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Augmentation of AAV-mediated cardiac gene transfer after systemic administration in adult rats

Gene Therapy volume 15, pages 1558–1565 (2008)Cite this article

Abstract

Adeno-associated virus (AAV)-6 or -9-pseudotyped vectors are suitable for efficient cardiac gene transfer after intravenous injection in mice. However, a systemic application in larger animals or humans would require very high doses of viral particles. Therefore, the aim of our study was to test if ultrasound-targeted microbubble destruction could augment cardiac transduction of AAV vectors after intravenous administration in rats. To analyze efficiency and specificity of gene transfer, microbubbles loaded with AAV-6 or -9 harboring a luciferase or enhanced green fluorescent protein (EGFP) reporter gene were infused into the jugular vein of adult Sprague–Dawley rats. During the infusion, high mechanical index ultrasound was administered to the heart. Control rats received the same amount of virus without microbubbles, but with ultrasound. After 4 weeks, organs were harvested and analyzed for reporter gene expression. In contrast to low cardiac expression after systemic transfer of the vector solution without microbubbles, ultrasound-targeted destruction of microbubbles significantly increased cardiac reporter activities between 6- and 20-fold. Analysis of spatial distribution of transgene expression using an AAV-9 vector encoding for EGFP revealed transmural expression predominantly in the left ventricular anterior wall. In conclusion, ultrasound targeted microbubble destruction augments cardiac transduction of AAV vectors in rats. This approach may be suitable for efficient, specific and noninvasive AAV-mediated gene transfer in larger animals or humans.

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Acknowledgements

We thank Barbara Leuchs, Renate Eudenbach and the DKFZ vector core production unit for generating high titer AAV vector stocks. We also thank the Nikon Center Heidelberg for their support in microscopy. This work was supported by a grant of the Deutsche Forschungsgemeinschaft (MU 1654/3-2), the Bundesministerium für Bildung und Forschung (01GU0527) to OM and by the BioFuture grant of the Bundesministerium für Bildung und Forschung to RB.

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Authors and Affiliations

  1. Department of Cardiology, Internal Medicine III, University Hospital Heidelberg, Heidelberg, Germany

    O J Müller, S Schinkel, H A Katus & R Bekeredjian

  2. Applied Tumorvirology F010, German Cancer Research Center, Heidelberg, Germany

    J A Kleinschmidt

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Correspondence to R Bekeredjian.

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Müller, O., Schinkel, S., Kleinschmidt, J. et al. Augmentation of AAV-mediated cardiac gene transfer after systemic administration in adult rats. Gene Ther 15, 1558–1565 (2008). https://doi.org/10.1038/gt.2008.111

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