title: Correction of acid beta-galactosidase deficiency in GM1 gangliosidosis human fibroblasts by retrovirus vector-mediated gene transfer: higher efficiency of release and cross-correction by the murine enzyme.
abstract: Mutations in the lysosomal acid beta-galactosidase (EC 3.2.1.23) underlie two different disorders: GM1 gangliosidosis, which involves the nervous system and visceral organs to varying extents, and Morquio's syndrome type B (Morquio B disease), which is a skeletal-connective tissue disease without any CNS symptoms. This article shows that transduction of human GM1 gangliosidosis fibroblasts with retrovirus vectors encoding the human acid beta-galactosidase cDNA leads to complete correction of the enzymatic deficiency. The newly synthesized enzyme is correctly processed and targeted to the lysosomes in transduced cells. Cross-correction experiments using retrovirus-modified cells as enzyme donors showed, however, that the human enzyme is transferred at low efficiencies. Experiments using a different retrovirus vector carrying the human cDNA confirmed this observation. Transduction of human GM1 fibroblasts and mouse NIH 3T3 cells with a retrovirus vector encoding the mouse beta-galactosidase cDNA resulted in high levels of enzymatic activity. Furthermore, the mouse enzyme was found to be transferred to human cells at high efficiency. Enzyme activity measurements in medium conditioned by genetically modified cells suggest that the human beta-galactosidase enzyme is less efficiently released to the extracellular space than its mouse counterpart. This study suggests that lysosomal enzymes, contrary to the generalized perception in the field of gene therapy, may differ significantly in their properties and provides insights for design of future gene therapy interventions in acid beta-galactosidase deficiency.
authors.affiliation: Molecular Neurogenetics Unit, Massachusetts General Hospital, Harvard Medical School, Boston 02129, USA.
authors.forename: M
authors.initials: M
authors.lastname: Sena-Esteves
authors.forename: S M
authors.initials: SM
authors.lastname: Camp
authors.forename: J
authors.initials: J
authors.lastname: Alroy
authors.forename: X O
authors.initials: XO
authors.lastname: Breakefield
authors.forename: E M
authors.initials: EM
authors.lastname: Kaye
mesh_terms: D016475:3T3 Cells / Q000201:enzymology / Q000821:virology; D000818:Animals; D017077:Culture Media, Conditioned; D005347:Fibroblasts / Q000201:enzymology / Q000821:virology; D016537:Gangliosidosis, GM1 / Q000201:enzymology* / Q000235:genetics; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D008247:Lysosomes / Q000378:metabolism; D051379:Mice; D012190:Retroviridae / Q000235:genetics*; D001616:beta-Galactosidase / Q000172:deficiency* / Q000235:genetics
chemical_list: D017077:Culture Media, Conditioned; D001616:beta-Galactosidase