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AAV vectors transduce hepatocytes in vivo as efficiently in cirrhotic as in healthy rat livers

Gene Therapy volume 19, pages 411–417 (2012)Cite this article

Abstract

In liver cirrhosis, abnormal liver architecture impairs efficient transduction of hepatocytes with large viral vectors such as adenoviruses. Here we evaluated the ability of adeno-associated virus (AAV) vectors, small viral vectors, to transduce normal and cirrhotic rat livers. Using AAV serotype-1 (AAV1) encoding luciferase (AAV1Luc) we analyzed luciferase expression with a CCD camera. AAV1Luc was injected through the hepatic artery (intra-arterial (IA)), the portal vein (intra-portal (IP)), directly into the liver (intra-hepatic (IH)) or infused into the biliary tree (intra-biliar). We found that AAV1Luc allows long-term and constant luciferase expression in rat livers. Interestingly, IP administration leads to higher expression levels in healthy than in cirrhotic livers, whereas the opposite occurs when using IA injection. IH administration leads to similar transgene expression in cirrhotic and healthy rats, whereas intra-biliar infusion is the least effective route. After 70% partial hepatectomy, luciferase expression decreased in the regenerating liver, suggesting lack of efficient integration of AAV1 DNA into the host genome. AAV1Luc transduced mainly the liver but also the testes and spleen. Within the liver, transgene expression was found mainly in hepatocytes. Using a liver-specific promoter, transgene expression was detected in hepatocytes but not in other organs. Our results indicate that AAVs are convenient vectors for the treatment of liver cirrhosis.

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Acknowledgements

We thank Gabor Gondi, Erkuden Casales, Sara Arcelus, Nerea Juanarena and Cristina Olagüe for technical assistance, and Pilar Peréz and Uxue Latasa (CIMA) for help with liver cell isolation. We also thank Gloria González-Aseguinolaza for advice with AAV vectors, AMT for production of ssAAV1EalbAATLuc and Paul Miller for English editorial work. This work was supported by MICINN (BIO2006-13225 and 2009/09295), Fundación Echevano, Instituto de Salud Carlos III, through the ‘UTE project CIMA,’ and by the project RNAREG (CSD2009-00080), funded by the Ministry of Science and Innovation under the programme CONSOLIDER INGENIO 2010. LS was an FPI fellow.

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  1. L Sobrevals

    Present address: 3Current address: Programme Genes and Disease, Centre for Genomic Regulation, Barcelona 08003, Spain.,

Authors and Affiliations

  1. Department of Hepatology and Gene Therapy, Centre for Applied Medical Research (CIMA), University of Navarra, Pamplona, Spain

    L Sobrevals, M Enguita, C Rodriguez, J Gonzalez-Rojas, P Alzaguren, N Razquin, J Prieto & P Fortes

  2. CIBERehd Liver Unit, University Clinic, School of Medicine, University of Navarra, Pamplona, Spain

    C Rodriguez & J Prieto

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  1. L Sobrevals
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Correspondence to P Fortes.

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Sobrevals, L., Enguita, M., Rodriguez, C. et al. AAV vectors transduce hepatocytes in vivo as efficiently in cirrhotic as in healthy rat livers. Gene Ther 19, 411–417 (2012). https://doi.org/10.1038/gt.2011.119

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