Glycoproteomics in Ischemic Stroke Related Hyperglycemia

项目摘要/摘要 急性高血糖是缺血性卒中患者常见的临床问题，并经常与 与tPA溶栓效果受损和卒中患者功能预后不良有关。 然而，急性高血糖的临床治疗仍然不确定。最近的研究表明， 急性高血糖与蛋白质糖基化的快速改变有关，这与广泛的 一系列细胞外信号通路和生物功能。因此，急性高血糖症有可能 依赖蛋白糖基化可能在调节血栓溶解和神经血管损伤中起关键作用 中风患者。我们建议使用临床蛋白质组学方法来检测这些细胞的蛋白质糖基化。 进而探讨细胞培养的作用机制。第一个目标是系统地识别血液中的变化 蛋白质糖基化与tPA治疗卒中患者急性高血糖相关的MS研究 了解临床糖蛋白信号的广阔前景的方法。然后，我们验证了结果在 一个独立的大队列缺血性中风患者的血液样本，以选择具有高水平的新的生物标志物 翻译影响。最后，由于脑微血管内皮细胞在调节 血栓溶解和血管功能通过合成和分泌调节因子，我们的目的是检查 脑内皮细胞在高血糖状态下分泌的蛋白质的糖基化。这可能会有帮助 了解脑内皮细胞在调节缺血性卒中相关细胞外反应中的作用 高血糖症。在这次重新提交中，我们根据审查者的意见进行了广泛的修改，包括 增加新的专家、新的数据和出版物以及实验细节和替代方案。

Project Summary/Abstract Acute hyperglycemia is a common clinical problem in ischemic stroke patients, and is frequently associated with impaired thrombolytic efficacy of tPA and predictive of poor functional outcome of stroke patients. However, the clinical management of acute hyperglycemia remains uncertain. Recent studies showed that acute hyperglycemia is associated with rapid alteration in protein glycosylation, which is implicated in a wide range of extracellular signaling pathways and biological functions. It is thus possible that acute hyperglycemia dependent protein glycosylation might play a critical role in regulating thrombolysis and neurovascular injury in stroke patients. We propose to use clinical proteomic approaches to examine protein glycosylation in these patients and then probe mechanism in cell culture. The first aim is to systematically identify changes in blood protein glycosylation associated with acute hyperglycemia in tPA-treated stroke patients using MS-based approach to understand a broad landscape of clinical glyco-protein signals. Then, we validate the results in blood samples of an independent, large cohort of ischemic stroke patients, to select novel biomarkers with high translational impact. Finally, as brain microvascular endothelial cells play a critical role in regulating thrombolysis and blood vessel function via synthesizing and secreting regulatory factors, we aim to examine the glycosylation of proteins secreted by brain endothelial cells under hyperglycemic condition. This may help to understand the role of brain endothelium in regulating extracellular response to ischemic stroke related hyperglycemia. In this resubmission, we have revised extensively according to reviewers’ comments, including adding new experts, new data and publications and experimental details and alternatives.