Towards unravelling biological mechanisms behind radiation-induced oral mucositis via mass spectrometry-based proteomics
ObjectiveHead and neck cancer (HNC) accounts for almost 890,000 new cases per year. Radiotherapy (RT) is used to treat the majority of these patients. A common side-effect of RT is the onset of oral mucositis, which decreases the quality of life and represents the major dose-limiting factor in RT. T...
| Main Authors: | , , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2023-06-01
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| Series: | Frontiers in Oncology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fonc.2023.1180642/full |
| _version_ | 1797805486955823104 |
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| author | Prabal Subedi Katharina Huber Christoph Sterr Anne Dietz Lukas Strasser Felix Kaestle Stefanie M. Hauck Lukas Duchrow Christine Aldrian Elsa Beatriz Monroy Ordonez Benedikt Luka Andreas R. Thomsen Andreas R. Thomsen Michael Henke Michael Henke Maria Gomolka Ute Rößler Omid Azimzadeh Simone Moertl Sabine Hornhardt |
| author_facet | Prabal Subedi Katharina Huber Christoph Sterr Anne Dietz Lukas Strasser Felix Kaestle Stefanie M. Hauck Lukas Duchrow Christine Aldrian Elsa Beatriz Monroy Ordonez Benedikt Luka Andreas R. Thomsen Andreas R. Thomsen Michael Henke Michael Henke Maria Gomolka Ute Rößler Omid Azimzadeh Simone Moertl Sabine Hornhardt |
| author_sort | Prabal Subedi |
| collection | DOAJ |
| description | ObjectiveHead and neck cancer (HNC) accounts for almost 890,000 new cases per year. Radiotherapy (RT) is used to treat the majority of these patients. A common side-effect of RT is the onset of oral mucositis, which decreases the quality of life and represents the major dose-limiting factor in RT. To understand the origin of oral mucositis, the biological mechanisms post-ionizing radiation (IR) need to be clarified. Such knowledge is valuable to develop new treatment targets for oral mucositis and markers for the early identification of “at-risk” patients.MethodsPrimary keratinocytes from healthy volunteers were biopsied, irradiated in vitro (0 and 6 Gy), and subjected to mass spectrometry-based analyses 96 h after irradiation. Web-based tools were used to predict triggered biological pathways. The results were validated in the OKF6 cell culture model. Immunoblotting and mRNA validation was performed and cytokines present in cell culture media post-IR were quantified.ResultsMass spectrometry-based proteomics identified 5879 proteins in primary keratinocytes and 4597 proteins in OKF6 cells. Amongst them, 212 proteins in primary keratinocytes and 169 proteins in OKF6 cells were differentially abundant 96 h after 6 Gy irradiation compared to sham-irradiated controls. In silico pathway enrichment analysis predicted interferon (IFN) response and DNA strand elongation pathways as mostly affected pathways in both cell systems. Immunoblot validations showed a decrease in minichromosome maintenance (MCM) complex proteins 2-7 and an increase in IFN-associated proteins STAT1 and ISG15. In line with affected IFN signalling, mRNA levels of IFNβ and interleukin 6 (IL-6) increased significantly following irradiation and also levels of secreted IL-1β, IL-6, IP-10, and ISG15 were elevated.ConclusionThis study has investigated biological mechanisms in keratinocytes post-in vitro ionizing radiation. A common radiation signature in keratinocytes was identified. The role of IFN response in keratinocytes along with increased levels of pro-inflammatory cytokines and proteins could hint towards a possible mechanism for oral mucositis. |
| first_indexed | 2024-03-13T05:51:48Z |
| format | Article |
| id | doaj.art-54557a50150149be9e67c2ae864cb7e6 |
| institution | Directory Open Access Journal |
| issn | 2234-943X |
| language | English |
| last_indexed | 2024-03-13T05:51:48Z |
| publishDate | 2023-06-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Oncology |
| spelling | doaj.art-54557a50150149be9e67c2ae864cb7e62023-06-13T11:25:19ZengFrontiers Media S.A.Frontiers in Oncology2234-943X2023-06-011310.3389/fonc.2023.11806421180642Towards unravelling biological mechanisms behind radiation-induced oral mucositis via mass spectrometry-based proteomicsPrabal Subedi0Katharina Huber1Christoph Sterr2Anne Dietz3Lukas Strasser4Felix Kaestle5Stefanie M. Hauck6Lukas Duchrow7Christine Aldrian8Elsa Beatriz Monroy Ordonez9Benedikt Luka10Andreas R. Thomsen11Andreas R. Thomsen12Michael Henke13Michael Henke14Maria Gomolka15Ute Rößler16Omid Azimzadeh17Simone Moertl18Sabine Hornhardt19Bundesamt für Strahlenschutz/Federal Office for Radiation Protection, Neuherberg, GermanyBundesamt für Strahlenschutz/Federal Office for Radiation Protection, Neuherberg, GermanyBundesamt für Strahlenschutz/Federal Office for Radiation Protection, Neuherberg, GermanyBundesamt für Strahlenschutz/Federal Office for Radiation Protection, Neuherberg, GermanyBundesamt für Strahlenschutz/Federal Office for Radiation Protection, Neuherberg, GermanyBundesamt für Strahlenschutz/Federal Office for Radiation Protection, Neuherberg, GermanyHelmholtz Zentrum München, German Research Centre for Environmental Health, Metabolomics and Proteomics Core, Munich, GermanyBundesamt für Strahlenschutz/Federal Office for Radiation Protection, Neuherberg, GermanyDepartment of Radiation Oncology, Medical Center, Faculty of Medicine, University of Freiburg, German Cancer Consortium (DKTK) partner site Freiburg, Freiburg, GermanyDepartment of Radiation Oncology, Medical Center, Faculty of Medicine, University of Freiburg, German Cancer Consortium (DKTK) partner site Freiburg, Freiburg, GermanyDepartment of Conservative Dentistry Periodontology and Preventive Dentistry, Hannover Medical School (MHH), Hannover, GermanyDepartment of Radiation Oncology, Medical Center, Faculty of Medicine, University of Freiburg, German Cancer Consortium (DKTK) partner site Freiburg, Freiburg, GermanyGerman Cancer Consortium (DKTK) Partner Site Freiburg, German Cancer Research Center (dkfz), Heidelberg, GermanyDepartment of Radiation Oncology, Medical Center, Faculty of Medicine, University of Freiburg, German Cancer Consortium (DKTK) partner site Freiburg, Freiburg, GermanyGerman Cancer Consortium (DKTK) Partner Site Freiburg, German Cancer Research Center (dkfz), Heidelberg, GermanyBundesamt für Strahlenschutz/Federal Office for Radiation Protection, Neuherberg, GermanyBundesamt für Strahlenschutz/Federal Office for Radiation Protection, Neuherberg, GermanyBundesamt für Strahlenschutz/Federal Office for Radiation Protection, Neuherberg, GermanyBundesamt für Strahlenschutz/Federal Office for Radiation Protection, Neuherberg, GermanyBundesamt für Strahlenschutz/Federal Office for Radiation Protection, Neuherberg, GermanyObjectiveHead and neck cancer (HNC) accounts for almost 890,000 new cases per year. Radiotherapy (RT) is used to treat the majority of these patients. A common side-effect of RT is the onset of oral mucositis, which decreases the quality of life and represents the major dose-limiting factor in RT. To understand the origin of oral mucositis, the biological mechanisms post-ionizing radiation (IR) need to be clarified. Such knowledge is valuable to develop new treatment targets for oral mucositis and markers for the early identification of “at-risk” patients.MethodsPrimary keratinocytes from healthy volunteers were biopsied, irradiated in vitro (0 and 6 Gy), and subjected to mass spectrometry-based analyses 96 h after irradiation. Web-based tools were used to predict triggered biological pathways. The results were validated in the OKF6 cell culture model. Immunoblotting and mRNA validation was performed and cytokines present in cell culture media post-IR were quantified.ResultsMass spectrometry-based proteomics identified 5879 proteins in primary keratinocytes and 4597 proteins in OKF6 cells. Amongst them, 212 proteins in primary keratinocytes and 169 proteins in OKF6 cells were differentially abundant 96 h after 6 Gy irradiation compared to sham-irradiated controls. In silico pathway enrichment analysis predicted interferon (IFN) response and DNA strand elongation pathways as mostly affected pathways in both cell systems. Immunoblot validations showed a decrease in minichromosome maintenance (MCM) complex proteins 2-7 and an increase in IFN-associated proteins STAT1 and ISG15. In line with affected IFN signalling, mRNA levels of IFNβ and interleukin 6 (IL-6) increased significantly following irradiation and also levels of secreted IL-1β, IL-6, IP-10, and ISG15 were elevated.ConclusionThis study has investigated biological mechanisms in keratinocytes post-in vitro ionizing radiation. A common radiation signature in keratinocytes was identified. The role of IFN response in keratinocytes along with increased levels of pro-inflammatory cytokines and proteins could hint towards a possible mechanism for oral mucositis.https://www.frontiersin.org/articles/10.3389/fonc.2023.1180642/fullradiotherapykeratinocytesbiomarkermass spectrometry-based proteomicsinterferon responseSTAT phosphorylation |
| spellingShingle | Prabal Subedi Katharina Huber Christoph Sterr Anne Dietz Lukas Strasser Felix Kaestle Stefanie M. Hauck Lukas Duchrow Christine Aldrian Elsa Beatriz Monroy Ordonez Benedikt Luka Andreas R. Thomsen Andreas R. Thomsen Michael Henke Michael Henke Maria Gomolka Ute Rößler Omid Azimzadeh Simone Moertl Sabine Hornhardt Towards unravelling biological mechanisms behind radiation-induced oral mucositis via mass spectrometry-based proteomics Frontiers in Oncology radiotherapy keratinocytes biomarker mass spectrometry-based proteomics interferon response STAT phosphorylation |
| title | Towards unravelling biological mechanisms behind radiation-induced oral mucositis via mass spectrometry-based proteomics |
| title_full | Towards unravelling biological mechanisms behind radiation-induced oral mucositis via mass spectrometry-based proteomics |
| title_fullStr | Towards unravelling biological mechanisms behind radiation-induced oral mucositis via mass spectrometry-based proteomics |
| title_full_unstemmed | Towards unravelling biological mechanisms behind radiation-induced oral mucositis via mass spectrometry-based proteomics |
| title_short | Towards unravelling biological mechanisms behind radiation-induced oral mucositis via mass spectrometry-based proteomics |
| title_sort | towards unravelling biological mechanisms behind radiation induced oral mucositis via mass spectrometry based proteomics |
| topic | radiotherapy keratinocytes biomarker mass spectrometry-based proteomics interferon response STAT phosphorylation |
| url | https://www.frontiersin.org/articles/10.3389/fonc.2023.1180642/full |
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