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

Oncogene. 2012 Jan 19;31(3):269-81. doi: 10.1038/onc.2011.245. Epub 2011 Jun 27.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / antagonists & inhibitors
  • Adamantane / analogs & derivatives*
  • Adamantane / chemistry
  • Adamantane / pharmacology
  • Animals
  • Antineoplastic Agents / chemistry
  • Antineoplastic Agents / pharmacology*
  • Bone Neoplasms / pathology*
  • Cell Line, Tumor
  • Cell Movement / drug effects
  • Cytoskeletal Proteins / antagonists & inhibitors*
  • Female
  • Lung Neoplasms / secondary*
  • Mice
  • Mice, Inbred BALB C
  • Neoplasm Invasiveness / pathology
  • Organ Culture Techniques
  • Osteosarcoma / metabolism
  • Osteosarcoma / secondary*
  • Phenols / chemistry
  • Phenols / pharmacology*
  • Phosphorylation / drug effects
  • Quinolines / chemistry
  • Quinolines / pharmacology*
  • Quinolones / chemistry
  • Quinolones / pharmacology*
  • Surface Plasmon Resonance
  • Xenopus
  • Zebrafish

Substances

  • Actins
  • Antineoplastic Agents
  • Cytoskeletal Proteins
  • NSC305787
  • NSC668394
  • Phenols
  • Quinolines
  • Quinolones
  • ezrin
  • Adamantane