Abstract
Biodistribution, magnitude and duration of a therapeutic transgene's expression may be assessed by linking it to the expression of a positron emission tomography (PET) reporter gene (PRG) and then imaging the PRG's expression by a PET reporter probe (PRP) in living animals. We validate the simple approach of co-administering two distinct but otherwise identical adenoviruses, one expressing a therapeutic transgene and the other expressing the PRG, to track the therapeutic gene's expression. Two PET reporter genes, a mutant herpes simplex virus type 1 thymidine kinase (HSV1-sr39tk) and dopamine-2 receptor (D2R), each regulated by the same cytomegalovirus (CMV) promoter, have been inserted into separate adenoviral vectors (Ad). We demonstrate that cells co-infected with equivalent titers of Ad-CMV-HSV1-sr39tk and Ad-CMV-D2R express both reporter genes with good correlation (r2 = 0.93). Similarly, a high correlation (r2 = 0.97) was observed between the expression of both PRGs in the livers of mice co-infected via tail-vein injection with equivalent titers of these two adenoviruses. Finally, microPET imaging of HSV1-sr39tk and D2R expression with 9-(4-[18F]fluoro-3-hydroxymethylbutyl) guanine ([18F]FHBG) and 3-(2-[18F]fluoroethyl)spiperone ([18F]FESP), utilizing several adenovirus-mediated delivery routes, illustrates the feasibility of evaluating relative levels of transgene expression in living animals, using this approach.
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Acknowledgements
We thank the members of the UCLA Gene Imaging Consortium, K Nguyen, E Bauer and Y Yu for various technical assistance, R Sumida, W Ladno, J Edwards, DJ Liu and V Dominguez for assistance with microPET imaging, M Ho for the synthesis of [18F]FESP and [18F]FHBG and the UCLA cyclotron crew for outstanding support. We are grateful for J Matherly's assistance with adenovirus replication. We thank A Green for helpful discussions. Grants: this work was partially supported by funding from NIH RO1 CA82214–01, DOE contract DE-FC03–87ER60615, University of California Biotechnology Grant, Dana Foundation, and the UCLA-Jonsson Comprehensive Cancer Center.
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Yaghoubi, S., Wu, L., Liang, Q. et al. Direct correlation between positron emission tomographic images of two reporter genes delivered by two distinct adenoviral vectors. Gene Ther 8, 1072–1080 (2001). https://doi.org/10.1038/sj.gt.3301490
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DOI: https://doi.org/10.1038/sj.gt.3301490
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