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High and long-term von Willebrand factor expression after Sleeping Beauty transposon-mediated gene therapy in a mouse model of severe von Willebrand disease

Item Type:Article
Title:High and long-term von Willebrand factor expression after Sleeping Beauty transposon-mediated gene therapy in a mouse model of severe von Willebrand disease
Creators Name:Portier, I. and Vanhoorelbeke, K. and Verhenne, S. and Pareyn, I. and Vandeputte, N. and Deckmyn, H. and Goldenberg, D.S. and Samal, H.B. and Singh, M. and Ivics, Z. and Izsvák, Z. and De Meyer, S.F.
Abstract:BACKGROUND: Type 3 von Willebrand disease (VWD) is characterized by complete absence of von Willebrand factor (VWF). Current therapy is limited to treatment with exogenous VWF/FVIII products, which only provide a short-term solution. Gene therapy offers the potential for a long-term treatment for VWD. OBJECTIVES: To develop an integrative Sleeping Beauty (SB) transposon-mediated VWF gene transfer approach in a preclinical mouse model of severe VWD. METHODS: We established a robust platform for sustained transgene murine (m)VWF expression in the liver of Vwf(-/-) mice by combining a liver-specific promoter with a sandwich transposon design and the SB100X transposase via hydrodynamic gene delivery. RESULTS: The sandwich SB transposon was suitable to deliver the full-length mVWF cDNA (8.4 kb) and supported supra-physiological expression that remained stable for up to 1.5 year after gene transfer. The sandwich vector stayed episomal (~60 weeks) or integrated in the host genome, respectively in the absence or presence of the transposase. Transgene integration was confirmed using carbon tetrachloride-induced liver regeneration. Analysis of integration sites by high-throughput analysis revealed random integration of the sandwich vector. While the SB vector supported long-term expression of supraphysiological mVWF levels, the bleeding phenotype was not corrected in all mice. Long-term expression of VWF by hepatocytes resulted in relatively reduced amounts of high molecular weight multimers, potentially limiting its hemostatic efficacy. CONCLUSIONS: While this integrative platform for VWF gene transfer is an important milestone of VWD gene therapy, cell type specific targeting is yet to be solved.
Keywords:Genetic Therapy, Phenotype, Transposases, Von Willebrand Diseases, Von Willebrand Factor, Animals, Mice
Source:Journal of Thrombosis and Haemostasis
ISSN:1538-7933
Publisher:Wiley-Blackwell (U.S.A.)
Volume:16
Number:3
Page Range:592-604
Date:March 2018
Official Publication:https://doi.org/10.1111/jth.13938
PubMed:View item in PubMed

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