Endothelial SCUBE2 interacts with VEGFR2 and regulates VEGF-induced angiogenesis

Yuh Charn Lin, Tsu Yi Chao, Chi Tai Yeh, Steve R. Roffler, Reiji Kannagi, Ruey Bing Yang

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Objective - Vascular endothelial growth factor (VEGF), a major mediator of angiogenesis, exerts its proangiogenic action by binding to VEGFR2 (VEGF receptor 2), the activity of which is further modulated by VEGFR2 coreceptors such as neuropilins. However, whether VEGFR2 is regulated by additional coreceptors is not clear. To investigate whether SCUBE2 (signal peptide-CUB-EGF domain-containing protein 2), a peripheral membrane protein expressed in vascular endothelial cells (ECs) known to bind other signaling receptors, functions as a VEGFR2 coreceptor and to verify the role of SCUBE2 in the VEGF-induced angiogenesis. Approach and Results - SCUBE2 lentiviral overexpression in human ECs increased and short hairpin RNA knockdown inhibited VEGF-induced EC growth and capillary-like network formation on Matrigel. Like VEGF, endothelial SCUBE2 was upregulated by hypoxia-inducible factor-1α at both mRNA and protein levels. EC-specific Scube2 knockout mice were not defective in vascular development but showed impaired VEGF-induced neovascularization in implanted Matrigel plugs and recovery of blood flow after hind-limb ischemia. Coimmunoprecipitation and ligand-binding assays showed that SCUBE2 forms a complex with VEGF and VEGFR2, thus acting as a coreceptor to facilitate VEGF binding and augment VEGFR2 signal activity. SCUBE2 knockdown or genetic knockout suppressed and its overexpression promoted the VEGF-induced activation of downstream proangiogenic and proliferating signals, including VEGFR2 phosphorylation and mitogen-activated protein kinase or AKT activation. Conclusions - Endothelial SCUBE2 may be a novel coreceptor for VEGFR2 and potentiate VEGF-induced signaling in adult angiogenesis.

Original languageEnglish
Pages (from-to)144-155
Number of pages12
JournalArteriosclerosis, Thrombosis, and Vascular Biology
Volume37
Issue number1
DOIs
Publication statusPublished - Jan 1 2017

Fingerprint

Vascular Endothelial Growth Factor Receptor
Vascular Endothelial Growth Factor A
Endothelial Cells
Vascular Endothelial Growth Factor Receptor-2
Neuropilins
Hypoxia-Inducible Factor 1
Mitogen-Activated Protein Kinase 1
Protein Sorting Signals
Epidermal Growth Factor
Knockout Mice
Small Interfering RNA
Blood Vessels
Membrane Proteins
Ischemia
Extremities
Phosphorylation
Ligands
Messenger RNA
Growth

Keywords

  • angiogenesis modulators
  • endothelial cells
  • HIF1A protein, human
  • SCUBE2 protein, human
  • vascular endothelial growth factor A

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine

Cite this

Endothelial SCUBE2 interacts with VEGFR2 and regulates VEGF-induced angiogenesis. / Lin, Yuh Charn; Chao, Tsu Yi; Yeh, Chi Tai; Roffler, Steve R.; Kannagi, Reiji; Yang, Ruey Bing.

In: Arteriosclerosis, Thrombosis, and Vascular Biology, Vol. 37, No. 1, 01.01.2017, p. 144-155.

Research output: Contribution to journalArticle

Lin, Yuh Charn ; Chao, Tsu Yi ; Yeh, Chi Tai ; Roffler, Steve R. ; Kannagi, Reiji ; Yang, Ruey Bing. / Endothelial SCUBE2 interacts with VEGFR2 and regulates VEGF-induced angiogenesis. In: Arteriosclerosis, Thrombosis, and Vascular Biology. 2017 ; Vol. 37, No. 1. pp. 144-155.
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AU - Chao, Tsu Yi

AU - Yeh, Chi Tai

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AU - Kannagi, Reiji

AU - Yang, Ruey Bing

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N2 - Objective - Vascular endothelial growth factor (VEGF), a major mediator of angiogenesis, exerts its proangiogenic action by binding to VEGFR2 (VEGF receptor 2), the activity of which is further modulated by VEGFR2 coreceptors such as neuropilins. However, whether VEGFR2 is regulated by additional coreceptors is not clear. To investigate whether SCUBE2 (signal peptide-CUB-EGF domain-containing protein 2), a peripheral membrane protein expressed in vascular endothelial cells (ECs) known to bind other signaling receptors, functions as a VEGFR2 coreceptor and to verify the role of SCUBE2 in the VEGF-induced angiogenesis. Approach and Results - SCUBE2 lentiviral overexpression in human ECs increased and short hairpin RNA knockdown inhibited VEGF-induced EC growth and capillary-like network formation on Matrigel. Like VEGF, endothelial SCUBE2 was upregulated by hypoxia-inducible factor-1α at both mRNA and protein levels. EC-specific Scube2 knockout mice were not defective in vascular development but showed impaired VEGF-induced neovascularization in implanted Matrigel plugs and recovery of blood flow after hind-limb ischemia. Coimmunoprecipitation and ligand-binding assays showed that SCUBE2 forms a complex with VEGF and VEGFR2, thus acting as a coreceptor to facilitate VEGF binding and augment VEGFR2 signal activity. SCUBE2 knockdown or genetic knockout suppressed and its overexpression promoted the VEGF-induced activation of downstream proangiogenic and proliferating signals, including VEGFR2 phosphorylation and mitogen-activated protein kinase or AKT activation. Conclusions - Endothelial SCUBE2 may be a novel coreceptor for VEGFR2 and potentiate VEGF-induced signaling in adult angiogenesis.

AB - Objective - Vascular endothelial growth factor (VEGF), a major mediator of angiogenesis, exerts its proangiogenic action by binding to VEGFR2 (VEGF receptor 2), the activity of which is further modulated by VEGFR2 coreceptors such as neuropilins. However, whether VEGFR2 is regulated by additional coreceptors is not clear. To investigate whether SCUBE2 (signal peptide-CUB-EGF domain-containing protein 2), a peripheral membrane protein expressed in vascular endothelial cells (ECs) known to bind other signaling receptors, functions as a VEGFR2 coreceptor and to verify the role of SCUBE2 in the VEGF-induced angiogenesis. Approach and Results - SCUBE2 lentiviral overexpression in human ECs increased and short hairpin RNA knockdown inhibited VEGF-induced EC growth and capillary-like network formation on Matrigel. Like VEGF, endothelial SCUBE2 was upregulated by hypoxia-inducible factor-1α at both mRNA and protein levels. EC-specific Scube2 knockout mice were not defective in vascular development but showed impaired VEGF-induced neovascularization in implanted Matrigel plugs and recovery of blood flow after hind-limb ischemia. Coimmunoprecipitation and ligand-binding assays showed that SCUBE2 forms a complex with VEGF and VEGFR2, thus acting as a coreceptor to facilitate VEGF binding and augment VEGFR2 signal activity. SCUBE2 knockdown or genetic knockout suppressed and its overexpression promoted the VEGF-induced activation of downstream proangiogenic and proliferating signals, including VEGFR2 phosphorylation and mitogen-activated protein kinase or AKT activation. Conclusions - Endothelial SCUBE2 may be a novel coreceptor for VEGFR2 and potentiate VEGF-induced signaling in adult angiogenesis.

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KW - vascular endothelial growth factor A

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