title: In vitro gene therapy of mucopolysaccharidosis type I by lentiviral vectors.
abstract: Mucopolysaccharidosis type I (MPS I) results from a deficiency in the enzyme alpha-L-iduronidase (IDUA), and is characterized by skeletal abnormalities, hepatosplenomegaly and neurological dysfunction. In this study, we used a late generation lentiviral vector to evaluate the utility of this vector system for the transfer and expression of the human IDUA cDNA in MPS I fibroblasts. We observed that the level of enzyme expression in transduced cells was 1.5-fold the level found in normal cells; the expression persisted for at least two months. In addition, transduced MPS I fibroblasts were capable of clearing intracellular radiolabeled glycosaminoglycan (GAG). Pulse-chase experiments on transduced fibroblasts showed that the recombinant enzyme was synthesized as a 76-kDa precursor form and processed to a 66-kDa mature form; it was released from transduced cells and was endocytosed into a second population of untreated MPS I fibroblasts via a mannose 6-phosphate receptor. These results suggest that the lentiviral vector may be used for the delivery and expression of the IDUA gene to cells in vivo for treatment of MPS I.
authors.affiliation: Department of Biochemistry and Medical Biotechnologies, University of Naples Federico II, Italy. dinatale@cds.unina.it
authors.forename: Paola
authors.initials: P
authors.lastname: Di Natale
authors.forename: Carmela
authors.initials: C
authors.lastname: Di Domenico
authors.forename: Guglielmo R D
authors.initials: GR
authors.lastname: Villani
authors.forename: Angelo
authors.initials: A
authors.lastname: Lombardo
authors.forename: Antonia
authors.initials: A
authors.lastname: Follenzi
authors.forename: Luigi
authors.initials: L
authors.lastname: Naldini
mesh_terms: D005347:Fibroblasts / Q000378:metabolism; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D007068:Iduronidase / Q000172:deficiency / Q000235:genetics* / Q000378:metabolism*; D016086:Lentivirus; D008059:Mucopolysaccharidosis I / Q000235:genetics / Q000628:therapy*; D014161:Transduction, Genetic
chemical_list: D007068:Iduronidase