Oxidation of human plasma fibronectin by inflammatory oxidants perturbs endothelial cell function

Siriluck Vanichkitrungruang; Christine Y. Chuang; Clare L. Hawkins; Astrid Hammer; Gerald Hoefler; Ernst Malle; Michael J. Davies

doi:10.1016/j.freeradbiomed.2019.04.003

Oxidation of human plasma fibronectin by inflammatory oxidants perturbs endothelial cell function

Siriluck Vanichkitrungruang, Christine Y. Chuang, Clare L. Hawkins, Astrid Hammer, Gerald Hoefler, Ernst Malle, Michael J. Davies

Inflammation, Metabolism and Oxidation

13 Citationer (Scopus)

Abstract

Dysfunction of endothelial cells of the artery wall is an early event in cardiovascular disease and atherosclerosis. The cause(s) of this dysfunction are unresolved, but accumulating evidence suggests that oxidants arising from chronic low-grade inflammation are contributory agents, with increasing data implicating myeloperoxidase (MPO, released by activated leukocytes), and the oxidants it generates (e.g. HOCl and HOSCN). As these are formed extracellularly and react rapidly with proteins, we hypothesized that MPO-mediated damage to the matrix glycoprotein fibronectin (FN) would modulate FN structure and function, and its interactions with human coronary artery endothelial cells (HCAEC). Exposure of human plasma FN to HOCl resulted in modifications to FN and its functional epitopes. A dose-dependent loss of methionine and tryptophan residues, together with increasing concentrations of methionine sulfoxide, and modification of the cell-binding fragment (CBF) and heparin-binding fragment (HBF) domains was detected with HOCl, but not HOSCN. FN modification resulted in a loss of HCAEC adhesion, impaired cell spreading and reduced cell proliferation. Exposure to HCAEC to HOCl-treated FN altered the expression of HCAEC genes associated with extracellular matrix (ECM) synthesis and adhesion. Modifications were detected on HCAEC-derived ECM pre-treated with HOCl, but not HOSCN, with a loss of antibody recognition of the CBF, HBF and extra-domain A. Co-localization of epitopes arising from MPO-generated HOCl and cell-derived FN was detected in human atherosclerotic lesions. Damage was also detected on FN extracted from lesions. These data support the hypothesis that HOCl, but not HOSCN, targets and modifies FN resulting in arterial wall endothelial cell dysfunction.

Originalsprog	Engelsk
Tidsskrift	Free Radical Biology and Medicine
Vol/bind	136
Sider (fra-til)	118-134
ISSN	0891-5849
DOI	https://doi.org/10.1016/j.freeradbiomed.2019.04.003
Status	Udgivet - 20 maj 2019

Adgang til dokumentet

10.1016/j.freeradbiomed.2019.04.003

Citationsformater

@article{5cd12f0dbbe6460c86e5e52525c9aefd,

title = "Oxidation of human plasma fibronectin by inflammatory oxidants perturbs endothelial cell function",

abstract = "Dysfunction of endothelial cells of the artery wall is an early event in cardiovascular disease and atherosclerosis. The cause(s) of this dysfunction are unresolved, but accumulating evidence suggests that oxidants arising from chronic low-grade inflammation are contributory agents, with increasing data implicating myeloperoxidase (MPO, released by activated leukocytes), and the oxidants it generates (e.g. HOCl and HOSCN). As these are formed extracellularly and react rapidly with proteins, we hypothesized that MPO-mediated damage to the matrix glycoprotein fibronectin (FN) would modulate FN structure and function, and its interactions with human coronary artery endothelial cells (HCAEC). Exposure of human plasma FN to HOCl resulted in modifications to FN and its functional epitopes. A dose-dependent loss of methionine and tryptophan residues, together with increasing concentrations of methionine sulfoxide, and modification of the cell-binding fragment (CBF) and heparin-binding fragment (HBF) domains was detected with HOCl, but not HOSCN. FN modification resulted in a loss of HCAEC adhesion, impaired cell spreading and reduced cell proliferation. Exposure to HCAEC to HOCl-treated FN altered the expression of HCAEC genes associated with extracellular matrix (ECM) synthesis and adhesion. Modifications were detected on HCAEC-derived ECM pre-treated with HOCl, but not HOSCN, with a loss of antibody recognition of the CBF, HBF and extra-domain A. Co-localization of epitopes arising from MPO-generated HOCl and cell-derived FN was detected in human atherosclerotic lesions. Damage was also detected on FN extracted from lesions. These data support the hypothesis that HOCl, but not HOSCN, targets and modifies FN resulting in arterial wall endothelial cell dysfunction.",

keywords = "Extracellular matrix, Hypochlorous acid, Myeloperoxidase, Hypothiocyanous acid, Fibronectin, Protein oxidation, Endothelial cell",

author = "Siriluck Vanichkitrungruang and Chuang, {Christine Y.} and Hawkins, {Clare L.} and Astrid Hammer and Gerald Hoefler and Ernst Malle and Davies, {Michael J.}",

year = "2019",

month = may,

day = "20",

doi = "10.1016/j.freeradbiomed.2019.04.003",

language = "English",

volume = "136",

pages = "118--134",

journal = "Free Radical Biology and Medicine",

issn = "0891-5849",

publisher = "Elsevier",

}

TY - JOUR

T1 - Oxidation of human plasma fibronectin by inflammatory oxidants perturbs endothelial cell function

AU - Vanichkitrungruang, Siriluck

AU - Chuang, Christine Y.

AU - Hawkins, Clare L.

AU - Hammer, Astrid

AU - Hoefler, Gerald

AU - Malle, Ernst

AU - Davies, Michael J.

PY - 2019/5/20

Y1 - 2019/5/20

N2 - Dysfunction of endothelial cells of the artery wall is an early event in cardiovascular disease and atherosclerosis. The cause(s) of this dysfunction are unresolved, but accumulating evidence suggests that oxidants arising from chronic low-grade inflammation are contributory agents, with increasing data implicating myeloperoxidase (MPO, released by activated leukocytes), and the oxidants it generates (e.g. HOCl and HOSCN). As these are formed extracellularly and react rapidly with proteins, we hypothesized that MPO-mediated damage to the matrix glycoprotein fibronectin (FN) would modulate FN structure and function, and its interactions with human coronary artery endothelial cells (HCAEC). Exposure of human plasma FN to HOCl resulted in modifications to FN and its functional epitopes. A dose-dependent loss of methionine and tryptophan residues, together with increasing concentrations of methionine sulfoxide, and modification of the cell-binding fragment (CBF) and heparin-binding fragment (HBF) domains was detected with HOCl, but not HOSCN. FN modification resulted in a loss of HCAEC adhesion, impaired cell spreading and reduced cell proliferation. Exposure to HCAEC to HOCl-treated FN altered the expression of HCAEC genes associated with extracellular matrix (ECM) synthesis and adhesion. Modifications were detected on HCAEC-derived ECM pre-treated with HOCl, but not HOSCN, with a loss of antibody recognition of the CBF, HBF and extra-domain A. Co-localization of epitopes arising from MPO-generated HOCl and cell-derived FN was detected in human atherosclerotic lesions. Damage was also detected on FN extracted from lesions. These data support the hypothesis that HOCl, but not HOSCN, targets and modifies FN resulting in arterial wall endothelial cell dysfunction.

AB - Dysfunction of endothelial cells of the artery wall is an early event in cardiovascular disease and atherosclerosis. The cause(s) of this dysfunction are unresolved, but accumulating evidence suggests that oxidants arising from chronic low-grade inflammation are contributory agents, with increasing data implicating myeloperoxidase (MPO, released by activated leukocytes), and the oxidants it generates (e.g. HOCl and HOSCN). As these are formed extracellularly and react rapidly with proteins, we hypothesized that MPO-mediated damage to the matrix glycoprotein fibronectin (FN) would modulate FN structure and function, and its interactions with human coronary artery endothelial cells (HCAEC). Exposure of human plasma FN to HOCl resulted in modifications to FN and its functional epitopes. A dose-dependent loss of methionine and tryptophan residues, together with increasing concentrations of methionine sulfoxide, and modification of the cell-binding fragment (CBF) and heparin-binding fragment (HBF) domains was detected with HOCl, but not HOSCN. FN modification resulted in a loss of HCAEC adhesion, impaired cell spreading and reduced cell proliferation. Exposure to HCAEC to HOCl-treated FN altered the expression of HCAEC genes associated with extracellular matrix (ECM) synthesis and adhesion. Modifications were detected on HCAEC-derived ECM pre-treated with HOCl, but not HOSCN, with a loss of antibody recognition of the CBF, HBF and extra-domain A. Co-localization of epitopes arising from MPO-generated HOCl and cell-derived FN was detected in human atherosclerotic lesions. Damage was also detected on FN extracted from lesions. These data support the hypothesis that HOCl, but not HOSCN, targets and modifies FN resulting in arterial wall endothelial cell dysfunction.

KW - Extracellular matrix

KW - Hypochlorous acid

KW - Myeloperoxidase

KW - Hypothiocyanous acid

KW - Fibronectin

KW - Protein oxidation

KW - Endothelial cell

U2 - 10.1016/j.freeradbiomed.2019.04.003

DO - 10.1016/j.freeradbiomed.2019.04.003

M3 - Journal article

C2 - 30959171

SN - 0891-5849

VL - 136

SP - 118

EP - 134

JO - Free Radical Biology and Medicine

JF - Free Radical Biology and Medicine

ER -