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Direct comparison of hepatocyte-specific expression cassettes following adenoviral and nonviral hydrodynamic gene transfer

Gene Therapy volume 15, pages 594–603 (2008)Cite this article

Abstract

Hepatocytes are a key target for treatment of inborn errors of metabolism, dyslipidemia and coagulation disorders. The development of potent expression cassettes is a critical target to improve the therapeutic index of gene transfer vectors. Here we evaluated 22 hepatocyte-specific expression cassettes containing a human apo A-I transgene following hydrodynamic transfer of plasmids or adenoviral transfer with E1E3E4-deleted vectors in C57BL/6 mice. The DC172 promoter consisting of a 890 bp human α1-antitrypsin promoter and two copies of the 160 bp α1-microglobulin enhancer results in superior expression levels compared to constructs containing the 1.5 kb human α1-antitrypsin promoter, the 790 bp synthetic liver-specific promoter or the DC190 promoter containing a 520 bp human albumin promoter and two copies of the 99 bp prothrombin enhancer. The most potent expression cassette consists of the DC172 promoter upstream of the transgene and two copies of the hepatic control region-1. Minicircles containing this expression cassette induce persistent physiological human apo A-I or human factor IX levels after hydrodynamic transfer. In conclusion, in this comparative study of 22 hepatocyte-specific expression cassettes, the DC172 promoter in combination with two copies of the hepatic control region-1 induces the highest expression levels following hydrodynamic and adenoviral transfer.

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Acknowledgements

This work was supported by grants G.0563.05 and G.0564.05 of the Fonds voor Wetenschappelijk Onderzoek-Vlaanderen. The Center for Molecular and Vascular Biology is supported by the Excellentiefinanciering KU Leuven (EF/05/013). Frank Jacobs is a Research Assistant of the Instituut voor de Aanmoediging van Innovatie door Wetenschap en Technologie in Vlaanderen. Eline Van Craeyveld is a Research Assistant of the Fonds voor Wetenschappelijk Onderzoek-Vlaanderen. We thank J Hendrix and Z Zhang for excellent technical assistance.

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  1. F Jacobs and J Snoeys: These authors contributed equally to this work.

Authors and Affiliations

  1. Molecular and Cardiovascular Medicine, Center for Molecular and Vascular Biology, University of Leuven, Leuven, Belgium

    F Jacobs, J Snoeys, Y Feng, E Van Craeyveld, J Lievens & B De Geest

  2. Genzyme, Framingham, MA, USA

    D Armentano & S H Cheng

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  1. F Jacobs
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Correspondence to B De Geest.

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Jacobs, F., Snoeys, J., Feng, Y. et al. Direct comparison of hepatocyte-specific expression cassettes following adenoviral and nonviral hydrodynamic gene transfer. Gene Ther 15, 594–603 (2008). https://doi.org/10.1038/sj.gt.3303096

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