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Intracellular disposition of fludarabine triphosphate in human natural killer cells

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Abstract

Purpose

Fludarabine is a key component of several reduced-intensity conditioning regimens for hematopoietic cell transplantation (HCT). Shortly after reduced-intensity conditioning, the percent of donor natural killer (NK) cells has been associated with progression-free survival. Insufficient suppression of the recipient’s NK cells by fludarabine may lead to lower donor chimerism; however, the effect of fludarabine upon NK cells is poorly understood. Thus, in purified human NK cells we evaluated the uptake and activation of fludarabine to its active metabolite, fludarabine triphosphate (F-ara-ATP), and assessed the degree of interindividual variability in F-ara-ATP accumulation.

Methods

Intracellular F-ara-ATP was measured in purified NK cells isolated from healthy volunteers (n = 6) after ex vivo exposure to fludarabine. Gene expression levels of the relevant transporters and enzymes involved in fludarabine uptake and activation were also measured in these cells.

Results

F-ara-ATP accumulation (mean ± SD) was 6.00 ± 3.67 pmol/1 × 106 cells/4 h, comparable to average levels previously observed in CD4+ and CD8+ T-lymphocytes. We observed considerable variability in F-ara-ATP accumulation and mRNA expression of transporters and enzymes relevant to F-ara-ATP accumulation in NK cells from different healthy volunteers.

Conclusions

Human NK cells have the ability to form F-ara-ATP intracellularly and large interindividual variability was observed in healthy volunteers. Further studies are needed to evaluate whether F-ara-ATP accumulation in NK cells are associated with apoptosis and clinical outcomes.

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Fig. 1

Abbreviations

NK:

Natural killer

HCT:

Hematopoietic cell transplantation

F-ara-ATP:

Fludarabine triphosphate

NBMPR:

Nitrobenzylthioinosine

CD73:

Ecto-5′-nucleotidase

hENT:

Human equilibrative nucleoside transporter

hCNT:

Human concentrative nucleoside transporter

dCK:

Deoxycytidine kinase

CN-II:

5′-nucleotidase

dNT-1:

Deoxynucleotidase-1

AK:

Adenylate kinase

NDK:

Nucleoside diphosphate kinase

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Acknowledgments

The analytical expertise of Brian Phillips and Linda Risler are greatly appreciated. Supported by grants from the National Institutes of Health (CA18029, CA15704, CA78902, DK56465, HL36444, HL91744). ELW was supported by the Elmer M. and Joy B. Plein Fellowship for Excellence in Pharmacy Education, School of Pharmacy, Seattle, WA.

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Author notes

  1. Erica L. Woodahl

    Present address: Department of Biomedical and Pharmaceutical Sciences, University of Montana, Missoula, MT, USA

Authors and Affiliations

  1. Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, WA, USA

    Erica L. Woodahl, Joanne Wang, Shelly Heimfeld, Brenda M. Sandmaier & Jeannine S. McCune

  2. School of Pharmacy, University of Washington, Box 357630, Seattle, WA, 98195, USA

    Erica L. Woodahl, Joanne Wang & Jeannine S. McCune

  3. School of Medicine, University of Washington, Seattle, WA, USA

    Brenda M. Sandmaier

Authors
  1. Erica L. Woodahl
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  2. Joanne Wang
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  3. Shelly Heimfeld
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  4. Brenda M. Sandmaier
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  5. Jeannine S. McCune
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Corresponding author

Correspondence to Jeannine S. McCune.

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Woodahl, E.L., Wang, J., Heimfeld, S. et al. Intracellular disposition of fludarabine triphosphate in human natural killer cells. Cancer Chemother Pharmacol 63, 959–964 (2009). https://doi.org/10.1007/s00280-008-0829-0

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  • DOI: https://doi.org/10.1007/s00280-008-0829-0

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