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A new transgene reporter for in vivo magnetic resonance imaging

Nature Medicine volume 11, pages 450–454 (2005)Cite this article

Abstract

We report a new platform technology for visualizing transgene expression in living subjects using magnetic resonance imaging (MRI). Using a vector, we introduced an MRI reporter, a metalloprotein from the ferritin family, into specific host tissues. The reporter is made superparamagnetic as the cell sequesters endogenous iron from the organism. In this new approach, the cells construct the MRI contrast agent in situ using genetic instructions introduced by the vector. No exogenous metal-complexed contrast agent is required, thereby simplifying intracellular delivery. We used a replication-defective adenovirus vector to deliver the ferritin transgenes. Following focal inoculation of the vector into the mouse brain, we monitored the reporter activity using in vivo time-lapse MRI. We observed robust contrast in virus-transduced neurons and glia for several weeks. This technology is adaptable to monitor transgene expression in vivo in many tissue types and has numerous biomedical applications, such as visualizing preclinical therapeutic gene delivery.

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Figure 1: Time course of AdV-FT expression in A549 cells.
Figure 2: In vitro 1/T2 in pelleted A549 cells.
Figure 3: In vitro cytotoxicity assays in A549 cells show no adverse effects resulting from AdV-FT expression.
Figure 4: In vivo longitudinal results of MRI reporter expression in the mouse brain.
Figure 5: Immunohistochemistry results showing ferritin expression in mouse brain.

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Acknowledgements

We thank C. Robison, J. Horner and K. Hitchens for their assistance. Also, we thank P. Arosio (University of Brescia, Italy) for the ferritin cDNA and P. Santambrogio (San Raffaele Scientific Institute, Milan, Italy) for the rH02 antibody. This work was funded by the Pittsburgh Life Sciences Greenhouse and the US National Institutes of Health (R01-EB003453, P41-EB001977 and P50-AR049617).

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Authors and Affiliations

  1. Department of Biological Sciences and Pittsburgh NMR Center for Biomedical Research, Carnegie Mellon University, 4400 Fifth Avenue, Pittsburgh, 15213, Pennsylvania, USA

    Guillem Genove, Ulrike DeMarco, Hongyan Xu & Eric T Ahrens

  2. Department of Molecular Genetics and Biochemistry, University of Pittsburgh School of Medicine, 200 Lothrop Street, Pittsburgh, 15213, Pennsylvania, USA

    William F Goins

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  1. Guillem Genove
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  2. Ulrike DeMarco
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  3. Hongyan Xu
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  4. William F Goins
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  5. Eric T Ahrens
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Correspondence to Eric T Ahrens.

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Supplementary information

Supplementary Fig. 1

In vitro uptake kinetics of 59Fe-enriched human transferring in A549 cells. (PDF 91 kb)

Supplementary Fig. 2

TfR-1 levels in AdV-FT transduced and control A549 cells. (PDF 90 kb)

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Genove, G., DeMarco, U., Xu, H. et al. A new transgene reporter for in vivo magnetic resonance imaging. Nat Med 11, 450–454 (2005). https://doi.org/10.1038/nm1208

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