Glomerular Capillary Wall: Development and Disease

描述：肾小球毛细血管壁由三部分组成：(1)内内皮细胞层；(2)肾小球基底膜（GBM）；(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，但是这种结构在肾脏器官形成过程中是如何发展的，在正常成熟过程中是如何维持的，在疾病中是如何受损的，还没有完全了解。我们申请的目的是建立一个集中和互补的研究项目小组，以确定GBM的发展和组装以及选定肾小球疾病的分子发病机制。具体来说，项目1将检测某些层粘连蛋白和胶原IV突变小鼠的异常肾小球表型，并通过后肾移植和可诱导的细胞选择性转基因表达策略来挽救这些表型。项目2将结合经典的分子和细胞方法，利用质谱、x射线晶体学和表面等离子体共振，研究IV型胶原网络的形成及其与肾小球细胞整合素的相互作用。项目3将绘制致病的Goodpasture和Alport同种抗原在IV型胶原蛋白上的分子位置，并在表达人源化IV型胶原蛋白的小鼠和Alport小鼠模型中检测抗gbm疾病的发展。项目4将使用反义敲除和实时成像技术等方法，研究水螅和斑马鱼胚胎作为简化模式生物体内细胞外基质组装和细胞-基质相互作用的分子调控。该计划旨在促进项目之间的广泛沟通和资源共享，并由行政核心(A)和分子识别核心(B)提供支持。我们预期这个项目将揭示健康和疾病中肾小球结构的基本新信息。

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.