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Efficient and stable transduction of dopaminergic neurons in rat substantia nigra by rAAV 2/1, 2/2, 2/5, 2/6.2, 2/7, 2/8 and 2/9

Gene Therapy volume 18, pages 517–527 (2011)Cite this article

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Abstract

Dysfunction of the nigrostriatal system is the major cause of Parkinson's disease (PD). This brain region is therefore an important target for gene delivery aiming at disease modeling and gene therapy. Recombinant adeno-associated viral (rAAV) vectors have been developed as efficient vehicles for gene transfer into the central nervous system. Recently, several serotypes have been described, with varying tropism for brain transduction. In light of the further development of a viral vector-mediated rat model for PD, we performed a comprehensive comparison of the transduction and tropism for dopaminergic neurons (DNs) in the adult Wistar rat substantia nigra (SN) of seven rAAV vector serotypes (rAAV 2/1, 2/2, 2/5, 2/6.2, 2/7, 2/8 and 2/9). All vectors were normalized by titer and volume, and stereotactically injected into the SN. Gene expression was assessed non-invasively and quantitatively in vivo by bioluminescence imaging at 2 and 5 weeks after injection, and was found to be stable over time. Immunohistochemistry at 6 weeks following injection revealed the most widespread enhanced green fluorescence protein expression and the highest number of positive nigral cells using rAAV 2/7, 2/9 and 2/1. The area transduced by rAAV 2/8 was smaller, but nevertheless almost equal numbers of nigral cells were targeted. Detailed confocal analysis revealed that serotype 2/7, 2/9, 2/1 and 2/8 transduced at least 70% of the DNs. In conclusion, these results show that various rAAV serotypes efficiently transduce nigral DNs, but significant differences in transgene expression pattern and level were observed.

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Acknowledgements

We thank Martine Michiels and Phebe van Wijk for their excellent technical assistance. AVdP, JT and MC are doctoral fellows supported by grants from the Institute for the Promotion of Innovation through Science and Technology in Flanders (IWT-Vlaanderen). BH is a doctoral fellow supported by a grant from the Flemish Fund for Scientific Research (FWO-Vlaanderen). Research was funded by the IWT-Vlaanderen (IWT SBO/060838 and IWT SBO/80020), by the EC-FP6 program ‘DiMI’ (LSHB-CT-2005-512146), the FP7 RTD project MEFOPA (HEALTH-2009-241791), the KULeuven (IOF-KP/07/001 and OT/08/052A), the Interuniversity Attraction Pole programme NiMI (P6/38) and the KULeuven Center of Excellence ‘MoSAIC’ (EF/05/08). Confocal images were taken in the cell imaging core of the KULeuven.

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Authors and Affiliations

  1. Neurobiology and Gene Therapy, Molecular Medicine, Katholieke Universiteit Leuven, Flanders, Belgium

    A Van der Perren, C Van den Haute, F Coun, B Heeman, V Reumers & V Baekelandt

  2. Molecular Virology and Gene Therapy, Molecular Medicine, Katholieke Universiteit Leuven, Flanders, Belgium

    J Toelen, M Carlon & Z Debyser

  3. Department of Pathology and Laboratory Medicine, Division of Transfusion Medicine, Gene Therapy Program, University of Pennsylvania, Philadelphia, PA, USA

    L H Vandenberghe & J M Wilson

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  1. A Van der Perren
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Correspondence to V Baekelandt.

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LHV holds patents on the technology described within. JMW is an inventor on patents licensed to various biopharmaceutical companies, including ReGenX, for which he has equity in, consults for and receives a grant from.

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Van der Perren, A., Toelen, J., Carlon, M. et al. Efficient and stable transduction of dopaminergic neurons in rat substantia nigra by rAAV 2/1, 2/2, 2/5, 2/6.2, 2/7, 2/8 and 2/9. Gene Ther 18, 517–527 (2011). https://doi.org/10.1038/gt.2010.179

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