title: Long-term transfer and expression of the human beta-globin gene in a mouse transplant model.
abstract: Somatic gene therapy of hemoglobinopathies depends initially on the demonstration of safe, efficient gene transfer and long-term, high-level expression of the transferred human beta-globin gene in animal models. We have used a beta-globin gene/beta-locus control region retroviral vector containing several modifications to optimize gene transfer and expression in a mouse transplant model. In this report we show that transplantation of beta-globin-transduced hematopoietic cells into lethally irradiated mice leads to the continued presence of the gene up to 8 months posttransplantation. The transferred human beta-globin gene is detected in 3 of 5 mice surviving long term (>4 months) transplanted with bone marrow cells transduced with high-titer virus. Southern blotting confirms the presence of the unrearranged 5.1-kb human beta-globin gene-containing provirus in 2 of these mice. In addition, long-term expression of the transferred gene is seen in 2 mice at levels of 5% and 20% that of endogenous murine beta-globin at 6 and 8 months posttransplantation. We further document stem cell transduction by the successful transfer and high-level expression of the human beta-globin gene from mice transduced 9 months earlier into irradiated secondary recipient mice. These results demonstrate high-level, long-term somatic human beta-globin gene transfer into the hematopoietic stem cells of an animal for the first time, and suggest the potential feasibility of a retroviral gene therapy approach to sickle cell disease and the beta thalassemias.
authors.affiliation: Columbia University, College of Physicians and Surgeons, Department of Genetics and Development, New York, New York, USA.
authors.forename: H
authors.initials: H
authors.lastname: Raftopoulos
authors.forename: M
authors.initials: M
authors.lastname: Ward
authors.forename: P
authors.initials: P
authors.lastname: Leboulch
authors.forename: A
authors.initials: A
authors.lastname: Bank
mesh_terms: D000818:Animals; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D005914:Globins / Q000235:genetics*; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000502:physiology*; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D012190:Retroviridae; D013997:Time Factors; D014184:Transplantation, Homologous
chemical_list: D005914:Globins