title: Evi1 is essential for hematopoietic stem cell self-renewal, and its expression marks hematopoietic cells with long-term multilineage repopulating activity.
abstract: Ecotropic viral integration site 1 (Evi1), a transcription factor of the SET/PR domain protein family, is essential for the maintenance of hematopoietic stem cells (HSCs) in mice and is overexpressed in several myeloid malignancies. Here, we generate reporter mice in which an internal ribosome entry site (IRES)-GFP cassette is knocked-in to the Evi1 locus. Using these mice, we find that Evi1 is predominantly expressed in long-term HSCs (LT-HSCs) in adult bone marrow, and in the hematopoietic stem/progenitor fraction in the aorta-gonad-mesonephros, placenta, and fetal liver of embryos. In both fetal and adult hematopoietic systems, Evi1 expression marks cells with long-term multilineage repopulating activity. When combined with conventional HSC surface markers, sorting according to Evi1 expression markedly enhances purification of cells with HSC activity. Evi1 heterozygosity leads to marked impairment of the self-renewal capacity of LT-HSCs, whereas overexpression of Evi1 suppresses differentiation and boosts self-renewal activity. Reintroduction of Evi1, but not Mds1-Evi1, rescues the HSC defects caused by Evi1 heterozygosity. Thus, in addition to documenting a specific relationship between Evi1 expression and HSC self-renewal activity, these findings highlight the utility of Evi1-IRES-GFP reporter mice for the identification and sorting of functional HSCs.
authors.affiliation: Department of Hematology and Oncology, Graduate School of Medicine, University of Tokyo, Bunkyo-ku, Tokyo, 113-8655, Japan.
authors.forename: Keisuke
authors.initials: K
authors.lastname: Kataoka
authors.forename: Tomohiko
authors.initials: T
authors.lastname: Sato
authors.forename: Akihide
authors.initials: A
authors.lastname: Yoshimi
authors.forename: Susumu
authors.initials: S
authors.lastname: Goyama
authors.forename: Takako
authors.initials: T
authors.lastname: Tsuruta
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kobayashi
authors.forename: Munetake
authors.initials: M
authors.lastname: Shimabe
authors.forename: Shunya
authors.initials: S
authors.lastname: Arai
authors.forename: Masahiro
authors.initials: M
authors.lastname: Nakagawa
authors.forename: Yoichi
authors.initials: Y
authors.lastname: Imai
authors.forename: Keiki
authors.initials: K
authors.lastname: Kumano
authors.forename: Katsuyoshi
authors.initials: K
authors.lastname: Kumagai
authors.forename: Naoto
authors.initials: N
authors.lastname: Kubota
authors.forename: Takashi
authors.initials: T
authors.lastname: Kadowaki
authors.forename: Mineo
authors.initials: M
authors.lastname: Kurokawa
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002454:Cell Differentiation / Q000502:physiology; D017931:DNA Primers / Q000235:genetics; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004622:Embryo, Mammalian / Q000378:metabolism*; D005260:Female; D005434:Flow Cytometry; D055879:Gene Knock-In Techniques; D005838:Genotype; D006410:Hematopoiesis / Q000502:physiology*; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism; D008099:Liver / Q000378:metabolism; D000074008:MDS1 and EVI1 Complex Locus Protein; D008650:Mesonephros / Q000378:metabolism; D051379:Mice; D010920:Placenta / Q000378:metabolism; D011247:Pregnancy; D011519:Proto-Oncogenes; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014157:Transcription Factors / Q000378:metabolism*
chemical_list: D017931:DNA Primers; D004268:DNA-Binding Proteins; D000074008:MDS1 and EVI1 Complex Locus Protein; C499305:Mecom protein, mouse; D014157:Transcription Factors
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