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A cell-based screen identifies ATR inhibitors with synthetic lethal properties for cancer-associated mutations

Nature Structural & Molecular Biology volume 18, pages 721–727 (2011)Cite this article

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Abstract

Oncogene activation has been shown to generate replication-born DNA damage, also known as replicative stress. The primary responder to replicative stress is not Ataxia-Telangiectasia Mutated (ATM) but rather the kinase ATM and Rad3-related (ATR). One limitation for the study of ATR is the lack of potent inhibitors. We here describe a cell-based screening strategy that has allowed us to identify compounds with ATR inhibitory activity in the nanomolar range. Pharmacological inhibition of ATR generates replicative stress, leading to chromosomal breakage in the presence of conditions that stall replication forks. Moreover, ATR inhibition is particularly toxic for p53-deficient cells, this toxicity being exacerbated by replicative stress–generating conditions such as the overexpression of cyclin E. Notably, one of the compounds we identified is NVP-BEZ235, a dual phosphatidylinositol-3-OH kinase (PI3K) and mTOR inhibitor that is being tested for cancer chemotherapy but that we now show is also very potent against ATM, ATR and the catalytic subunit of DNA-dependent protein kinase (DNA-PKcs).

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Figure 1: Screening strategy for the identification of ATR inhibitors.
Figure 2: Effect of ATRi and DDRi on the G2/M checkpoint.
Figure 3: ATM-, ATR- and DNA-PKcs-dependent phosphorylation in vivo.
Figure 4: ATRi and DDRi promote the breakage of stalled replication forks.
Figure 5: Inhibition of ATR generates replicative stress.
Figure 6: Synthetic lethality of ATR inhibition with cyclin E overexpression and/or p53 loss.

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Change history

  • 15 May 2011

    In the version of this article initially published online, the structures of the compounds in Figure 2c were incorrect. The error has been corrected for all versions of this article.

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Acknowledgements

We thank M. Barbacid (Spanish National Cancer Research Centre) for providing reagents, M. Serrano for critical comments on the manuscript, and M.I. Albarran and P. Alfonso for help with the kinase assays. We also thank J.M. Silva (Irving Cancer Research Centre, Columbia University) for providing reagents. M.M. is supported by a grant from Fondo de Investigaciones Sanitarias (PI05945). Work in O.F.-C's laboratory is supported by grants from the Spanish Ministry of Science (CSD2007-00017 and SAF2008-01596), a Pfizer Foundation Award, the European Molecular Biology Organization Young Investigator Programme and the European Research Council (ERC-210520).

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Author notes

  1. Julen Oyarzabal

    Present address: Present address: Center for Applied Medical Research, University of Navarra, Pamplona, Spain.,

Authors and Affiliations

  1. Genomic Instability Group, Spanish National Cancer Research Centre, Madrid, Spain

    Luis I Toledo, Matilde Murga, Rafal Zur, Rebeca Soria & Oscar Fernandez-Capetillo

  2. Experimental Therapeutics Programme, Spanish National Cancer Research Centre, Madrid, Spain

    Antonio Rodriguez, Sonia Martinez, Julen Oyarzabal, Joaquin Pastor & James R Bischoff

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Contributions

O.F.-C. designed the study and experiments and wrote the paper. L.I.T. conducted most of the experiments presented. M.M. helped in the work with oncogenes and CDC25A. R.Z. and R.S. provided technical help. J.O., A.R., S.M., J.P. and J.R.B. provided the chemicals and helped in the development of the small molecule screening.

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Correspondence to Oscar Fernandez-Capetillo.

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Toledo, L., Murga, M., Zur, R. et al. A cell-based screen identifies ATR inhibitors with synthetic lethal properties for cancer-associated mutations. Nat Struct Mol Biol 18, 721–727 (2011). https://doi.org/10.1038/nsmb.2076

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