MicroRNA-126 is a regulator of platelet-supported thrombin generation

Abstract

Circulating microRNA (miRNA) expression profiles correlate with platelet reactivity. MiR-126 is a promising candidates in this regard. We generated a transgenic zebrafish line with thrombocyte-specific overexpression of miR-126. Laser injury of the posterior cardinal vein of 5 day-old larvae was performed with or without antithrombotic pre-treatment. Platelet-like structures (PLS) derived from human megakaryocytes transfected with miR-126 were also evaluated for procoagulant activity. Finally, we studied the correlation between miR-126 level and thrombin generation markers in a cohort of stable cardiovascular patients. Control zebrafish developed small thrombocyte-rich thrombi at the site of vessel injury, without vessel occlusion. The miR-126 transgenic line developed an occluding thrombus in 75% (95% CI: 51–91%) of larvae. Pre-treatment with the direct thrombin inhibitor argatroban, but not aspirin, prevented vessel occlusion in the transgenic line (0% occlusion, 95%CI: 0–18%). Upon activation, human PLS showed an increased procoagulant profile after miR-126 transfection compared to control. Finally, the plasma levels of miR-126, but not a control platelet-derived miRNA, correlated with markers of in vivo thrombin generation in a cohort of 185 cardiovascular patients. Our results from three complementary approaches support a key role for miR-126 in platelet-supported thrombin generation and open new avenues in the tailoring of antithrombotic treatment.

Keywords:

• Platelet
• platelet reactivity
• thrombin
• thrombosis
• zebrafish

Acknowledgements

The authors wish to thank Catherine Strassel and Christian Gachet for sharing protocols regarding the generation of CD34⁺-derived megakaryocytes. The authors are grateful to Séverine Nolli for excellent technical assistance. This work was supported by the Swiss National Science Foundation under grant 310030_162567; the Novartis Foundation under grant 15B097; the Swiss Heart Foundation and the Private Foundation of the University Hospitals of Geneva under grant RC04-05.

Authorship Contributions

V.Z., R.J.F, J.-L.R and M.N-A. designed the study and analysed the data. V.Z, A.G., S.D-G performed the experiments and analysed the data. All authors revised the intellectual content of the manuscript and approved the final version.

Declaration of interest statement

P. Fontana reports a travel grant from Sobi, outside the submitted work. J.-L. Reny reports non-financial support from Bayer, unrestricted research grant from Daichii-Sankyo, outside the submitted work. R. J. Fish reports travel support from Novo Nordisk, outside the submitted work. The other authors have no conflict of interest to report.

Supplementary material

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