Single-cell RNA-sequencing analysis of aortic valve interstitial cells demonstrates the regulation of integrin signaling by nitric oxide
Calcific aortic valve disease (CAVD) is an increasingly prevalent condition among the elderly population that is associated with significant morbidity and mortality. Insufficient understanding of the underlying disease mechanisms has hindered the development of pharmacologic therapies for CAVD. Rece...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-10-01
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| Series: | Frontiers in Cardiovascular Medicine |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fcvm.2022.742850/full |
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| author | Uddalak Majumdar Uddalak Majumdar Talita Z. Choudhury Talita Z. Choudhury Sathiyanarayanan Manivannan Sathiyanarayanan Manivannan Yukie Ueyama Yukie Ueyama Madhumita Basu Madhumita Basu Madhumita Basu Vidu Garg Vidu Garg Vidu Garg Vidu Garg |
| author_facet | Uddalak Majumdar Uddalak Majumdar Talita Z. Choudhury Talita Z. Choudhury Sathiyanarayanan Manivannan Sathiyanarayanan Manivannan Yukie Ueyama Yukie Ueyama Madhumita Basu Madhumita Basu Madhumita Basu Vidu Garg Vidu Garg Vidu Garg Vidu Garg |
| author_sort | Uddalak Majumdar |
| collection | DOAJ |
| description | Calcific aortic valve disease (CAVD) is an increasingly prevalent condition among the elderly population that is associated with significant morbidity and mortality. Insufficient understanding of the underlying disease mechanisms has hindered the development of pharmacologic therapies for CAVD. Recently, we described nitric oxide (NO) mediated S-nitrosylation as a novel mechanism for preventing the calcific process. We demonstrated that NO donor or an S-nitrosylating agent, S-nitrosoglutathione (GSNO), inhibits spontaneous calcification in porcine aortic valve interstitial cells (pAVICs) and this was supported by single-cell RNA sequencing (scRNAseq) that demonstrated NO donor and GSNO inhibited myofibroblast activation of pAVICs. Here, we investigated novel signaling pathways that are critical for the calcification of pAVICs that are altered by NO and GSNO by performing an in-depth analysis of the scRNA-seq dataset. Transcriptomic analysis revealed 1,247 differentially expressed genes in pAVICs after NO donor or GSNO treatment compared to untreated cells. Pathway-based analysis of the differentially expressed genes revealed an overrepresentation of the integrin signaling pathway, along with the Rho GTPase, Wnt, TGF-β, and p53 signaling pathways. We demonstrate that ITGA8 and VCL, two of the identified genes from the integrin signaling pathway, which are known to regulate cell-extracellular matrix (ECM) communication and focal adhesion, were upregulated in both in vitro and in vivo calcific conditions. Reduced expression of these genes after treatment with NO donor suggests that NO inhibits calcification by targeting myofibroblast adhesion and ECM remodeling. In addition, withdrawal of NO donor after 3 days of exposure revealed that NO-mediated transcriptional and translational regulation is a transient event and requires continuous NO exposure to inhibit calcification. Overall, our data suggest that NO and S-nitrosylation regulate the integrin signaling pathway to maintain healthy cell-ECM interaction and prevent CAVD. |
| first_indexed | 2024-04-13T18:14:18Z |
| format | Article |
| id | doaj.art-8db773e75a59473ebe8022861c469e6c |
| institution | Directory Open Access Journal |
| issn | 2297-055X |
| language | English |
| last_indexed | 2024-04-13T18:14:18Z |
| publishDate | 2022-10-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cardiovascular Medicine |
| spelling | doaj.art-8db773e75a59473ebe8022861c469e6c2022-12-22T02:35:46ZengFrontiers Media S.A.Frontiers in Cardiovascular Medicine2297-055X2022-10-01910.3389/fcvm.2022.742850742850Single-cell RNA-sequencing analysis of aortic valve interstitial cells demonstrates the regulation of integrin signaling by nitric oxideUddalak Majumdar0Uddalak Majumdar1Talita Z. Choudhury2Talita Z. Choudhury3Sathiyanarayanan Manivannan4Sathiyanarayanan Manivannan5Yukie Ueyama6Yukie Ueyama7Madhumita Basu8Madhumita Basu9Madhumita Basu10Vidu Garg11Vidu Garg12Vidu Garg13Vidu Garg14Center for Cardiovascular Research, Nationwide Children’s Hospital, Columbus, OH, United StatesThe Heart Center, Nationwide Children’s Hospital, Columbus, OH, United StatesCenter for Cardiovascular Research, Nationwide Children’s Hospital, Columbus, OH, United StatesThe Heart Center, Nationwide Children’s Hospital, Columbus, OH, United StatesCenter for Cardiovascular Research, Nationwide Children’s Hospital, Columbus, OH, United StatesThe Heart Center, Nationwide Children’s Hospital, Columbus, OH, United StatesCenter for Cardiovascular Research, Nationwide Children’s Hospital, Columbus, OH, United StatesThe Heart Center, Nationwide Children’s Hospital, Columbus, OH, United StatesCenter for Cardiovascular Research, Nationwide Children’s Hospital, Columbus, OH, United StatesThe Heart Center, Nationwide Children’s Hospital, Columbus, OH, United StatesDepartment of Pediatrics, The Ohio State University, Columbus, OH, United StatesCenter for Cardiovascular Research, Nationwide Children’s Hospital, Columbus, OH, United StatesThe Heart Center, Nationwide Children’s Hospital, Columbus, OH, United StatesDepartment of Pediatrics, The Ohio State University, Columbus, OH, United StatesDepartment of Molecular Genetics, The Ohio State University, Columbus, OH, United StatesCalcific aortic valve disease (CAVD) is an increasingly prevalent condition among the elderly population that is associated with significant morbidity and mortality. Insufficient understanding of the underlying disease mechanisms has hindered the development of pharmacologic therapies for CAVD. Recently, we described nitric oxide (NO) mediated S-nitrosylation as a novel mechanism for preventing the calcific process. We demonstrated that NO donor or an S-nitrosylating agent, S-nitrosoglutathione (GSNO), inhibits spontaneous calcification in porcine aortic valve interstitial cells (pAVICs) and this was supported by single-cell RNA sequencing (scRNAseq) that demonstrated NO donor and GSNO inhibited myofibroblast activation of pAVICs. Here, we investigated novel signaling pathways that are critical for the calcification of pAVICs that are altered by NO and GSNO by performing an in-depth analysis of the scRNA-seq dataset. Transcriptomic analysis revealed 1,247 differentially expressed genes in pAVICs after NO donor or GSNO treatment compared to untreated cells. Pathway-based analysis of the differentially expressed genes revealed an overrepresentation of the integrin signaling pathway, along with the Rho GTPase, Wnt, TGF-β, and p53 signaling pathways. We demonstrate that ITGA8 and VCL, two of the identified genes from the integrin signaling pathway, which are known to regulate cell-extracellular matrix (ECM) communication and focal adhesion, were upregulated in both in vitro and in vivo calcific conditions. Reduced expression of these genes after treatment with NO donor suggests that NO inhibits calcification by targeting myofibroblast adhesion and ECM remodeling. In addition, withdrawal of NO donor after 3 days of exposure revealed that NO-mediated transcriptional and translational regulation is a transient event and requires continuous NO exposure to inhibit calcification. Overall, our data suggest that NO and S-nitrosylation regulate the integrin signaling pathway to maintain healthy cell-ECM interaction and prevent CAVD.https://www.frontiersin.org/articles/10.3389/fcvm.2022.742850/fullcalcific aortic valve diseasenitric oxideS-nitrosylationsingle cell RNA-sequencingintegrin signalingextracellular matrix (ECM) |
| spellingShingle | Uddalak Majumdar Uddalak Majumdar Talita Z. Choudhury Talita Z. Choudhury Sathiyanarayanan Manivannan Sathiyanarayanan Manivannan Yukie Ueyama Yukie Ueyama Madhumita Basu Madhumita Basu Madhumita Basu Vidu Garg Vidu Garg Vidu Garg Vidu Garg Single-cell RNA-sequencing analysis of aortic valve interstitial cells demonstrates the regulation of integrin signaling by nitric oxide Frontiers in Cardiovascular Medicine calcific aortic valve disease nitric oxide S-nitrosylation single cell RNA-sequencing integrin signaling extracellular matrix (ECM) |
| title | Single-cell RNA-sequencing analysis of aortic valve interstitial cells demonstrates the regulation of integrin signaling by nitric oxide |
| title_full | Single-cell RNA-sequencing analysis of aortic valve interstitial cells demonstrates the regulation of integrin signaling by nitric oxide |
| title_fullStr | Single-cell RNA-sequencing analysis of aortic valve interstitial cells demonstrates the regulation of integrin signaling by nitric oxide |
| title_full_unstemmed | Single-cell RNA-sequencing analysis of aortic valve interstitial cells demonstrates the regulation of integrin signaling by nitric oxide |
| title_short | Single-cell RNA-sequencing analysis of aortic valve interstitial cells demonstrates the regulation of integrin signaling by nitric oxide |
| title_sort | single cell rna sequencing analysis of aortic valve interstitial cells demonstrates the regulation of integrin signaling by nitric oxide |
| topic | calcific aortic valve disease nitric oxide S-nitrosylation single cell RNA-sequencing integrin signaling extracellular matrix (ECM) |
| url | https://www.frontiersin.org/articles/10.3389/fcvm.2022.742850/full |
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