Orai1 and CRAC channel dependence of VEGF-activated Ca2+ entry and endothelial tube formation

Jing Li, Richard M. Cubbon, Lesley A. Wilson, Mohamed S. Amer, Lynn McKeown, Bing Hou, Yasser Majeed, Sarka Tumova, Victoria A L Seymour, Hilary Taylor, Martin Stacey, David O'Regan, Richard Foster, Karen E. Porter, Mark T. Kearney, David J. Beech

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Abstract

Rationale: Orai1 and the associated calcium release-activated calcium (CRAC) channel were discovered in the immune system. Existence also in endothelial cells has been suggested, but the relevance to endothelial biology is mostly unknown. Objective: The aim of this study was to investigate the relevance of Orai1 and CRAC channels to vascular endothelial growth factor (VEGF) and endothelial tube formation. Methods and Results: In human umbilical vein endothelial cells, Orai1 disruption by short-interfering RNA or dominant-negative mutant Orai1 inhibited calcium release-activated (store-operated) calcium entry, VEGF-evoked calcium entry, cell migration, and in vitro tube formation. Expression of exogenous wild-type Orai1 rescued the tube formation. VEGF receptor-2 and Orai1 partially colocalized. Orai1 disruption also inhibited calcium entry and tube formation in endothelial progenitor cells from human blood. A known blocker of the immune cell CRAC channel (3-fluoropyridine-4-carboxylic acid (2′,5′- dimethoxybiphenyl-4-yl)amide) was a strong blocker of store-operated calcium entry in endothelial cells and inhibited calcium entry evoked by VEGF in 3 types of human endothelial cell. The compound lacked effect on VEGF-evoked calcium-release, STIM1 clustering, and 2 types of transient receptor potential channels, TRPC6 and TRPV4. Without effect on cell viability, the compound inhibited human endothelial cell migration and tube formation in vitro and suppressed angiogenesis in vivo in the chick chorioallantoic membrane. The compound showed 100-fold greater potency for endothelial compared with immune cell calcium entry. Conclusions: The data suggest positive roles for Orai1 and CRAC channels in VEGF-evoked calcium entry and new opportunity for chemical modulation of angiogenesis.

Original languageEnglish
Pages (from-to)1190-1198
Number of pages9
JournalCirculation Research
Volume108
Issue number10
DOIs
Publication statusPublished - 13 May 2011
Externally publishedYes

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Keywords

  • angiogenesis
  • calcium channels
  • endothelium
  • growth factors

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine

Cite this

Li, J., Cubbon, R. M., Wilson, L. A., Amer, M. S., McKeown, L., Hou, B., Majeed, Y., Tumova, S., Seymour, V. A. L., Taylor, H., Stacey, M., O'Regan, D., Foster, R., Porter, K. E., Kearney, M. T., & Beech, D. J. (2011). Orai1 and CRAC channel dependence of VEGF-activated Ca2+ entry and endothelial tube formation. Circulation Research, 108(10), 1190-1198. https://doi.org/10.1161/CIRCRESAHA.111.243352