Constitutive Store-Operated Ca2+ Entry Leads to Enhanced Nitric Oxide Production and Proliferation in Infantile Hemangioma-Derived Endothelial Colony-Forming Cells

Estella Zuccolo, Cinzia Bottino, Federica Diofano, Valentina Poletto, Alessia Claudia Codazzi, Savina Mannarino, Rita Campanelli, Gabriella Fois, Gian Luigi Marseglia, Germano Guerra, Daniela Montagna, Umberto Laforenza, Vittorio Rosti, Margherita Massa, Francesco Moccia

Research output: Contribution to journalArticle

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 Ca2+ toolkit in tumor-derived patients to acquire an adaptive advantage. Particularly, they upregulate the proangiogenic store-operated Ca2+ 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 Ca2+ signalosome is altered in IH-ECFCs by employing Ca2+ and nitric oxide (NO) imaging, real-time polymerase chain reaction, western blotting, and functional assays. IH-ECFCs display a lower intracellular Ca2+ 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, La3+, and Pyr6 and recharges the intracellular Ca2+ 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.

Original languageEnglish
Pages (from-to)301-319
Number of pages19
JournalStem Cells and Development
Volume25
Issue number4
DOIs
Publication statusPublished - Feb 15 2016

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ASJC Scopus subject areas

  • Cell Biology
  • Developmental Biology
  • Hematology

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