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Ferrata Storti Foundation
Haematologica 2019 Volume 104(6):1244-1255
Platelet Biology & Its Disorders
Downregulation of TREM-like transcript-1
and collagen receptor α2 subunit, two
novel RUNX1-targets, contributes to platelet dysfunction in familial platelet disorder with predisposition to acute myelogenous leukemia
Ana C. Glembotsky,1* Dominika Sliwa,2* Dominique Bluteau,2,3*
Nathalie Balayn,2 Cecilia P. Marin Oyarzún,1 Anna Raimbault,2 Marie Bordas,2 Nathalie Droin,2,4 Iryna Pirozhkova,5 Valance Washington,6 Nora P. Goette,1 Rosana F. Marta,1 Rémi Favier,2,7 Hana Raslova2** and Paula G. Heller1**
1Hematología Investigación, Instituto de Investigaciones Médicas “Dr. Alfredo Lanari”, Facultad de Medicina, Universidad de Buenos Aires, Consejo Nacional de Investigaciones Científicas y Tecnológicas (CONICET), Argentina; 2INSERM UMR 1170, Gustave Roussy, Université Paris-Saclay, Equipe Labellisée par la Ligue Nationale Contre le Cancer, Villejuif, France; 3Ecole Pratique des Hautes Etudes (EPHE), Paris, France; 4Gustave Roussy, Université Paris-Saclay, Genomic Platform UMS AMMICA, Villejuif, France; 5CNRS UMR 8126, Gustave Roussy, Université Paris-Saclay, Villejuif, France; 6Department of Biology, University of Puerto Rico-Rio Piedras, San Juan, Puerto Rico and 7Assistance Publique-Hôpitaux de Paris, Hôpital Trousseau, CRPP, Services d’Hématologie Biologique et Clinique, Paris, France
*ACG, DS and DB contributed equally to this work. **HR and PGH contributed equally to this work as senior co-authors.
ABSTRACT
Germline RUNX1 mutations lead to thrombocytopenia and platelet dysfunction in familial platelet disorder with predisposi- tion to acute myelogenous leukemia (AML). Multiple aspects of platelet function are impaired in these patients, associated with altered expression of genes regulated by RUNX1. We aimed to identify RUNX1- targets involved in platelet function by combining transcriptome analysis of patient and shRUNX1-transduced megakaryocytes (MK). Down-reg- ulated genes included TREM-like transcript (TLT)-1 (TREML1) and the integrin subunit alpha (α)-2 (ITGA2) of collagen receptor α2-beta (b)-1, which are involved in platelet aggregation and adhesion, respectively. RUNX1 binding to regions enriched for H3K27Ac marks was demon- strated for both genes using chromatin immunoprecipitation. Cloning of these regions upstream of the respective promoters in lentivirus allowing mCherry reporter expression showed that RUNX1 positively regulates TREML1 and ITGA2, and this regulation was abrogated after deletion of RUNX1 sites. TLT-1 content was reduced in patient MK and platelets. A blocking anti-TLT-1 antibody was able to block aggregation of normal but not patient platelets, whereas recombinant soluble TLT-1 potentiat- ed fibrinogen binding to patient platelets, pointing to a role for TLT-1 deficiency in the platelet function defect. Low levels of α2 integrin sub- unit were demonstrated in patient platelets and MK, coupled with reduced platelet and MK adhesion to collagen, both under static and flow conditions. In conclusion, we show that gene expression profiling of RUNX1 knock-down or mutated MK provides a suitable approach to identify novel RUNX1 targets, among which downregulation of TREML1 and ITGA2 clearly contribute to the platelet phenotype of familial platelet disorder with predisposition to AML.
Correspondence:
PAULA HELLER
paulaheller@hotmail.com
HANA RASLOVA
hana.raslova@gustaveroussy.fr
Received: January 20, 2018. Accepted: December 10, 2018. Pre-published: December 13, 2018.
doi:10.3324/haematol.2018.188904
Check the online version for the most updated information on this article, online supplements, and information on authorship & disclosures: www.haematologica.org/content/104/6/1244
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