Vascular endothelial growth factor stimulates endothelial colony forming cells proliferation and tubulogenesis by inducing oscillations in intracellular Ca 2+ concentration

Silvia Dragoni, Umberto Laforenza, Elisa Bonetti, Francesco Lodola, Cinzia Bottino, Roberto Berra-Romani, Giacomo Carlo Bongio, Maria Pia Cinelli, Germano Guerra, Paolo Pedrazzoli, Vittorio Rosti, Franco Tanzi, Francesco Moccia

Research output: Contribution to journalArticle

Abstract

Endothelial progenitor cells (EPCs) home from the bone marrow to the site of tissue regeneration and sustain neovascularization after acute vascular injury and upon the angiogenic switch in solid tumors. Therefore, they represent a suitable tool for cell-based therapy (CBT) in regenerative medicine and provide a novel promising target in the fight against cancer. Intracellular Ca 2+ signals regulate numerous endothelial functions, such as proliferation and tubulogenesis. The growth of endothelial colony forming cells (ECFCs), which are EPCs capable of acquiring a mature endothelial phenotype, is governed by storedependent Ca 2+ entry (SOCE). This study aimed at investigating the nature and the role of VEGF-elicited Ca 2+ signals in ECFCs. VEGF induced asynchronous Ca 2+ oscillations, whose latency, amplitude, and frequency were correlated to the growth factor dose. Removal of external Ca 2+ (0Ca 2+) and SOCE inhibition with N-(4-[3,5-bis(trifluoromethyl)-1H-pyrazol-1-yl]phenyl)-4-methyl-1,2, 3-thiadiazole-5-carboxamide (BTP-2) reduced the duration of the oscillatory signal. Blockade of phospholipase C-γ with U73122, emptying the inositol-1,4,5-trisphosphate (InsP3)-sensitive Ca 2+ pools with cyclopiazonic acid (CPA), and inhibition of InsP 3 receptors with 2-APB prevented the Ca 2+ response to VEGF. VEGF-induced ECFC proliferation and tubulogenesis were inhibited by the Ca 2+-chelant, BAPTA, and BTP-2. NF-κB activation by VEGF was impaired by BAPTA, BTP-2, and its selective blocker, thymoquinone. Thymoquinone, in turn, suppressed VEGF-dependent ECFC proliferation and tubulogenesis. These data indicate that VEGF-induced Ca 2+oscillations require the interplay between InsP3-dependent Ca 2+ release and SOCE, and promote ECFC growth and tubulogenesis by engaging NF-κB. This novel signaling pathway might be exploited to enhance the outcome of CBT and chemotherapy.

Original languageEnglish
Pages (from-to)1898-1907
Number of pages10
JournalStem Cells
Volume29
Issue number11
DOIs
Publication statusPublished - Nov 2011

Keywords

  • Calcium signalling
  • Cell proliferation
  • Endothelium
  • Vascular endothelial growth factor-A

ASJC Scopus subject areas

  • Cell Biology
  • Developmental Biology
  • Molecular Medicine

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    Dragoni, S., Laforenza, U., Bonetti, E., Lodola, F., Bottino, C., Berra-Romani, R., Bongio, G. C., Cinelli, M. P., Guerra, G., Pedrazzoli, P., Rosti, V., Tanzi, F., & Moccia, F. (2011). Vascular endothelial growth factor stimulates endothelial colony forming cells proliferation and tubulogenesis by inducing oscillations in intracellular Ca 2+ concentration. Stem Cells, 29(11), 1898-1907. https://doi.org/10.1002/stem.734