The Regulatory Roles of MicroRNA-146b-5p and Its Target Platelet-derived Growth Factor Receptor α (PDGFRA) in Erythropoiesis and Megakaryocytopoiesis

• Published in: The Journal of biological chemistry 2014-08, Vol.289 (33), p.22600-22613
• Main Authors: Zhai, Peng-Fei, Wang, Fang, Su, Rui, Lin, Hai-Shuang, Jiang, Chong-Liang, Yang, Gui-Hua, Yu, Jia, Zhang, Jun-Wu
• Format: Article
• Language: English
• Subjects: Animals; Basic Helix-Loop-Helix Transcription Factors - genetics; Basic Helix-Loop-Helix Transcription Factors - metabolism; Cell Biology; Cell Differentiation - physiology; Erythroid Cells - cytology; Erythroid Cells - metabolism; Erythropoiesis - physiology; GATA1 Transcription Factor - genetics; GATA1 Transcription Factor - metabolism; Hematopoietic Stem Cells - cytology; Hematopoietic Stem Cells - metabolism; Humans; K562 Cells; Megakaryocytes - cytology; Megakaryocytes - metabolism; Mice; MicroRNAs - genetics; MicroRNAs - metabolism; Promoter Regions, Genetic - physiology; Receptor, Platelet-Derived Growth Factor alpha - genetics; Receptor, Platelet-Derived Growth Factor alpha - metabolism; Signal Transduction - physiology; Thrombopoiesis - physiology; Up-Regulation - physiology
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M114.547380

Emerging evidence has shown that microRNAs have key roles in regulating various normal physiological processes, whereas their deregulated expression is correlated with various diseases. The miR-146 family includes miR-146a and miR-146b, with a distinct expression spectrum in different hematopoietic cells. Recent work indicated that miR-146a has a close relationship with inflammation and autoimmune diseases. miR-146-deficient mice have developed some abnormal hematopoietic phenotypes, suggesting the potential functions of miR-146 in hematopoietic development. In this study, we found that miR-146b was consistently up-regulated in both K562 and CD34+ hematopoietic stem/progenitor cells (HSPCs) undergoing either erythroid or megakaryocytic differentiation. Remarkably, erythroid and megakaryocytic maturation of K562 cells was induced by excess miR-146b but inhibited by decreased miR-146b levels. More importantly, an mRNA encoding receptor tyrosine kinase, namely platelet-derived growth factor receptor α (PDGFRA), was identified and validated as a direct target of miR-146b in hematopoietic cells. Gain-of-function and loss-of-function assays showed that PDGFRA functioned as a negative regulator in erythroid and megakaryocytic differentiation. miR-146b could ultimately affect the expression of the GATA-1 gene, which is regulated by HEY1 (Hairy/enhancer-of-split related with YRPW motif protein 1), a transcriptional repressor, via inhibition of the PDGFRA/JNK/JUN/HEY1 pathway. Lentivirus-mediated gene transfer also demonstrated that the overexpression of miR-146b promoted erythropoiesis and megakaryocytopoiesis of HSPCs via its regulation on the PDGFRA gene and effects on GATA-1 expression. Moreover, we confirmed that the binding of GATA-1 to the miR-146b promoter and induction of miR-146b during hematopoietic maturation were dependent on GATA-1. Therefore, miR-146b, PDGFRA, and GATA-1 formed a regulatory circuit to promote erythroid and megakaryocytic differentiation.