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Tumor-specific imaging through progression elevated gene-3 promoter-driven gene expression

Abstract

Molecular-genetic imaging is advancing from a valuable preclinical tool to a guide for patient management. The strategy involves pairing an imaging reporter gene with a complementary imaging agent in a system that can be used to measure gene expression or protein interaction or track gene-tagged cells in vivo. Tissue-specific promoters can be used to delineate gene expression in certain tissues, particularly when coupled with an appropriate amplification mechanism. Here we show that the progression elevated gene-3 (PEG-3) promoter, derived from a rodent gene mediating tumor progression and metastatic phenotypes, can be used to drive imaging reporters selectively to enable detection of micrometastatic disease in mouse models of human melanoma and breast cancer using bioluminescence and radionuclide-based molecular imaging techniques. Because of its strong promoter activity, tumor specificity and capacity for clinical translation, PEG-3 promoter–driven gene expression may represent a practical, new system for facilitating cancer imaging and therapy.

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Figure 1: Cancer-specific PEG-3 promoter activity shown by bioluminescence imaging in experimental metastasis models of human melanoma (Mel) and breast cancer (BCa).
Figure 2: Correlation between PEG-3 promoter–driven Luc expression and microscopic metastatic sites shown by histopathological analysis in a mouse model of melanoma metastasis.
Figure 3: Histological confirmation of the microscopic metastatic sites detected by the PEG-3 promoter–driven bioluminescence imaging system in a human breast cancer metastasis model.
Figure 4: Cancer-specific expression of HSV1-TK driven by PEG-3 promoter shown by SPECT-CT imaging in an experimental model of human melanoma metastasis.
Figure 5: Detection and localization of metastatic masses by SPECT-CT imaging after the systemic administration of pPEG-HSV1tk.

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Acknowledgements

We appreciate the technical support provided by S. Dhara, S. Nimmagadda, M. Goggins and M. Griffith. We are grateful to J. Fox and G. Green for providing [125I]FIAU and assisting in the SPECT-CT and PET-CT imaging studies. We also thank C. Endres, B. Tsui, J. Yu and J. Fox for help with SPECT and PET data processing. The MDA-MB-231 cell line and pCMV-Tri construct were generous gifts from Z. Bhujwalla (Johns Hopkins University) and S. Gambhir (Stanford University), respectively. Funding was provided by US National Institutes of Health grant U24 CA92871 (to M.G.P.), by the Predoctoral Molecular Imaging Scholar Program from the Society of Nuclear Medicine and the Korea Science and Engineering Foundation Fellowship Program (to H.C.B.) and by the US National Institutes of Health grant P01 CA104177 and the US National Foundation for Cancer Research (to P.B.F.). P.B.F. holds the Thelma Newmeyer Corman Chair in Cancer Research at the Virginia Commonwealth University Massey Cancer Center.

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Authors

Contributions

H.-e.C.B. designed and performed the experiments, analyzed data and prepared the manuscript. K.L.G. provided technical support in histopathology. J.L., K.L.G. and P.B.F. gave conceptual advice and edited the manuscript. M.G.P. and P.B.F. conceived of the project. M.G.P. supervised the project and prepared the manuscript.

Corresponding author

Correspondence to Martin G Pomper.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Methods and Supplementary Figures 1–8 (PDF 1603 kb)

Supplementary Video 1

Detection of metastatic melanoma after systemic administration of pPEG-HSV1tk/PEI polyplex using SPECT-CT. Movie of a representative melanoma metastasis model, Mel-2 from Figure 4b and Figure 5a,b. The image was acquired at 24 h post injection of [125I]FIAU (1.4 mCi), which was 70 h after the pPEG-HSV1tk delivery. Multiple metastatic sites are predicted in the lung and upper dorsal area of the animal by [125I]FIAU SPECT. Melanoma masses were confirmed under the brown adipose tissue in the corresponding area (Fig. 5b) as well as in its lung by the gross pathological analysis. (MOV 3009 kb)

Supplementary Video 2

Movie of a representative control animal, Ctrl-3 from Figure 4a. This whole body SPECT-CT image was acquired at 24 h post injection of 1.4 mCi of [125I]FIAU, which was 70 h after the IV injection of pPEG-HSV1tk/PEI polyplex. Accumulated radioactivity was only detected in the urinary bladder and intestines of the animal. The same pseudo color scale used for Mel-2 in Supplementary Video 1 was applied. (MOV 2835 kb)

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Bhang, He., Gabrielson, K., Laterra, J. et al. Tumor-specific imaging through progression elevated gene-3 promoter-driven gene expression. Nat Med 17, 123–129 (2011). https://doi.org/10.1038/nm.2269

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