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Tracking cellular stress with labeled FMAU reflects changes in mitochondrial TK2

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European Journal of Nuclear Medicine and Molecular Imaging Aims and scope Submit manuscript

Abstract

Purpose

Fluoropyrimidines like 1-(2′-deoxy-2′-fluoro-β-d-arabinofuranosyl)-thymine (FMAU) and 3′-deoxy-3′-fluorothymidine (FLT) accumulate in tumors and are being used as positron emission tomography tumor-imaging tracers. Proliferating tissues with high thymidine kinase 1 (TK1) activity retain FLT; however, the mechanism of selective accumulation of FMAU in tumors and certain other tissues requires further study.

Methods

Retention of [3H]FLT and [3H]FMAU was measured in prostate cancer cell lines PC3, LNCaP, DU145, and the breast cancer cell line MD-MBA-231, and the tracer metabolites were analyzed by high-performance liquid chromatography (HPLC). FMAU retention, thymidine kinase 2 (TK2) activity, and mitochondrial mass were determined in cells stressed by depleted cell culture medium or by treating with oxidative, reductive, and energy stress, or specific adenosine monophosphate-activated protein kinase activator, or eIF2 inhibitor. TK1 and TK2 activities and mitochondrial mass were determined by FLT phosphorylation, 1-β-d-arabinofuranosylthymine (Ara-T) phosphorylation, and flow cytometry, respectively.

Results

FMAU retention in rapidly proliferating cancer cell lines was five to ten times lower than FLT after 10 min incubation. HPLC analysis of the cellular extracts showed that phosphorylated tracers are the main retained metabolites. Nutritional stress decreased TK1 activity and FLT retention but increased retained FMAU. TK2 inhibition decreased FMAU retention and phosphorylation with negligible effects on FLT. Oxidative, reductive, or energy stress increased FMAU retention and correlated with mitochondrial mass (r 2 = 0.88, p = 0.006). FMAU phosphorylation correlated with increased TK2 activity (r 2 = 0.87, p = 0.0002).

Conclusion

FMAU is preferably phosphorylated by TK2 and can track TK2 activity and mitochondrial mass in cellular stress. FMAU may provide an early marker of treatment effects.

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Acknowledgments

The authors want to express their deep appreciation to Mr. Eric Van Buren at Karmanos Flow Cytometry Core and Dr. Kristin L. Piwowar, who helped us with their valuable skills. This work was partially supported by funding from the National Cancer Institute CA 39566, CA 22453 and the US Department of Defense Award W81XWH-04-1-0140.

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Authors and Affiliations

  1. Karmanos Cancer Institute, Wayne State University, 4100 John R, 4 HWCRC, Detroit, MI, 48201-2013, USA

    Omid S. Tehrani, Kirk A. Douglas, Jawana M. Lawhorn-Crews & Anthony F. Shields

  2. Department of Medicine, Wayne State University, Detroit, MI, 48201, USA

    Omid S. Tehrani, Kirk A. Douglas, Jawana M. Lawhorn-Crews & Anthony F. Shields

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  1. Omid S. Tehrani
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  2. Kirk A. Douglas
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Correspondence to Anthony F. Shields.

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Tehrani, O.S., Douglas, K.A., Lawhorn-Crews, J.M. et al. Tracking cellular stress with labeled FMAU reflects changes in mitochondrial TK2. Eur J Nucl Med Mol Imaging 35, 1480–1488 (2008). https://doi.org/10.1007/s00259-008-0738-9

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  • DOI: https://doi.org/10.1007/s00259-008-0738-9

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