Characterization of the Cellular Microenvironment and Novel Specific Biomarkers in Pterygia Using RNA Sequencing
With a worldwide prevalence of ~12%, pterygium is a common degenerative and environmentally triggered ocular surface disorder characterized by wing-shaped growth of conjunctival tissue onto the cornea that can lead to blindness if left untreated. This study characterizes the transcriptional profile...
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Frontiers Media S.A.
2022-01-01
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| Series: | Frontiers in Medicine |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmed.2021.714458/full |
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| author | Julian Wolf Rozina Ida Hajdu Rozina Ida Hajdu Stefaniya Boneva Anja Schlecht Anja Schlecht Thabo Lapp Katrin Wacker Hansjürgen Agostini Thomas Reinhard Claudia Auw-Hädrich Günther Schlunck Clemens Lange Clemens Lange |
| author_facet | Julian Wolf Rozina Ida Hajdu Rozina Ida Hajdu Stefaniya Boneva Anja Schlecht Anja Schlecht Thabo Lapp Katrin Wacker Hansjürgen Agostini Thomas Reinhard Claudia Auw-Hädrich Günther Schlunck Clemens Lange Clemens Lange |
| author_sort | Julian Wolf |
| collection | DOAJ |
| description | With a worldwide prevalence of ~12%, pterygium is a common degenerative and environmentally triggered ocular surface disorder characterized by wing-shaped growth of conjunctival tissue onto the cornea that can lead to blindness if left untreated. This study characterizes the transcriptional profile and the cellular microenvironment of conjunctival pterygia and identifies novel pterygia-specific biomarkers. Formalin-fixed and paraffin-embedded pterygia as well as healthy conjunctival specimens were analyzed using MACE RNA sequencing (n = 8 each) and immunohistochemistry (pterygia n = 7, control n = 3). According to the bioinformatic cell type enrichment analysis using xCell, the cellular microenvironment of pterygia was characterized by an enrichment of myofibroblasts, T-lymphocytes and various antigen-presenting cells, including dendritic cells and macrophages. Differentially expressed genes that were increased in pterygia compared to control tissue were mainly involved in autophagy (including DCN, TMBIM6), cellular response to stress (including TPT1, DDX5) as well as fibroblast proliferation and epithelial to mesenchymal transition (including CTNNB1, TGFBR1, and FN1). Immunohistochemical analysis confirmed a significantly increased FN1 stromal immunoreactivity in pterygia when compared to control tissue. In addition, a variety of factors involved in apoptosis were significantly downregulated in pterygia, including LCN2, CTSD, and NISCH. Furthermore, 450 pterygia-specific biomarkers were identified by including transcriptional data of different ocular surface pathologies serving as controls (training group), which were then validated using transcriptional data of cultured human pterygium cells. Among the most pterygia-specific factors were transcripts such as AHNAK, RTN4, TPT1, FSTL1, and SPARC. Immunohistochemical validation of SPARC revealed a significantly increased stromal immunoreactivity in pterygia when compared to controls, most notably in vessels and intravascular vessel wall-adherent mononuclear cells. Taken together, the present study provides new insights into the cellular microenvironment and the transcriptional profile of pterygia, identifies new and specific biomarkers and in addition to fibrosis-related genes, uncovers autophagy, stress response and apoptosis modulation as pterygium-associated processes. These findings expand our understanding of the pathophysiology of pterygia, provide new diagnostic tools, and may enable new targeted therapeutic options for this common and sight-threatening ocular surface disease. |
| first_indexed | 2024-12-20T06:25:07Z |
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| id | doaj.art-907a6eef25cd4843b9014c4ee31c215d |
| institution | Directory Open Access Journal |
| issn | 2296-858X |
| language | English |
| last_indexed | 2024-12-20T06:25:07Z |
| publishDate | 2022-01-01 |
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| series | Frontiers in Medicine |
| spelling | doaj.art-907a6eef25cd4843b9014c4ee31c215d2022-12-21T19:50:20ZengFrontiers Media S.A.Frontiers in Medicine2296-858X2022-01-01810.3389/fmed.2021.714458714458Characterization of the Cellular Microenvironment and Novel Specific Biomarkers in Pterygia Using RNA SequencingJulian Wolf0Rozina Ida Hajdu1Rozina Ida Hajdu2Stefaniya Boneva3Anja Schlecht4Anja Schlecht5Thabo Lapp6Katrin Wacker7Hansjürgen Agostini8Thomas Reinhard9Claudia Auw-Hädrich10Günther Schlunck11Clemens Lange12Clemens Lange13Eye Center, Medical Center, Faculty of Medicine, University of Freiburg, Freiburg im Breisgau, GermanyEye Center, Medical Center, Faculty of Medicine, University of Freiburg, Freiburg im Breisgau, GermanyDepartment of Ophthalmology, Semmelweis University, Budapest, HungaryEye Center, Medical Center, Faculty of Medicine, University of Freiburg, Freiburg im Breisgau, GermanyEye Center, Medical Center, Faculty of Medicine, University of Freiburg, Freiburg im Breisgau, GermanyInstitute of Anatomy and Cell Biology, Wuerzburg University, Wuerzburg, GermanyEye Center, Medical Center, Faculty of Medicine, University of Freiburg, Freiburg im Breisgau, GermanyEye Center, Medical Center, Faculty of Medicine, University of Freiburg, Freiburg im Breisgau, GermanyEye Center, Medical Center, Faculty of Medicine, University of Freiburg, Freiburg im Breisgau, GermanyEye Center, Medical Center, Faculty of Medicine, University of Freiburg, Freiburg im Breisgau, GermanyEye Center, Medical Center, Faculty of Medicine, University of Freiburg, Freiburg im Breisgau, GermanyEye Center, Medical Center, Faculty of Medicine, University of Freiburg, Freiburg im Breisgau, GermanyEye Center, Medical Center, Faculty of Medicine, University of Freiburg, Freiburg im Breisgau, GermanyOphtha-Lab, Department of Ophthalmology, St. Franziskus Hospital, Münster, GermanyWith a worldwide prevalence of ~12%, pterygium is a common degenerative and environmentally triggered ocular surface disorder characterized by wing-shaped growth of conjunctival tissue onto the cornea that can lead to blindness if left untreated. This study characterizes the transcriptional profile and the cellular microenvironment of conjunctival pterygia and identifies novel pterygia-specific biomarkers. Formalin-fixed and paraffin-embedded pterygia as well as healthy conjunctival specimens were analyzed using MACE RNA sequencing (n = 8 each) and immunohistochemistry (pterygia n = 7, control n = 3). According to the bioinformatic cell type enrichment analysis using xCell, the cellular microenvironment of pterygia was characterized by an enrichment of myofibroblasts, T-lymphocytes and various antigen-presenting cells, including dendritic cells and macrophages. Differentially expressed genes that were increased in pterygia compared to control tissue were mainly involved in autophagy (including DCN, TMBIM6), cellular response to stress (including TPT1, DDX5) as well as fibroblast proliferation and epithelial to mesenchymal transition (including CTNNB1, TGFBR1, and FN1). Immunohistochemical analysis confirmed a significantly increased FN1 stromal immunoreactivity in pterygia when compared to control tissue. In addition, a variety of factors involved in apoptosis were significantly downregulated in pterygia, including LCN2, CTSD, and NISCH. Furthermore, 450 pterygia-specific biomarkers were identified by including transcriptional data of different ocular surface pathologies serving as controls (training group), which were then validated using transcriptional data of cultured human pterygium cells. Among the most pterygia-specific factors were transcripts such as AHNAK, RTN4, TPT1, FSTL1, and SPARC. Immunohistochemical validation of SPARC revealed a significantly increased stromal immunoreactivity in pterygia when compared to controls, most notably in vessels and intravascular vessel wall-adherent mononuclear cells. Taken together, the present study provides new insights into the cellular microenvironment and the transcriptional profile of pterygia, identifies new and specific biomarkers and in addition to fibrosis-related genes, uncovers autophagy, stress response and apoptosis modulation as pterygium-associated processes. These findings expand our understanding of the pathophysiology of pterygia, provide new diagnostic tools, and may enable new targeted therapeutic options for this common and sight-threatening ocular surface disease.https://www.frontiersin.org/articles/10.3389/fmed.2021.714458/fullconjunctival pterygiumRNA sequencingFFPExCellcellular microenvironment |
| spellingShingle | Julian Wolf Rozina Ida Hajdu Rozina Ida Hajdu Stefaniya Boneva Anja Schlecht Anja Schlecht Thabo Lapp Katrin Wacker Hansjürgen Agostini Thomas Reinhard Claudia Auw-Hädrich Günther Schlunck Clemens Lange Clemens Lange Characterization of the Cellular Microenvironment and Novel Specific Biomarkers in Pterygia Using RNA Sequencing Frontiers in Medicine conjunctival pterygium RNA sequencing FFPE xCell cellular microenvironment |
| title | Characterization of the Cellular Microenvironment and Novel Specific Biomarkers in Pterygia Using RNA Sequencing |
| title_full | Characterization of the Cellular Microenvironment and Novel Specific Biomarkers in Pterygia Using RNA Sequencing |
| title_fullStr | Characterization of the Cellular Microenvironment and Novel Specific Biomarkers in Pterygia Using RNA Sequencing |
| title_full_unstemmed | Characterization of the Cellular Microenvironment and Novel Specific Biomarkers in Pterygia Using RNA Sequencing |
| title_short | Characterization of the Cellular Microenvironment and Novel Specific Biomarkers in Pterygia Using RNA Sequencing |
| title_sort | characterization of the cellular microenvironment and novel specific biomarkers in pterygia using rna sequencing |
| topic | conjunctival pterygium RNA sequencing FFPE xCell cellular microenvironment |
| url | https://www.frontiersin.org/articles/10.3389/fmed.2021.714458/full |
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