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.

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

Genova, Tullio;Camillo, Chiara;Bernardini, Michela;Scianna, Marco;Valdembri, Donatella;Munaron, Luca Maria;Serini, Guido;Fiorio Pla, Alessandra
Co-last
2017-01-01

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.
2017
216
7
2107
2130
EMTREE drug terms: guanosine triphosphataseintegrinRap1 proteintransient receptor potential channel M8beta1 integrinprotein bindingRap1 proteintransient receptor potential channel MTRPM8 protein, human EMTREE medical terms: Articlecell adhesioncell membranecell migrationconformational transitioncontrolled studycytoplasmendothelium cellhumanhuman cellmembrane transportpriority journalprotein expressionprotein functionprotein protein interactionsignal transductionangiogenesisbiological modelcell motionconfocal microscopyendothelium cellenzymologyfluorescence microscopygenetic transfectiongeneticsHEK293 cell linemetabolismprotein transportRNA interferencetime factorumbilical vein endothelial cellvideo microscopy MeSH: Antigens, CD29Cell AdhesionCell MovementEndothelial CellsHEK293 CellsHuman Umbilical Vein Endothelial CellsHumansMicroscopy, ConfocalMicroscopy, FluorescenceMicroscopy, VideoModels, CardiovascularNeovascularization, PhysiologicProtein BindingProtein Transportrap1 GTP-Binding ProteinsRNA InterferenceSignal TransductionTime FactorsTransfectionTRPM Cation Channels
Genova, Tullio; Grolez, Guillaume P; Camillo, Chiara; Bernardini, Michela; Bokhobza, Alexandre; Richard, Elodie; Scianna, Marco; Lemonnier, Loic; Valdembri, Donatella; Munaron, Luca Maria; Philips, Mark R; Mattot, Virginie; Serini, Guido; Prevarskaya, Natalia; Gkika, Dimitra; FIORIO PLA, Alessandra
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2318/1639774
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