title: Canonical transient receptor potential 3 channel triggers vascular endothelial growth factor-induced intracellular Ca2+ oscillations in endothelial progenitor cells isolated from umbilical cord blood.
abstract: Endothelial colony-forming cells (ECFCs) are the only endothelial progenitor cells (EPCs) that are capable of acquiring a mature endothelial phenotype. ECFCs are mainly mobilized from bone marrow to promote vascularization and represent a promising tool for cell-based therapy of severe ischemic diseases. Vascular endothelial growth factor (VEGF) stimulates the proliferation of peripheral blood-derived ECFCs (PB-ECFCs) through oscillations in intracellular Ca(2+) concentration ([Ca(2+)]i). VEGF-induced Ca(2+) spikes are driven by the interplay between inositol-1,4,5-trisphosphate (InsP3)-dependent Ca(2+) release and store-operated Ca(2+) entry (SOCE). The therapeutic potential of umbilical cord blood-derived ECFCs (UCB-ECFCs) has also been shown in recent studies. However, VEGF-induced proliferation of UCB-ECFCs is faster compared with their peripheral counterpart. Unlike PB-ECFCs, UCB-ECFCs express canonical transient receptor potential channel 3 (TRPC3) that mediates diacylglycerol-dependent Ca(2+) entry. The present study aimed at investigating whether the higher proliferative potential of UCB-ECFCs was associated to any difference in the molecular underpinnings of their Ca(2+) response to VEGF. We found that VEGF induces oscillations in [Ca(2+)]i that are patterned by the interaction between InsP3-dependent Ca(2+) release and SOCE. Unlike PB-ECFCs, VEGF-evoked Ca(2+) oscillations do not arise in the absence of extracellular Ca(2+) entry and after pharmacological (with Pyr3 and flufenamic acid) and genetic (by employing selective small interference RNA) suppression of TRPC3. VEGF-induced UCB-ECFC proliferation is abrogated on inhibition of the intracellular Ca(2+) spikes. Therefore, the Ca(2+) response to VEGF in UCB-ECFCs is shaped by a different Ca(2+) machinery as compared with PB-ECFCs, and TRPC3 stands out as a promising target in EPC-based treatment of ischemic pathologies.
authors.affiliation: 1 Department of Biology and Biotechnology "Lazzaro Spallanzani,", University of Pavia , Pavia, Italy .
authors.forename: Silvia
authors.initials: S
authors.lastname: Dragoni
authors.forename: Umberto
authors.initials: U
authors.lastname: Laforenza
authors.forename: Elisa
authors.initials: E
authors.lastname: Bonetti
authors.forename: Francesco
authors.initials: F
authors.lastname: Lodola
authors.forename: Cinzia
authors.initials: C
authors.lastname: Bottino
authors.forename: Germano
authors.initials: G
authors.lastname: Guerra
authors.forename: Alessandro
authors.initials: A
authors.lastname: Borghesi
authors.forename: Mauro
authors.initials: M
authors.lastname: Stronati
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: D000328:Adult; D000893:Anti-Inflammatory Agents / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D049109:Cell Proliferation; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000378:metabolism; D005260:Female; D005312:Fetal Blood / Q000166:cytology / Q000378:metabolism; D005439:Flufenamic Acid / Q000494:pharmacology; D006801:Humans; D015544:Inositol 1,4,5-Trisphosphate / Q000378:metabolism; D017136:Ion Transport / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D008875:Middle Aged; D018919:Neovascularization, Physiologic; D011720:Pyrazoles / Q000494:pharmacology; D034622:RNA Interference; D034741:RNA, Small Interfering; D015398:Signal Transduction / Q000187:drug effects; D013234:Stem Cells / Q000378:metabolism*; D050052:TRPC Cation Channels / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*; D055815:Young Adult
chemical_list: D000893:Anti-Inflammatory Agents; D011720:Pyrazoles; D034741:RNA, Small Interfering; D050052:TRPC Cation Channels; C104603:TRPC3 cation channel; D042461:Vascular Endothelial Growth Factor A; C559943:ethyl-1-(4-(2*3*3-trichloroacrylamide)phenyl)-5-(trifluoromethyl)-1H-pyrazole-4-carboxylate; D005439:Flufenamic Acid; D015544:Inositol 1,4,5-Trisphosphate; D002118:Calcium