Dissecting the biochemical architecture and morphological release pathways of the human platelet extracellular vesiculome
Dissecting the biochemical architecture and morphological release pathways of the human platelet extracellular vesiculome
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DOI：
10.1007/s00018-018-2771-6
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发表时间：
2018-10-01
影响因子：
8
通讯作者：
Simak, Jan
中科院分区：
文献类型：
--
作者：
De Paoli, Silvia H.;Tegegn, Tseday Z.;Simak, Jan
血小板细胞外囊泡（PEVs）已成为细胞间通讯的潜在介质。PEV表现出几种具有病理生理学重要性的活性，并可作为诊断生物标志物。在这里，成像和分析技术被用来揭示形态学途径的释放，结构，组成，和表面特性的PEV衍生自人血小板（PLT）激活的凝血酶受体活化肽（TRAP）。基于广泛的电子显微镜分析，我们提出了TRAP激活的PLT释放PEV的四种形态学途径：（1）质膜出芽，（2）多泡α颗粒和细胞质空泡的挤出，（3）质膜起泡和（4）PLT伪足的“珍珠状”。PLT细胞外囊泡组包括外泌体、外泌体、游离线粒体、含线粒体的囊泡、“足状体”和PLT“血影”。有趣的是，流式细胞术显示了TOM 20（+）LC 3（+）PEV的群体，可能是血小板线粒体自噬的产物。我们发现，脂质体和蛋白质组学的配置文件是不同的小PEV（S-PEVs;平均直径103 nm）和大囊泡（L-PEVs;平均直径350 nm）的差速离心分离的馏分之间。此外，由活化的PLT释放的大多数PEV由S-PEV组成，与L-PEV相比，S-PEV具有显著更高的每单位PEV表面积的凝血酶生成活性，并且贡献约60%的PLT囊泡体促凝血效力。
Platelet extracellular vesicles (PEVs) have emerged as potential mediators in intercellular communication. PEVs exhibit several activities with pathophysiological importance and may serve as diagnostic biomarkers. Here, imaging and analytical techniques were employed to unveil morphological pathways of the release, structure, composition, and surface properties of PEVs derived from human platelets (PLTs) activated with the thrombin receptor activating peptide (TRAP). Based on extensive electron microscopy analysis, we propose four morphological pathways for PEVs release from TRAP-activated PLTs: (1) plasma membrane budding, (2) extrusion of multivesicular alpha-granules and cytoplasmic vacuoles, (3) plasma membrane blistering and (4) "pearling" of PLT pseudopodia. The PLT extracellular vesiculome encompasses ectosomes, exosomes, free mitochondria, mitochondria-containing vesicles, "podiasomes" and PLT "ghosts". Interestingly, a flow cytometry showed a population of TOM20(+)LC3(+) PEVs, likely products of platelet mitophagy. We found that lipidomic and proteomic profiles were different between the small PEV (S-PEVs; mean diameter 103 nm) and the large vesicle (L-PEVs; mean diameter 350 nm) fractions separated by differential centrifugation. In addition, the majority of PEVs released by activated PLTs was composed of S-PEVs which have markedly higher thrombin generation activity per unit of PEV surface area compared to L-PEVs, and contribute approximately 60% of the PLT vesiculome procoagulant potency.