title: {gamma}-Globin gene expression in chemical inducer of dimerization (CID)-dependent multipotential cells established from human {beta}-globin locus yeast artificial chromosome ({beta}-YAC) transgenic mice.
abstract: Identification of trans-acting factors or drugs capable of reactivating gamma-globin gene expression is complicated by the lack of suitable cell lines. Human K562 cells co-express epsilon- and gamma-globin but not beta-globin; transgenic mouse erythroleukemia 585 cells express predominantly human beta-globin but also gamma-globin; and transgenic murine GM979 cells co-express human gamma-and beta-globin. Human beta-globin locus yeast artificial chromosome transgenic mice display correct developmental regulation of beta-like globin gene expression. We rationalized that cells established from the adult bone marrow of these mice might express exclusively beta-globin and therefore could be employed to select or screen inducers of gamma-globin expression. A thrombopoietin receptor derivative that brings the proliferative status of primary mouse bone marrow cells under control of a chemical inducer of dimerization was employed to institute and maintain these cell populations. Human beta-globin was expressed, but gamma-globin was not; a similar expression pattern was observed in cells derived from fetal liver. gamma-Globin expression was induced upon exposure to 5-azacytidine, in cells derived from -117 Greek hereditary persistence of fetal hemoglobin human beta-globin locus yeast artificial chromosome (beta-YAC) mice, showing that the hereditary persistence of fetal hemoglobin (HPFH) phenotype was maintained in these cells or was reactivated by an artificial zinc finger-gamma-globin transcription factor and the previously identified fetal globin transactivators fetal Krüppel-like factor (FKLF) and fetal globin-increasing factor (FGIF). These cells may be useful for identifying transcription factors that reactivate gamma-globin synthesis or screening gamma-globin inducers for the treatment of sickle cell disease or beta-thalassemia.
authors.affiliation: Division of Hematology, Department of Medicine, University of Washington Medical Center, Seattle, Washington 98195, USA.
authors.forename: C Anthony
authors.initials: CA
authors.lastname: Blau
authors.forename: Carlos F
authors.initials: CF
authors.lastname: Barbas
authors.forename: Anna L
authors.initials: AL
authors.lastname: Bomhoff
authors.forename: Renee
authors.initials: R
authors.lastname: Neades
authors.forename: James
authors.initials: J
authors.lastname: Yan
authors.forename: Patrick A
authors.initials: PA
authors.lastname: Navas
authors.forename: Kenneth R
authors.initials: KR
authors.lastname: Peterson
mesh_terms: D000818:Animals; D001374:Azacitidine / Q000494:pharmacology*; D001854:Bone Marrow Cells / Q000378:metabolism; D018244:Chromosomes, Artificial, Yeast; D019281:Dimerization; D005319:Fetal Hemoglobin / Q000235:genetics*; D018507:Gene Expression Regulation, Developmental; D005914:Globins / Q000235:genetics*; D006801:Humans; D008099:Liver; D051379:Mice; D008822:Mice, Transgenic; D039902:Multipotent Stem Cells / Q000378:metabolism*; D010641:Phenotype
chemical_list: D005914:Globins; D005319:Fetal Hemoglobin; D001374:Azacitidine