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microRNA122-regulated transgene expression increases specificity of cardiac gene transfer upon intravenous delivery of AAV9 vectors

Gene Therapy volume 18, pages 199–209 (2011)Cite this article

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Abstract

Adeno-associated virus (AAV) vectors with capsids of AAV serotype 9 enable an efficient transduction of the heart upon intravenous injection of adult mice but also transduce the liver. The aim of this study was to improve specificity of AAV9 vector-mediated cardiac gene transfer by microRNA (miR)-dependent control of transgene expression. We constructed plasmids and AAV vectors containing target sites (TSs) of liver-specific miR122, miR192 and miR148a in the 3′ untranslated region (3′UTR) of a luciferase expression cassette. Luciferase expression was efficiently suppressed in liver cell lines expressing high levels of the corresponding miRs, whereas luciferase expression was unaffected in cardiac myocytes. Intravenous injections of AAV9 vectors bearing three repeats of miR122 TS in the 3′UTR of an enhanced green fluorescent expression (EGFP) expression cassette resulted in the absence of EGFP expression in the liver of adult mice, whereas the control vectors without miR TS displayed significant hepatic EGFP expression. EGFP expression levels in the heart, however, were comparable between miR122-regulated and control vectors. The liver-specific de-targeting in vivo using miR122 was even more efficient than transcriptional targeting with a cardiac cytomegalovirus (CMV)-enhanced myosin light chain (MLC) promoter. These data indicate that miR-regulated targeting is a powerful new tool to further improve cardiospecificity of AAV9 vectors.

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Abbreviations

miR:

microRNA

AAV:

adeno-associated virus

TS:

target site

nt:

nucleotides

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Acknowledgements

We thank Barbara Leuchs and the DKFZ vector core production unit for their support in generating high titer AAV vector stocks and Stefanie Schinkel and Georg Zingler for support in histological analyses. In addition, we thank Ulrike Gärtner for skillful intravenous injections and the Nikon Center Heidelberg for support in microscopy. This work was supported through SFB Transregio 19 by project grants C1 to HF and RV and by a grants of the Deutsche Forschungsgemeinschaft (FE785/2-1 and FE785/3-1 to HF, KU1436/6-1 to JK, MU 1654/3-2 to OJM); and the Bundesministerium für Bildung und Forschung (01GU0527 to OJM and HAK).

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Author notes

  1. A Geisler, A Jungmann, H Fechner and O J Müller: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Cardiology and Pneumology, Charité-Universitätsmedizin Berlin, Campus Benjamin Franklin, Berlin, Germany

    A Geisler, W Poller & H Fechner

  2. Department of Internal Medicine III, University Hospital Heidelberg, Heidelberg, Germany

    A Jungmann, H A Katus & O J Müller

  3. Institute of Biotechnology, University of Technology, Berlin, Germany

    J Kurreck & H Fechner

  4. Institute of Clinical Pharmacology and Toxicology, Charité-Universitätsmedizin Berlin, Campus Benjamin Franklin, Berlin, Germany

    R Vetter

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  1. A Geisler
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Correspondence to H Fechner.

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Geisler, A., Jungmann, A., Kurreck, J. et al. microRNA122-regulated transgene expression increases specificity of cardiac gene transfer upon intravenous delivery of AAV9 vectors. Gene Ther 18, 199–209 (2011). https://doi.org/10.1038/gt.2010.141

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