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Increased expression of HIF2α during iron deficiency–associated megakaryocytic differentiation
Author(s) -
Jimenez K.,
Khare V.,
Evstatiev R.,
KulniggDabsch S.,
Jambrich M.,
Strobl H.,
Gasche C.
Publication year - 2015
Publication title -
journal of thrombosis and haemostasis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.947
H-Index - 178
eISSN - 1538-7836
pISSN - 1538-7933
DOI - 10.1111/jth.12884
Subject(s) - thrombopoietin , megakaryocyte , haematopoiesis , biology , microbiology and biotechnology , cancer research , chemistry , stem cell
Summary Background Iron deficiency is associated with reactive thrombocytosis; however, the mechanisms driving this phenomenon remain unclear. We previously demonstrated that this occurs alongside enhanced megakaryopoiesis in iron‐deficient rats, without alterations in the megakaryopoietic growth factors thrombopoietin, interleukin‐6, or interleukin‐11. Objectives The aim of this study was to evaluate megakaryocyte differentiation under iron deficiency in an in vitro model and to investigate potential genes involved in this process. Methods Human erythroleukemia and megakaryoblastic leukemia cell lines, as well as cord‐blood derived hematopoietic stem cells were cultured under iron deficiency. Cell morphology, ploidy, expression of CD 41, CD 61, and CD 42b, and proplatelet formation were assessed in iron‐deficient cultures. Polymerase chain reaction arrays were used to identify candidate genes that were verified using real‐time polymerase chain reaction. Hypoxia‐inducible factor 1, α subunit ( HIF 2α) protein expression was assessed in bone marrow sections from iron‐deficient rats and vascular endothelial growth factor ( VEGF)‐A in culture supernatants. Results and Conclusions Iron deficiency enhanced megakaryoid features in cell lines, increasing ploidy and initiating formation of proplatelet‐like structures. In cord blood cell cultures, iron deficiency increased the percentage of cells expressing megakaryopoietic markers and enhanced proplatelet formation. HIF 2α and VEGF were identified as potential pathways involved in this process. HIF 2α protein expression was increased in megakaryocytes from iron‐deficient rats, and VEGF‐A concentration was higher in iron‐deficient culture supernatants. Addition of VEGF‐A to cell cultures increased percentage expression of megakaryocyte CD 41. In conclusion, the data demonstrate that iron deficiency augments megakaryocytic differentiation and proplatelet formation and a potential role of HIF 2α in megakaryopoiesis.