Macrothrombocytopenia and dense granule deficiency associated with FLI1 variants: ultrastructural and pathogenic features.

Fiche publication


Date publication

juin 2017

Journal

Haematologica

Auteurs

Membres identifiés du Cancéropôle Est :
Dr GACHET Christian


Tous les auteurs :
Saultier P, Vidal L, Canault M, Bernot D, Falaise C, Pouymayou C, Bordet JC, Saut N, Rostan A, Baccini V, Peiretti F, Favier M, Lucca P, Deleuze JF, Olaso R, Boland A, Morange PE, Gachet C, Malergue F, Fauré S, Eckly A, Trégouët DA, Poggi M, Alessi MC

Résumé

Congenital macrothrombocytopenia is a family of rare diseases, of which a significant fraction remains to be genetically characterized. To analyze cases of unexplained thrombocytopenia, 27 individuals from a patient cohort of the Bleeding and Thrombosis Exploration Center of the University Hospital of Marseille were recruited for a high-throughput gene sequencing study. This strategy led to the identification of two novel variants (c.1010G>A and c.1033A>G) responsible for macrothrombocytopenia. The variant carriers' platelets exhibited a defect in aggregation induced by low-dose adenosine diphosphate (ADP), collagen and thrombin receptor-activating peptide (TRAP), a defect in adenosine triphosphate (ATP) secretion, a reduced mepacrine uptake and release and a reduced CD63 expression upon TRAP stimulation. Precise ultrastructural analysis of platelet content was performed using transmission electron microscopy and focused ion beam scanning electron microscopy. Remarkably, dense granules were nearly absent in the carriers' platelets, presumably due to a biogenesis defect. Additionally, 25-29% of the platelets displayed giant α-granules, while a smaller proportion displayed vacuoles (7-9%) and autophagosome-like structures (0-3%). study of megakaryocytes derived from circulating CD34 cells of the carriers revealed a maturation defect and reduced proplatelet formation potential. The study of the variants revealed a significant reduction in protein nuclear accumulation and transcriptional activity properties. Intraplatelet flow cytometry efficiently detected the biomarker MYH10 in variant carriers. Overall, this study provides new insights into the phenotype, pathophysiology and diagnosis of variant-associated thrombocytopenia.

Mots clés

Adult, Blood Platelets, pathology, Cell Nucleus, chemistry, Cytoplasmic Granules, metabolism, Genetic Variation, Humans, Male, Megakaryocytes, pathology, Middle Aged, Platelet Aggregation, genetics, Proto-Oncogene Protein c-fli-1, genetics, Thrombocytopenia, congenital, Transcription, Genetic, Young Adult

Référence

Haematologica. 2017 06;102(6):1006-1016