title: Transduction of CD34+ enriched cord blood and Gaucher bone marrow cells by a retroviral vector carrying the glucocerebrosidase gene.
abstract: One promising strategy for gene therapy of Gaucher disease involves ex vivo retroviral transduction of autologous hematopoietic stem cells. Studies in small animals have demonstrated that this approach provides a life-long supply of the glucocerebrosidase (GC) enzyme. Human application has developed to the stage of a clinical trial. In this study, we describe development of a high titer amphotropic producer line for the vector, MFG-GC, and explore transduction of CD34+ cells from various human sources. Higher than three times the normal levels of glucocerebrosidase activity in non-transduced cells were achieved following transduction of CD34+ cells obtained from bone marrow or cord blood from normal donors. The improvement in enzyme activity in Gaucher marrow was about 40-fold above deficient levels. We examined the timing and stepwise effect of multiple rounds of infection and evaluated post-infection expansion of cells in two different cytokine mixtures. Transduction efficiency was determined using immunocytochemistry and Southern blot hybridization.
authors.affiliation: Department of Human Genetics, Graduate School of Public Health, University of Pittsburgh, USA.
authors.forename: A B
authors.initials: AB
authors.lastname: Bahnson
authors.forename: M
authors.initials: M
authors.lastname: Nimgaonkar
authors.forename: Y
authors.initials: Y
authors.lastname: Fei
authors.forename: S S
authors.initials: SS
authors.lastname: Boggs
authors.forename: P D
authors.initials: PD
authors.lastname: Robbins
authors.forename: T
authors.initials: T
authors.lastname: Ohashi
authors.forename: J
authors.initials: J
authors.lastname: Dunigan
authors.forename: J
authors.initials: J
authors.lastname: Li
authors.forename: E D
authors.initials: ED
authors.lastname: Ball
authors.forename: J A
authors.initials: JA
authors.lastname: Barranger
mesh_terms: D018952:Antigens, CD34 / Q000378:metabolism; D001853:Bone Marrow / Q000201:enzymology / Q000473:pathology; D005312:Fetal Blood / Q000166:cytology / Q000201:enzymology* / Q000276:immunology; D005776:Gaucher Disease / Q000201:enzymology / Q000473:pathology / Q000628:therapy*; D015316:Genetic Therapy; D005822:Genetic Vectors; D005962:Glucosylceramidase / Q000235:genetics*; D006801:Humans; D066298:In Vitro Techniques; D007231:Infant, Newborn; D012190:Retroviridae / Q000235:genetics*; D014161:Transduction, Genetic
chemical_list: D018952:Antigens, CD34; D005962:Glucosylceramidase