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Enhancing transduction of the liver by adeno-associated viral vectors

Gene Therapy volume 16, pages 60–69 (2009)Cite this article

Abstract

A number of distinct factors acting at different stages of the adeno-associated virus vector (AAV)-mediated gene transfer process were found to influence murine hepatocyte transduction. Foremost among these was the viral capsid protein. Self-complementary (sc) AAV pseudotyped with capsid from serotype 8 or rh.10 mediated fourfold greater hepatocyte transduction for a given vector dose when compared with vector packaged with AAV7 capsid. An almost linear relationship between vector dose and transgene expression was noted for all serotypes with vector doses as low as 1 × 107 vg per mouse (4 × 108 vg kg−1) mediating therapeutic levels of human FIX (hFIX) expression. Gender significantly influenced scAAV-mediated transgene expression, with twofold higher levels of expression observed in male compared with female mice. Pretreatment of mice with the proteasome inhibitor bortezomib increased scAAV-mediated hFIX expression from 4±0.6 to 9±2 μg ml−1 in female mice, although the effect of this agent was less profound in males. Exposure of mice to adenovirus 10–20 weeks after gene transfer with AAV vectors augmented AAV transgene expression twofold by increasing the level of proviral mRNA. Hence, optimization of individual steps in the AAV gene transfer process can further enhance the potency of AAV-mediated transgene expression, thus increasing the probability of successful gene therapy.

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Acknowledgements

We thank Dr Arthur Nienhuis and Professor David Linch for critically reviewing this paper. We also thank Dr James Wilson for providing the AAV packaging plasmids. This work was supported by The Katharine Dormandy Trust, UK; Medical Research Council, UK; Department of Health's NIHR Biomedical Research Centres funding award to UCLH/UCL, UK; The ASSISI Foundation of Memphis; the American Lebanese Syrian Associated Charities (ALSAC); National Heart, Lung, and Blood Institute (NHLBI) grant HL073838.

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Authors and Affiliations

  1. Department of Haematology, UCL Cancer Institute, London, UK

    A C Nathwani, M Cochrane & J McIntosh

  2. National Blood Service, London, UK

    A C Nathwani

  3. Department of Surgery, St Jude Children's Research Hospital, Memphis, TN, USA

    C Y C Ng, J Zhou & A M Davidoff

  4. Division of Experimental Hematology, St Jude Children's Research Hospital, Memphis, TN, USA

    J T Gray

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  1. A C Nathwani
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Correspondence to A C Nathwani.

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Nathwani, A., Cochrane, M., McIntosh, J. et al. Enhancing transduction of the liver by adeno-associated viral vectors. Gene Ther 16, 60–69 (2009). https://doi.org/10.1038/gt.2008.137

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