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Enhanced store-operated Ca(2+) influx and ORAI1 expression in ventricular fibroblasts from human failing heart

Excessive cardiac fibrosis, characterized by increased collagen-rich extracellular matrix (ECM) deposition, is a major predisposing factor for mechanical and electrical dysfunction in heart failure (HF). The human ventricular fibroblast (hVF) remodeling mechanisms that cause excessive collagen depos...

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Autores principales: Ross, Gracious R., Bajwa, Tanvir, Edwards, Stacie, Emelyanova, Larisa, Rizvi, Farhan, Holmuhamedov, Ekhson L., Werner, Paul, Downey, Francis X., Tajik, A. Jamil, Jahangir, Arshad
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Company of Biologists Ltd 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5374400/
https://www.ncbi.nlm.nih.gov/pubmed/28126709
http://dx.doi.org/10.1242/bio.022632
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author Ross, Gracious R.
Bajwa, Tanvir
Edwards, Stacie
Emelyanova, Larisa
Rizvi, Farhan
Holmuhamedov, Ekhson L.
Werner, Paul
Downey, Francis X.
Tajik, A. Jamil
Jahangir, Arshad
author_facet Ross, Gracious R.
Bajwa, Tanvir
Edwards, Stacie
Emelyanova, Larisa
Rizvi, Farhan
Holmuhamedov, Ekhson L.
Werner, Paul
Downey, Francis X.
Tajik, A. Jamil
Jahangir, Arshad
author_sort Ross, Gracious R.
collection PubMed
description Excessive cardiac fibrosis, characterized by increased collagen-rich extracellular matrix (ECM) deposition, is a major predisposing factor for mechanical and electrical dysfunction in heart failure (HF). The human ventricular fibroblast (hVF) remodeling mechanisms that cause excessive collagen deposition in HF are unclear, although reports suggest a role for intracellular free Ca(2+) in fibrosis. Therefore, we determined the association of differences in cellular Ca(2+) dynamics and collagen secretion/deposition between hVFs from failing and normal (control) hearts. Histology of left ventricle sections (Masson trichrome) confirmed excessive fibrosis in HF versus normal. In vitro, hVFs from HF showed increased secretion/deposition of soluble collagen in 48 h of culture compared with control [85.9±7.4 µg/10(6) cells vs 58.5±8.8 µg/10(6) cells, P<0.05; (Sircol™ assay)]. However, collagen gene expressions (COL1A1 and COL1A2; RT-PCR) were not different. Ca(2+) imaging (fluo-3) of isolated hVFs showed no difference in the thapsigargin-induced intracellular Ca(2+) release capacity (control 16±1.4% vs HF 17±1.1%); however, Ca(2+) influx via store-operated Ca(2+) entry/Ca(2+) release-activated channels (SOCE/CRAC) was significantly (P≤0.05) greater in HF-hVFs (47±3%) compared with non-failing (35±5%). Immunoblotting for I(CRAC) channel components showed increased ORAI1 expression in HF-hVFs compared with normal without any difference in STIM1 expression. The Pearson's correlation coefficient for co-localization of STIM1/ORAI1 was significantly (P<0.01) greater in HF (0.5±0.01) than control (0.4±0.01) hVFs. The increase in collagen secretion of HF versus control hVFs was eliminated by incubation of hVFs with YM58483 (10 µM), a selective I(CRAC) inhibitor, for 48 h (66.78±5.87 µg/10(6) cells vs 55.81±7.09 µg/10(6) cells, P=0.27). In conclusion, hVFs from HF have increased collagen secretion capacity versus non-failing hearts and this is related to increase in Ca(2+) entry via SOCE and enhanced expression of ORAI, the pore-forming subunit. Therapeutic inhibition of SOCE may reduce the progression of cardiac fibrosis/HF.
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spelling pubmed-53744002017-04-03 Enhanced store-operated Ca(2+) influx and ORAI1 expression in ventricular fibroblasts from human failing heart Ross, Gracious R. Bajwa, Tanvir Edwards, Stacie Emelyanova, Larisa Rizvi, Farhan Holmuhamedov, Ekhson L. Werner, Paul Downey, Francis X. Tajik, A. Jamil Jahangir, Arshad Biol Open Research Article Excessive cardiac fibrosis, characterized by increased collagen-rich extracellular matrix (ECM) deposition, is a major predisposing factor for mechanical and electrical dysfunction in heart failure (HF). The human ventricular fibroblast (hVF) remodeling mechanisms that cause excessive collagen deposition in HF are unclear, although reports suggest a role for intracellular free Ca(2+) in fibrosis. Therefore, we determined the association of differences in cellular Ca(2+) dynamics and collagen secretion/deposition between hVFs from failing and normal (control) hearts. Histology of left ventricle sections (Masson trichrome) confirmed excessive fibrosis in HF versus normal. In vitro, hVFs from HF showed increased secretion/deposition of soluble collagen in 48 h of culture compared with control [85.9±7.4 µg/10(6) cells vs 58.5±8.8 µg/10(6) cells, P<0.05; (Sircol™ assay)]. However, collagen gene expressions (COL1A1 and COL1A2; RT-PCR) were not different. Ca(2+) imaging (fluo-3) of isolated hVFs showed no difference in the thapsigargin-induced intracellular Ca(2+) release capacity (control 16±1.4% vs HF 17±1.1%); however, Ca(2+) influx via store-operated Ca(2+) entry/Ca(2+) release-activated channels (SOCE/CRAC) was significantly (P≤0.05) greater in HF-hVFs (47±3%) compared with non-failing (35±5%). Immunoblotting for I(CRAC) channel components showed increased ORAI1 expression in HF-hVFs compared with normal without any difference in STIM1 expression. The Pearson's correlation coefficient for co-localization of STIM1/ORAI1 was significantly (P<0.01) greater in HF (0.5±0.01) than control (0.4±0.01) hVFs. The increase in collagen secretion of HF versus control hVFs was eliminated by incubation of hVFs with YM58483 (10 µM), a selective I(CRAC) inhibitor, for 48 h (66.78±5.87 µg/10(6) cells vs 55.81±7.09 µg/10(6) cells, P=0.27). In conclusion, hVFs from HF have increased collagen secretion capacity versus non-failing hearts and this is related to increase in Ca(2+) entry via SOCE and enhanced expression of ORAI, the pore-forming subunit. Therapeutic inhibition of SOCE may reduce the progression of cardiac fibrosis/HF. The Company of Biologists Ltd 2017-01-26 /pmc/articles/PMC5374400/ /pubmed/28126709 http://dx.doi.org/10.1242/bio.022632 Text en © 2017. Published by The Company of Biologists Ltd http://creativecommons.org/licenses/by/3.0This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.
spellingShingle Research Article
Ross, Gracious R.
Bajwa, Tanvir
Edwards, Stacie
Emelyanova, Larisa
Rizvi, Farhan
Holmuhamedov, Ekhson L.
Werner, Paul
Downey, Francis X.
Tajik, A. Jamil
Jahangir, Arshad
Enhanced store-operated Ca(2+) influx and ORAI1 expression in ventricular fibroblasts from human failing heart
title Enhanced store-operated Ca(2+) influx and ORAI1 expression in ventricular fibroblasts from human failing heart
title_full Enhanced store-operated Ca(2+) influx and ORAI1 expression in ventricular fibroblasts from human failing heart
title_fullStr Enhanced store-operated Ca(2+) influx and ORAI1 expression in ventricular fibroblasts from human failing heart
title_full_unstemmed Enhanced store-operated Ca(2+) influx and ORAI1 expression in ventricular fibroblasts from human failing heart
title_short Enhanced store-operated Ca(2+) influx and ORAI1 expression in ventricular fibroblasts from human failing heart
title_sort enhanced store-operated ca(2+) influx and orai1 expression in ventricular fibroblasts from human failing heart
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5374400/
https://www.ncbi.nlm.nih.gov/pubmed/28126709
http://dx.doi.org/10.1242/bio.022632
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