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In vitro and in vivo transfection of plasmid DNA in the Dunning prostate tumor R3327-AT1 is enhanced by focused ultrasound

Gene Therapy volume 7, pages 1516–1525 (2000)Cite this article

Abstract

Gene therapy as a form of molecular medicine is expected to have a major impact on medical treatments in the future. However, the clinical use of gene therapy today is hampered by inadequate gene delivering systems to ensure sufficient, accurate and safe DNA uptake in the target cells in vivo. Nonviral transfection methods might have the advantage of safe application, but it would be helpful to increase their transfection rates, especially in vivo. In this study, we show that focused ultrasound provides an enhanced transfer of DNA plasmids in vitro and in vivo. In vitro, the β-galactosidase and luciferase DNA reporter plasmid were transfected into four cell lines (NIH 3T3 fibroblasts, malignant melanoma Mewo, HeLa, Dunning prostate tumor R3327-AT1). Ultrasound induced a 55- (Mewo) to 220-fold (AT1) stimulation resulting in transfection efficiencies in vitro between 2% (Mewo) and 12% (AT1). The in vivo stimulation was assessed in the Dunning prostate tumor R3327-AT1 implanted subcutaneously in Copenhagen rats using the β-galactosidase reporter. After intratumoral DNA injection, focused ultrasound induced a 10-fold increase of β-galactosidase positive cells in histology and a 15-fold increase of β-galactosidase protein expression in the ELISA assay. In contrast, ultrasound was not found to enhance reporter gene expression after intravenous plasmid application. Because ultrasound waves can be focused on different anatomical locations in the human body without significant adverse effects, the control of DNA transfer by focused ultrasound is a promising in vivo method for spatial regulation of gene-based medical treatments.

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Acknowledgements

We wish to thank Juergen Jenne for support in physical ultrasound measurements, Peter Peschke for providing the AT1 cells and support in histological analysis, Klaus Weber for providing the Mewo cells, and Juergen Debus for support of the studies. We thank Alexandra Tietze for excellent technical assistance with animal experiments. We thank Thomas Wirth for discussing the project and providing reporter constructs and the NIH 3T3 and HeLa cells. This work was supported in part by the Deutsche Forschungsgemeinschaft (Hu 798/1-1).

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Author notes

  1. P E Huber

    Present address: Department of Radiology, Brigham and Women's Hospital, Harvard Medical School, 75 Francis Street, Boston, 02115, MA, USA

  2. P Pfisterer

    Present address: Forschungszentrum Karlsruhe, Institut für Genetik, Karlsruhe, 76133, Germany

Authors and Affiliations

  1. Department of Radiation Oncology, University of Heidelberg, INF 400, Heidelberg, Germany

    P E Huber

  2. Department of Radiation Oncology, German Cancer Research Center (DKFZ), INF 280, Heidelberg, Germany

    P E Huber & P Pfisterer

  3. Zentrum für Molekulare Biologie Heidelberg (ZMBH), University of Heidelberg, INF 282, Heidelberg, Germany

    P Pfisterer

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  1. P E Huber
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Huber, P., Pfisterer, P. In vitro and in vivo transfection of plasmid DNA in the Dunning prostate tumor R3327-AT1 is enhanced by focused ultrasound. Gene Ther 7, 1516–1525 (2000). https://doi.org/10.1038/sj.gt.3301242

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  • DOI: https://doi.org/10.1038/sj.gt.3301242

Keywords

  • gene therapy
  • nonviral gene transfer
  • focused ultrasound
  • ultrasound therapy
  • Dunning prostate tumor

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