Cellular/Molecular Mechanisms of Vascular Permeability

描述（由申请人提供）：血管内皮的主要功能是介导和控制血浆和间质液之间的水和溶质（小分子和大分子）的跨内皮交换，简言之，控制血管通透性。这种功能显然是“至关重要的”，对于所有组织和器官的所有细胞的正常生长，发育和存活至关重要。虽然已经确定了跨内皮交换中涉及的形态结构（即小窝、内皮特异性细胞器，如跨内皮通道、窗孔、囊泡-空泡细胞器以及细胞间连接），但关于所涉及结构的分子组成、其生物起源和调节的生化证据很少或没有。本研究的主要目的是阐明内皮细胞小窝及其相关的气孔隔膜在跨内皮运输生理中的作用。这些发现还将记录跨内皮转运的一个新方面，即其调节的可能性和方式（在速率和转运组分中）。除了它们对理解跨内皮运输的正常生理过程的影响之外，这些数据还可以进一步用于研究几种人类疾病的病理生理学，例如肿瘤血管生成、动脉粥样硬化、急性和慢性炎症（病毒和细菌感染、移植排斥、过敏性脑脊髓炎、阿尔茨海默病），其中已经显示发生这种调节。这些研究还可以提供新的运输相关的内皮特异性分子标记物，可用于设计药物和基因靶向选定的微血管床的策略。所采用的技术是细胞分级分离、生化测定、无细胞测定、体内转运研究、转基因动物模型、细胞培养、转染、光学和电子显微镜。

DESCRIPTION (provided by applicant): The main function of the vascular endothelium is the mediation and control of the transendothelial exchanges of water and solutes (both small and large molecules) between blood plasma and theinterstitial fluid, in short the control of vascular permeability. This function is obviously "vital", being critical for normal growth, maintainance and survival of all the cells from all tissues and organs . While the morphological structures involved in transendothelial exchanges have been identified (i.e. caveolae, endothelial specific organelles such as transendothelial channels, fenestrae, vesiculo-vacuolar organelles as well as intercellular junctions), there is very little to no biochemical evidence on the molecular composition of the structures involved, their biogenesis and regulation. The major goals of this research proposal are to elucidate the role of endothelial caveolae and their associated stomatal diaphragms in the transendothelial transport physiology. The findings will also document a novel aspect of the transendothelial transport namely the possibility and ways of its modulation (in rate and components transported). Besides their impact on the understanding of the normal physiological process of the transendothelial transport, the data could be used further in the study of the pathophysiology of several human diseases such as tumor angiogenesis, atherosclerosis, acute and chronic inflammation (both viral and bacterial infections, transplant rejection, allergic encephalomyelytis, Alzheimer disease) where such modulations have been shown to occur. These studies could also provide novel transport related endothelial specific molecular markers that could be used in designing strategies for drugs and gene targeting to selected microvascular beds. The techniques employed are cell fractionation, biochemical assays, cell free-assays, in-vivo transport studies, genetically modified animal models, cell culture, transfections, light and electron microscopy.