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

Research output: Contribution to journalArticle

114 Citations (Scopus)

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

Fingerprint

Vascular Endothelial Growth Factor A
Calcium
Endothelial Cells
Cell Movement
Transient Receptor Potential Channels
Calcium Release Activated Calcium Channels
Chorioallantoic Membrane
Vascular Endothelial Growth Factor Receptor-2
Human Umbilical Vein Endothelial Cells
Amides
Small Interfering RNA
Cluster Analysis
Immune System
Cell Survival

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., ... 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

Orai1 and CRAC channel dependence of VEGF-activated Ca2+ entry and endothelial tube formation. / Li, Jing; Cubbon, Richard M.; Wilson, Lesley A.; Amer, Mohamed S.; McKeown, Lynn; Hou, Bing; Majeed, Yasser; Tumova, Sarka; Seymour, Victoria A L; Taylor, Hilary; Stacey, Martin; O'Regan, David; Foster, Richard; Porter, Karen E.; Kearney, Mark T.; Beech, David J.

In: Circulation Research, Vol. 108, No. 10, 13.05.2011, p. 1190-1198.

Research output: Contribution to journalArticle

Li, J, Cubbon, RM, Wilson, LA, Amer, MS, McKeown, L, Hou, B, Majeed, Y, Tumova, S, Seymour, VAL, Taylor, H, Stacey, M, O'Regan, D, Foster, R, Porter, KE, Kearney, MT & Beech, DJ 2011, 'Orai1 and CRAC channel dependence of VEGF-activated Ca2+ entry and endothelial tube formation', Circulation Research, vol. 108, no. 10, pp. 1190-1198. https://doi.org/10.1161/CIRCRESAHA.111.243352
Li, Jing ; Cubbon, Richard M. ; Wilson, Lesley A. ; Amer, Mohamed S. ; McKeown, Lynn ; Hou, Bing ; Majeed, Yasser ; Tumova, Sarka ; Seymour, Victoria A L ; Taylor, Hilary ; Stacey, Martin ; O'Regan, David ; Foster, Richard ; Porter, Karen E. ; Kearney, Mark T. ; Beech, David J. / Orai1 and CRAC channel dependence of VEGF-activated Ca2+ entry and endothelial tube formation. In: Circulation Research. 2011 ; Vol. 108, No. 10. pp. 1190-1198.
@article{24dd6475a08d411b95165b6f62cf9025,
title = "Orai1 and CRAC channel dependence of VEGF-activated Ca2+ entry and endothelial tube formation",
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.",
keywords = "angiogenesis, calcium channels, endothelium, growth factors",
author = "Jing Li and Cubbon, {Richard M.} and Wilson, {Lesley A.} and Amer, {Mohamed S.} and Lynn McKeown and Bing Hou and Yasser Majeed and Sarka Tumova and Seymour, {Victoria A L} and Hilary Taylor and Martin Stacey and David O'Regan and Richard Foster and Porter, {Karen E.} and Kearney, {Mark T.} and Beech, {David J.}",
year = "2011",
month = "5",
day = "13",
doi = "10.1161/CIRCRESAHA.111.243352",
language = "English",
volume = "108",
pages = "1190--1198",
journal = "Circulation Research",
issn = "0009-7330",
publisher = "Lippincott Williams and Wilkins",
number = "10",

}

TY - JOUR

T1 - Orai1 and CRAC channel dependence of VEGF-activated Ca2+ entry and endothelial tube formation

AU - Li, Jing

AU - Cubbon, Richard M.

AU - Wilson, Lesley A.

AU - Amer, Mohamed S.

AU - McKeown, Lynn

AU - Hou, Bing

AU - Majeed, Yasser

AU - Tumova, Sarka

AU - Seymour, Victoria A L

AU - Taylor, Hilary

AU - Stacey, Martin

AU - O'Regan, David

AU - Foster, Richard

AU - Porter, Karen E.

AU - Kearney, Mark T.

AU - Beech, David J.

PY - 2011/5/13

Y1 - 2011/5/13

N2 - 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.

AB - 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.

KW - angiogenesis

KW - calcium channels

KW - endothelium

KW - growth factors

UR - http://www.scopus.com/inward/record.url?scp=79957444479&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79957444479&partnerID=8YFLogxK

U2 - 10.1161/CIRCRESAHA.111.243352

DO - 10.1161/CIRCRESAHA.111.243352

M3 - Article

VL - 108

SP - 1190

EP - 1198

JO - Circulation Research

JF - Circulation Research

SN - 0009-7330

IS - 10

ER -