[HTML][HTML] Hyperactive sleeping beauty transposase enables persistent phenotypic correction in mice and a canine model for hemophilia B

MA Hausl, W Zhang, N Müther, C Rauschhuber… - Molecular Therapy, 2010 - cell.com
MA Hausl, W Zhang, N Müther, C Rauschhuber, HG Franck, EP Merricks, TC Nichols…
Molecular Therapy, 2010cell.com
Sleeping Beauty (SB) transposase enables somatic integration of exogenous DNA in
mammalian cells, but potency as a gene transfer vector especially in large mammals has
been lacking. Herein, we show that hyperactive transposase system delivered by high-
capacity adenoviral vectors (HC-AdVs) can result in somatic integration of a canine factor IX
(cFIX) expression-cassette in canine liver, facilitating stabilized transgene expression and
persistent haemostatic correction of canine hemophilia B with negligible toxicity. We …
Sleeping Beauty (SB) transposase enables somatic integration of exogenous DNA in mammalian cells, but potency as a gene transfer vector especially in large mammals has been lacking. Herein, we show that hyperactive transposase system delivered by high-capacity adenoviral vectors (HC-AdVs) can result in somatic integration of a canine factor IX (cFIX) expression-cassette in canine liver, facilitating stabilized transgene expression and persistent haemostatic correction of canine hemophilia B with negligible toxicity. We observed stabilized cFIX expression levels during rapid cell cycling in mice and phenotypic correction of the bleeding diathesis in hemophilia B dogs for up to 960 days. In contrast, systemic administration of an inactive transposase system resulted in rapid loss of transgene expression and transient phenotypic correction. Notably, in dogs a higher viral dose of the active SB transposase system resulted into transient phenotypic correction accompanied by transient increase of liver enzymes. Molecular analysis of liver samples revealed SB-mediated integration and provide evidence that transgene expression was derived mainly from integrated vector forms. Demonstrating that a viral vector system can deliver clinically relevant levels of a therapeutic protein in a large animal model of human disease paves a new path toward the possible cure of genetic diseases.
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