title: Retrovirus-mediated transfer of human alpha-galactosidase A gene to human CD34+ hematopoietic progenitor cells.
abstract: Fabry disease, caused by a deficiency of lysosomal enzyme alpha-galactosidase A (alpha-gal A), is one of the inherited disorders potentially treatable by gene transfer to hematopoietic stem cells. In this study, a high-titer amphotropic retroviral producer cell line, MFG-alpha-gal A, was established. CD34+ cells from normal umbilical cord blood were transduced by centrifugal enhancement. The alpha-gal A activity in transduced cells increased 3.6-fold above the activity in nontransduced cells. Transduction efficiency measured by PCR for the integrated alpha-gal A cDNA in CFU-GM colonies was in the range of 42-88% (average, 63%). The expression of functional enzyme in TFI erythroleukemia was sustained for as long as cells remained in culture (84 days) and for 28 days in LTC-IC cultures of CD34+ cells. The ability of the transduced CD34+ cells to secrete the enzyme and to correct enzyme-deficient Fabry fibroblasts was assessed by cocultivation of these cells. The enzyme was secreted into the medium from transduced CD34+ cells and taken up by Fabry fibroblasts through mannose 6-phosphate receptors. These findings suggest that genetically corrected hematopoietic stem/progenitor cells can be an enzymatic source for neighboring enzyme-deficient cells, and can potentially be useful for gene therapy of Fabry disease.
authors.affiliation: Department of Human Genetics, Graduate School of Public Health, University of Pittsburgh, PA 15261, USA.
authors.forename: N
authors.initials: N
authors.lastname: Takiyama
authors.forename: J T
authors.initials: JT
authors.lastname: Dunigan
authors.forename: M J
authors.initials: MJ
authors.lastname: Vallor
authors.forename: R
authors.initials: R
authors.lastname: Kase
authors.forename: H
authors.initials: H
authors.lastname: Sakuraba
authors.forename: J A
authors.initials: JA
authors.lastname: Barranger
mesh_terms: D018952:Antigens, CD34 / Q000032:analysis*; D001483:Base Sequence; D002478:Cells, Cultured; D017931:DNA Primers; D000795:Fabry Disease / Q000201:enzymology / Q000628:therapy; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors; D006412:Hematopoietic Stem Cells / Q000201:enzymology / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D012190:Retroviridae / Q000235:genetics*; D000519:alpha-Galactosidase / Q000235:genetics*
chemical_list: D018952:Antigens, CD34; D017931:DNA Primers; D000519:alpha-Galactosidase