Cytotoxic response of tumor-infiltrating lymphocytes of head and neck cancer slice cultures under mitochondrial dysfunction
BackgroundHead and neck squamous cell carcinomas (HNSCC) are highly heterogeneous tumors. In the harsh tumor microenvironment (TME), metabolic reprogramming and mitochondrial dysfunction may lead to immunosuppressive phenotypes. Aerobic glycolysis is needed for the activation of cytotoxic T-cells an...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2024-03-01
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| Series: | Frontiers in Oncology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fonc.2024.1364577/full |
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| author | Maria do Carmo Greier Annette Runge Jozsef Dudas Roland Hartl Matthias Santer Daniel Dejaco Teresa Bernadette Steinbichler Julia Federspiel Christof Seifarth Marko Konschake Susanne Sprung Sieghart Sopper Avneet Randhawa Melissa Mayr Benedikt Gabriel Hofauer Herbert Riechelmann |
| author_facet | Maria do Carmo Greier Annette Runge Jozsef Dudas Roland Hartl Matthias Santer Daniel Dejaco Teresa Bernadette Steinbichler Julia Federspiel Christof Seifarth Marko Konschake Susanne Sprung Sieghart Sopper Avneet Randhawa Melissa Mayr Benedikt Gabriel Hofauer Herbert Riechelmann |
| author_sort | Maria do Carmo Greier |
| collection | DOAJ |
| description | BackgroundHead and neck squamous cell carcinomas (HNSCC) are highly heterogeneous tumors. In the harsh tumor microenvironment (TME), metabolic reprogramming and mitochondrial dysfunction may lead to immunosuppressive phenotypes. Aerobic glycolysis is needed for the activation of cytotoxic T-cells and the absence of glucose may hamper the full effector functions of cytotoxic T-cells. To test the effect of mitochondrial dysfunction on cytotoxic T cell function, slice cultures (SC) of HNSCC cancer were cultivated under different metabolic conditions.MethodsTumor samples from 21 patients with HNSCC were collected, from which, SC were established and cultivated under six different conditions. These conditions included high glucose, T cell stimulation, and temporarily induced mitochondrial dysfunction (MitoDys) using FCCP and oligomycin A with or without additional T cell stimulation, high glucose and finally, a control medium. Over three days of cultivation, sequential T cell stimulation and MitoDys treatments were performed. Supernatant was collected, and SC were fixed and embedded. Granzyme B was measured in the supernatant and in the SC via immunohistochemistry (IHC). Staining of PD1, CD8/Ki67, and cleavedcaspase3 (CC3) were performed in SC.ResultsHematoxylin eosin stains showed that overall SC quality remained stable over 3 days of cultivation. T cell stimulation, both alone and combined with MitoDys, led to significantly increased granzyme levels in SC and in supernatant. Apoptosis following T cell stimulation was observed in tumor and stroma. Mitochondrial dysfunction alone increased apoptosis in tumor cell aggregates. High glucose concentration alone had no impact on T cell activity and apoptosis. Apoptosis rates were significantly lower under conditions with high glucose and MitoDys (p=0.03).ConclusionStimulation of tumor-infiltrating lymphocytes in SC was feasible, which led to increased apoptosis in tumor cells. Induced mitochondrial dysfunction did not play a significant role in the activation and function of TILs in SC of HNSCC. Moreover, high glucose concentration did not promote cytotoxic T cell activity in HNSCC SC. |
| first_indexed | 2024-03-07T14:03:07Z |
| format | Article |
| id | doaj.art-7e69a60e5eab45c68f556a77a7892fb6 |
| institution | Directory Open Access Journal |
| issn | 2234-943X |
| language | English |
| last_indexed | 2024-03-07T14:03:07Z |
| publishDate | 2024-03-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Oncology |
| spelling | doaj.art-7e69a60e5eab45c68f556a77a7892fb62024-03-07T04:52:04ZengFrontiers Media S.A.Frontiers in Oncology2234-943X2024-03-011410.3389/fonc.2024.13645771364577Cytotoxic response of tumor-infiltrating lymphocytes of head and neck cancer slice cultures under mitochondrial dysfunctionMaria do Carmo Greier0Annette Runge1Jozsef Dudas2Roland Hartl3Matthias Santer4Daniel Dejaco5Teresa Bernadette Steinbichler6Julia Federspiel7Christof Seifarth8Marko Konschake9Susanne Sprung10Sieghart Sopper11Avneet Randhawa12Melissa Mayr13Benedikt Gabriel Hofauer14Herbert Riechelmann15Department of Otorhinolaryngology, Head and Neck Surgery, Medical University of Innsbruck, Innsbruck, AustriaDepartment of Otorhinolaryngology, Head and Neck Surgery, Medical University of Innsbruck, Innsbruck, AustriaDepartment of Otorhinolaryngology, Head and Neck Surgery, Medical University of Innsbruck, Innsbruck, AustriaDepartment of Otorhinolaryngology, Head and Neck Surgery, Medical University of Innsbruck, Innsbruck, AustriaDepartment of Otorhinolaryngology, Head and Neck Surgery, Medical University of Innsbruck, Innsbruck, AustriaDepartment of Otorhinolaryngology, Head and Neck Surgery, Medical University of Innsbruck, Innsbruck, AustriaDepartment of Otorhinolaryngology, Head and Neck Surgery, Medical University of Innsbruck, Innsbruck, AustriaDepartment of Otorhinolaryngology, Head and Neck Surgery, Medical University of Innsbruck, Innsbruck, AustriaInstitute for Clinical and Functional Anatomy, Medical University Innsbruck (MUI), Innsbruck, AustriaInstitute for Clinical and Functional Anatomy, Medical University Innsbruck (MUI), Innsbruck, AustriaINNPATH GmbH, Institute for Pathology, Innsbruck, AustriaClinic for Internal Medicine V, Medical University Innsbruck, Innsbruck, AustriaDepartment of Otolaryngology, Rutgers University Medical School, Newark, NJ, United StatesViraTherapeutics GmbH, Rum, AustriaDepartment of Otorhinolaryngology, Head and Neck Surgery, Medical University of Innsbruck, Innsbruck, AustriaDepartment of Otorhinolaryngology, Head and Neck Surgery, Medical University of Innsbruck, Innsbruck, AustriaBackgroundHead and neck squamous cell carcinomas (HNSCC) are highly heterogeneous tumors. In the harsh tumor microenvironment (TME), metabolic reprogramming and mitochondrial dysfunction may lead to immunosuppressive phenotypes. Aerobic glycolysis is needed for the activation of cytotoxic T-cells and the absence of glucose may hamper the full effector functions of cytotoxic T-cells. To test the effect of mitochondrial dysfunction on cytotoxic T cell function, slice cultures (SC) of HNSCC cancer were cultivated under different metabolic conditions.MethodsTumor samples from 21 patients with HNSCC were collected, from which, SC were established and cultivated under six different conditions. These conditions included high glucose, T cell stimulation, and temporarily induced mitochondrial dysfunction (MitoDys) using FCCP and oligomycin A with or without additional T cell stimulation, high glucose and finally, a control medium. Over three days of cultivation, sequential T cell stimulation and MitoDys treatments were performed. Supernatant was collected, and SC were fixed and embedded. Granzyme B was measured in the supernatant and in the SC via immunohistochemistry (IHC). Staining of PD1, CD8/Ki67, and cleavedcaspase3 (CC3) were performed in SC.ResultsHematoxylin eosin stains showed that overall SC quality remained stable over 3 days of cultivation. T cell stimulation, both alone and combined with MitoDys, led to significantly increased granzyme levels in SC and in supernatant. Apoptosis following T cell stimulation was observed in tumor and stroma. Mitochondrial dysfunction alone increased apoptosis in tumor cell aggregates. High glucose concentration alone had no impact on T cell activity and apoptosis. Apoptosis rates were significantly lower under conditions with high glucose and MitoDys (p=0.03).ConclusionStimulation of tumor-infiltrating lymphocytes in SC was feasible, which led to increased apoptosis in tumor cells. Induced mitochondrial dysfunction did not play a significant role in the activation and function of TILs in SC of HNSCC. Moreover, high glucose concentration did not promote cytotoxic T cell activity in HNSCC SC.https://www.frontiersin.org/articles/10.3389/fonc.2024.1364577/fullimmune responsecytotoxic T-cellsmitochondrial electron transport chainhead and neck carcinomamitochondrial dysfunction |
| spellingShingle | Maria do Carmo Greier Annette Runge Jozsef Dudas Roland Hartl Matthias Santer Daniel Dejaco Teresa Bernadette Steinbichler Julia Federspiel Christof Seifarth Marko Konschake Susanne Sprung Sieghart Sopper Avneet Randhawa Melissa Mayr Benedikt Gabriel Hofauer Herbert Riechelmann Cytotoxic response of tumor-infiltrating lymphocytes of head and neck cancer slice cultures under mitochondrial dysfunction Frontiers in Oncology immune response cytotoxic T-cells mitochondrial electron transport chain head and neck carcinoma mitochondrial dysfunction |
| title | Cytotoxic response of tumor-infiltrating lymphocytes of head and neck cancer slice cultures under mitochondrial dysfunction |
| title_full | Cytotoxic response of tumor-infiltrating lymphocytes of head and neck cancer slice cultures under mitochondrial dysfunction |
| title_fullStr | Cytotoxic response of tumor-infiltrating lymphocytes of head and neck cancer slice cultures under mitochondrial dysfunction |
| title_full_unstemmed | Cytotoxic response of tumor-infiltrating lymphocytes of head and neck cancer slice cultures under mitochondrial dysfunction |
| title_short | Cytotoxic response of tumor-infiltrating lymphocytes of head and neck cancer slice cultures under mitochondrial dysfunction |
| title_sort | cytotoxic response of tumor infiltrating lymphocytes of head and neck cancer slice cultures under mitochondrial dysfunction |
| topic | immune response cytotoxic T-cells mitochondrial electron transport chain head and neck carcinoma mitochondrial dysfunction |
| url | https://www.frontiersin.org/articles/10.3389/fonc.2024.1364577/full |
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