Margaux Aize, Arthur Boilève, Benoit D Roussel, Laura Brard, Harlyne Mpweme Bangando, Corentin Kerevel, Alexandre Lebrun, Hind Messaoudi, Vladimir Saplacan, Alain Manrique, Romain Guinamard, Christophe Simard
{"title":"TRPM4通道在人主动脉瓣间质细胞成骨分化中的作用。","authors":"Margaux Aize, Arthur Boilève, Benoit D Roussel, Laura Brard, Harlyne Mpweme Bangando, Corentin Kerevel, Alexandre Lebrun, Hind Messaoudi, Vladimir Saplacan, Alain Manrique, Romain Guinamard, Christophe Simard","doi":"10.1161/JAHA.124.038542","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>Aortic stenosis due to deleterious remodeling of the aortic valve is a health concern since it can be corrected only by valve replacement due to the poor knowledge of cellular mechanisms involved. Fibroblastic valvular interstitial cells (VICs) play a central role in valve leaflet stiffness by trans-differentiation into osteoblast-like cells leading to calcification. The TRPM4 (transient receptor potential melastatin 4) cation channel was shown to participate in cardiac fibroblast remodeling. It is also involved in radiation-induced aortic valve remodeling in vivo in mice. We hypothesized that TRPM4 might participate in human VIC transition to osteoblastic phenotype.</p><p><strong>Methods: </strong>Human aortic valves were collected from patients undergoing surgical valve replacement. Isolated VICs were maintained 14 days in culture in standard or pro-calcifying media and submitted to the TRPM4 inhibitor 9-phenanthrol, or small hairpin RNA-TRPM4. Osteogenic differentiation was evaluated by measuring hydroxyapatite crystals by Alizarin red staining and protein expression of osteogenic markers.</p><p><strong>Results: </strong>Western blot on VICs revealed TRPM4 protein expression and channel functionality was confirmed by patch-clamp recordings exhibiting a cationic current sensitive to voltage and internal Ca<sup>2+</sup>. VICs maintained in pro-calcifying media exhibited a higher mineralization than in standard media, with an increase in osteogenic markers. Mineralization and osteogenic markers (bone morphogenetic protein 2, runt-related transcription factor 2) were decreased when pro-calcifying media contained 9-phenanthrol or small hairpin RNA-TRPM4. Similarly, SMAD1/5 and nuclear factor of activated T-cell pathways were stimulated in pro-calcifying media conditions compared with standard media but reduced by 9-phenanthrol or small hairpin RNA-TRPM4.</p><p><strong>Conclusions: </strong>TRPM4 participates in osteogenic differentiation of human VICs and thus appears as a target to prevent aortic valve remodeling.</p>","PeriodicalId":54370,"journal":{"name":"Journal of the American Heart Association","volume":" ","pages":"e038542"},"PeriodicalIF":5.0000,"publicationDate":"2025-04-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Contribution of the TRPM4 Channel to Osteogenic Differentiation of Human Aortic Valve Interstitial Cells.\",\"authors\":\"Margaux Aize, Arthur Boilève, Benoit D Roussel, Laura Brard, Harlyne Mpweme Bangando, Corentin Kerevel, Alexandre Lebrun, Hind Messaoudi, Vladimir Saplacan, Alain Manrique, Romain Guinamard, Christophe Simard\",\"doi\":\"10.1161/JAHA.124.038542\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Background: </strong>Aortic stenosis due to deleterious remodeling of the aortic valve is a health concern since it can be corrected only by valve replacement due to the poor knowledge of cellular mechanisms involved. Fibroblastic valvular interstitial cells (VICs) play a central role in valve leaflet stiffness by trans-differentiation into osteoblast-like cells leading to calcification. The TRPM4 (transient receptor potential melastatin 4) cation channel was shown to participate in cardiac fibroblast remodeling. It is also involved in radiation-induced aortic valve remodeling in vivo in mice. We hypothesized that TRPM4 might participate in human VIC transition to osteoblastic phenotype.</p><p><strong>Methods: </strong>Human aortic valves were collected from patients undergoing surgical valve replacement. Isolated VICs were maintained 14 days in culture in standard or pro-calcifying media and submitted to the TRPM4 inhibitor 9-phenanthrol, or small hairpin RNA-TRPM4. Osteogenic differentiation was evaluated by measuring hydroxyapatite crystals by Alizarin red staining and protein expression of osteogenic markers.</p><p><strong>Results: </strong>Western blot on VICs revealed TRPM4 protein expression and channel functionality was confirmed by patch-clamp recordings exhibiting a cationic current sensitive to voltage and internal Ca<sup>2+</sup>. VICs maintained in pro-calcifying media exhibited a higher mineralization than in standard media, with an increase in osteogenic markers. Mineralization and osteogenic markers (bone morphogenetic protein 2, runt-related transcription factor 2) were decreased when pro-calcifying media contained 9-phenanthrol or small hairpin RNA-TRPM4. Similarly, SMAD1/5 and nuclear factor of activated T-cell pathways were stimulated in pro-calcifying media conditions compared with standard media but reduced by 9-phenanthrol or small hairpin RNA-TRPM4.</p><p><strong>Conclusions: </strong>TRPM4 participates in osteogenic differentiation of human VICs and thus appears as a target to prevent aortic valve remodeling.</p>\",\"PeriodicalId\":54370,\"journal\":{\"name\":\"Journal of the American Heart Association\",\"volume\":\" \",\"pages\":\"e038542\"},\"PeriodicalIF\":5.0000,\"publicationDate\":\"2025-04-07\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of the American Heart Association\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.1161/JAHA.124.038542\",\"RegionNum\":1,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"CARDIAC & CARDIOVASCULAR SYSTEMS\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of the American Heart Association","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1161/JAHA.124.038542","RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"CARDIAC & CARDIOVASCULAR SYSTEMS","Score":null,"Total":0}
Contribution of the TRPM4 Channel to Osteogenic Differentiation of Human Aortic Valve Interstitial Cells.
Background: Aortic stenosis due to deleterious remodeling of the aortic valve is a health concern since it can be corrected only by valve replacement due to the poor knowledge of cellular mechanisms involved. Fibroblastic valvular interstitial cells (VICs) play a central role in valve leaflet stiffness by trans-differentiation into osteoblast-like cells leading to calcification. The TRPM4 (transient receptor potential melastatin 4) cation channel was shown to participate in cardiac fibroblast remodeling. It is also involved in radiation-induced aortic valve remodeling in vivo in mice. We hypothesized that TRPM4 might participate in human VIC transition to osteoblastic phenotype.
Methods: Human aortic valves were collected from patients undergoing surgical valve replacement. Isolated VICs were maintained 14 days in culture in standard or pro-calcifying media and submitted to the TRPM4 inhibitor 9-phenanthrol, or small hairpin RNA-TRPM4. Osteogenic differentiation was evaluated by measuring hydroxyapatite crystals by Alizarin red staining and protein expression of osteogenic markers.
Results: Western blot on VICs revealed TRPM4 protein expression and channel functionality was confirmed by patch-clamp recordings exhibiting a cationic current sensitive to voltage and internal Ca2+. VICs maintained in pro-calcifying media exhibited a higher mineralization than in standard media, with an increase in osteogenic markers. Mineralization and osteogenic markers (bone morphogenetic protein 2, runt-related transcription factor 2) were decreased when pro-calcifying media contained 9-phenanthrol or small hairpin RNA-TRPM4. Similarly, SMAD1/5 and nuclear factor of activated T-cell pathways were stimulated in pro-calcifying media conditions compared with standard media but reduced by 9-phenanthrol or small hairpin RNA-TRPM4.
Conclusions: TRPM4 participates in osteogenic differentiation of human VICs and thus appears as a target to prevent aortic valve remodeling.
期刊介绍：
As an Open Access journal, JAHA - Journal of the American Heart Association is rapidly and freely available, accelerating the translation of strong science into effective practice.
JAHA is an authoritative, peer-reviewed Open Access journal focusing on cardiovascular and cerebrovascular disease. JAHA provides a global forum for basic and clinical research and timely reviews on cardiovascular disease and stroke. As an Open Access journal, its content is free on publication to read, download, and share, accelerating the translation of strong science into effective practice.
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