Development of hematopoietic populations through the process of differentiation is critical for proper hematopoiesis. The transcription factor CCAAT/enhancer binding protein alpha (C/EBPα) is a master regulator of myeloid differentiation, and the identification of C/EBPα target genes is key to understand this process. Here we identified the Ecotropic Viral Integration Site 2B (EVI2B) gene as a direct target of C/EBPα. We showed that the product of the gene, the transmembrane glycoprotein EVI2B (CD361), is abundantly expressed on the surface of primary hematopoietic cells, the highest levels of expression being reached in mature granulocytes. Using shRNA-mediated downregulation of EVI2B in human and murine cell lines and in primary hematopoietic stem and progenitor cells, we demonstrated impaired myeloid lineage development and altered progenitor functions in EVI2B-silenced cells. We showed that the compromised progenitor functionality in Evi2b-depleted cells can be in part explained by deregulation of cell proliferation and apoptosis. In addition, we generated an Evi2b knockout murine model and demonstrated altered properties of hematopoietic progenitors, as well as impaired G-CSF dependent myeloid colony formation in the knockout cells. Remarkably, we found that EVI2B is significantly downregulated in human acute myeloid leukemia samples characterized by defects in CEBPA. Altogether, our data demonstrate that EVI2B is a downstream target of C/EBPα, which regulates myeloid differentiation and functionality of hematopoietic progenitors.

Cancer Science Institute National University of Singapore 14 Medical Drive Singapore 117599 Singapore

Centre d'Immunologie de Marseille Luminy Aix Marseille Université Inserm CNRS Marseille F 13288 France

Centre d'Immunophénomique Aix Marseille Université Inserm CNRS Marseille F 13288 France

Childhood Leukaemia Investigation Prague Department of Paediatric Haematology and Oncology 2nd Faculty of Medicine Charles University Prague University Hospital Motol 5 Uvalu 84 Praha 150 06 Czech Republic

Department Hematology Oncology Harvard Stem Cell Institute Harvard Medical School 3 Blackfan Circle Boston 02115 MA USA

Department of Hemato oncology Institute of Molecular Genetics of the ASCR Videnska 1083 Prague 142 20 Prague 4 Czech Republic

Department of Hematology Erasmus University Medical Center Wytemaweg 80 Rotterdam 3015 CN The Netherlands

Department of Leukocyte Signalling Institute of Molecular Genetics of the ASCR Videnska 1083 Prague 142 20 Czech Republic

Department of Molecular Immunology Institute of Molecular Genetics of the ASCR Videnska 1083 Prague 142 20 Czech Republic

Department of Molecular Neurobiology Institute of Physiology of the ASCR Videnska 1083 Prague 142 20 Czech Republic

Division of Hematology and Oncology Leipzig University Hospital Leipzig Germany

Cell Death Differ. 2019 Jan;26(1):198. doi: 10.1038/s41418-018-0146-z. PubMed

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