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Thromb Haemost 2015; 114(06): 1218-1229
DOI: 10.1160/TH14-10-0872
Blood Cells, Inflammation and Infection
Schattauer GmbH

Abnormal megakaryopoiesis and platelet function in cyclooxygenase-2-deficient mice

Silvia S. Barbieri
1   Centro Cardiologico Monzino, IRCCS, Milan, Italy
,
Giovanna Petrucci   *
2   Institute of Pharmacology, Catholic University School of Medicine, Rome, Italy
,
Eva Tarantino   *
3   Department of Pharmacological and Biomolecular Sciences, University of Milan, Milan, Italy
,
Patrizia Amadio
1   Centro Cardiologico Monzino, IRCCS, Milan, Italy
,
Bianca Rocca
2   Institute of Pharmacology, Catholic University School of Medicine, Rome, Italy
,
Maurizio Pesce
1   Centro Cardiologico Monzino, IRCCS, Milan, Italy
,
Kellie R. Machlus
4   Hematology Division, Department of Medicine, Brigham and Women’s Hospital, Boston, Massachusetts, USA
5   Harvard Medical School, Boston, Massachusetts, USA
,
Franco O. Ranelletti
6   Department of Histology, Catholic University School of Medicine, Rome, Italy
,
Sara Gianellini
1   Centro Cardiologico Monzino, IRCCS, Milan, Italy
,
Babette B. Weksler
7   Division of Hematology-Medical Oncology, Weill Cornell Medical College, New York, New York, USA
,
Joseph E. Italiano Jr.
4   Hematology Division, Department of Medicine, Brigham and Women’s Hospital, Boston, Massachusetts, USA
5   Harvard Medical School, Boston, Massachusetts, USA
8   Vascular Biology Program, Department of Surgery, Children’s Hospital, Boston, Massachusetts, USA
,
Elena Tremoli
1   Centro Cardiologico Monzino, IRCCS, Milan, Italy
3   Department of Pharmacological and Biomolecular Sciences, University of Milan, Milan, Italy
› Author Affiliations Financial support: This work was supported by the Italian Ministry of Health (Ricerca Corrente BIO52–2011; BIO55–2011; BIO36–2012; BIO39–2012; BIO32–2013).
Further Information

Publication History

Received: 22 October 2014

Accepted after major revision: 29 June 2015

Publication Date:
30 November 2017 (online)

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Summary

Previous studies suggest that cyclooxygenase-2 (COX-2) might influence megakaryocyte (MK) maturation and platelet production in vitro. Using a gene deletion model, we analysed the effect of COX-2 deficiency on megakaryopoiesis and platelet function. COX-2-/- mice (10–12 weeks old) have hyper-responsive platelets as suggested by their enhanced aggregation, TXA2 biosynthesis, CD62P and CD41/CD61 expression, platelet-fibrinogen binding, and increased thromboembolic death after collagen/epinephrine injection compared to wild-type (WT). Moreover, increased platelet COX-1 expression and reticulated platelet fraction were observed in COX-2-/- mice while platelet count was similar to WT. MKs were significantly reduced in COX-2-/- bone marrows (BMs), with high nuclear/cytoplasmic ratios, low ploidy and poor expression of lineage markers of maturation (CD42d, CD49b). However, MKs were significantly increased in COX-2-/- spleens, with features of MK maturation markers which were not observed in MKs of WT spleens. Interestingly, the expression of COX-1, prostacyclin and PGE2 synthases and prostanoid pattern were modified in BMs and spleens of COX-2-/- mice. Moreover, COX-2 ablation reduced the percentage of CD49b+ cells, the platelet formation and the haematopoietic stem cells in bone marrow and increased their accumulation in the spleen. Splenectomy decreased peripheral platelet number, reverted their hyper-responsive phenotype and protected COX-2-/- mice from thromboembolism. Interestingly, fibrosis was observed in spleens of old COX-2-/- mice (28 weeks old). In conclusion, COX-2 deletion delays BM megakaryopoiesis promoting a compensatory splenic MK hyperplasia, with a release of hyper-responsive platelets and increased thrombogenicity in vivo. COX-2 seems to contribute to physiological MK maturation and pro-platelet formation.

Keywords

COX-2 - platelets - thrombosis - megakaryocytes

* The authors contributed equally to this article.


 

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