Peroxynitrous acid-modified extracellular matrix alters gene and protein expression in human coronary artery smooth muscle cells and induces a pro-inflammatory phenotype

Sara M Jørgensen; Lasse G Lorentzen; Christine Y Chuang; Michael J Davies

doi:10.1016/j.freeradbiomed.2022.05.001

Peroxynitrous acid-modified extracellular matrix alters gene and protein expression in human coronary artery smooth muscle cells and induces a pro-inflammatory phenotype

Free Radic Biol Med. 2022 Jun:186:43-52. doi: 10.1016/j.freeradbiomed.2022.05.001. Epub 2022 May 6.

Authors

Sara M Jørgensen¹, Lasse G Lorentzen¹, Christine Y Chuang¹, Michael J Davies²

Affiliations

¹ Department of Biomedical Sciences, Panum Institute, University of Copenhagen, Copenhagen, 2200, Denmark.
² Department of Biomedical Sciences, Panum Institute, University of Copenhagen, Copenhagen, 2200, Denmark. Electronic address: davies@sund.ku.dk.

PMID: 35526806
DOI: 10.1016/j.freeradbiomed.2022.05.001

Abstract

Leukocytes produce oxidants at inflammatory sites, including within the artery wall during the development of atherosclerosis. Developing lesions contain high numbers of activated leukocytes that generate reactive nitrogen species, including peroxynitrite/peroxynitrous acid (ONOO^-/ONOOH), as evidenced by the presence of oxidized/nitrated molecules including extracellular matrix (ECM) proteins. ECM materials are critical for arterial wall integrity, function, and determine cell phenotype, with smooth muscle cells undergoing a phenotypic switch from quiescent/contractile to proliferative/synthetic during disease development. We hypothesized that ECM modification by ONOO^-/ONOOH might drive this switch, and thereby potentially contribute to atherogenesis. ECM generated by primary human coronary artery smooth muscle cells (HCASMCs) was treated with increasing ONOO^-/ONOOH concentrations (1-1000 μM). This generated significant damage on laminin, fibronectin and versican, and lower levels on collagens and glycosaminoglycans, together with the increasing concentrations of the damage biomarker 3-nitrotyrosine. Adhesion of naïve HCASMC to ECM modified by 1 μM ONOO^-/ONOOH was enhanced, but significantly diminished by higher ONOO^-/ONOOH treatment. Cell proliferation and metabolic activity were significantly enhanced by 100 μM ONOO^-/ONOOH pre-treatment. These changes were accompanied by increased expression of genes involved in mitosis (PCNA, CCNA1, CCNB1), ECM (LAMA4, LAMB1, VCAN, FN1) and inflammation (IL-1B, IL-6, VCAM-1), and corresponding protein secretion (except VCAM-1) into the medium. These changes induced by modified ECM are consistent with HCASMC switching to a synthetic/proliferative/pro-inflammatory phenotype, together with ECM remodelling. These changes model those in atherosclerosis, suggesting a link between oxidant-modified ECM and disease progression, and highlight the potential of targeting oxidant generation as a therapeutic strategy.

Keywords: 3-Nitrotyrosine; Extracellular matrix; Peroxynitrite; Protein oxidation; Smooth muscle cell; Versican.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Atherosclerosis* / metabolism
Coronary Vessels / metabolism
Extracellular Matrix / metabolism
Humans
Myocytes, Smooth Muscle / metabolism
Oxidants / metabolism
Oxidation-Reduction
Peroxynitrous Acid* / metabolism
Phenotype
Vascular Cell Adhesion Molecule-1 / metabolism

Substances

Oxidants
Vascular Cell Adhesion Molecule-1
Peroxynitrous Acid