Retroviral gene transfer and sustained expression of human arylsulfatase A.
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.
Learish
R
R
Department of Molecular Genetics and Biochemistry, University of Pittsburgh, PA, USA.
Ohashi
T
T
Robbins
P A
PA
Bahnson
A
A
Boggs
S S
SS
Patrene
K
K
Schwartz
B E
BE
Gieselmann
V
V
Barranger
J A
JA
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
D002553:Cerebroside-Sulfatase