Valve Interstitial Cell-Specific Cyclooxygenase-1 Associated With Calcification of Aortic Valves.
Aged
Aged, 80 and over
Aortic Valve
/ cytology
Aortic Valve Stenosis
/ enzymology
Calcinosis
/ enzymology
Calcium
/ metabolism
Cells, Cultured
Culture Media
/ pharmacology
Cyclooxygenase 1
/ biosynthesis
Female
Fibroblasts
/ enzymology
Gene Expression Profiling
Heart Valve Prosthesis Implantation
Humans
Male
Middle Aged
Osteoblasts
/ pathology
Osteogenesis
RNA Interference
RNA, Messenger
/ biosynthesis
RNA, Small Interfering
/ genetics
Vimentin
/ analysis
Journal
The Annals of thoracic surgery
ISSN: 1552-6259
Titre abrégé: Ann Thorac Surg
Pays: Netherlands
ID NLM: 15030100R
Informations de publication
Date de publication:
07 2020
07 2020
Historique:
received:
08
02
2019
revised:
10
09
2019
accepted:
24
09
2019
pubmed:
25
11
2019
medline:
28
8
2020
entrez:
25
11
2019
Statut:
ppublish
Résumé
The molecular mechanisms underlying aortic valve calcification are poorly understood. Here, we aimed to identify the master regulators of calcification by comparison of genes in valve interstitial cells (VICs) with calcified and noncalcified aortic valves. Calcified aortic valves were surgically excised from patients with aortic valve stenosis who required aortic valve replacements. Noncalcified and calcified sections were obtained from aortic valve leaflets. Collagenase-digested tissues were seeded into dishes, and VICs adhering to the dishes were cultured for 3 weeks, followed by comprehensive gene expression analysis. Functional analyses of identified proteins were performed by in vitro calcification assays. Tissue localization was determined by immunohistochemical staining for normal (n = 11) and stenotic valves (n = 30). We found 87 genes showing greater than a twofold change in calcified tissues. Among these genes, 68 were downregulated and 19 were upregulated. Cyclooxygenase-1 (COX1) messenger RNA and protein levels were upregulated in VICs from calcified tissues. The COX1 messenger RNA and protein levels in VICs were also strongly increased by stimulation with osteoblast differentiation medium. These were VIC-specific phenotypes and were not observed in other cell types. Immunohistochemical staining revealed that COX1-positive VICs were specifically localized in the calcified area of aortic valve tissues. The VIC-specific COX1 overexpression played a crucial role in calcification by promoting osteoblast differentiation in aortic valve tissues.
Sections du résumé
BACKGROUND
The molecular mechanisms underlying aortic valve calcification are poorly understood. Here, we aimed to identify the master regulators of calcification by comparison of genes in valve interstitial cells (VICs) with calcified and noncalcified aortic valves.
METHODS
Calcified aortic valves were surgically excised from patients with aortic valve stenosis who required aortic valve replacements. Noncalcified and calcified sections were obtained from aortic valve leaflets. Collagenase-digested tissues were seeded into dishes, and VICs adhering to the dishes were cultured for 3 weeks, followed by comprehensive gene expression analysis. Functional analyses of identified proteins were performed by in vitro calcification assays. Tissue localization was determined by immunohistochemical staining for normal (n = 11) and stenotic valves (n = 30).
RESULTS
We found 87 genes showing greater than a twofold change in calcified tissues. Among these genes, 68 were downregulated and 19 were upregulated. Cyclooxygenase-1 (COX1) messenger RNA and protein levels were upregulated in VICs from calcified tissues. The COX1 messenger RNA and protein levels in VICs were also strongly increased by stimulation with osteoblast differentiation medium. These were VIC-specific phenotypes and were not observed in other cell types. Immunohistochemical staining revealed that COX1-positive VICs were specifically localized in the calcified area of aortic valve tissues.
CONCLUSIONS
The VIC-specific COX1 overexpression played a crucial role in calcification by promoting osteoblast differentiation in aortic valve tissues.
Identifiants
pubmed: 31760051
pii: S0003-4975(19)31717-5
doi: 10.1016/j.athoracsur.2019.09.085
pii:
doi:
Substances chimiques
Culture Media
0
RNA, Messenger
0
RNA, Small Interfering
0
VIM protein, human
0
Vimentin
0
Cyclooxygenase 1
EC 1.14.99.1
PTGS1 protein, human
EC 1.14.99.1
Calcium
SY7Q814VUP
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
40-49Informations de copyright
Copyright © 2020 The Society of Thoracic Surgeons. Published by Elsevier Inc. All rights reserved.