Characterisation of extracellular vesicle-subsets derived from brain endothelial cells and analysis of their protein cargo modulation after TNF exposure.
Characterisation of extracellular vesicle-subsets derived from brain endothelial cells and analysis of their protein cargo modulation after TNF exposure.
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DOI：
10.1080/20013078.2017.1302705
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发表时间：
2017
影响因子：
16
通讯作者：
Sanchez JC
中科院分区：
文献类型：
--
作者：
Dozio V;Sanchez JC
关于细胞外囊泡（EV）子集（例如微囊泡（MV）和外来体（EXO））之间的组成和功能差异知之甚少，也不知道它们的货物在多大程度上反映了起源细胞的表型状态。脑内皮细胞是血脑屏障（BBB）的组成部分，血脑屏障是维持脑内环境稳定的选择性屏障。血脑屏障损伤与几种神经炎性疾病有关，其中促炎细胞因子肿瘤坏死因子（TNF）通常起关键作用。在本研究中，鸟枪蛋白质组学和基于平行反应监测（PRM）的靶向质谱法被用来检测脑内皮细胞释放的EV，并研究TNF暴露如何调制EV蛋白货物。发现MV富含线粒体和细胞骨架蛋白，而EXO富含粘附、组蛋白和核糖体蛋白。TNF刺激后，发现在细胞中差异表达的TNF和NF-κB信号通路中涉及的几种蛋白在MV和EXO中也差异表达。因此，我们的研究结果揭示了一些新的蛋白质作为区分MV和EXO的潜在有用的候选物，以及细胞系统地并根据其表型状态在EV中“包装”蛋白质的额外证据。
Little is known about the composition and functional differences between extracellular vesicle (EV) subsets, such as microvesicles (MVs) and exosomes (EXOs), nor to what extent their cargo reflects the phenotypic state of the cell of origin. Brain endothelial cells are the constitutive part of the blood–brain barrier (BBB), a selective barrier that maintains brain homeostasis. BBB impairment is associated with several neuroinflammatory diseases with the pro-inflammatory cytokine tumour necrosis factor (TNF) often playing a key role. In the present study, shotgun proteomics and parallel reaction monitoring (PRM)-based targeted mass spectrometry were used to characterise brain endothelial cell-released EVs, and to study how TNF exposure modulated EV protein cargoes. MVs were found to be enriched in mitochondrial and cytoskeletal proteins, whereas EXOs were enriched in adhesion, histone and ribosomal proteins. After stimulation with TNF, several proteins involved in TNF and NF-κB signalling pathways, that were found to be differentially expressed in cells, were also differentially expressed in both MVs and EXOs. Thus, our results revealed some novel proteins as potentially useful candidates for discriminating between MVs and EXOs, together with additional evidence that cells “package” proteins in EVs systematically and according to their phenotypic state.