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No tumour-initiating risk associated with scAAV transduction in newborn rat liver

Gene Therapy volume 20, pages 779–784 (2013)Cite this article

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Abstract

Delivery of recombinant adeno-associated virus (rAAV) vectors to the newborn liver is followed by a rapid loss of episomal vector copies because of hepatocyte proliferation. In selected hepatocytes, integration of rAAV genomes can lead to a sustained expression of the transgene. The safety of in vivo gene therapy with single-stranded AAV vectors has been questioned in a study reporting a high incidence of hepatocellular carcinoma, associated with provirus integration events in mice that receive an single-stranded AAV injection at birth. To investigate the tumour-initiating potential of the newly established self-complementary AAV (scAAV) vectors in the liver, groups of newborn rats received intravenous injection of a scAAV vector encoding the green fluorescent protein (GFP), or were injected with phosphate-buffered saline (PBS) or diethylnitrosamine (DEN), a well-known liver tumour initiator. The rats were fed on a diet containing 2-acetylaminofluorene, a potent liver tumour-promoting agent to accelerate the carcinogenic process. After 2 months, the animals were killed and their livers analysed. Preneoplastic nodules were identified by glutathion S-transferase-p (GSTp) staining, and GFP expression was detected by immunohistochemistry. Vector genome integration events were analysed. The numbers of GSTp-positive foci were comparable in the PBS and the scAAV–GFP groups and significantly higher in the DEN group. The proportion of GSTp-positive foci that also expressed GFP was low and in the range expected for random occurrence. No specific integration hot spots were detected by linear amplification-mediated-PCR in transduced liver. In conclusion, scAAV transduction of newborn rat liver does not trigger preneoplastic lesions suggesting an absence of liver tumourigenesis.

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Acknowledgements

This work was supported by the Association Française contre les Myopathies (AFM). V Gauttier was funded by INSERM and la Région Pays de la Loire.

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Authors and Affiliations

  1. INSERM UMR1064, Centre de Recherche en Transplantation et en Immunologie, Université de Nantes, CHU Hôtel Dieu, Nantes, France

    V Gauttier, V Pichard, N Ferry & S Conchon

  2. INSERM UMR1089, Laboratoire de Thérapie Génique, Université de Nantes, IRT-UN, Nantes, France

    D Aubert

  3. Abteilung für Translationale Onkologie (G100), Nationales Zentrum für Tumorerkrankungen, Deutsches Krebsforschungszentrum, Heidelberg, Germany

    C Kaeppel & M Schmidt

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  1. V Gauttier
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Correspondence to S Conchon.

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Gauttier, V., Pichard, V., Aubert, D. et al. No tumour-initiating risk associated with scAAV transduction in newborn rat liver. Gene Ther 20, 779–784 (2013). https://doi.org/10.1038/gt.2013.7

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