Systematic analysis of microRNA fingerprints in thrombocythemic platelets using integrated platforms

• Published in: Blood Vol. 120; no. 17; pp. 3575 - 3585
• Main Authors: Xu, Xiao, Gnatenko, Dmitri V., Ju, Jingfang, Hitchcock, Ian S., Martin, Dwight W., Zhu, Wei, Bahou, Wadie F.
• Format: Journal Article
• Language: English
• Published: Washington, DC Elsevier Inc 25.10.2012; Americain Society of Hematology; American Society of Hematology
• Subjects: Adaptor Proteins, Signal Transducing - genetics; Adaptor Proteins, Signal Transducing - metabolism; Binding Sites; Biological and medical sciences; Blood Platelets - metabolism; Blood Platelets - pathology; Cell Differentiation; Cell Lineage - genetics; Cells, Cultured; Female; Gene Expression Profiling; Gene Expression Regulation, Developmental; Genes, Reporter; Hematologic and hematopoietic diseases; Humans; Lentivirus; Luciferases; Male; Mass Spectrometry; Medical sciences; Megakaryocytes - metabolism; Megakaryocytes - pathology; Microfilament Proteins; MicroRNAs - genetics; MicroRNAs - metabolism; Oligonucleotide Array Sequence Analysis; Platelets and Thrombopoiesis; Protein Binding; Proteomics; rho GTP-Binding Proteins; Thrombocythemia, Essential - genetics; Thrombocythemia, Essential - metabolism; Thrombocythemia, Essential - pathology; Thrombopoiesis - genetics; Gene silencing; Platelet; RNA interference; Hematology; Fingerprint method; Micro RNA
• ISSN: 0006-4971; 1528-0020; 1528-0020
• DOI: 10.1182/blood-2012-02-411264

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.