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Small molecule inhibitors of ezrin inhibit the invasive phenotype of osteosarcoma cells

Oncogene volume 31, pages 269–281 (2012)Cite this article

Abstract

Ezrin is a multifunctional protein that connects the actin cytoskeleton to the extracellular matrix through transmembrane proteins. High ezrin expression is associated with lung metastasis and poor survival in cancer. We screened small molecule libraries for compounds that directly interact with ezrin protein using surface plasmon resonance to identify lead compounds. The secondary functional assays used for lead compound selection included ezrin phosphorylation as measured by immunoprecipitation and in vitro kinase assays, actin binding, chemotaxis, invasion into an endothelial cell monolayer, zebrafish and Xenopus embryonic development, mouse lung organ culture and an in vivo lung metastasis model. Two molecules, NSC305787 and NSC668394, that directly bind to ezrin with low micromolar affinity were selected based on inhibition of ezrin function in multiple assays. They inhibited ezrin phosphorylation, ezrin–actin interaction and ezrin-mediated motility of osteosarcoma (OS) cells in culture. NSC305787 mimicked the ezrin morpholino phenotype, and NSC668394 caused a unique developmental defect consistent with reduced cell motility in zebrafish. Following tail vein injection of OS cells into mice, both molecules inhibited lung metastasis of ezrin-sensitive cells, but not ezrin-resistant cells. The small molecule inhibitors NSC305787 and NSC668394 demonstrate a novel targeted therapy that directly inhibits ezrin protein as an approach to prevent tumor metastasis.

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Acknowledgements

Support for our work came from the Children's Cancer Foundation of Baltimore (AÜ), US Department of Defense (W81XWH-10-1-0137, AÜ), Brandon Carrington Lee Foundation (G.K., AÜ), Go4theGoal, Dani's Foundation, Alex's Lemonade Stand Foundation, Liddy Shriver Sarcoma Initiative, Burroughs Wellcome Clinical Scientist Award in Translational Research (JT), and the NIH R01CA133662 (JT), R01CA138212 (JT) and Cancer Center Support Grant P30 CA051008 for use of Biacore Molecular Interaction Shared Resource. We would like to thank to Dr Anthony Bretscher (Cornell University) for providing the WT Ezrin construct, Developmental Therapeutics Program, NCI for providing the small molecule libraries.

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Authors and Affiliations

  1. Georgetown University Medical Center, Lombardi Comprehensive Cancer Center, Washington, DC, USA

    G Bulut, K Chen, E M Beauchamp, S Rahim, G W Kosturko, E Glasgow, S Dakshanamurthy, J A Toretsky & A Üren

  2. Tumor and Metastasis Biology Section, Center for Cancer Research, National Cancer Institute, Bethesda, MD, USA

    S-H Hong & C Khanna

  3. Laboratory of Cell and Developmental Signaling, National Cancer Institute-Frederick, Frederick, MD, USA

    H-S Lee & I Daar

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  1. G Bulut
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Correspondence to A Üren.

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Bulut, G., Hong, SH., Chen, K. et al. Small molecule inhibitors of ezrin inhibit the invasive phenotype of osteosarcoma cells. Oncogene 31, 269–281 (2012). https://doi.org/10.1038/onc.2011.245

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