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Self-inactivating MLV vectors have a reduced genotoxic profile in human epidermal keratinocytes

Gene Therapy volume 20, pages 949–957 (2013)Cite this article

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Abstract

Transplantation of epithelia derived from keratinocyte stem cells transduced by retroviral vectors is a potential therapy for epidermolysis bullosa (EB), a family of inherited skin adhesion defects. The biosafety characteristics of retroviral vectors in keratinocytes are, however, poorly defined. We developed self-inactivating (SIN) vectors derived from the Moloney murine leukemia (MLV) and the human immunodeficiency (HIV) viruses expressing therapeutic levels of LAMB3, a transgene defective in junctional EB, and tested their integration profile in human primary keratinocytes. The SIN–HIV vector showed the expected preference for transcribed genes while the SIN–MLV vector integrated preferentially in regulatory elements, but showed a significantly lower tendency to target cell growth-related genes, transcription start sites and epigenetically defined promoters compared with a wild-type MLV vector in an epithelial cell context. A quantitative gene expression assay in individual keratinocyte clones showed that MLV-derived vectors deregulate expression of targeted genes at a lower frequency than in hematopoietic cells, and that the SIN–MLV design has the lowest activity compared to both MLV and SIN–HIV vectors. This study indicates that SIN–MLV vectors may have a better safety profile in keratinocyte than in hematopoietic cells, and be a reasonable alternative to lentiviral vectors for gene therapy of inherited skin disorders.

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Acknowledgements

This work was supported by grants from the European Research Council (GT-SKIN) and the Italian National Research Council (EPIGEN). FL was supported by grants PI11/01225 from ISCIII and S2010/BMD-2359 from CM.

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Author notes

  1. A Cavazza

    Present address: 6Current address: Department of Medical Oncology, Dana Farber Cancer Institute, Harvard Medical School, Boston, MA, USA.,

Authors and Affiliations

  1. Department of Life Sciences, University of Modena and Reggio Emilia, Modena, Italy

    A Cavazza, F Cocchiarella & F Mavilio

  2. Department of Translational Oncology, National Center for Tumor Diseases and German Cancer Research Center (DKFZ), Heidelberg, Germany

    C Bartholomae & M Schmidt

  3. Department of Medicine, Emergency Medicine and Medical Specialties, University of Modena and Reggio Emilia, Modena, Italy

    C Pincelli

  4. Epithelial Biomedicine Division, CIEMAT-CIBERER, Madrid, Spain

    F Larcher

  5. Genethon, Evry, France

    F Mavilio

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Correspondence to F Mavilio.

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Cavazza, A., Cocchiarella, F., Bartholomae, C. et al. Self-inactivating MLV vectors have a reduced genotoxic profile in human epidermal keratinocytes. Gene Ther 20, 949–957 (2013). https://doi.org/10.1038/gt.2013.18

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