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Evaluation of the specificity and sensitivity of ferritin as an MRI reporter gene in the mouse brain using lentiviral and adeno-associated viral vectors

Gene Therapy volume 18, pages 594–605 (2011)Cite this article

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Abstract

The development of in vivo imaging protocols to reliably track transplanted cells or to report on gene expression is critical for treatment monitoring in (pre)clinical cell and gene therapy protocols. Therefore, we evaluated the potential of lentiviral vectors (LVs) and adeno-associated viral vectors (AAVs) to express the magnetic resonance imaging (MRI) reporter gene ferritin in the rodent brain. First, we compared the induction of background MRI contrast for both vector systems in immune-deficient and immune-competent mice. LV injection resulted in hypointense (that is, dark) changes of T2/T2* (spin–spin relaxation time)-weighted MRI contrast at the injection site, which can be partially explained by an inflammatory response against the vector injection. In contrast to LVs, AAV injection resulted in reduced background contrast. Moreover, AAV-mediated ferritin overexpression resulted in significantly enhanced contrast to background on T2*-weighted MRI. Although sensitivity associated with the ferritin reporter remains modest, AAVs seem to be the most promising vector system for in vivo MRI reporter gene imaging.

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Abbreviations

AAV:

adeno-associated viral vector

BLI:

bioluminescence imaging

CNS:

central nervous system

eGFP:

enhanced green fluorescent protein

FerrH:

ferritin heavy subunit

fLuc:

firefly luciferase

LV:

lentiviral vector

MRI:

magnetic resonance imaging

PBS:

phosphate-buffered saline

T2(*):

spin–spin relaxation time

3D:

three-dimensional

VOI:

volume of interest

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Acknowledgements

We acknowledge the KU Leuven Cell Imaging Core and vector core and thank Frea Coun, Martine Michiels, Sylvie De Swaef, Marly Balcer and Ann Van Santvoort for their excellent technical assistance. We also thank Gaëlle Jestin (Guerbet Biological Research Department, Aulnay-sous-Bois, France) for performing ICP-MS analyses. We are grateful to Paolo Arosio (Department of Biomedical Science and Technology, San Raffaele Hospital, Milano, Italy) for kindly providing us with the rH02 antibody. This work was supported by the EC-FP6 program ‘DiMI’ (LSHB-CT-2005-512146), the EC-FP6 project ‘StrokeMAP’ (LSHC-CT-2006-037186), the KU Leuven Center of Excellence ‘MoSAIC’ (EF/05/08) and the Flemish government IWT SBO/060838 ‘Brainstim’, IWT SBO ‘iMAGiNe’ (IWT 80017), IWT SBO 60819 (Quantiviam) and the EC-FP7 network ‘European Network for Cell Imaging and Tracking Expertise’ (ENCITE, 2007-201842). GVV was funded by the Interuniversity Attraction Pole programme NiMI (P6/38) and by the European FP6 project StrokeMAP.

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Authors and Affiliations

  1. Laboratory for Neurobiology and Gene Therapy, Katholieke Universiteit Leuven, Leuven, Flanders, Belgium

    G Vande Velde, O Krylychkina & V Baekelandt

  2. Molecular Small Animal Imaging Center (MoSAIC), Katholieke Universiteit Leuven, Leuven, Flanders, Belgium

    G Vande Velde, J R Rangarajan, T Dresselaers, A Ibrahimi, F Maes, Z Debyser, U Himmelreich & V Baekelandt

  3. Biomedical NMR Unit, Katholieke Universiteit Leuven, Leuven, Flanders, Belgium

    G Vande Velde, T Dresselaers & U Himmelreich

  4. Medical Image Computing (ESAT/PSI), Katholieke Universiteit Leuven and Medical Imaging Research Center, University Hospital Leuven, Leuven, Flanders, Belgium

    J R Rangarajan & F Maes

  5. Laboratory for Molecular Virology and Gene Therapy, Katholieke Universiteit Leuven, Leuven, Flanders, Belgium

    J Toelen, A Ibrahimi & Z Debyser

  6. Department of Biomedical Sciences, Bio-Imaging Laboratory, University of Antwerp, Antwerp, Flanders, Belgium

    R Vreys & A Van der Linden

  7. Interdisciplinary Research Center, KU Leuven Campus Kortrijk, Kortrijk, Flanders, Belgium

    Z Debyser

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  1. G Vande Velde
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Correspondence to V Baekelandt.

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Vande Velde, G., Rangarajan, J., Toelen, J. et al. Evaluation of the specificity and sensitivity of ferritin as an MRI reporter gene in the mouse brain using lentiviral and adeno-associated viral vectors. Gene Ther 18, 594–605 (2011). https://doi.org/10.1038/gt.2011.2

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