title: Regulated high-level human beta-globin gene expression in erythroid cells following recombinant adeno-associated virus-mediated gene transfer.
abstract: Gene therapy approaches for beta-thalassemia and sickle cell anemia focus on the transfer of a human beta-globin gene into the patient's hematopoietic stem cells (HSC). Expression of the transferred sequences should be erythroid specific and match the expression of the endogenous alpha-globin genes in adult erythropoiesis. Here we explore the potential of recombinant adeno-associated virus (AAV) vectors for human beta-globin gene transfer. We have constructed a recombinant AAV-vector containing a human beta-globin gene together with the DNasel hypersensitive sites 4, 3 and 2 of the human beta-globin locus control region. The vector replicates to high titers and can efficiently transduce hematopoietic and non-hematopoietic cells. In transduced and G418 selected murine erythroleukemia (MEL) cell clones, human beta-globin gene expression was regulated and reached levels comparable to endogenous murine beta maj. These data show that AAV-vectors are promising tools in gene therapy approaches for the haemoglobinopathies.
authors.affiliation: Department of Medical Biochemistry, University of Leiden, Rijswijk, The Netherlands.
authors.forename: M P
authors.initials: MP
authors.lastname: Einerhand
authors.forename: M
authors.initials: M
authors.lastname: Antoniou
authors.forename: S
authors.initials: S
authors.lastname: Zolotukhin
authors.forename: N
authors.initials: N
authors.lastname: Muzyczka
authors.forename: K I
authors.initials: KI
authors.lastname: Berns
authors.forename: F
authors.initials: F
authors.lastname: Grosveld
authors.forename: D
authors.initials: D
authors.lastname: Valerio
mesh_terms: D000818:Animals; D002461:Cell Line, Transformed; D002999:Clone Cells; D003001:Cloning, Molecular; D000229:Dependovirus / Q000235:genetics*; D015672:Erythroid Precursor Cells / Q000378:metabolism*; D015967:Gene Expression Regulation, Viral; D018014:Gene Transfer Techniques; D005914:Globins / Q000235:genetics*; D006801:Humans; D004915:Leukemia, Erythroblastic, Acute / Q000235:genetics / Q000473:pathology; D051379:Mice; D011995:Recombination, Genetic; D014161:Transduction, Genetic
chemical_list: D005914:Globins