title: Systematic analysis of microRNA fingerprints in thrombocythemic platelets using integrated platforms.
abstract: Posttranscriptional and translational controls mediated by microRNAs (miRNA) regulate diverse biologic processes. We dissected regulatory effects of miRNAs relevant to megakaryocytopoiesis and platelet biology by analyzing expression patterns in 79 subjects with thrombocytosis and controls, and integrated data with transcriptomic and proteomic platforms. We validated a unique 21-miRNA genetic fingerprint associated with thrombocytosis, and demonstrated that a 3-member subset defines essential thrombocythemia (ET). The genetic signature includes functional guide and passenger strands of the previously uncharacterized miR 490 (5p and 3p), which displayed restricted, low-level expression in megakaryocytes/platelets (compared with leukocytes), and aberrant expression during thrombocytosis, most profound in ET. Overexpression of miR 490 in a bilineage differentiation model of megakaryocyte/erythroid progenitor formation was insufficient for hematopoietic colony differentiation and/or lineage specification. Integration of transcriptomic and mass spectrometric datasets with functional reporter assays identified dishevelled associated activator of morphogenesis 1 (DAAM1) as a miR 490 5p protein target demonstrating decreased expression in ET platelets, putatively by translational control (and not by mRNA target degradation). Our data define a dysregulated miRNA fingerprint in thrombocytosis and support a developmentally restricted function of miR 490 (and its putative DAAM1 target) to conditions associated with exaggerated megakaryocytopoiesis and/or proplatelet formation.
authors.affiliation: Departments of Medicine, Stony Brook University, Stony Brook, NY, USA.
authors.forename: Xiao
authors.initials: X
authors.lastname: Xu
authors.forename: Dmitri V
authors.initials: DV
authors.lastname: Gnatenko
authors.forename: Jingfang
authors.initials: J
authors.lastname: Ju
authors.forename: Ian S
authors.initials: IS
authors.lastname: Hitchcock
authors.forename: Dwight W
authors.initials: DW
authors.lastname: Martin
authors.forename: Wei
authors.initials: W
authors.lastname: Zhu
authors.forename: Wadie F
authors.initials: WF
authors.lastname: Bahou
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics* / Q000378:metabolism; D001665:Binding Sites; D001792:Blood Platelets / Q000378:metabolism* / Q000473:pathology; D002454:Cell Differentiation; D019070:Cell Lineage / Q000235:genetics; D002478:Cells, Cultured; D005260:Female; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D017930:Genes, Reporter; D006801:Humans; D016086:Lentivirus; D008156:Luciferases; D008297:Male; D013058:Mass Spectrometry; D008533:Megakaryocytes / Q000378:metabolism* / Q000473:pathology; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D008840:Microfilament Proteins; D020411:Oligonucleotide Array Sequence Analysis; D011485:Protein Binding; D040901:Proteomics; D013920:Thrombocythemia, Essential / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D034061:Thrombopoiesis / Q000235:genetics; D020741:rho GTP-Binding Proteins
chemical_list: D048868:Adaptor Proteins, Signal Transducing; C499094:DAAM1 protein, human; C574862:MIRN490 microRNA, human; D035683:MicroRNAs; D008840:Microfilament Proteins; D008156:Luciferases; D020741:rho GTP-Binding Proteins