title: Store-operated Ca(2+) entry is expressed in human endothelial progenitor cells.
abstract: Endothelial progenitor cells (EPCs) may be recruited from the bone marrow to sites of tissue regeneration to sustain neovascularization and reendothelialization after acute vascular injury. This feature makes them particularly suitable for cell-based therapy. In mature endothelium, store-operated Ca(2+) entry (SOCE) is activated following emptying of inositol-1,4,5-trisphosphate-sensitive stores, and controls a wide number of functions, including proliferation, nitric oxide synthesis, and vascular permeability. The present work aimed at investigating SOCE expression in EPCs harvested from both peripheral blood (PB-EPCs) and umbilical cord blood (UCB-EPCs) by employing both Ca(2+) imaging and molecular biology techniques. SOCE was induced upon either pharmacological (ie, cyclopiazonic acid) or physiological (ie, ATP) depletion of the intracellular Ca(2+) pool. Further, store-dependent Ca(2+) entry was inhibited by the SOCE inhibitor, N-(4-[3,5-bis(trifluoromethyl)-1H-pyrazol-1-yl]phenyl)-4-methyl-1,2,3-thiadiazole-5-carboxamide (BTP-2). Real-time reverse transcription-polymerase chain reaction and western blot analyses showed that both PB-EPCs and UCB-EPCs express all the molecular candidates to mediate SOCE in differentiated cells, including TRPC1, TRPC4, Orai1, and Stim1. Moreover, pharmacological maneuvers demonstrated that, as well as in differentiated endothelial cells, the signal transduction pathway leading to depletion of the intracellular Ca(2+) pool impinged on the phospholipase C/inositol-1,4,5-trisphosphate pathway. Finally, blockage of SOCE with BTP-2 impaired PB-EPC proliferation. These findings provide the first evidence that EPCs express SOCE, which might thus be regarded as a novel target to enhance the regenerative outcome of cell-based therapy.
authors.affiliation: Department of Physiology, University of Pavia, Pavia, Italy.
authors.forename: Yuly
authors.initials: Y
authors.lastname: Sánchez-Hernández
authors.forename: Umberto
authors.initials: U
authors.lastname: Laforenza
authors.forename: Elisa
authors.initials: E
authors.lastname: Bonetti
authors.forename: Jacopo
authors.initials: J
authors.lastname: Fontana
authors.forename: Silvia
authors.initials: S
authors.lastname: Dragoni
authors.forename: Marika
authors.initials: M
authors.lastname: Russo
authors.forename: José Everardo
authors.initials: JE
authors.lastname: Avelino-Cruz
authors.forename: Sergio
authors.initials: S
authors.lastname: Schinelli
authors.forename: Domenico
authors.initials: D
authors.lastname: Testa
authors.forename: Germano
authors.initials: G
authors.lastname: Guerra
authors.forename: Vittorio
authors.initials: V
authors.lastname: Rosti
authors.forename: Franco
authors.initials: F
authors.lastname: Tanzi
authors.forename: Francesco
authors.initials: F
authors.lastname: Moccia
mesh_terms: D000813:Anilides / Q000378:metabolism / Q000494:pharmacology; D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000235:genetics / Q000378:metabolism*; D042783:Endothelial Cells / Q000166:cytology* / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D015544:Inositol 1,4,5-Trisphosphate / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism*; D013830:Thiadiazoles / Q000378:metabolism / Q000494:pharmacology; D010738:Type C Phospholipases / Q000378:metabolism; D014470:Umbilical Cord / Q000166:cytology
chemical_list: C476308:4-methyl-4'-(3,5-bis(trifluoromethyl)-1H-pyrazol-1-yl)-1,2,3-thiadiazole-5-carboxanilide; D000813:Anilides; D015220:Calcium Channels; D013830:Thiadiazoles; D015544:Inositol 1,4,5-Trisphosphate; D010738:Type C Phospholipases; D002118:Calcium