Transcriptomic and phospho-proteomic analyzes of erythroblasts expanded in vitro from normal donors and from patients with polycythemia vera Journal Article
| Authors: | Hricik, T.; Federici, G.; Zeuner, A.; Alimena, G.; Tafuri, A.; Tirelli, V.; Varricchio, L.; Masiello, F.; Ciaffoni, F.; Vaglio, S.; Petricoin, E. F.; Girelli, G.; Levine, R. L.; Migliaccio, A. R. F. |
| Article Title: | Transcriptomic and phospho-proteomic analyzes of erythroblasts expanded in vitro from normal donors and from patients with polycythemia vera |
| Abstract: | Erythropoiesis is a tightly regulated process which becomes decoupled from its normal differentiation program in patients with polycythemia vera (PV). Somatic mutations in JAK2 are commonly associated with this myeloid proliferative disorder. To gain insight into the molecular events that are required for abnormally developing erythroid cells to escape dependence on normal growth signals, we performed in vitro expansion of mature erythroblasts (ERY) from seven normal healthy donors and from seven polycythemic patients in the presence of IL3, EPO, SCF for 10, 11, or 13 days. Normal ERYs required exposure to the glucocorticoid dexamethasone (Dex) for expansion, while PV-derived ERYs expanded in the absence of dexamethasone. RNA expression profiling revealed enrichment of two known oncogenes, GPR56 and RAB4a, in PV-derived ERYs along with reduced expression levels of transcription factor TAL1 (ANOVA FDR < 0.05). While both normal and polycythemic-derived ERYs integrated signaling cascades for growth, they did so via different signaling pathways which are represented by their differential phospho-profiles. Our results show that normal ERYs displayed greater levels of phosphorylation of EGFR, PDGFRβ, TGFβ, and cKit, while PV-derived ERYs were characterized by increased phosphorylation of cytoplasmic kinases in the JAK/STAT, PI3K, and GATA1 pathways. Together these data suggest that PV erythroblast expansion and maturation may be maintained and enriched in the absence of dexamethasone through reduced TAL1 expression and by accessing additional signaling cascades. Members of this acquired repertoire may provide important insight into the pathogenesis of aberrant erythropoiesis in myeloproliferative neoplasms such as polycythemia vera. © 2013 Wiley Periodicals, Inc. |
| Keywords: | signal transduction; adult; clinical article; controlled study; human tissue; aged; middle aged; human cell; somatic mutation; case-control studies; proto-oncogene proteins; janus kinase 2; cells, cultured; stem cell factor; gene expression profiling; erythropoietin; cell maturation; erythroblast; erythropoiesis; basic helix-loop-helix transcription factors; dexamethasone; in vitro study; phosphorylation; proteomics; transcriptomics; phosphatidylinositol 3 kinase; rna; gene expression regulation; erythroid cell; phosphoproteins; erythroblasts; receptors, g-protein-coupled; normal human; polycythemia vera; transcription factor gata 1; intracellular signaling; stat protein; stem cell expansion; lymphotoxin; blood donor; interleukin 3; interleukin-3; transcription factor tal1; enrichment culture; escape behavior; rab4 gtp-binding proteins |
| Journal Title: | American Journal of Hematology |
| Volume: | 88 |
| Issue: | 9 |
| ISSN: | 0361-8609 |
| Publisher: | John Wiley & Sons, Inc. |
| Date Published: | 2013-01-01 |
| Start Page: | 723 |
| End Page: | 729 |
| Language: | English |
| DOI: | 10.1002/ajh.23487 |
| PROVIDER: | scopus |
| PMCID: | PMC3771389 |
| PUBMED: | 23720412 |
| DOI/URL: | |
| Notes: | --- - "Export Date: 1 October 2013" - "CODEN: AJHED" - "Source: Scopus" |
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