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Further characterisation of the cellular activity of the DNA-PK inhibitor, NU7441, reveals potential cross-talk with homologous recombination

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Cancer Chemotherapy and Pharmacology Aims and scope Submit manuscript

Abstract

Purpose

Inhibition of DNA repair is emerging as a new therapeutic strategy for cancer treatment. One promising target is DNA-PK, a pivotal kinase in double-strand break repair. The purpose of this study was to further characterise the activity of the DNA-PK inhibitor NU7441, giving some new insights into the biology of DNA-PK.

Methods

We used NU7441, a potent DNA-PK inhibitor, to evaluate potential pharmacodynamic markers of DNA-PK inhibition, inhibition of DNA repair and chemo- and radio-potentiation in isogenic human cancer cells proficient (M059-Fus1) and deficient (M059 J) in DNA-PK.

Results

NU7441 strongly inhibited DNA-PK in cell lines (IC50 = 0.3 μM) but only weakly inhibited PI3 K (IC50 = 7 μM). The only available anti-phospho-DNA-PK antibody also recognised some phosphoprotein targets of ATM. NU7441 caused doxorubicin- and IR-induced DNA DSBs (measured by γ-H2AX foci) to persist and also slightly decreased homologous recombination activity, as assessed by Rad51 foci. Chemo- and radio-potentiation were induced by NU7441 in M059-Fus-1, but not in DNA-PK-deficient M059 J cells. DNA-PK was highly expressed in a chronic lymphocytic leukaemia sample but undetectable in resting normal human lymphocytes, although it could be induced by PHA-P treatment. In K652 cells, DNA-PK expression was not related to cell cycle phase.

Conclusion

These data confirm NU7441 not only as a potent chemo- and radio-sensitiser clinical candidate but also as a powerful tool to study the biology of DNA-PK.

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Acknowledgments

We gratefully acknowledge the financial support of CR UK (NJC, CC and DRN) and AstraZeneca (MT and JMM), Newcastle Cancer Centre-Medicinal Chemistry, Newcastle University, for providing NU7441 and ZSTK474 and Dr GCM Smith, AstraZeneca for providing KU55933.

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Authors and Affiliations

  1. Northern Institute for Cancer Research, School of Medical Sciences, Newcastle University, Paul O’ Gorman Building, Framlington Place, NE2 4HH, Newcastle upon Tyne, UK

    Michele Tavecchio, Joanne M. Munck, Celine Cano, David R. Newell & Nicola J. Curtin

Authors
  1. Michele Tavecchio
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  2. Joanne M. Munck
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  3. Celine Cano
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  4. David R. Newell
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  5. Nicola J. Curtin
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Corresponding author

Correspondence to Nicola J. Curtin.

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Tavecchio, M., Munck, J.M., Cano, C. et al. Further characterisation of the cellular activity of the DNA-PK inhibitor, NU7441, reveals potential cross-talk with homologous recombination. Cancer Chemother Pharmacol 69, 155–164 (2012). https://doi.org/10.1007/s00280-011-1662-4

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  • DOI: https://doi.org/10.1007/s00280-011-1662-4

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