title: Correction of Sanfilippo A skin fibroblasts by retroviral vector-mediated gene transfer.
abstract: The recent cloning of the sulfamidase gene has made possible the consideration of gene-based therapies for Sanfilippo A syndrome (mucopolysaccharidosis type IIIA), one of the most common of the mucopolysaccharidoses. In this paper, we present the construction of a retroviral vector in which a sulfamidase cDNA is under the transcriptional control of the Moloney murine leukemia virus long terminal repeat. This construct was used to make a high-titer (4 x 10(5) colony-forming units/ml) producer cell line, PA317/LNSSN#19, in the amphotropic packaging cell line PA317. This producer cell line was shown to be helper virus free using an assay for horizontal spread of virus. Virus supernatant from PA317/LNSSN#19 was used to transduce Sanfilippo A fibroblasts, resulting in complete correction of both the enzymatic defect and the storage phenotype as assessed by intracellular accumulation of 35SO4(-)-labeled material. Phenotypic correction was seen even when the levels of viral transduction were low. These results show that gene therapy of the Sanfilippo A syndrome is practicable, although the nature of the disorder suggests that careful consideration needs to be given to the choice of the cellular target for gene transfer.
authors.affiliation: Department of Chemical Pathology, Women's and Children's Hospital, North Adelaide, South Australia.
authors.forename: J
authors.initials: J
authors.lastname: Bielicki
authors.forename: J J
authors.initials: JJ
authors.lastname: Hopwood
authors.forename: D S
authors.initials: DS
authors.lastname: Anson
mesh_terms: D016475:3T3 Cells; D000818:Animals; D002460:Cell Line; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism*; D022761:Gene Transfer, Horizontal; D005822:Genetic Vectors; D006025:Glycosaminoglycans / Q000378:metabolism; D006867:Hydrolases / Q000235:genetics*; D051379:Mice; D008979:Moloney murine leukemia virus / Q000235:genetics*; D009084:Mucopolysaccharidosis III / Q000628:therapy; D012867:Skin / Q000166:cytology; D014162:Transfection
chemical_list: D006025:Glycosaminoglycans; D006867:Hydrolases; C022708:N-sulfoglucosamine sulfohydrolase