title: A human beta-globin gene fused to the human beta-globin locus control region is expressed at high levels in erythroid cells of mice engrafted with retrovirus-transduced hematopoietic stem cells.
abstract: Retroviral-mediated gene transfer of human beta-globin provides a model system for the development of somatic gene therapy for hemoglobinopathies. Previous work has shown that mice receiving a transplant of bone marrow cells infected with a retroviral vector containing the human beta-globin gene can express human beta-globin specifically in erythroid cells; however, the level of expression of the transduced globin gene was low (1% to 2% per gene copy as compared with that of the endogenous mouse beta-globin gene). We report here the construction of a recombinant retrovirus vector encoding a human beta-globin gene fused to the 4 major regulatory elements of the human beta-globin locus control region (LCR). The LCR cassette increases the level of expression of the globin gene in murine erythroleukemia cells by 10-fold. To study the level of expression in vivo, mouse bone marrow cells were infected with virus-producing cells and the transduced cells were injected into lethally irradiated recipients. In the majority of provirus-containing mice (up to 75%), expression of human beta-globin in peripheral blood was detected at least 3 to 6 months after transplantation. Twelve animals representative of the level of expression of the transduced gene in blood (0.04% to 3.2% of the endogenous mouse beta-globin RNA) were selected for further analysis. A range of 0.4% to 12% of circulating erythrocytes stained positive for human beta-globin protein. Based on these values, the level of expression of the transduced gene per cell was estimated to be 10% to 39% of the endogenous mouse beta-globin gene. These data demonstrate that fusion of the LCR to the beta-globin gene in a retroviral vector increases the level of beta-globin expression in murine erythroleukemia cells and suggest that high-level expression can be obtained in erythroid cells in vivo after transduction into hematopoietic stem cells.
authors.affiliation: Laboratoire d'Oncologie Virale UPR 274, Centre National de la Recherche Scientifique, Villejuif, France.
authors.forename: I
authors.initials: I
authors.lastname: Plavec
authors.forename: T
authors.initials: T
authors.lastname: Papayannopoulou
authors.forename: C
authors.initials: C
authors.lastname: Maury
authors.forename: F
authors.initials: F
authors.lastname: Meyer
mesh_terms: D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D015672:Erythroid Precursor Cells / Q000378:metabolism*; D015870:Gene Expression; D005809:Genes, Regulator; D005822:Genetic Vectors; D005914:Globins / Q000235:genetics*; D006801:Humans; D004915:Leukemia, Erythroblastic, Acute / Q000235:genetics*; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D008979:Moloney murine leukemia virus / Q000235:genetics*; D012091:Repetitive Sequences, Nucleic Acid; D014162:Transfection; D014407:Tumor Cells, Cultured
chemical_list: D005914:Globins