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Transcriptional targeting of adenoviral gene delivery into migrating wound keratinocytes using FiRE, a growth factor-inducible regulatory element

  • Acquired Diseases
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Abstract

Impaired cutaneous wound healing is a common complication in diabetes, ischemia and venous insufficiency of lower extremities, and in long-term treatment with corticosteroids or other immunosuppressive agents. In development of gene therapy for wound repair, expression of therapeutic transgenes should be precisely targeted and controlled. Here, we describe a recombinant adenovirus RAdFiRE-EGFP, in which a growth factor inducible element (FiRE) of the murine syndecan-1 gene controls the expression of enhanced green fluorescent protein (EGFP) reporter gene. Treatment of RAdFiRE-EGFP-transduced murine epidermal keratinocytes in culture with FiRE-activating growth factor markedly enhanced the expression of EGFP. In ex vivo organ culture of wounded murine skin transduced with RAdFiRE-EGFP, the EGFP expression was specifically detected in wound margin keratinocytes, but not in intact skin. Activity of EGFP was first detected 2 days after a single application of RAdFiRE-EGFP and persisted up to 10 days. Similarly, FiRE-driven EGFP expression was detected specifically in epidermal keratinocytes in the edge of incisional wounds in murine skin transduced with RAdFiRE-EGFP. In contrast, adenovirus-mediated lacZ expression driven by CMV promoter was detected scattered in epidermal, dermal and subcutaneous layers in ex vivo and in vivo wounds, as well as in intact skin. These data demonstrate the feasibility of FiRE as a tool for transcriptional targeting of adenovirus-mediated transgene expression to cutaneous wound edge keratinocytes.

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Acknowledgements

The authors are grateful to Mrs Hanna Haavisto, Mrs Taina Kalevo-Mattila and Mrs Anni Kieksi for expert technical help. Professor Seppo Ylä-Herttuala is acknowledged for fruitful discussions. This work was financially supported by the Academy of Finland, the Technical Research Center of Finland (TEKES), the Sigrid Jusélius Foundation, the Finnish Cancer Union, Turku University Central Hospital, Diabetes Research Foundation of Finland, Turku Graduate School of Biomedical Sciences, the Maud Kuistila Memorial Foundation (to PJ). PJ is Junior Research Fellow of the Academy of Finland.

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  1. P Jaakkola & M Ahonen

    Present address: Wellcome Trust Centre, University of Oxford, Roosevelt Drive, Headington, Oxford, OX3 7BN, UK

Authors and Affiliations

  1. Turku Centre for Biotechnology, University of Turku and Åbo Akademi University, Finland

    P Jaakkola, M Ahonen, V-M Kähäri & M Jalkanen

  2. Departments of Medical Biochemistry, and Dermatology, University of Turku, Turku, Finland

    V-M Kähäri

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  1. P Jaakkola
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  2. M Ahonen
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  4. M Jalkanen
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Jaakkola, P., Ahonen, M., Kähäri, VM. et al. Transcriptional targeting of adenoviral gene delivery into migrating wound keratinocytes using FiRE, a growth factor-inducible regulatory element. Gene Ther 7, 1640–1647 (2000). https://doi.org/10.1038/sj.gt.3301293

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  • DOI: https://doi.org/10.1038/sj.gt.3301293

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