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Application of a Rapid, Simple, and Accurate Adenovirus-Based Method to Compare PET Reporter Gene/PET Reporter Probe Systems

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Abstract

Purpose

This study aims to use a simple, quantitative method to compare the HSV1sr39TK/18 F-FHBG PET reporter gene/PET reporter probe (PRG/PRP) system with PRGs derived from human nucleoside kinases.

Procedures

The same adenovirus vector is used to express alternative PRGs. Equal numbers of vectors are injected intravenously into mice. After PRP imaging, quantitative hepatic PET signals are normalized for transduction by measuring hepatic viral genomes.

Results

The same adenovirus vector was used to express equivalent amounts of HSV1sr39TK, mutant human thymidine kinase 2 (TK2-DM), and mutant human deoxycytidine kinase (dCK-A100VTM) in mouse liver. HSV1sr39TK expression was measured with 18 F-FHBG, TK2-DM and dCK-A100VTM with 18 F-L-FMAU. TK2-DM/18 F-L-FMAU and HSV1sr39TK/18 F-FHBG had equivalent sensitivities; dCK-A100VTM/18 F-L-FMAU was twice as sensitive as HSV1sr39TK/18 F-FHBG.

Conclusions

The human PRG/PRP sensitivities are comparable and/or better than HSV1sr39TK/18 F-FHBG. However, for clinical use, identification of the best PRP substrate for each enzyme, characterization of probe distribution, and consequences of overexpressing nucleoside kinases must be evaluated.

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Acknowledgments

We thank Arthur Catapang for technical help; David Stout, Liu Wei, and Waldemar Ladno for advice and assistance with imaging experiments; and members of the Herschman, Radu, and Witte laboratories for helpful discussions. This study was funded by the National Cancer Institute In Vivo Cellular and Molecular Imaging Center (ICMIC) award P50 CA086306 (HRH), the National Cancer Institute grant 5U54 CA119347 (CGR), and the National Institute of Biomedical Imaging and Bioengineering grant 1R01CA16077001 (CGR). JG was supported by a Scholars in Oncologic Medical Imaging (SOMI) fellowship from the National Cancer Institute (Award R25T CA098010).

Conflict of interest

DOC and CR have filed a patent application for the TK2-DM/18 F-L-FMAU PRG/PRP system. The remaining authors declare they have no conflict of interest.

Author information

Authors and Affiliations

  1. Department of Biological Chemistry, David Geffen School of Medicine, UCLA, 341 Boyer Hall, 611 Charles E Young Drive, Los Angeles, CA, 90095-1570, USA

    Jose S. Gil, Hidevaldo B. Machado & Harvey R. Herschman

  2. Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, UCLA, Los Angeles, CA, USA

    Jose S. Gil, Hidevaldo B. Machado, Dean O. Campbell, Melissa McCracken, Caius Radu, Owen N. Witte & Harvey R. Herschman

  3. Department of Microbiology, Immunology and Molecular Genetics, David Geffen School of Medicine, UCLA, Los Angeles, CA, USA

    Owen N. Witte

Authors
  1. Jose S. Gil
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  2. Hidevaldo B. Machado
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  3. Dean O. Campbell
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  4. Melissa McCracken
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  5. Caius Radu
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  6. Owen N. Witte
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  7. Harvey R. Herschman
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Corresponding author

Correspondence to Harvey R. Herschman.

Additional information

Owen N. Witte is an Investigator of the Howard Hughes Medical Institute.

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Gil, J.S., Machado, H.B., Campbell, D.O. et al. Application of a Rapid, Simple, and Accurate Adenovirus-Based Method to Compare PET Reporter Gene/PET Reporter Probe Systems. Mol Imaging Biol 15, 273–281 (2013). https://doi.org/10.1007/s11307-012-0596-5

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  • DOI: https://doi.org/10.1007/s11307-012-0596-5

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