying selected glomerular diseases. Specifically, Project 1 will examine abnormal glomerular phenotypes in certain laminin and collagen IV mutant mice and rescue these phenotypes through metanephric grafting and inducible, cell-selective transgene expression strategies. Project 2 will study type IV collagen network formation and interaction with glomerular cell integrins using mass spectrometry, x-ray crystallography, and surface plasmon resonance, in combination with classic molecular and cellular methodologies. Project 3 will map the molecular location of pathogenic Goodpasture and Alport alloantigens on type IV collagen, examine the development of anti-GBM disease in a mouse expressing humanized type IV collagen, and in an Alport mouse model. Project 4 will examine the molecular regulation of extracellular matrix assembly and cell-matrix interactions in vivo in hydra and Zebrafish embryos as simplified model organisms, using antisense knock down and real-time imaging techniques, among other approaches. The Program is designed to promote extensive communication and sharing of resources among the Projects, which are also supported by an Administrative Core (A) and a Molecular Recognition Core (B). We anticipate that this Program will reveal fundamentally new information regarding glomerular structure in health and disease.