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Coexpression of p21WAF1/CIP1 in adenovirus vector transfected human primary hepatocytes prevents apoptosis resulting in improved transgene expression

Gene Therapy volume 10, pages 668–677 (2003)Cite this article

Abstract

Replication-deficient adenovirus (Ad vector) is one of the most effective gene transfer systems. However, its employment in human gene therapy trials is hampered by Ad vector associated cytotoxicity and induction of apoptosis of the infected cells. Here, we identify one underlying mechanism as uncoupling of S phase and mitosis of the cell cycle leading to apoptosis and decline of transgene expression. Moreover, we demonstrate a strategy to avoid Ad vector associated cytotoxicity and induction of apoptosis in human primary hepatocytes by coinfection of Ad vector carrying the cDNA of choice and the cell cycle regulator p21WAF1/CIP1 (p21). In addition, animal experiments were performed using Ad vector directed coexpression of p21 and human α 1-antitrypsin. As serum analysis of α 1-antitrypsin after Ad vector mediated gene transfer to the liver of mice revealed, this strategy resulted also in the improvement of transgene expression by two orders of magnitude. These data suggest that coexpression of p21 and Ad vector carrying a therapeutic gene may be a promising strategy to avoid cytotoxicity and induction of apoptosis leading to improved safety in human gene therapy.

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Acknowledgements

The authors would like to thank Dr Mark Kay (Stanford University, Stanford, USA), Dr Ronald Crystal (Cornell University, New York, USA), and Dr Michael Strauss (Max Delbrueck Center for Molecular Medicine, Berlin, Germany) for generously providing the adenoviral vectors. Furthermore, we would like to thank Mrs B Durieux, M Gries, K Jäger and P Pierschalek for expert technical assistance, C Woischwill for critically reading the manuscript, and M Dittmar for help in preparing the manuscript.

This study was partly funded by BMBF grant No. 0311710 and QBP A.3. (Max Delbrueck Center grant).

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Authors and Affiliations

  1. Department of Hematology, Humboldt University of Berlin, Berlin, Germany

    G Wolff, V Ruppert, L Karawajew, E Wehnes & B Dörken

  2. Max Delbrueck Center for Molecular Medicine, Berlin, Germany

    G Wolff & B Dörken

  3. Department of Cell Biology, Institute of Biology, Humboldt University of Berlin, Berlin, Germany

    A Schumacher

  4. Department of General, Visceral, and Transplantation Surgery, Humboldt University of Berlin, Berlin, Germany

    A K Nuessler & P Neuhaus

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Wolff, G., Schumacher, A., Nuessler, A. et al. Coexpression of p21WAF1/CIP1 in adenovirus vector transfected human primary hepatocytes prevents apoptosis resulting in improved transgene expression. Gene Ther 10, 668–677 (2003). https://doi.org/10.1038/sj.gt.3301864

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