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Stable therapeutic serum levels of human alpha-1 antitrypsin (AAT) after portal vein injection of recombinant adeno-associated virus (rAAV) vectors

Gene Therapy volume 8, pages 1299–1306 (2001)Cite this article

Abstract

Previous work from our group showed that recombinant adeno-associated virus (rAAV) vectors mediated long-term secretion of therapeutic serum levels of human alpha-1 antitrypsin (hAAT) after a single injection in murine muscle. We hypothesized that hepatocyte transduction could be even more efficient, since these cells represent the natural site of AAT production and secretion. To test this hypothesis, rAAV vectors containing the hAAT cDNA driven by either the human elongation factor 1 alpha promoter, the human cytomegalovirus immediate–early promoter (CMV), or the CMV-chicken beta actin hybrid (CB) promoter were injected into the portal or tail veins of adult C57Bl/6 mice. Potentially therapeutic serum levels of hAAT (600 μg/ml) were achieved after portal vein injection of doses of 4 × 109 infectious units (IU), a 10-fold lower dose than that required for similar levels of expression via the i.m. route. Serum levels greater than 1 mg/ml were achieved at doses of 3 × 1010 IU. Southern blotting of liver DNA revealed the presence of circular episomal vector genomes. Immunostaining showed that transgene expression was scattered throughout the liver parenchyma. Similar results were obtained with a rAAV-CB-green fluorescent protein (GFP) vector. There was no evidence of hepatic toxicity. These data indicate that liver-directed rAAV-based gene therapy is effective in the murine model, and hence might be feasible for treatment of human AAT deficiency.

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Acknowledgements

We wish to thanks Marda Scott Jorgensen and Trevor Jorgensen for technical assistance and Ed Mason, Michael Morgan, Kye Chesnut and the UF vector core facility for rAAV production. This work was supported by grants from the NHLBI (P50-HL-59412, P01-HL-51811), the NIDDK (R01-DK-51809, R01-DK-53512, and P01-DK-58327), and the Alpha One Foundation. Some of the authors (SS and TRF) could be entitled to patent royalties for products described in this manuscript.

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Authors and Affiliations

  1. Department of Pediatrics of the University of Florida Genetics Institute, University of Florida, Gainesville, FL, USA

    S Song, KI Berns & TR Flotte

  2. Department of Biochemistry and Molecular Biology of the University of Florida Genetics Institute, University of Florida, Gainesville, FL, USA

    J Embury, PJ Laipis & JM Crawford

  3. Department of Powell Gene Therapy Centre of the University of Florida Genetics Institute, University of Florida, Gainesville, FL, USA

    S Song, J Embury, PJ Laipis, KI Berns & TR Flotte

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Song, S., Embury, J., Laipis, P. et al. Stable therapeutic serum levels of human alpha-1 antitrypsin (AAT) after portal vein injection of recombinant adeno-associated virus (rAAV) vectors. Gene Ther 8, 1299–1306 (2001). https://doi.org/10.1038/sj.gt.3301422

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