CELL AND MOLECULAR PATHOBIOLOGY OF RENAL DISEASE

本申请寻求对肾脏研究的广泛项目的支持，该项目旨在了解肾脏疾病的细胞分子机制。该计划的重点是了解肾脏对急性缺血损伤的反应的病理生理学基础。对这一理解至关重要的是关于控制肾上皮和内皮细胞极性建立的分子细胞生物学过程的基础知识，维持上皮和内皮细胞屏障功能的机制，以及这些组织对急性缺血损伤应激的反应。这项调查将在四个互补和综合项目的支持下进行，并得到三个核心计划的支持。研究的焦点包括：1)在黏附分子ZO-1、PECAM和VE-钙粘素的指导下，肾微血管内皮细胞的分化和通透性控制；2)皮质和高尔基光谱蛋白-肌动蛋白细胞骨架结构的调节，以及它们在建立和维持肾上皮细胞Na，K-ATPase和E-钙粘素极性中的作用；3)肾肌球蛋白的特性和作用，它们与肾细胞肌动蛋白骨架的相互作用，以及它们在膜运输和极性建立中的作用；4)建立和维持紧密连接的极性和完整性的机制，重点是肾封闭蛋白作为黏附分子的作用及其与细胞骨架的相互作用。实验将在肾细胞培养系统中进行，包括内皮细胞培养，以及已建立的肾脏系，如MDCK和LLC-PK1细胞。野生型(WT)和转基因小鼠的肾脏损伤也将被研究。支持这一计划的核心设施包括一个复杂的成像核心和一个活体肾损伤核心。集团内部的协同作用、对基础知识的关注、计划的外部审查计划、每月进行中的研究会议以及强大的研讨会系列共同确保了最大的发现机会。对肾脏疾病有重要相关性的理解是有保证的。

DESCRIPTION (Abstract of the application) This application seeks support for a broad program in renal research directed at understanding the cellular molecular mechanisms of renal disease. The emphasis of the program is on understanding the pathophysiologic basis of the kidney's response to acute ischemic injury. Crucial to this understanding is the fundamental knowledge of the molecular cell biological processes governing the establishment of polarity in both renal epithelial and endothelial cells, the mechanisms by which the barrier functions of both epithelium and endothelium are maintained, and the response of these tissues to the stress of acute ischemic injury. This investigation will be carried out under the aegis of four complementary and integrated projects, supported by three Core programs. The areas of focus include: 1) endothelial cell differentiation and permeability control in the renal microvasculature, as guided by the adhesion molecules ZO-1, PECAM and VE-cadherin; 2) the regulation of cortical and Golgi spectrin-actin cytoskeletal organization, and their role in establishing and maintaining Na,K-ATPase and E-cadherin polarity in renal epithelial cells; 3) the identity and role of the renal myosins, their interaction with the renal cell actin skeleton and their role in membrane trafficking and the establishment of polarity; and 4) the mechanisms establishing and maintaining tight-junctional polarity and integrity, focusing on the role of renal occludin as an adhesion molecule and its interactions with the cytoskeleton. Experiments will be carried out both in renal cell culture systems, including cultures of endothelial cells, and in established renal lines such as MDCK and LLC-PK1 cells. Renal injury in both wild type (WT) and genetically modified mice will also be studied. The core facilities, which will support this program, include a sophisticated imaging core and an in vivo renal injury core. Synergies within the group, a focus on fundamental knowledge, a planned program of external review, monthly research in progress meetings, and a robust seminar series collectively assure maximal opportunities for discovery. Understandings of significant relevance to renal disease are assured.