title: Development of a stable retrovirus vector capable of long-term expression of gamma-globin mRNA in mouse erythrocytes.
abstract: Gene therapy for patients with hemoglobin disorders such has been hampered by the inability of retrovirus vectors to transfer globin genes and the locus control region (LCR) into hematopoietic stem cells without rearrangement. In addition, the expression from intact globin gene vectors has been variable in red blood cells as a result of position effects and retrovirus silencing. We hypothesized that by substituting the globin gene promoter for the promoter of another gene expressed in red blood cells, we could generate stable retrovirus vectors that would express globin at sufficient levels to treat hemoglobinopathies. Transgenic mice containing the human ankyrin (Ank) gene promoter fused to the human gamma-globin gene showed position-independent, copy number-dependent expression of a linked gamma-globin mRNA. We generated a "double-copy" Ank/A gamma-globin retrovirus vector that transferred two copies of the Ank/A gamma-globin gene into target cells. Stable gene transfer was observed in primary primary mouse progenitor cells and long-term repopulating hematopoietic stem cells. Expression of Ank/A gamma-globin mRNA in mature red blood cells was approximately 8% of the level of mouse alpha-globin mRNA. We conclude that this novel retrovirus vector may be valuable for treating a variety of hemoglobinopathies by gene therapy if the level of expression can be further increased.
authors.affiliation: Hematopoiesis Section, Genetics and Molecular Biology Branch, National Human Genome Research Institute, National Institutes of Health, Building 49, Room 3A14 MSC 4442, Bethesda, Maryland 20892-4442, USA.
authors.forename: D E
authors.initials: DE
authors.lastname: Sabatino
authors.forename: N E
authors.initials: NE
authors.lastname: Seidel
authors.forename: A P
authors.initials: AP
authors.lastname: Cline
authors.forename: S M
authors.initials: SM
authors.lastname: Anderson
authors.forename: P G
authors.initials: PG
authors.lastname: Gallagher
authors.forename: D M
authors.initials: DM
authors.lastname: Bodine
mesh_terms: D016475:3T3 Cells; D000740:Anemia / Q000235:genetics / Q000628:therapy; D000818:Animals; D017487:Ankyrins / Q000235:genetics; D004912:Erythrocytes / Q000378:metabolism*; D005434:Flow Cytometry; D015870:Gene Expression; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics*; D018380:Hematopoietic Stem Cell Transplantation; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D011401:Promoter Regions, Genetic / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D012190:Retroviridae / Q000235:genetics*; D005719:gamma-Globulins / Q000235:genetics*
chemical_list: D017487:Ankyrins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D005719:gamma-Globulins