title: Constitutive Store-Operated Ca(2+) Entry Leads to Enhanced Nitric Oxide Production and Proliferation in Infantile Hemangioma-Derived Endothelial Colony-Forming Cells.
abstract: Clonal endothelial progenitor cells (EPCs) have been implicated in the aberrant vascular growth that features infantile hemangioma (IH), the most common benign vascular tumor in childhood that may cause ulceration, bleeding, and/or permanent disfigurement. Endothelial colony-forming cells (ECFCs), truly endothelial EPCs endowed with clonal ability and capable of forming patent vessels in vivo, remodel their Ca(2+) toolkit in tumor-derived patients to acquire an adaptive advantage. Particularly, they upregulate the proangiogenic store-operated Ca(2+) entry (SOCE) pathway due to the overexpression of its underlying components, that is, stromal interaction molecule 1 (Stim1), Orai1, and transient receptor potential canonical 1 (TRPC1). The present work was undertaken to assess whether and how the Ca(2+) signalosome is altered in IH-ECFCs by employing Ca(2+) and nitric oxide (NO) imaging, real-time polymerase chain reaction, western blotting, and functional assays. IH-ECFCs display a lower intracellular Ca(2+) release in response to either pharmacological (i.e., cyclopiazonic acid) or physiological (i.e., ATP and vascular endothelial growth factor) stimulation. Conversely, Stim1, Orai1, and TRPC1 transcripts and proteins are normally expressed in these cells and mediate a constitutive SOCE, which is sensitive to BTP-2, La(3+), and Pyr6 and recharges the intracellular Ca(2+) pool. The resting SOCE in IH-ECFCs is also associated to an increase in their proliferation rate and the basal production of NO compared to normal cells. Likewise, the pharmacological blockade of SOCE and NO synthesis block IH-ECFC growth. Collectively, these data indicate that the constitutive SOCE activation enhances IH-ECFC proliferation by augmenting basal NO production and sheds novel light on the molecular mechanisms of IH.
authors.affiliation: 1 Laboratory of General Physiology, Department of Biology and Biotechnology "Lazzaro Spallanzani," University of Pavia , Pavia, Italy .
authors.forename: Estella
authors.initials: E
authors.lastname: Zuccolo
authors.affiliation: 2 Department of Molecular Medicine, University of Pavia , Pavia, Italy .
authors.forename: Cinzia
authors.initials: C
authors.lastname: Bottino
authors.affiliation: 1 Laboratory of General Physiology, Department of Biology and Biotechnology "Lazzaro Spallanzani," University of Pavia , Pavia, Italy .
authors.forename: Federica
authors.initials: F
authors.lastname: Diofano
authors.affiliation: 3 Research Laboratory of Biotechnology, Center for the Study of Myelofibrosis , Foundation IRCCS Policlinico San Matteo, Pavia, Italy .
authors.forename: Valentina
authors.initials: V
authors.lastname: Poletto
authors.affiliation: 4 Department of Pediatrics, Foundation IRCCS Policlinico San Matteo , Pavia, Italy .
authors.forename: Alessia Claudia
authors.initials: AC
authors.lastname: Codazzi
authors.affiliation: 4 Department of Pediatrics, Foundation IRCCS Policlinico San Matteo , Pavia, Italy .
authors.forename: Savina
authors.initials: S
authors.lastname: Mannarino
authors.affiliation: 3 Research Laboratory of Biotechnology, Center for the Study of Myelofibrosis , Foundation IRCCS Policlinico San Matteo, Pavia, Italy .
authors.forename: Rita
authors.initials: R
authors.lastname: Campanelli
authors.affiliation: 3 Research Laboratory of Biotechnology, Center for the Study of Myelofibrosis , Foundation IRCCS Policlinico San Matteo, Pavia, Italy .
authors.forename: Gabriella
authors.initials: G
authors.lastname: Fois
authors.affiliation: 4 Department of Pediatrics, Foundation IRCCS Policlinico San Matteo , Pavia, Italy .
authors.forename: Gian Luigi
authors.initials: GL
authors.lastname: Marseglia
authors.affiliation: 5 Department of Medicine and Health Sciences "Vincenzo Tiberio," University of Molise , Campobasso, Italy .
authors.forename: Germano
authors.initials: G
authors.lastname: Guerra
authors.affiliation: 6 Laboratory of Immunology Transplantation , Foundation IRCCS Policlinico San Matteo, Pavia, Italy .
authors.forename: Daniela
authors.initials: D
authors.lastname: Montagna
authors.affiliation: 2 Department of Molecular Medicine, University of Pavia , Pavia, Italy .
authors.forename: Umberto
authors.initials: U
authors.lastname: Laforenza
authors.affiliation: 3 Research Laboratory of Biotechnology, Center for the Study of Myelofibrosis , Foundation IRCCS Policlinico San Matteo, Pavia, Italy .
authors.forename: Vittorio
authors.initials: V
authors.lastname: Rosti
authors.affiliation: 7 Laboratory of Biotechnology , Foundation IRCCS Policlinico San Matteo, Pavia, Italy .
authors.forename: Margherita
authors.initials: M
authors.lastname: Massa
authors.affiliation: 1 Laboratory of General Physiology, Department of Biology and Biotechnology "Lazzaro Spallanzani," University of Pavia , Pavia, Italy .
authors.forename: Francesco
authors.initials: F
authors.lastname: Moccia
mesh_terms: D000813:Anilides / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D049109:Cell Proliferation / Q000187:drug effects; D002648:Child; D002675:Child, Preschool; D003114:Colony-Forming Units Assay; D003710:Demography; D042783:Endothelial Cells / Q000187:drug effects / Q000473:pathology*; D066026:Endothelial Progenitor Cells / Q000187:drug effects / Q000473:pathology*; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects; D005839:Gentamicins / Q000494:pharmacology; D006391:Hemangioma / Q000473:pathology*; D006801:Humans; D007211:Indoles / Q000494:pharmacology; D042541:Intracellular Space / Q000378:metabolism; D007811:Lanthanum / Q000494:pharmacology; D008297:Male; D009569:Nitric Oxide / Q000096:biosynthesis*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013830:Thiadiazoles / Q000494:pharmacology
chemical_list: C476308:4-methyl-4'-(3,5-bis(trifluoromethyl)-1H-pyrazol-1-yl)-1,2,3-thiadiazole-5-carboxanilide; D000813:Anilides; D005839:Gentamicins; D007211:Indoles; D012333:RNA, Messenger; D013830:Thiadiazoles; D009569:Nitric Oxide; D007811:Lanthanum; C010680:antibiotic G 418; D002118:Calcium; C000543:cyclopiazonic acid