title: Hyperactive sleeping beauty transposase enables persistent phenotypic correction in mice and a canine model for hemophilia B.
abstract: 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.
authors: Max von Pettenkofer-Institut, Department of Virology, Ludwig-Maximilians-Universität Munich, Munich, Germany.
authors: Martin A
authors: MA
authors: Hausl
authors: Wenli
authors: W
authors: Zhang
authors: Nadine
authors: N
authors: Müther
authors: Christina
authors: C
authors: Rauschhuber
authors: Helen G
authors: HG
authors: Franck
authors: Elizabeth P
authors: EP
authors: Merricks
authors: Timothy C
authors: TC
authors: Nichols
authors: Mark A
authors: MA
authors: Kay
authors: Anja
authors: A
authors: Ehrhardt
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D004251:DNA Transposable Elements / Q000235:genetics; D004195:Disease Models, Animal; D004285:Dogs; D005164:Factor IX / Q000276:immunology / Q000378:metabolism; D015316:Genetic Therapy; D005822:Genetic Vectors; D002836:Hemophilia B / Q000235:genetics / Q000628:therapy*; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D010641:Phenotype; D012689:Sequence Homology, Nucleic Acid; D019076:Transgenes / Q000502:physiology; D019895:Transposases / Q000502:physiology*
chemical_list: D004251:DNA Transposable Elements; D005164:Factor IX; D019895:Transposases; C494471:sleeping beauty transposase, human