Thromb Haemost 2019; 119(11): 1795-1806
DOI: 10.1055/s-0039-1695009
Cellular Haemostasis and Platelets
Georg Thieme Verlag KG Stuttgart · New York

Transcriptome Analysis of Reticulated Platelets Reveals a Prothrombotic Profile

Dario Bongiovanni*
1  Medical Department 1, Cardiology, Klinikum rechts der Isar, Technische Universität, Munich, Germany
2  DZHK (German Center for Cardiovascular Research), Partner site Munich Heart Alliance, Munich, Germany
,
1  Medical Department 1, Cardiology, Klinikum rechts der Isar, Technische Universität, Munich, Germany
,
Melissa Klug
1  Medical Department 1, Cardiology, Klinikum rechts der Isar, Technische Universität, Munich, Germany
,
Donato Santovito
3  Institute for Cardiovascular Prevention (IPEK), Ludwig-Maximilians-University Munich, Germany
,
Arianna Felicetta
4  Department of Cardiovascular Medicine, Humanitas Research Hospital, Humanitas University, Milan, Italy
,
Michael Hristov
3  Institute for Cardiovascular Prevention (IPEK), Ludwig-Maximilians-University Munich, Germany
,
Moritz von Scheidt
2  DZHK (German Center for Cardiovascular Research), Partner site Munich Heart Alliance, Munich, Germany
5  Department of Cardiology, Deutsches Herzzentrum München, Technische Universität, Munich, Germany
,
Maria Aslani
3  Institute for Cardiovascular Prevention (IPEK), Ludwig-Maximilians-University Munich, Germany
,
Javier Cibella
6  Genomic Unit, Humanitas Clinical and Research Center, Rozzano, Milan, Italy
,
Christian Weber
2  DZHK (German Center for Cardiovascular Research), Partner site Munich Heart Alliance, Munich, Germany
3  Institute for Cardiovascular Prevention (IPEK), Ludwig-Maximilians-University Munich, Germany
,
Alessandra Moretti
1  Medical Department 1, Cardiology, Klinikum rechts der Isar, Technische Universität, Munich, Germany
2  DZHK (German Center for Cardiovascular Research), Partner site Munich Heart Alliance, Munich, Germany
,
Karl-Ludwig Laugwitz
1  Medical Department 1, Cardiology, Klinikum rechts der Isar, Technische Universität, Munich, Germany
2  DZHK (German Center for Cardiovascular Research), Partner site Munich Heart Alliance, Munich, Germany
,
Clelia Peano**
6  Genomic Unit, Humanitas Clinical and Research Center, Rozzano, Milan, Italy
7  Institute of Genetic and Biomedical Research, National Research Council, UoS Milan, Italy
,
Isabell Bernlochner**
1  Medical Department 1, Cardiology, Klinikum rechts der Isar, Technische Universität, Munich, Germany
2  DZHK (German Center for Cardiovascular Research), Partner site Munich Heart Alliance, Munich, Germany
› Author Affiliations
Funding The financial support from the German society of cardiology (grant DGK, Forschungsstipendium, DGK102018 to D.B.) and the European society of cardiology (ESC first contact initiative grant 2018 to D.B.) is gratefully acknowledged.
Further Information

Publication History

15 April 2019

04 July 2019

Publication Date:
01 September 2019 (online)

Abstract

Reticulated platelets (RPs) are larger, hyperreactive platelets that contain significantly more ribonucleic acid (RNA) compared with mature platelets (MPs). High levels of RPs in peripheral blood are predictors of an insufficient response to dual antiplatelet therapy in cardiovascular patients and of adverse cardiovascular events. However, the mechanisms underlying these correlations remain widely unknown and the biology of RPs has not been investigated yet. Here, we compared for the first time the transcriptomic profiles of RPs and MPs isolated from peripheral blood of healthy donors. Total RNA sequencing revealed 1,744 differentially expressed genes (670 downregulated, 1,074 upregulated) in RPs compared with MPs. In particular, transcripts for the collagen receptor GP6, thromboxane receptor A2 (TBXA2R), thrombin receptor PAR4 (F2RL3), and adenosine triphosphate receptors P2RX1, ORAI2, and STIM1 (both involved in calcium signaling) were significantly upregulated in RPs, whereas several RNA regulators as the ribonuclease PARN, the RISC-component TNRC6A, and the splicing factor LUC7L3 were downregulated in RPs. Gene ontology analysis revealed an enrichment of relevant biological categories in RPs including platelet activation and blood coagulation. Gene Set Enrichment Analysis showed an overrepresentation of several platelet activation pathways like thrombin, thromboxane, and glycoprotein IIb/IIIa signaling in RPs. Small-RNA sequencing reported 9 micro-RNAs significantly downregulated in RPs with targets involved in platelet reactivity. Our data show for the first time an enrichment of several prothrombotic transcripts in RPs providing a first biological explanation for their hyperreactive phenotype.

* These authors contributed equally.


** These are co-last authors.


Authors' Contributions

D.B. and I.B. designed the research and wrote the paper. D.B., M.K., M.H., D.S., and M.S. performed the experiments. A.F. performed the qPCRs. J.C. and C.P. prepared the library and performed the RNA-sequencing. G.S., D.S., M.A., M.K., and D.B. analyzed the results and made the figures. C.W., D.S., A.M., K.L.L., and I.B. interpreted the results and reviewed the paper.


Supplementary Material