TRPM8 inhibits endothelial cell migration via a non-channel function by trapping the small GTPase Rap1

J Cell Biol. 2017 Jul 3;216(7):2107-2130. doi: 10.1083/jcb.201506024. Epub 2017 May 26.

Abstract

Endothelial cell adhesion and migration are critical steps of the angiogenic process, whose dysfunction is associated with tumor growth and metastasis. The TRPM8 channel has recently been proposed to play a protective role in prostate cancer by impairing cell motility. However, the mechanisms by which it could influence vascular behavior are unknown. Here, we reveal a novel non-channel function for TRPM8 that unexpectedly acts as a Rap1 GTPase inhibitor, thereby inhibiting endothelial cell motility, independently of pore function. TRPM8 retains Rap1 intracellularly through direct protein-protein interaction, thus preventing its cytoplasm-plasma membrane trafficking. In turn, this mechanism impairs the activation of a major inside-out signaling pathway that triggers the conformational activation of integrin and, consequently, cell adhesion, migration, in vitro endothelial tube formation, and spheroid sprouting. Our results bring to light a novel, pore-independent molecular mechanism by which endogenous TRPM8 expression inhibits Rap1 GTPase and thus plays a critical role in the behavior of vascular endothelial cells by inhibiting migration.

Publication types

  • Video-Audio Media
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Adhesion
  • Cell Movement*
  • Endothelial Cells / enzymology*
  • HEK293 Cells
  • Human Umbilical Vein Endothelial Cells / enzymology
  • Humans
  • Integrin beta1 / metabolism
  • Microscopy, Confocal
  • Microscopy, Fluorescence
  • Microscopy, Video
  • Models, Cardiovascular
  • Neovascularization, Physiologic*
  • Protein Binding
  • Protein Transport
  • RNA Interference
  • Signal Transduction
  • TRPM Cation Channels / genetics
  • TRPM Cation Channels / metabolism*
  • Time Factors
  • Transfection
  • rap1 GTP-Binding Proteins / genetics
  • rap1 GTP-Binding Proteins / metabolism*

Substances

  • Integrin beta1
  • TRPM Cation Channels
  • TRPM8 protein, human
  • rap1 GTP-Binding Proteins

Associated data

  • GENBANK/AY328400