Lung endothelial cell phenotypes

项目总结 血管内皮细胞连接血管和所有器官系统。然而，内皮细胞表现出丰富的 结构和功能的多样性。这种异质性在不同类型的血管内皮细胞之间表现明显 器官，沿着器官内单个血管段的内皮细胞，以及立即 相邻单元格。这项计划项目拨款建立在内皮衬里的总体假设基础上 肺动脉、毛细血管和静脉在表型上是不同的，每种细胞类型都是特化的。 以满足其血管利基的独特需求。这些单元之间的专门化由离散编码 第二信使信令网络的组织。因此，在这场竞争性的更新中，我们每一个人 PROJECTS研究内皮细胞信号网络的组织和功能。我们评估了原产地 第二信使信号，这些信号如何在细胞内传播，是什么限制了 信号，以及哪些效应蛋白主要由信号激活。我们第一次可以测量 第二信使在真实的细胞几何形状中随时间的三维传播。细菌，如 铜绿假单胞菌在感染过程中损害内皮细胞功能，最终导致 急性肺损伤。铜绿假单胞菌改变第二信使信号的组织和功能，并在 在某些情况下，细菌利用在宿主期间产生第二信使信号的酶- 病原体相互作用。铜绿假单胞菌及其外酶破坏内皮细胞屏障，因此，我们的 项目使用这种细菌及其毒素来探索内皮异质性和功能的决定因素， 包括屏障的完整性。所有项目都利用垂直集成的方法，从使用 培养的细胞(例如通过控制底物硬度)到各种原位(例如分离的器官，肺切片， 分离的血管和无细胞的肺支架)和体内制剂。在每一种情况下，注意 不同细胞表型的相似之处和不同之处。总而言之，项目 系统地解决三个具体目标或目标：(1)识别和测试新的分子机制 (例如，信号)负责部位特定的内皮细胞功能，特别是在微循环中， (2)确定这些机制在疾病的临床前模型中的重要性，以及(3)翻译小说 临床前疾病模型的治疗方法。项目具有很强的互动性。定义机制 第二信使信号网络不仅会告诉我们内皮细胞的性质 异质性，但对破坏内皮细胞的宿主-病原体相互作用提供了前所未有的洞察力 细胞屏障并导致组织浮肿。对内皮细胞信号转导网络的机械性洞察将 揭示改善内皮功能障碍的新治疗方法。

PROJECT SUMMARY Endothelium lines blood vessels and interconnects all organ systems. However, endothelial cells exhibit a rich diversity in structure and function. Such heterogeneity is apparent between endothelial cells in different organs, in endothelial cells along a single vascular segment within an organ, and between immediately adjacent cells. This program project grant is founded on the overall hypothesis that endothelium lining pulmonary arteries, capillaries, and veins is phenotypically distinct, where each cell type is specialized to fulfill the unique demands of its vascular niche. Specialization among these cells is encoded by discrete organization of second messenger signaling networks. Therefore, in this competitive renewal each of our projects examines the organization and function of endothelial cell signaling networks. We evaluate the origin of second messenger signals, how these signals spread inside of the cell, what limits the lifespan of the signals, and which effector proteins are principally activated by the signals. For the first time we can measure three-dimensional spread of second messengers in realistic cellular geometries over time. Bacteria such as Pseudomonas aeruginosa impair endothelial cell function during the course of infection that culminates in acute lung injury. P. aeruginosa alters the organization and function of second messenger signaling, and in some instances, the bacterium utilizes enzymes that generate second messenger signals during the host- pathogen interaction. P. aeruginosa and its exoenzymes disrupt the endothelial cell barrier, and hence, our projects use this bacterium and its toxins to probe determinants of endothelial heterogeneity and function, including barrier integrity. All projects take advantage of vertically integrated approaches, ranging from the use of cultured cells (e.g. with control for substrate stiffness) to various in situ (e.g. isolated organs, lung slices, isolated blood vessels, and cell-free lung scaffolds) and in vivo preparations. In each of these cases, attention is paid to what is similar, and what is dissimilar, about the respective cell phenotypes. Altogether, projects systematically address three specific aims or objectives, to: (1) identify and test novel molecular mechanisms (e.g. signatures) that are responsible for site-specific endothelial cell function, especially in the microcirculation, (2) determine the importance of these mechanisms in preclinical models of disease, and (3) translate novel therapeutic approaches to preclinical models of disease. Projects are highly interactive. Defining mechanisms that underlie second messenger signaling networks will not only inform us as to the nature of endothelial cell heterogeneity, but provide unprecedented insight into host-pathogen interactions that disrupt the endothelial cell barrier and cause tissue edema. Mechanistic insight into endothelial signal transduction networks will reveal novel therapeutic approaches to improve endothelial dysfunction.