title: Enzymatic and functional correction along with long-term enzyme secretion from transduced bone marrow hematopoietic stem/progenitor and stromal cells derived from patients with Fabry disease.
abstract: Fabry disease is a lysosomal storage disorder that is due to a deficiency in alpha-galactosidase A (alpha-gal A). Previously we have shown that a recombinant retrovirus synthesized for the transfer of the human alpha-gal A coding sequence was able to engineer enzymatic correction of the hydrolase deficiency in fibroblasts and lymphoblasts from Fabry patients. The corrected cells secreted alpha-gal A that was taken up and utilized by uncorrected bystander cells, thus demonstrating metabolic cooperativity. In separate experiments we used transduced murine bone marrow cells and successfully tested and quantitated this phenomenon in vivo. In the present studies, which were designed to bring this therapeutic approach closer to clinical utility, we establish that cells originating from the bone marrow of numerous Fabry patients and normal volunteers can be effectively transduced and that these target cells demonstrate metabolic cooperativity. Both isolated CD34+-enriched cells and long-term bone marrow culture cells, including nonadherent hematopoietic cells and adherent stromal cells, were transduced. The transferred gene generates increased intracellular alpha-gal A enzyme activity in these cells. Further, it causes functional correction of lipid accumulation and provides for long-term alpha-gal A secretion. Collectively, these results indicate that a multifaceted gene transfer approach to bone marrow cells may be of therapeutic benefit for patients with Fabry disease.
authors.affiliation: Developmental and Metabolic Neurology Branch, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD, USA.
authors.forename: T
authors.initials: T
authors.lastname: Takenaka
authors.forename: C S
authors.initials: CS
authors.lastname: Hendrickson
authors.forename: D M
authors.initials: DM
authors.lastname: Tworek
authors.forename: M
authors.initials: M
authors.lastname: Tudor
authors.forename: R
authors.initials: R
authors.lastname: Schiffmann
authors.forename: R O
authors.initials: RO
authors.lastname: Brady
authors.forename: J A
authors.initials: JA
authors.lastname: Medin
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000201:enzymology* / Q000378:metabolism / Q000473:pathology; D002448:Cell Adhesion; D002478:Cells, Cultured; D004279:DNA, Viral / Q000032:analysis; D000795:Fabry Disease / Q000201:enzymology / Q000235:genetics / Q000473:pathology* / Q000628:therapy; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics; D006028:Glycosphingolipids / Q000378:metabolism; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000201:enzymology* / Q000378:metabolism / Q000473:pathology; D006801:Humans; D008247:Lysosomes / Q000201:enzymology; D051379:Mice; D011533:Proviruses / Q000302:isolation & purification; D012190:Retroviridae / Q000235:genetics / Q000302:isolation & purification; D017154:Stromal Cells / Q000201:enzymology / Q000378:metabolism / Q000473:pathology; D014162:Transfection; D000519:alpha-Galactosidase / Q000235:genetics / Q000378:metabolism*
chemical_list: D004279:DNA, Viral; D006028:Glycosphingolipids; D000519:alpha-Galactosidase