EZH2 as a major histone methyltransferase in PDGF-BB-activated orbital fibroblast in the pathogenesis of Graves’ ophthalmopathy
Abstract Graves’ ophthalmopathy (GO) is an extra-thyroidal complication of Graves’ disease which can lead to vision loss in severe cases. Currently, treatments of GO are not sufficiently effective, so novel therapeutic strategies are needed. As platelet-derived growth factor (PDGF)-BB induces severa...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2024-04-01
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| Series: | Scientific Reports |
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| Online Access: | https://doi.org/10.1038/s41598-024-57926-x |
| _version_ | 1827291592796405760 |
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| author | Sopita Visamol Tanapat Palaga Preamjit Saonanon Vannakorn Pruksakorn Nattiya Hirankarn P. Martin van Hagen Willem A. Dik Sita Virakul |
| author_facet | Sopita Visamol Tanapat Palaga Preamjit Saonanon Vannakorn Pruksakorn Nattiya Hirankarn P. Martin van Hagen Willem A. Dik Sita Virakul |
| author_sort | Sopita Visamol |
| collection | DOAJ |
| description | Abstract Graves’ ophthalmopathy (GO) is an extra-thyroidal complication of Graves’ disease which can lead to vision loss in severe cases. Currently, treatments of GO are not sufficiently effective, so novel therapeutic strategies are needed. As platelet-derived growth factor (PDGF)-BB induces several effector mechanisms in GO orbital fibroblasts including cytokine production and myofibroblast activation, this study aims to investigate the roles of histone lysine methyltransferases (HKMTs) in PDGF-BB-activated GO orbital fibroblasts by screening with HKMTs inhibitors library. From the total of twelve selective HKMT inhibitors in the library, EZH2, G9a and DOT1L inhibitors, DZNeP, BIX01294 and Pinometostat, respectively, prevented PDGF-BB-induced proliferation and hyaluronan production by GO orbital fibroblasts. However, only EZH2 inhibitor, DZNeP, significantly blocked pro-inflammatory cytokine production. For the HKMTs expression in GO orbital fibroblasts, PDGF-BB significantly and time-dependently induced EZH2, G9a and DOT1L mRNA expression. To confirm the role of EZH2 in PDGF-BB-induced orbital fibroblast activation, EZH2 silencing experiments revealed suppression of PDGF-BB-induced collagen type I and α-SMA expression along with decreasing histone H3 lysine 27 trimethylation (H3K27me3) level. In a more clinically relevant model than orbital fibroblast culture experiments, DZNeP treated GO orbital tissues significantly reduced pro-inflammatory cytokine production while slightly reduced ACTA2 mRNA expression. Our data is the first to demonstrate that among all HKMTs EZH2 dominantly involved in the expression of myofibroblast markers in PDGF-BB-activated orbital fibroblast from GO presumably via H3K27me3. Thus, EZH2 may represent a novel therapeutics target for GO. |
| first_indexed | 2024-04-24T12:40:39Z |
| format | Article |
| id | doaj.art-8dd7fca722344457ab37162a3d76d16f |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-04-24T12:40:39Z |
| publishDate | 2024-04-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-8dd7fca722344457ab37162a3d76d16f2024-04-07T11:17:07ZengNature PortfolioScientific Reports2045-23222024-04-0114111010.1038/s41598-024-57926-xEZH2 as a major histone methyltransferase in PDGF-BB-activated orbital fibroblast in the pathogenesis of Graves’ ophthalmopathySopita Visamol0Tanapat Palaga1Preamjit Saonanon2Vannakorn Pruksakorn3Nattiya Hirankarn4P. Martin van Hagen5Willem A. Dik6Sita Virakul7Medical Microbiology, Interdisciplinary Program, Graduate School, Chulalongkorn UniversityDepartment of Microbiology, Faculty of Science, Chulalongkorn UniversityDepartment of Ophthalmology, Faculty of Medicine, Chulalongkorn UniversityDepartment of Ophthalmology, Faculty of Medicine, Chulalongkorn UniversityDepartment of Microbiology, Faculty of Medicine, Center of Excellence in Immunology and Immune Mediated Disease, Chulalongkorn UniversityDepartment of Microbiology, Faculty of Medicine, Center of Excellence in Immunology and Immune Mediated Disease, Chulalongkorn UniversityLaboratory Medical Immunology, Department of Immunology, Erasmus MC, University Medical CenterDepartment of Microbiology, Faculty of Science, Chulalongkorn UniversityAbstract Graves’ ophthalmopathy (GO) is an extra-thyroidal complication of Graves’ disease which can lead to vision loss in severe cases. Currently, treatments of GO are not sufficiently effective, so novel therapeutic strategies are needed. As platelet-derived growth factor (PDGF)-BB induces several effector mechanisms in GO orbital fibroblasts including cytokine production and myofibroblast activation, this study aims to investigate the roles of histone lysine methyltransferases (HKMTs) in PDGF-BB-activated GO orbital fibroblasts by screening with HKMTs inhibitors library. From the total of twelve selective HKMT inhibitors in the library, EZH2, G9a and DOT1L inhibitors, DZNeP, BIX01294 and Pinometostat, respectively, prevented PDGF-BB-induced proliferation and hyaluronan production by GO orbital fibroblasts. However, only EZH2 inhibitor, DZNeP, significantly blocked pro-inflammatory cytokine production. For the HKMTs expression in GO orbital fibroblasts, PDGF-BB significantly and time-dependently induced EZH2, G9a and DOT1L mRNA expression. To confirm the role of EZH2 in PDGF-BB-induced orbital fibroblast activation, EZH2 silencing experiments revealed suppression of PDGF-BB-induced collagen type I and α-SMA expression along with decreasing histone H3 lysine 27 trimethylation (H3K27me3) level. In a more clinically relevant model than orbital fibroblast culture experiments, DZNeP treated GO orbital tissues significantly reduced pro-inflammatory cytokine production while slightly reduced ACTA2 mRNA expression. Our data is the first to demonstrate that among all HKMTs EZH2 dominantly involved in the expression of myofibroblast markers in PDGF-BB-activated orbital fibroblast from GO presumably via H3K27me3. Thus, EZH2 may represent a novel therapeutics target for GO.https://doi.org/10.1038/s41598-024-57926-xHistone lysine methyltransferasesEZH2H3K27me3Graves’ ophthalmopathyOrbital fibroblast |
| spellingShingle | Sopita Visamol Tanapat Palaga Preamjit Saonanon Vannakorn Pruksakorn Nattiya Hirankarn P. Martin van Hagen Willem A. Dik Sita Virakul EZH2 as a major histone methyltransferase in PDGF-BB-activated orbital fibroblast in the pathogenesis of Graves’ ophthalmopathy Scientific Reports Histone lysine methyltransferases EZH2 H3K27me3 Graves’ ophthalmopathy Orbital fibroblast |
| title | EZH2 as a major histone methyltransferase in PDGF-BB-activated orbital fibroblast in the pathogenesis of Graves’ ophthalmopathy |
| title_full | EZH2 as a major histone methyltransferase in PDGF-BB-activated orbital fibroblast in the pathogenesis of Graves’ ophthalmopathy |
| title_fullStr | EZH2 as a major histone methyltransferase in PDGF-BB-activated orbital fibroblast in the pathogenesis of Graves’ ophthalmopathy |
| title_full_unstemmed | EZH2 as a major histone methyltransferase in PDGF-BB-activated orbital fibroblast in the pathogenesis of Graves’ ophthalmopathy |
| title_short | EZH2 as a major histone methyltransferase in PDGF-BB-activated orbital fibroblast in the pathogenesis of Graves’ ophthalmopathy |
| title_sort | ezh2 as a major histone methyltransferase in pdgf bb activated orbital fibroblast in the pathogenesis of graves ophthalmopathy |
| topic | Histone lysine methyltransferases EZH2 H3K27me3 Graves’ ophthalmopathy Orbital fibroblast |
| url | https://doi.org/10.1038/s41598-024-57926-x |
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