title: Retrovirus-mediated enzymatic correction of Tay-Sachs defect in transduced and non-transduced cells.
abstract: Tay-Sachs disease is a severe neurodegenerative disorder due to mutations in the HEXA gene coding for the alpha-chain of the alpha-beta heterodimeric lysosomal enzyme beta-hexosaminidase A (HexA). Because no treatment is available for this disease, we have investigated the possibility of enzymatic correction of HexA-deficient cells by HEXA gene transfer. Human HEXA cDNA was subcloned into a retroviral plasmid generating to G.HEXA vector. The best Psi-CRIP producer clone of G.HEXA retroviral particles was isolated, and murine HexA-deficient fibroblasts derived from hexa -/- mice were transduced with the G.HEXA vector. Transduced cells overexpressed the alpha-chain, resulting in the synthesis of interspecific HexA (human alpha-chain/murine beta-chain) and in a total correction of HexA deficiency. The alpha-chain was secreted in the culture medium and taken up by HexA-deficient cells via mannose-6-phosphate receptor binding, allowing for the restoration of intracellular HexA activity in non-transduced cells.
authors.affiliation: Institut Cochin de Génétique Moléculaire (ICGM), U129 INSERM, Université René Descartes-Paris V, CHU Cochin-Port Royal, Paris, France. guidotti@cochin.inserm.fr
authors.forename: J
authors.initials: J
authors.lastname: Guidotti
authors.forename: S
authors.initials: S
authors.lastname: Akli
authors.forename: L
authors.initials: L
authors.lastname: Castelnau-Ptakhine
authors.forename: A
authors.initials: A
authors.lastname: Kahn
authors.forename: L
authors.initials: L
authors.lastname: Poenaru
mesh_terms: D016475:3T3 Cells; D000818:Animals; D003001:Cloning, Molecular; D004789:Enzyme Activation / Q000187:drug effects / Q000235:genetics; D005347:Fibroblasts / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics; D054818:Hexosaminidase A; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D012190:Retroviridae / Q000235:genetics; D013661:Tay-Sachs Disease / Q000201:enzymology* / Q000235:genetics*; D014162:Transfection; D001619:beta-N-Acetylhexosaminidases / Q000235:genetics* / Q000378:metabolism
chemical_list: D007527:Isoenzymes; D054818:Hexosaminidase A; D001619:beta-N-Acetylhexosaminidases