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Comparison of [14C]FMAU, [3H]FEAU, [14C]FIAU, and [3H]PCV for Monitoring Reporter Gene Expression of Wild Type and Mutant Herpes Simplex Virus Type 1 Thymidine Kinase in Cell Culture

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Abstract

Purpose

To assess the optimal reporter probe/reporter gene combination for monitoring herpes simplex virus type 1 thymidine kinase (HSV1-tk) gene expression, we compared the cellular uptake of 1-(2′-fluoro-2′-deoxy-d-arabinofuranosyl)-5-methyluracil (FMAU), 2′-fluoro-2′-deoxyarabinofuranosyl-5-ethyluracil (FEAU), 2′-fluoro-2′-deoxy-β-d-arabinofuranosyl-5-iodouracil (FIAU) and penciclovir (PCV) in both HSV1-tk and HSV1-sr39tk expressing cells.

Procedures

For stably transfected cell studies, C6 rat glioma cells, C6 HSV1-tk transfectant, C6 mutant HSV1-sr39tk transfectant, rat Morris hepatoma cells (MH3924A), and MH3924A HSV1-tk transfectant cells were used. For adenoviral infection studies, C6 rat glioma cells were exposed to serial titers of AdCMV–HSV1-tk, AdCMV–HSV1-sr39tk, or AdCMV–fluc for 24 hours. These cells were incubated with [14C]FMAU, [3H]FEAU, [14C]FIAU, and [3H]PCV, and cellular uptake of radioactivity was measured.

Results

[3H]FEAU exhibited the highest or second highest accumulation and the most selectivity regardless of the mode of gene transfer for both HSV1-tk and mutant HSV1-sr39tk reporter genes.

Conclusion

This combination of high accumulation and high selectivity for both HSV1-tk and HSV1-sr39tk makes suitably radiolabeled FEAU a promising candidate as a radiotracer for imaging HSV1-tk/HSV1-sr39tk gene expression in living subjects.

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Acknowledgment

We thank Dr. G. Sundaresan for technical assistance. This work funded in part by ICMIC P50 (SSG), and NIH R05 CA 082214 (SSG).

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Authors and Affiliations

  1. Department of Radiology, Bio-X Program, Stanford University, Stanford, CA, USA

    Keon Wook Kang MD, PhD, Xiaoyuan Chen PhD & Sanjiv S. Gambhir MD, PhD

  2. Department of Nuclear Medicine, National Cancer Center, Goyang, South Korea

    Keon Wook Kang MD, PhD

  3. Department of Nuclear Medicine, Chonnam National University Medical School, Gwangju, South Korea

    Jung-Joon Min MD, PhD

  4. Molecular Imaging Program at Stanford (MIPS), Stanford University, 318 Campus Drive, Stanford, CA, 94305, USA

    Sanjiv S. Gambhir MD, PhD

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  1. Keon Wook Kang MD, PhD
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  2. Jung-Joon Min MD, PhD
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  4. Sanjiv S. Gambhir MD, PhD
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Correspondence to Sanjiv S. Gambhir MD, PhD.

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Kang, K.W., Min, JJ., Chen, X. et al. Comparison of [14C]FMAU, [3H]FEAU, [14C]FIAU, and [3H]PCV for Monitoring Reporter Gene Expression of Wild Type and Mutant Herpes Simplex Virus Type 1 Thymidine Kinase in Cell Culture. Mol Imaging Biol 7, 296–303 (2005). https://doi.org/10.1007/s11307-005-0010-7

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