title: Retroviral gene transfer and sustained expression of human arylsulfatase A.
abstract: Transduction of mouse hematopoietic stem cells and their progeny was studied using a recombinant retroviral vector (MFG-ASA) which incorporates the human arylsulfatase A gene (ASA; EC 3.1.6.8). Successful transduction was demonstrated in spleen colonies of mice that received bone marrow transplantation, cultured bone marrow-derived macrophages, visceral tissues and brain of long-term reconstituted mice, and also the spleen colonies of secondarily transplanted mice. The efficiency of transduction in primary spleen colonies was 90%. Expression of the ASA transgene exceeded endogenous levels in spleen colonies and in cultured macrophages by 50-100%. Enzyme activity in the visceral tissues of long-term reconstituted mice consistently showed elevated ASA activity, greater than three-fold in the spleen and lung of one animal. Increased activity of ASA also could be detected in secondary spleen colonies. These data demonstrate the usefulness of the MFG-ASA vector for efficient gene transfer and expression in mouse hematopoietic stem cells and their differentiated progeny. The presence of vector DNA in the brain 4 months after transplantation suggests a role for gene transfer and stem cell transplantation in the treatment strategies for metachromatic leukodystrophy.
authors.affiliation: Department of Molecular Genetics and Biochemistry, University of Pittsburgh, PA, USA.
authors.forename: R
authors.initials: R
authors.lastname: Learish
authors.forename: T
authors.initials: T
authors.lastname: Ohashi
authors.forename: P A
authors.initials: PA
authors.lastname: Robbins
authors.forename: A
authors.initials: A
authors.lastname: Bahnson
authors.forename: S S
authors.initials: SS
authors.lastname: Boggs
authors.forename: K
authors.initials: K
authors.lastname: Patrene
authors.forename: B E
authors.initials: BE
authors.lastname: Schwartz
authors.forename: V
authors.initials: V
authors.lastname: Gieselmann
authors.forename: J A
authors.initials: JA
authors.lastname: Barranger
mesh_terms: D000818:Animals; D016026:Bone Marrow Transplantation; D001921:Brain / Q000201:enzymology; D002553:Cerebroside-Sulfatase / Q000235:genetics*; D015870:Gene Expression; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors; D006378:Helper Viruses / Q000235:genetics; D006412:Hematopoietic Stem Cells / Q000201:enzymology; D006801:Humans; D007966:Leukodystrophy, Metachromatic / Q000201:enzymology / Q000628:therapy; D051379:Mice; D012190:Retroviridae / Q000235:genetics*; D013154:Spleen / Q000201:enzymology; D013997:Time Factors; D014161:Transduction, Genetic
chemical_list: D002553:Cerebroside-Sulfatase