title: EVI2B is a C/EBPα target gene required for granulocytic differentiation and functionality of hematopoietic progenitors.
abstract: 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.
authors.affiliation: Department of Hemato-oncology, Institute of Molecular Genetics of the ASCR, Videnska 1083, Prague 142 20 Prague 4, Czech Republic.|Childhood Leukaemia Investigation Prague, Department of Paediatric Haematology and Oncology, 2nd Faculty of Medicine, Charles University in Prague, University Hospital Motol, V Uvalu 84, Praha 150 06, Czech Republic.
authors.forename: Polina
authors.initials: P
authors.lastname: Zjablovskaja
authors.affiliation: Department of Hemato-oncology, Institute of Molecular Genetics of the ASCR, Videnska 1083, Prague 142 20 Prague 4, Czech Republic.
authors.forename: Miroslava
authors.initials: M
authors.lastname: Kardosova
authors.affiliation: Department of Hemato-oncology, Institute of Molecular Genetics of the ASCR, Videnska 1083, Prague 142 20 Prague 4, Czech Republic.
authors.forename: Petr
authors.initials: P
authors.lastname: Danek
authors.affiliation: Department of Molecular Immunology, Institute of Molecular Genetics of the ASCR, Videnska 1083, Prague 142 20, Czech Republic.
authors.forename: Pavla
authors.initials: P
authors.lastname: Angelisova
authors.affiliation: Cancer Science Institute, National University of Singapore, 14 Medical Drive, Singapore 117599, Singapore.
authors.forename: Touati
authors.initials: T
authors.lastname: Benoukraf
authors.affiliation: Division of Hematology and Oncology, Leipzig University Hospital, Leipzig, Germany.
authors.forename: Alexander A
authors.initials: AA
authors.lastname: Wurm
authors.affiliation: Childhood Leukaemia Investigation Prague, Department of Paediatric Haematology and Oncology, 2nd Faculty of Medicine, Charles University in Prague, University Hospital Motol, V Uvalu 84, Praha 150 06, Czech Republic.
authors.forename: Tomas
authors.initials: T
authors.lastname: Kalina
authors.affiliation: Cancer Science Institute, National University of Singapore, 14 Medical Drive, Singapore 117599, Singapore.
authors.forename: Stephanie
authors.initials: S
authors.lastname: Sian
authors.affiliation: Department of Molecular Neurobiology, Institute of Physiology of the ASCR, Videnska 1083, Prague 142 20, Czech Republic.
authors.forename: Martin
authors.initials: M
authors.lastname: Balastik
authors.affiliation: Department of Hematology, Erasmus University Medical Center, Wytemaweg 80, Rotterdam 3015 CN, The Netherlands.
authors.forename: Ruud
authors.initials: R
authors.lastname: Delwel
authors.affiliation: Department of Leukocyte Signalling, Institute of Molecular Genetics of the ASCR, Videnska 1083, Prague 142 20, Czech Republic.
authors.forename: Tomas
authors.initials: T
authors.lastname: Brdicka
authors.affiliation: Cancer Science Institute, National University of Singapore, 14 Medical Drive, Singapore 117599, Singapore.|Department Hematology/Oncology, Harvard Stem Cell Institute, Harvard Medical School, 3 Blackfan Circle, Boston 02115, MA, USA.
authors.forename: Daniel G
authors.initials: DG
authors.lastname: Tenen
authors.affiliation: Division of Hematology and Oncology, Leipzig University Hospital, Leipzig, Germany.
authors.forename: Gerhard
authors.initials: G
authors.lastname: Behre
authors.affiliation: Centre d'Immunophénomique, Aix Marseille Université, Inserm, CNRS, Marseille F-13288, France.
authors.forename: Fréderic
authors.initials: F
authors.lastname: Fiore
authors.affiliation: Centre d'Immunologie de Marseille-Luminy, Aix Marseille Université, Inserm, CNRS, Marseille F-13288, France.
authors.forename: Bernard
authors.initials: B
authors.lastname: Malissen
authors.affiliation: Department of Molecular Immunology, Institute of Molecular Genetics of the ASCR, Videnska 1083, Prague 142 20, Czech Republic.
authors.forename: Vaclav
authors.initials: V
authors.lastname: Horejsi
authors.affiliation: Department of Hemato-oncology, Institute of Molecular Genetics of the ASCR, Videnska 1083, Prague 142 20 Prague 4, Czech Republic.
authors.forename: Meritxell
authors.initials: M
authors.lastname: Alberich-Jorda
mesh_terms: D000818:Animals; D017209:Apoptosis; D001854:Bone Marrow Cells / Q000166:cytology; D022763:CCAAT-Enhancer-Binding Protein-alpha / Q000235:genetics / Q000378:metabolism*; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D015536:Down-Regulation / Q000187:drug effects; D004958:Estradiol / Q000494:pharmacology; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology; D006098:Granulocytes / Q000166:cytology / Q000378:metabolism; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000378:metabolism / Q000473:pathology*; D008562:Membrane Glycoproteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D008565:Membrane Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D011401:Promoter Regions, Genetic; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism
chemical_list: D022763:CCAAT-Enhancer-Binding Protein-alpha; C000626077:EVI2B protein, human; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D034741:RNA, Small Interfering; D016179:Granulocyte Colony-Stimulating Factor; D004958:Estradiol