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An shRNA barcode screen provides insight into cancer cell vulnerability to MDM2 inhibitors

Abstract

The identification of the cellular targets of small molecules with anticancer activity is crucial to their further development as drug candidates. Here, we present the application of a large-scale RNA interference–based short hairpin RNA (shRNA) barcode screen to gain insight in the mechanism of action of nutlin-3 (1). Nutlin-3 is a small-molecule inhibitor of MDM2, which can activate the p53 pathway. Nutlin-3 shows strong antitumor effects in mice, with surprisingly few side effects on normal tissues1. Aside from p53, we here identify 53BP1 as a critical mediator of nutlin-3–induced cytotoxicity. 53BP1 is part of a signaling network induced by DNA damage that is frequently activated in cancer but not in healthy tissues2. Our results suggest that nutlin-3's tumor specificity may result from its ability to turn a cancer cell–specific property (activated DNA damage signaling3) into a weakness that can be exploited therapeutically.

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Figure 1: shRNA barcode screen identifies 53BP1 as a mediator of the antiproliferative effects of nutlin-3.
Figure 2: 53BP1 modulates the cellular cytotoxicity induced by nutlin-3 in MCF-7 cells.
Figure 3: Intrinsic DNA damage signaling contributes to the cytotoxic effect of nutlin-3 in MCF-7 cells.
Figure 4: The cytotoxic effect of nutlin-3 in tumor cells is enhanced by intrinsic DNA damage signaling.

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Acknowledgements

We thank the Netherlands Cancer Institute microarray facility group for assistance; K. Berns, S. Nijman, H. Ovaa and members of the Bernards and Beijersbergen laboratories for help and discussions; L. Oomen for help with confocal microscopy; and R. Kortlever for critically reading the manuscript. This work was supported by grants from the Dutch Cancer Society (KWF) and the European Union integrated project INTACT.

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Correspondence to René Bernards or Roderick L Beijersbergen.

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Brummelkamp, T., Fabius, A., Mullenders, J. et al. An shRNA barcode screen provides insight into cancer cell vulnerability to MDM2 inhibitors. Nat Chem Biol 2, 202–206 (2006). https://doi.org/10.1038/nchembio774

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