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Repeated AAV-mediated gene transfer by serotype switching enables long-lasting therapeutic levels of hUgt1a1 enzyme in a mouse model of Crigler–Najjar Syndrome Type I

Gene Therapy volume 24, pages 649–660 (2017)Cite this article

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Abstract

Adeno-associated virus (AAV) -mediated gene therapy is a promising strategy to treat liver-based monogenic diseases. However, two major obstacles limit its success: first, vector dilution in actively dividing cells, such as hepatocytes in neonates/children, due to the non-integrating nature of the vector; second, development of an immune response against the transgene and/or viral vector. Crigler–Najjar Syndrome Type I is a rare monogenic disease with neonatal onset, caused by mutations in the liver-specific UGT1 gene, with toxic accumulation of unconjugated bilirubin in plasma, tissues and brain. To establish an effective and long lasting cure, we applied AAV-mediated liver gene therapy to a relevant mouse model of the disease. Repeated gene transfer to adults by AAV-serotype switching, upon neonatal administration, resulted in lifelong correction of total bilirubin (TB) levels in both genders. In contrast, vector loss over time was observed after a single neonatal administration. Adult administration resulted in lifelong TB levels correction in male, but not female Ugt1−/− mice. Our findings demonstrate that neonatal AAV-mediated gene transfer to the liver supports a second transfer of the therapeutic vector, by preventing the induction of an immune response and supporting the possibility to improve AAV-therapeutic efficacy by repeated administration.

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Acknowledgements

We thank Prof E Tongiorgi from University of Trieste, Italy for the microscope facility resources; the ICGEB bioExperimentation Facility for help with animal care; to Dr M Bestagno for help with the FACS; M Dapas and M Zotti for AAV production; B Boziglav for the help in histological preparations; Dr Gloria Gonzalez Aseguinolaza from CIMA, Spain who kindly provided the authors with the construct containing Albumin promoter and Albumin enhancer element-AAT promoter. This work was supported by Telethon (GGP10051) and by Beneficentia Stiftung to AFM (ICGEB).

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  1. L Bočkor and G Bortolussi: Both authors contributed equally and should be considered joint first authors.

Authors and Affiliations

  1. Mouse Molecular Genetics, Molecular Medicine and Cellular Immunology Groups, International Centre for Genetic Engineering and Biotechnology (ICGEB), Trieste, Italy

    L Bočkor, G Bortolussi, A Iaconcig, G Chiaruttini, M Giacca, F Benvenuti, L Zentilin & A F Muro

  2. Centro Studi Fegato, Fondazione Italiana Fegato, AREA Science Park, Trieste, Italy

    C Tiribelli

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  1. L Bočkor
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Bočkor, L., Bortolussi, G., Iaconcig, A. et al. Repeated AAV-mediated gene transfer by serotype switching enables long-lasting therapeutic levels of hUgt1a1 enzyme in a mouse model of Crigler–Najjar Syndrome Type I. Gene Ther 24, 649–660 (2017). https://doi.org/10.1038/gt.2017.75

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