Activating stimuli induce platelet microRNA modulation and proteome reorganisation
Activating stimuli induce platelet microRNA modulation and proteome reorganisation
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DOI：
10.1160/th14-09-0726
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发表时间：
2015-07-01
影响因子：
6.7
通讯作者：
Golino, Paolo
中科院分区：
文献类型：
--
作者：
Cimmino, Giovanni;Tarallo, Roberta;Golino, Paolo
血小板携带巨核细胞来源的mrna，其在激活前和激活过程中的翻译效率尚不清楚，尽管这可以极大地影响血小板功能，无论是在基础条件下还是在对生理和病理刺激（如急性冠状动脉综合征）的反应中。本研究的目的是确定microRNA （miRNA）表达的变化是否发生在对激活刺激的反应中，以及这是否影响血小板转录组和蛋白质组的活性和组成。收集健康志愿者纯化的富血小板血浆，用ADP、胶原蛋白或凝血酶受体激活肽激活。RNA-Seq转录组分析显示，在刺激的前2小时内，血小板转录组基本未受影响。相比之下，定量蛋白质组学显示，在相同的条件下，几乎一半的bbb700蛋白被定量调节。全球miRNA分析表明，激活过程中血小板蛋白质组的重组反映了成熟miRNA表达的变化，因此，这似乎是观察到的转录组和蛋白质组变化之间差异的主要驱动因素。受调节的mirna显著影响的血小板功能包括整合素/细胞骨架、凝血和炎症免疫反应途径。这些结果表明，血小板mirna在活化过程中发生了显著的重编程，从而导致血小板蛋白质组发生显著变化，并首次提供了大量证据，证明mirna对常驻mRNA翻译的微调是血小板病理生理中的一个关键事件。
Platelets carry megakaryocyte-derived mRNAs whose translation efficiency before and during activation is not known, although this can greatly affect platelet functions, both under basal conditions and in response to physiological and pathological stimuli, such as those involved in acute coronary syndromes. Aim of the present study was to determine whether changes in microRNA (miRNA) expression occur in response to activating stimuli and whether this affects activity and composition of platelet transcriptome and proteome. Purified platelet-rich plasmas from healthy volunteers were collected and activated with ADP, collagen, or thrombin receptor activating peptide. Transcriptome analysis by RNA-Seq revealed that platelet transcriptome remained largely unaffected within the first 2 hours of stimulation. In contrast, quantitative proteomics showed that almost half of >700 proteins quantified were modulated under the same conditions. Global miRNA analysis indicated that reorganisation of platelet proteome occurring during activation reflected changes in mature miRNA expression, which therefore, appears to be the main driver of the observed discrepancy between transcriptome and proteome changes. Platelet functions significantly affected by modulated miRNAs include, among others, the integrin/cytoskeletal, coagulation and inflammatory-immune response pathways. These results demonstrate a significant reprogramming of the platelet miRNome during activation, with consequent significant changes in platelet proteome and provide for 1 the first time substantial evidence that fine-tuning of resident mRNA translation by miRNAs is a key event in platelet pathophysiology.