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Thromb Haemost 2015; 114(01): 96-108
DOI: 10.1160/TH14-09-0726
Cellular Haemostasis and Platelets
Schattauer GmbH

Activating stimuli induce platelet microRNA modulation and proteome reorganisation

Giovanni Cimmino   *
1   Department of Cardiothoracic and Respiratory Sciences, Second University of Naples, Naples, Italy
,
Roberta Tarallo   *
2   Laboratory of Molecular Medicine and Genomics, Department of Medicine and Surgery, University of Salerno, Baronissi (SA), Italy
,
Giovanni Nassa
2   Laboratory of Molecular Medicine and Genomics, Department of Medicine and Surgery, University of Salerno, Baronissi (SA), Italy
,
Maria Rosaria De Filippo
2   Laboratory of Molecular Medicine and Genomics, Department of Medicine and Surgery, University of Salerno, Baronissi (SA), Italy
3   Department of Pathology, Memorial Sloan Kettering Cancer Center, New York, USA
,
Giorgio Giurato
2   Laboratory of Molecular Medicine and Genomics, Department of Medicine and Surgery, University of Salerno, Baronissi (SA), Italy
,
Maria Ravo
2   Laboratory of Molecular Medicine and Genomics, Department of Medicine and Surgery, University of Salerno, Baronissi (SA), Italy
,
Francesca Rizzo
2   Laboratory of Molecular Medicine and Genomics, Department of Medicine and Surgery, University of Salerno, Baronissi (SA), Italy
,
Stefano Conte
1   Department of Cardiothoracic and Respiratory Sciences, Second University of Naples, Naples, Italy
,
Grazia Pellegrino
4   Department of Advanced Biomedical Sciences, University of Naples Federico II, Naples, Italy
,
Plinio Cirillo
4   Department of Advanced Biomedical Sciences, University of Naples Federico II, Naples, Italy
,
Paolo Calabrò
1   Department of Cardiothoracic and Respiratory Sciences, Second University of Naples, Naples, Italy
,
Tiina Öhman
5   Institute of Biotechnology, University of Helsinki, Helsinki, Finland
,
Tuula A. Nyman
1   Department of Cardiothoracic and Respiratory Sciences, Second University of Naples, Naples, Italy
,
Alessandro Weisz
2   Laboratory of Molecular Medicine and Genomics, Department of Medicine and Surgery, University of Salerno, Baronissi (SA), Italy
6   Molecular Pathology and Medical Genomics Unit, ‘SS. Giovanni di Dio e Ruggi d’Aragona – Schola Medica Salernitana’ University Hospital, Salerno, Italy
,
Paolo Golino
1   Department of Cardiothoracic and Respiratory Sciences, Second University of Naples, Naples, Italy
› Institutsangaben
Weitere Informationen

Publikationsverlauf

Received: 03. September 2014

Accepted after major revision: 27. Januar 2015

Publikationsdatum:
22. November 2017 (online)

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Summary

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 the first time substantial evidence that fine-tuning of resident mRNA translation by miRNAs is a key event in platelet pathophysiology.

Keywords

Platelet activation - microRNA - RNA-Seq - quantitative proteomics - platelet system biology

* G. C. and R. T. equally contributed to this work.


 

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