Live Cell Confocal Microscopy for Real-Time Imaging of Endothelial Glycocalyx Damage and Repair

项目总结/摘要 微血管功能失调是创伤后间接器官损伤的病理生理学基础。 特别是，内皮糖萼的损伤发生在创伤性损伤的几分钟内， 微血管渗透性增加导致器官衰竭和死亡率增加。战略 通过防止糖萼分裂和/或促进其修复来减弱糖萼不完整性具有重要的前景 用于最小化微血管功能障碍和创伤后器官损伤。因此， 建议的工作是建立一个基础科学和转化研究计划，重点是识别 新的治疗靶点，将（1）防止糖萼损伤，（2）修复糖萼完整性或（3）抑制 由于糖萼不完整而发生的内皮细胞渗透性失调。具体 重点项目领域将包括确定乙酰肝素酶在调节糖萼中的作用的研究。 创伤-出血后的完整性和介导糖萼合成的机制。此外，我们的 拟议的研究将确定调节内皮屏障功能的信号通路， 细胞表面糖胺聚糖、硫酸乙酰肝素和透明质酸的损失，它们是 糖萼层。我们的研究计划将集中在内皮细胞对创伤的特异性反应上， 最易受到继发性损伤的器官系统中的出血（例如，肾、肺和肠） 其过度的目标是确定复苏策略如何介导糖萼依赖性 每个器官的机制。这些研究项目领域有望对人类产生重大影响 通过帮助发现新的创伤失血性休克患者复苏模式， 可用于抑制糖萼功能障碍、促进其修复或逆转糖萼功能障碍的治疗靶点。 糖萼不完整的下游后果。

PROJECT SUMMARY/ABSTRACT Dysregulation of microvascular function contributes to the pathophysiology of indirect organ injury after trauma. In particular, damage to the endothelial glycocalyx occurs within minutes of traumatic injury and is associated with increased microvascular permeability resulting in organ failure and increased mortality. Strategies that attenuate glycocalyx disintegrity by preventing its cleavage and/or facilitating its repair hold significant promise for minimizing microvascular dysfunction and post-traumatic organ injury. Thus, the overall objective of the proposed work is to establish a basic science and translational research program focused on the identification of novel therapeutic targets that will (1) prevent glycocalyx damage, (2) repair glycocalyx integrity or (3) inhibit dysregulation of endothelial cell permeability that occurs as a result of glycocalyx disintegrity. The specific programmatic areas of focus will include studies to identify the role of heparanase in regulating glycocalyx (dis)integrity after trauma-hemorrhage and on mechanisms that mediate glycocalyx synthesis. Additionally, our proposed studies will identify signaling pathways that regulate endothelial barrier function that are effected by loss of cell surface glycosaminoglycans, heparan sulfate and hyaluronic acid, which are primary constituents of the glycocalyx layer. Our research program will focus on the endothelial cell-specific response to trauma- hemorrhage in organ systems that are most susceptible to secondary injury (e.g., kidney, lung and intestines) with the over-arching goal of determining how resuscitation strategies mediate glycocalyx-dependent mechanisms in each organ. These programmatic areas of research hold promise for significantly impacting the current resuscitation paradigm for trauma patients in hemorrhagic shock by aiding in the discovery of novel therapeutic targets that can be used to inhibit glycocalyx dysfunction, facilitate its repair or reverse the downstream consequences of glycocalyx disintegrity.