Thrombospondin-1 regulates blood flow via CD47 receptor-mediated activation of NADPH oxidase 1

Gabor Csanyi, Mingyi Yao, Andrés I. Rodríguez, Imad Al Ghouleh, Maryam Sharifi-Sanjani, Giovanna Frazziano, Xiaojun Huang, Eric E. Kelley, Jeffrey S. Isenberg, Patrick J. Pagano

Research output: Contribution to journalArticle

56 Citations (Scopus)

Abstract

OBJECTIVE-: Although the matricellular protein thrombospondin-1 (TSP1) is highly expressed in the vessel wall in response to injury, its pathophysiological role in the development of vascular disease is poorly understood. This study was designed to test the hypothesis that TSP1 stimulates reactive oxygen species production in vascular smooth muscle cells and induces vascular dysfunction by promoting oxidative stress. METHODS AND RESULTS-: Nanomolar concentrations of TSP1 found in human vascular disease robustly stimulated superoxide (O2) levels in vascular smooth muscle cells at both cellular and tissue level as measured by cytochrome c and electron paramagnetic resonance. A peptide mimicking the C terminus of TSP1 known to specifically bind CD47 recapitulated this response. Transcriptional knockdown of CD47 and a monoclonal inhibitory CD47 antibody abrogated TSP1-triggered O2 in vitro and ex vivo. TSP1 treatment of vascular smooth muscle cells activated phospholipase C and protein kinase C, resulting in phosphorylation of the NADPH oxidase organizer subunit p47 and subsequent Nox1 activation, leading to impairment of arterial vasodilatation ex vivo. Further, we observed that blockade of CD47 and NADPH oxidase 1 gene silencing in vivo in rats improves TSP1-induced impairment of tissue blood flow after ischemia reperfusion. CONCLUSION-: Our data suggest a highly regulated process of reactive oxygen species stimulation and blood flow regulation promoted through a direct TSP1/CD47-mediated activation of Nox1. This is the first report, to our knowledge, of a matricellular protein acting as a ligand for NADPH oxidase activation and through specific engagement of integrin-associated protein CD47.

Original languageEnglish (US)
Pages (from-to)2966-2973
Number of pages8
JournalArteriosclerosis, thrombosis, and vascular biology
Volume32
Issue number12
DOIs
StatePublished - Dec 1 2012

Fingerprint

Thrombospondin 1
Vascular Smooth Muscle
Smooth Muscle Myocytes
NADPH Oxidase
Vascular Diseases
Reactive Oxygen Species
Proteins
Electron Spin Resonance Spectroscopy
Type C Phospholipases
Gene Silencing
Cytochromes c
NADPH oxidase 1
Vasodilation
Integrins
Superoxides
Protein Kinase C
Reperfusion
Blood Vessels
Oxidative Stress
Ischemia

Keywords

  • CD47
  • NADPH oxidase
  • reactive oxygen species
  • thrombospondin-1
  • vascular disease

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine

Cite this

Csanyi, G., Yao, M., Rodríguez, A. I., Ghouleh, I. A., Sharifi-Sanjani, M., Frazziano, G., ... Pagano, P. J. (2012). Thrombospondin-1 regulates blood flow via CD47 receptor-mediated activation of NADPH oxidase 1. Arteriosclerosis, thrombosis, and vascular biology, 32(12), 2966-2973. https://doi.org/10.1161/ATVBAHA.112.300031

Thrombospondin-1 regulates blood flow via CD47 receptor-mediated activation of NADPH oxidase 1. / Csanyi, Gabor; Yao, Mingyi; Rodríguez, Andrés I.; Ghouleh, Imad Al; Sharifi-Sanjani, Maryam; Frazziano, Giovanna; Huang, Xiaojun; Kelley, Eric E.; Isenberg, Jeffrey S.; Pagano, Patrick J.

In: Arteriosclerosis, thrombosis, and vascular biology, Vol. 32, No. 12, 01.12.2012, p. 2966-2973.

Research output: Contribution to journalArticle

Csanyi, G, Yao, M, Rodríguez, AI, Ghouleh, IA, Sharifi-Sanjani, M, Frazziano, G, Huang, X, Kelley, EE, Isenberg, JS & Pagano, PJ 2012, 'Thrombospondin-1 regulates blood flow via CD47 receptor-mediated activation of NADPH oxidase 1', Arteriosclerosis, thrombosis, and vascular biology, vol. 32, no. 12, pp. 2966-2973. https://doi.org/10.1161/ATVBAHA.112.300031
Csanyi, Gabor ; Yao, Mingyi ; Rodríguez, Andrés I. ; Ghouleh, Imad Al ; Sharifi-Sanjani, Maryam ; Frazziano, Giovanna ; Huang, Xiaojun ; Kelley, Eric E. ; Isenberg, Jeffrey S. ; Pagano, Patrick J. / Thrombospondin-1 regulates blood flow via CD47 receptor-mediated activation of NADPH oxidase 1. In: Arteriosclerosis, thrombosis, and vascular biology. 2012 ; Vol. 32, No. 12. pp. 2966-2973.
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AU - Csanyi, Gabor

AU - Yao, Mingyi

AU - Rodríguez, Andrés I.

AU - Ghouleh, Imad Al

AU - Sharifi-Sanjani, Maryam

AU - Frazziano, Giovanna

AU - Huang, Xiaojun

AU - Kelley, Eric E.

AU - Isenberg, Jeffrey S.

AU - Pagano, Patrick J.

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N2 - OBJECTIVE-: Although the matricellular protein thrombospondin-1 (TSP1) is highly expressed in the vessel wall in response to injury, its pathophysiological role in the development of vascular disease is poorly understood. This study was designed to test the hypothesis that TSP1 stimulates reactive oxygen species production in vascular smooth muscle cells and induces vascular dysfunction by promoting oxidative stress. METHODS AND RESULTS-: Nanomolar concentrations of TSP1 found in human vascular disease robustly stimulated superoxide (O2) levels in vascular smooth muscle cells at both cellular and tissue level as measured by cytochrome c and electron paramagnetic resonance. A peptide mimicking the C terminus of TSP1 known to specifically bind CD47 recapitulated this response. Transcriptional knockdown of CD47 and a monoclonal inhibitory CD47 antibody abrogated TSP1-triggered O2 in vitro and ex vivo. TSP1 treatment of vascular smooth muscle cells activated phospholipase C and protein kinase C, resulting in phosphorylation of the NADPH oxidase organizer subunit p47 and subsequent Nox1 activation, leading to impairment of arterial vasodilatation ex vivo. Further, we observed that blockade of CD47 and NADPH oxidase 1 gene silencing in vivo in rats improves TSP1-induced impairment of tissue blood flow after ischemia reperfusion. CONCLUSION-: Our data suggest a highly regulated process of reactive oxygen species stimulation and blood flow regulation promoted through a direct TSP1/CD47-mediated activation of Nox1. This is the first report, to our knowledge, of a matricellular protein acting as a ligand for NADPH oxidase activation and through specific engagement of integrin-associated protein CD47.

AB - OBJECTIVE-: Although the matricellular protein thrombospondin-1 (TSP1) is highly expressed in the vessel wall in response to injury, its pathophysiological role in the development of vascular disease is poorly understood. This study was designed to test the hypothesis that TSP1 stimulates reactive oxygen species production in vascular smooth muscle cells and induces vascular dysfunction by promoting oxidative stress. METHODS AND RESULTS-: Nanomolar concentrations of TSP1 found in human vascular disease robustly stimulated superoxide (O2) levels in vascular smooth muscle cells at both cellular and tissue level as measured by cytochrome c and electron paramagnetic resonance. A peptide mimicking the C terminus of TSP1 known to specifically bind CD47 recapitulated this response. Transcriptional knockdown of CD47 and a monoclonal inhibitory CD47 antibody abrogated TSP1-triggered O2 in vitro and ex vivo. TSP1 treatment of vascular smooth muscle cells activated phospholipase C and protein kinase C, resulting in phosphorylation of the NADPH oxidase organizer subunit p47 and subsequent Nox1 activation, leading to impairment of arterial vasodilatation ex vivo. Further, we observed that blockade of CD47 and NADPH oxidase 1 gene silencing in vivo in rats improves TSP1-induced impairment of tissue blood flow after ischemia reperfusion. CONCLUSION-: Our data suggest a highly regulated process of reactive oxygen species stimulation and blood flow regulation promoted through a direct TSP1/CD47-mediated activation of Nox1. This is the first report, to our knowledge, of a matricellular protein acting as a ligand for NADPH oxidase activation and through specific engagement of integrin-associated protein CD47.

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KW - thrombospondin-1

KW - vascular disease

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