Characterization of single microvesicles in plasma from glioblastoma patients
Characterization of single microvesicles in plasma from glioblastoma patients
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DOI：
10.1093/neuonc/noy187
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发表时间：
2019-05-01
期刊：
影响因子：
15.9
通讯作者：
Weissleder, Ralph
中科院分区：
文献类型：
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作者：
Fraser, Kyle;Jo, Ala;Weissleder, Ralph
背景细胞外囊泡（EV）是由肿瘤细胞脱落的，但对它们的个体分子表型和异质性知之甚少。虽然外来体已经受到了相当大的关注，但对更大的微泡知之甚少。在这里，我们分析了来自胶质母细胞瘤（GB）细胞的单个微泡（MV）和外来体以及来自患者血浆的MV。从小鼠胶质瘤GL 261和人原代GBM 8细胞系以及8名诊断为GB的患者和2名健康对照的血浆中分离并处理EV用于单囊泡分析。将EV固定在载玻片上，在单个囊泡水平上分析囊泡和肿瘤标志物的异质性。我们表明，（i）MV是丰富的，（ii）只有少数MV表达推定的MV标志物，和（iii）MV共享四跨膜蛋白生物标志物，以前被认为是外来体的诊断。使用MV捕获和染色技术，使宿主细胞和GB-衍生的MV的分化，我们进一步证明（i）肿瘤MV往往存在，
Background. Extracellular vesicles (EV) are shed by tumor cells but little is known about their individual molecular phenotypes and heterogeneity. While exosomes have received considerable attention, much less is known about larger microvesicles. Here we profile single microvesicles (MV) and exosomes from glioblastoma (GB) cells and MV from the plasma of patients.Methods. EV secreted from mouse glioma GL261 and human primary GBM8 cell lines as well as from the plasma of 8 patients with diagnoses of GB and 2 healthy controls were isolated and processed for single vesicle analysis. EV were immobilized on glass slides and the heterogeneity of vesicle and tumor markers were analyzed at the single vesicle level.Results. We show that (i) MV are abundant, (ii) only a minority of MV expresses putative MV markers, and (iii) MV share tetraspanin biomarkers previously thought to be diagnostic of exosomes. Using MV capture and staining techniques that allow differentiation of host cell and GB-derived MV we further demonstrate that (i) tumoral MV often present as