Enhanced store-operated Ca2+ 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...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
The Company of Biologists
2017-03-01
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| Series: | Biology Open |
| Subjects: | |
| Online Access: | http://bio.biologists.org/content/6/3/326 |
| _version_ | 1818667647834783744 |
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| author | Gracious R. Ross Tanvir Bajwa Stacie Edwards Larisa Emelyanova Farhan Rizvi Ekhson L. Holmuhamedov Paul Werner Francis X. Downey A. Jamil Tajik Arshad Jahangir |
| author_facet | Gracious R. Ross Tanvir Bajwa Stacie Edwards Larisa Emelyanova Farhan Rizvi Ekhson L. Holmuhamedov Paul Werner Francis X. Downey A. Jamil Tajik Arshad Jahangir |
| author_sort | Gracious R. Ross |
| collection | DOAJ |
| 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 Ca2+ in fibrosis. Therefore, we determined the association of differences in cellular Ca2+ 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/106 cells vs 58.5±8.8 µg/106 cells, P<0.05; (Sircol™ assay)]. However, collagen gene expressions (COL1A1 and COL1A2; RT-PCR) were not different. Ca2+ imaging (fluo-3) of isolated hVFs showed no difference in the thapsigargin-induced intracellular Ca2+ release capacity (control 16±1.4% vs HF 17±1.1%); however, Ca2+ influx via store-operated Ca2+ entry/Ca2+ release-activated channels (SOCE/CRAC) was significantly (P≤0.05) greater in HF-hVFs (47±3%) compared with non-failing (35±5%). Immunoblotting for ICRAC 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 ICRAC inhibitor, for 48 h (66.78±5.87 µg/106 cells vs 55.81±7.09 µg/106 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 Ca2+ entry via SOCE and enhanced expression of ORAI, the pore-forming subunit. Therapeutic inhibition of SOCE may reduce the progression of cardiac fibrosis/HF. |
| first_indexed | 2024-12-17T06:23:45Z |
| format | Article |
| id | doaj.art-3dcb7d0a7e4c4925913ded2133ab2440 |
| institution | Directory Open Access Journal |
| issn | 2046-6390 |
| language | English |
| last_indexed | 2024-12-17T06:23:45Z |
| publishDate | 2017-03-01 |
| publisher | The Company of Biologists |
| record_format | Article |
| series | Biology Open |
| spelling | doaj.art-3dcb7d0a7e4c4925913ded2133ab24402022-12-21T22:00:20ZengThe Company of BiologistsBiology Open2046-63902017-03-016332633210.1242/bio.022632022632Enhanced store-operated Ca2+ influx and ORAI1 expression in ventricular fibroblasts from human failing heartGracious R. Ross0Tanvir Bajwa1Stacie Edwards2Larisa Emelyanova3Farhan Rizvi4Ekhson L. Holmuhamedov5Paul Werner6Francis X. Downey7A. Jamil Tajik8Arshad Jahangir9 Center for Integrative Research on Cardiovascular Aging, Aurora Research Institute, Aurora Health Care, Milwaukee, WI 53215, USA Center for Integrative Research on Cardiovascular Aging, Aurora Research Institute, Aurora Health Care, Milwaukee, WI 53215, USA Center for Integrative Research on Cardiovascular Aging, Aurora Research Institute, Aurora Health Care, Milwaukee, WI 53215, USA Center for Integrative Research on Cardiovascular Aging, Aurora Research Institute, Aurora Health Care, Milwaukee, WI 53215, USA Center for Integrative Research on Cardiovascular Aging, Aurora Research Institute, Aurora Health Care, Milwaukee, WI 53215, USA Center for Integrative Research on Cardiovascular Aging, Aurora Research Institute, Aurora Health Care, Milwaukee, WI 53215, USA Aurora Cardiovascular Services, Aurora Sinai/Aurora St. Luke's Medical Centers, Milwaukee, WI 53215, USA Aurora Cardiovascular Services, Aurora Sinai/Aurora St. Luke's Medical Centers, Milwaukee, WI 53215, USA Aurora Cardiovascular Services, Aurora Sinai/Aurora St. Luke's Medical Centers, Milwaukee, WI 53215, USA Center for Integrative Research on Cardiovascular Aging, Aurora Research Institute, Aurora Health Care, Milwaukee, WI 53215, USA 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 Ca2+ in fibrosis. Therefore, we determined the association of differences in cellular Ca2+ 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/106 cells vs 58.5±8.8 µg/106 cells, P<0.05; (Sircol™ assay)]. However, collagen gene expressions (COL1A1 and COL1A2; RT-PCR) were not different. Ca2+ imaging (fluo-3) of isolated hVFs showed no difference in the thapsigargin-induced intracellular Ca2+ release capacity (control 16±1.4% vs HF 17±1.1%); however, Ca2+ influx via store-operated Ca2+ entry/Ca2+ release-activated channels (SOCE/CRAC) was significantly (P≤0.05) greater in HF-hVFs (47±3%) compared with non-failing (35±5%). Immunoblotting for ICRAC 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 ICRAC inhibitor, for 48 h (66.78±5.87 µg/106 cells vs 55.81±7.09 µg/106 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 Ca2+ entry via SOCE and enhanced expression of ORAI, the pore-forming subunit. Therapeutic inhibition of SOCE may reduce the progression of cardiac fibrosis/HF.http://bio.biologists.org/content/6/3/326CalciumCardiac fibrosisStore-operated Ca2+ entryHeart failure |
| spellingShingle | Gracious R. Ross Tanvir Bajwa Stacie Edwards Larisa Emelyanova Farhan Rizvi Ekhson L. Holmuhamedov Paul Werner Francis X. Downey A. Jamil Tajik Arshad Jahangir Enhanced store-operated Ca2+ influx and ORAI1 expression in ventricular fibroblasts from human failing heart Biology Open Calcium Cardiac fibrosis Store-operated Ca2+ entry Heart failure |
| title | Enhanced store-operated Ca2+ influx and ORAI1 expression in ventricular fibroblasts from human failing heart |
| title_full | Enhanced store-operated Ca2+ influx and ORAI1 expression in ventricular fibroblasts from human failing heart |
| title_fullStr | Enhanced store-operated Ca2+ influx and ORAI1 expression in ventricular fibroblasts from human failing heart |
| title_full_unstemmed | Enhanced store-operated Ca2+ influx and ORAI1 expression in ventricular fibroblasts from human failing heart |
| title_short | Enhanced store-operated Ca2+ influx and ORAI1 expression in ventricular fibroblasts from human failing heart |
| title_sort | enhanced store operated ca2 influx and orai1 expression in ventricular fibroblasts from human failing heart |
| topic | Calcium Cardiac fibrosis Store-operated Ca2+ entry Heart failure |
| url | http://bio.biologists.org/content/6/3/326 |
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