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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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).
The authors declare no conflict of interest.
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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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