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Variability of naked DNA expression after direct local injection: the influence of the injection speed

Gene Therapy volume 13, pages 1619–1627 (2006)Cite this article

Abstract

The simple injection of DNA into muscles is known to result in the expression of the injected genes, even though at low and variable levels. We report that this variability in DNA expression is partly dependent on the injection speed. The acceleration of the injection speed from values around 2 μl/s up to ones around 25 μl/s (depending on the tissue) results in a significant increase in gene expression in skeletal muscle (280 times on an average) and in liver (50 times) and a nonsignificant sevenfold increase in tumors. Heparin, which inhibits the spontaneous uptake of the injected DNA, also inhibits the increases related to the injection speed. However, at the highest injection speed, this inhibition is not total because very fast injections provoke a direct permeabilization of the cells. This ‘hydroporation’ could be similar to the permeabilization found in the hydrodynamics method based on the fast intravascular injection of a huge volume of DNA. Neither the ‘hydroporation’ nor the heparin-inhibitable uptake mechanism induces histologically detectable lesions. There is a limited muscle cell stress independent of the injection speed. Heterogeneity in the injection speed might thus be an explanation for the variability in DNA expression after simple injection.

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Abbreviations

ANOVA:

analysis of variance

GFP:

green fluorescent protein

HES:

hematoxylin–eosin–saffranin

Hsp70:

heat-shock protein 70

MEM:

minimum essential medium

MML:

mechanical massage of the liver

PBS:

phosphate-buffered saline

PI:

propidium iodide

RLU:

relative light units

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Acknowledgements

We acknowledge the excellent technical assistance of Elisabeth Connault (UMR 8121) for preparing and staining the material for the histological analysis; of Patrice Ardouin, Jennifer Huard and Désiré Challuau (SCEA IGR/CNRS) for their help with animal handling and housing and of Dorothée Peitzmann (UMR 8121) for the linguistic support. We acknowledge the financial support of the CNRS, IGR, AFM and the EU Commission through the Cliniporator project (QLK3-1999-00484) coordinated by LMM. We also acknowledge all our colleagues for fruitful discussions and collaborative work. FA is the recipient of an ‘aide aux études’ from the AFM (Association Française contre les Myopathies).

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Authors and Affiliations

  1. CNRS, UMR 8121, Laboratory of Vectorology and Gene Transfer, Institut Gustave-Roussy, Villejuif, France

    F M André, C Cournil-Henrionnet, P Opolon & L M Mir

  2. Univ Paris-Sud, UMR 8121, Laboratory of Vectorology and Gene Transfer, Institut Gustave-Roussy, Villejuif, France

    F M André, C Cournil-Henrionnet, P Opolon & L M Mir

  3. UMR7561 CNRS, Université de Nancy I, Vandœuvre Lès Nancy, France

    C Cournil-Henrionnet

  4. Biostatistics and Epidemiology Unit, Institut Gustave-Roussy, Villejuif, France

    D Vernerey

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  1. F M André
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  2. C Cournil-Henrionnet
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  4. P Opolon
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Correspondence to L M Mir.

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André, F., Cournil-Henrionnet, C., Vernerey, D. et al. Variability of naked DNA expression after direct local injection: the influence of the injection speed. Gene Ther 13, 1619–1627 (2006). https://doi.org/10.1038/sj.gt.3302827

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