title: Activating stimuli induce platelet microRNA modulation and proteome reorganisation.
abstract: 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.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Cimmino
authors.forename: Roberta
authors.initials: R
authors.lastname: Tarallo
authors.forename: Giovanni
authors.initials: G
authors.lastname: Nassa
authors.forename: Maria Rosaria
authors.initials: MR
authors.lastname: De Filippo
authors.forename: Giorgio
authors.initials: G
authors.lastname: Giurato
authors.forename: Maria
authors.initials: M
authors.lastname: Ravo
authors.forename: Francesca
authors.initials: F
authors.lastname: Rizzo
authors.forename: Stefano
authors.initials: S
authors.lastname: Conte
authors.forename: Grazia
authors.initials: G
authors.lastname: Pellegrino
authors.forename: Plinio
authors.initials: P
authors.lastname: Cirillo
authors.forename: Paolo
authors.initials: P
authors.lastname: Calabro
authors.forename: Tiina
authors.initials: T
authors.lastname: Öhman
authors.forename: Tuula A
authors.initials: TA
authors.lastname: Nyman
authors.affiliation: Prof. Alessandro Weisz, MD, Laboratory of Molecular Medicine and Genomics, University of Salerno, via S. Allende, 1, 84081 Baronissi (SA), Italy, Tel.: +39 089 965043, Fax: +39 089 969657, E-mail: aweisz@unisa.it.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Weisz
authors.affiliation: Prof. Paolo Golino, MD, Department of Cardiothoracic and Respiratory Sciences, Second University of Naples, Via L. Bianchi, 1, 80131 Naples, Italy, Tel.: +39 0823 306395, Fax: +39 0823 232395, E-mail: paolo.golino@unina2.it.
authors.forename: Paolo
authors.initials: P
authors.lastname: Golino
mesh_terms: D000244:Adenosine Diphosphate / Q000494:pharmacology*; D001792:Blood Platelets / Q000187:drug effects* / Q000378:metabolism; D003094:Collagen / Q000494:pharmacology*; D020869:Gene Expression Profiling / Q000379:methods; D005786:Gene Expression Regulation; D053263:Gene Regulatory Networks; D064368:Healthy Volunteers; D006801:Humans; D008297:Male; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D010455:Peptides / Q000494:pharmacology*; D015539:Platelet Activation / Q000187:drug effects*; D060066:Protein Interaction Maps; D020543:Proteome; D040901:Proteomics / Q000379:methods; D015398:Signal Transduction / Q000187:drug effects; D049490:Systems Biology; D013997:Time Factors; D059467:Transcriptome
chemical_list: D035683:MicroRNAs; D010455:Peptides; D020543:Proteome; D000244:Adenosine Diphosphate; D003094:Collagen
keywords: Platelet activation; RNA-Seq; microRNA; platelet system biology; quantitative proteomics