Onkološki inštitut Ljubljana (OILJ)
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  • Electroporation induced stress response and its effect on gene electrotransfer efficacy : in vivo imaging and numerical modelling
    Forjanič, Tadeja ...
    Skin is an attractive target tissue for gene transfer due to its size, accessibility and its immune competence. One of the promising delivery methods is gene delivery by means of electroporation ... (EP), i.e. gene electrotransfer (GET). To assess the importance of different effects of electroporation for successful GET we investigated: i) stress response; and ii) transfection efficacy of different pulse protocols. Moreover, numerical modelling was used to explain experimental results and to test the agreement of experimental results with current knowledge about GET. METHODS: A double transgenic mice Hspa1b-LucF (+/+) Hspa1b-mPlum (+/+) were used to determine the level of stress sensed by the cell in the tissue in vivo that was exposed to EP. The effect of five different pulse protocols on the stress levels sensed by the exposed cells and their efficacy for gene electrotransfer for two plasmids pEGFP-C1 (EGFP) and pCMV-tdTomato was tested. RESULTS: Quantification of the bioluminescence signal intensity showed that EP, regardless of the electric pulse parameters used, increased mean bioluminescence compared to the baseline bioluminescence signal of the non-exposed skin. The results of numerical modelling indicate that thermal stress alone is not sufficient to explain the measured bioluminescence signal. Of the tested pulse protocols, the highest expression of EGFP and tdTomato was achieved with HV-MV (high voltage - medium voltage) protocols, which agrees also with numerical model. SIGNIFICANCE: Although EP is widely used as a method for gene delivery, we show that the field could benefit from the use of mathematical modelling by introducing additional parameters such as EP induced stress and electrophoretic movement of plasmids.
    Vir: IEEE transactions on bio-medical engineering. - ISSN 0018-9294 (Vol. 66, no. 9, Sep. 2019, str. 2671-2683)
    Vrsta gradiva - članek, sestavni del
    Leto - 2019
    Jezik - angleški
    COBISS.SI-ID - 3142523
    DOI

vir: IEEE transactions on bio-medical engineering. - ISSN 0018-9294 (Vol. 66, no. 9, Sep. 2019, str. 2671-2683)

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