• Open Access

Hepatocyte-targeted expression by integrase-defective lentiviral vectors induces antigen-specific tolerance in mice with low genotoxic risk

Authors

  • Janka Mátrai,

    1. Free University of Brussels, Brussels, Belgium; Vesalius Research Center, Flanders Institute of Biotechnology; University of Leuven, Leuven, Belgium
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    • These authors contributed equally to this work.

      These authors share senior authorship.

  • Alessio Cantore,

    1. San Raffaele Telethon Institute for Gene Therapy, San Raffaele Scientific Institute, Milan, Italy
    2. Vita Salute San Raffaele University, Milan, Italy
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    • These authors contributed equally to this work.

      These authors share senior authorship.

  • Cynthia C. Bartholomae,

    1. National Center for Tumor Diseases, Department of Translational Oncology, German Research Cancer Center, Heidelberg, Germany
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    • These authors contributed equally to this work.

      These authors share senior authorship.

  • Andrea Annoni,

    1. San Raffaele Telethon Institute for Gene Therapy, San Raffaele Scientific Institute, Milan, Italy
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    • These authors contributed equally to this work.

      These authors share senior authorship.

  • Wei Wang,

    1. National Center for Tumor Diseases, Department of Translational Oncology, German Research Cancer Center, Heidelberg, Germany
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  • Abel Acosta-Sanchez,

    1. Free University of Brussels, Brussels, Belgium; Vesalius Research Center, Flanders Institute of Biotechnology; University of Leuven, Leuven, Belgium
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  • Ermira Samara-Kuko,

    1. Free University of Brussels, Brussels, Belgium; Vesalius Research Center, Flanders Institute of Biotechnology; University of Leuven, Leuven, Belgium
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  • Liesbeth De Waele,

    1. Free University of Brussels, Brussels, Belgium; Vesalius Research Center, Flanders Institute of Biotechnology; University of Leuven, Leuven, Belgium
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  • Ling Ma,

    1. Free University of Brussels, Brussels, Belgium; Vesalius Research Center, Flanders Institute of Biotechnology; University of Leuven, Leuven, Belgium
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  • Pietro Genovese,

    1. San Raffaele Telethon Institute for Gene Therapy, San Raffaele Scientific Institute, Milan, Italy
    2. Vita Salute San Raffaele University, Milan, Italy
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  • Martina Damo,

    1. San Raffaele Telethon Institute for Gene Therapy, San Raffaele Scientific Institute, Milan, Italy
    2. Vita Salute San Raffaele University, Milan, Italy
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  • Anne Arens,

    1. National Center for Tumor Diseases, Department of Translational Oncology, German Research Cancer Center, Heidelberg, Germany
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  • Kevin Goudy,

    1. San Raffaele Telethon Institute for Gene Therapy, San Raffaele Scientific Institute, Milan, Italy
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  • Timothy C. Nichols,

    1. University of North Carolina, Chapel Hill, NC
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  • Christof von Kalle,

    1. National Center for Tumor Diseases, Department of Translational Oncology, German Research Cancer Center, Heidelberg, Germany
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  • Marinee K. L. Chuah,

    1. Free University of Brussels, Brussels, Belgium; Vesalius Research Center, Flanders Institute of Biotechnology; University of Leuven, Leuven, Belgium
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  • Maria Grazia Roncarolo,

    1. San Raffaele Telethon Institute for Gene Therapy, San Raffaele Scientific Institute, Milan, Italy
    2. Vita Salute San Raffaele University, Milan, Italy
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  • Manfred Schmidt,

    Corresponding author
    1. National Center for Tumor Diseases, Department of Translational Oncology, German Research Cancer Center, Heidelberg, Germany
    • National Center for Tumor Diseases, Department of Translational Oncology, German Research Cancer Center, Im Neuenheimer Feld 350, Heidelberg, Germany 69120
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    • fax: +49 6221 421611

  • Thierry VandenDriessche,

    Corresponding author
    1. Free University of Brussels, Brussels, Belgium; Vesalius Research Center, Flanders Institute of Biotechnology; University of Leuven, Leuven, Belgium
    • Free University of Brussels, Laarbeeklaan 101, Brussels, Belgium 1090
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    • +32-2-4774159

  • Luigi Naldini

    Corresponding author
    1. San Raffaele Telethon Institute for Gene Therapy, San Raffaele Scientific Institute, Milan, Italy
    2. Vita Salute San Raffaele University, Milan, Italy
    • San Raffaele Telethon Institute for Gene Therapy, San Raffaele Scientific Institute, Via Olgettina 58, Milan, Italy 20132
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    • +32-2-4774159

    • fax: +39 02 2643 4621


Abstract

Lentiviral vectors are attractive tools for liver-directed gene therapy because of their capacity for stable gene expression and the lack of preexisting immunity in most human subjects. However, the use of integrating vectors may raise some concerns about the potential risk of insertional mutagenesis. Here we investigated liver gene transfer by integrase-defective lentiviral vectors (IDLVs) containing an inactivating mutation in the integrase (D64V). Hepatocyte-targeted expression using IDLVs resulted in the sustained and robust induction of immune tolerance to both intracellular and secreted proteins, despite the reduced transgene expression levels in comparison with their integrase-competent vector counterparts. IDLV-mediated and hepatocyte-targeted coagulation factor IX (FIX) expression prevented the induction of neutralizing antibodies to FIX even after antigen rechallenge in hemophilia B mice and accounted for relatively prolonged therapeutic FIX expression levels. Upon the delivery of intracellular model antigens, hepatocyte-targeted IDLVs induced transgene-specific regulatory T cells that contributed to the observed immune tolerance. Deep sequencing of IDLV-transduced livers showed only rare genomic integrations that had no preference for gene coding regions and occurred mostly by a mechanism inconsistent with residual integrase activity. Conclusion: IDLVs provide an attractive platform for the tolerogenic expression of intracellular or secreted proteins in the liver with a substantially reduced risk of insertional mutagenesis. (HEPATOLOGY 2011;)

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