Role of Hypoxia and the Adenosine System in Immune Evasion and Prognosis of Patients with Brain Metastases of Melanoma: A Multiplex Whole Slide Immunofluorescence Study
Following the introduction of immune checkpoint inhibitors, a substantial prolongation of the overall survival has been achieved for many patients with multiple brain metastases from melanoma. However, heterogeneity between individual tumor responses is incompletely understood. In order to determine...
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2020-12-01
|
| Series: | Cancers |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2072-6694/12/12/3753 |
| _version_ | 1797544798136041472 |
|---|---|
| author | Arnulf Mayer Maximilian Haist Carmen Loquai Stephan Grabbe Matthias Rapp Wilfried Roth Peter Vaupel Heinz Schmidberger |
| author_facet | Arnulf Mayer Maximilian Haist Carmen Loquai Stephan Grabbe Matthias Rapp Wilfried Roth Peter Vaupel Heinz Schmidberger |
| author_sort | Arnulf Mayer |
| collection | DOAJ |
| description | Following the introduction of immune checkpoint inhibitors, a substantial prolongation of the overall survival has been achieved for many patients with multiple brain metastases from melanoma. However, heterogeneity between individual tumor responses is incompletely understood. In order to determine the impact of the individual tumor phenotype on the prognosis of melanoma patients, we examined surgical sections from 33 patients who were treated with radiotherapy (whole-brain radiotherapy, WBRT, stereotactic radiotherapy, STX, or both) and Ipilimumab. We analyzed multiplex staining of the hypoxia marker GLUT-1, the adenosine (ADO)-associated enzymes CD73 and CD39, and CD8, a marker of cytotoxic T lymphocytes (CTL) on a single-cell basis using QuPath. Additionally, the MOSAIC interaction analysis algorithm was used to explore the hypothesis that CTL systematically avoid GLUT-1<sup>high</sup> tumor areas. Our results revealed, that a strong GLUT-1 expression, low numbers of CTL, or exclusion of CTL from the tumor were correlated with significant prognostic detriment. Hypoxic tumors overall have smaller amounts of CTL, and spatial analysis revealed a repellent effect of hypoxia on CTL. In contrast to in vitro studies, specific upregulation of ADO-related enzymes CD73 and CD39 in GLUT-1<sup>high</sup> tumor regions was never observed. In this study, we could show direct in vivo evidence for hypoxia-mediated immunosuppression in melanoma. Moreover, this study suggests a significant prognostic relevance of the tumor immune phenotype, the strength of CD8 infiltration in the tumor, and the expression of hypoxia marker GLUT-1 on melanoma cells. Last, our results suggest a temporal stability of the microenvironment-mediated immunosuppressive phenotype in melanoma. |
| first_indexed | 2024-03-10T14:05:36Z |
| format | Article |
| id | doaj.art-1b362069225d4b6993f60360834a2b7d |
| institution | Directory Open Access Journal |
| issn | 2072-6694 |
| language | English |
| last_indexed | 2024-03-10T14:05:36Z |
| publishDate | 2020-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cancers |
| spelling | doaj.art-1b362069225d4b6993f60360834a2b7d2023-11-21T00:38:12ZengMDPI AGCancers2072-66942020-12-011212375310.3390/cancers12123753Role of Hypoxia and the Adenosine System in Immune Evasion and Prognosis of Patients with Brain Metastases of Melanoma: A Multiplex Whole Slide Immunofluorescence StudyArnulf Mayer0Maximilian Haist1Carmen Loquai2Stephan Grabbe3Matthias Rapp4Wilfried Roth5Peter Vaupel6Heinz Schmidberger7Department of Radiation Oncology, University Medical Center of the Johannes Gutenberg University, 55131 Mainz, GermanyDepartment of Radiation Oncology, University Medical Center of the Johannes Gutenberg University, 55131 Mainz, GermanyDepartment of Dermatology, University Medical Center of the Johannes Gutenberg University, 55131 Mainz, GermanyDepartment of Dermatology, University Medical Center of the Johannes Gutenberg University, 55131 Mainz, GermanyDepartment of Radiation Oncology, University Medical Center of the Johannes Gutenberg University, 55131 Mainz, GermanyInstitute of Pathology, University Medical Center of the Johannes Gutenberg University, 55131 Mainz, GermanyDepartment of Radiation Oncology, University Medical Center of the Johannes Gutenberg University, 55131 Mainz, GermanyDepartment of Radiation Oncology, University Medical Center of the Johannes Gutenberg University, 55131 Mainz, GermanyFollowing the introduction of immune checkpoint inhibitors, a substantial prolongation of the overall survival has been achieved for many patients with multiple brain metastases from melanoma. However, heterogeneity between individual tumor responses is incompletely understood. In order to determine the impact of the individual tumor phenotype on the prognosis of melanoma patients, we examined surgical sections from 33 patients who were treated with radiotherapy (whole-brain radiotherapy, WBRT, stereotactic radiotherapy, STX, or both) and Ipilimumab. We analyzed multiplex staining of the hypoxia marker GLUT-1, the adenosine (ADO)-associated enzymes CD73 and CD39, and CD8, a marker of cytotoxic T lymphocytes (CTL) on a single-cell basis using QuPath. Additionally, the MOSAIC interaction analysis algorithm was used to explore the hypothesis that CTL systematically avoid GLUT-1<sup>high</sup> tumor areas. Our results revealed, that a strong GLUT-1 expression, low numbers of CTL, or exclusion of CTL from the tumor were correlated with significant prognostic detriment. Hypoxic tumors overall have smaller amounts of CTL, and spatial analysis revealed a repellent effect of hypoxia on CTL. In contrast to in vitro studies, specific upregulation of ADO-related enzymes CD73 and CD39 in GLUT-1<sup>high</sup> tumor regions was never observed. In this study, we could show direct in vivo evidence for hypoxia-mediated immunosuppression in melanoma. Moreover, this study suggests a significant prognostic relevance of the tumor immune phenotype, the strength of CD8 infiltration in the tumor, and the expression of hypoxia marker GLUT-1 on melanoma cells. Last, our results suggest a temporal stability of the microenvironment-mediated immunosuppressive phenotype in melanoma.https://www.mdpi.com/2072-6694/12/12/3753hypoxiaadenosinetumor microenvironmentimmunosuppressionmultiplex immunohistochemistryspatial statistics |
| spellingShingle | Arnulf Mayer Maximilian Haist Carmen Loquai Stephan Grabbe Matthias Rapp Wilfried Roth Peter Vaupel Heinz Schmidberger Role of Hypoxia and the Adenosine System in Immune Evasion and Prognosis of Patients with Brain Metastases of Melanoma: A Multiplex Whole Slide Immunofluorescence Study Cancers hypoxia adenosine tumor microenvironment immunosuppression multiplex immunohistochemistry spatial statistics |
| title | Role of Hypoxia and the Adenosine System in Immune Evasion and Prognosis of Patients with Brain Metastases of Melanoma: A Multiplex Whole Slide Immunofluorescence Study |
| title_full | Role of Hypoxia and the Adenosine System in Immune Evasion and Prognosis of Patients with Brain Metastases of Melanoma: A Multiplex Whole Slide Immunofluorescence Study |
| title_fullStr | Role of Hypoxia and the Adenosine System in Immune Evasion and Prognosis of Patients with Brain Metastases of Melanoma: A Multiplex Whole Slide Immunofluorescence Study |
| title_full_unstemmed | Role of Hypoxia and the Adenosine System in Immune Evasion and Prognosis of Patients with Brain Metastases of Melanoma: A Multiplex Whole Slide Immunofluorescence Study |
| title_short | Role of Hypoxia and the Adenosine System in Immune Evasion and Prognosis of Patients with Brain Metastases of Melanoma: A Multiplex Whole Slide Immunofluorescence Study |
| title_sort | role of hypoxia and the adenosine system in immune evasion and prognosis of patients with brain metastases of melanoma a multiplex whole slide immunofluorescence study |
| topic | hypoxia adenosine tumor microenvironment immunosuppression multiplex immunohistochemistry spatial statistics |
| url | https://www.mdpi.com/2072-6694/12/12/3753 |
| work_keys_str_mv | AT arnulfmayer roleofhypoxiaandtheadenosinesysteminimmuneevasionandprognosisofpatientswithbrainmetastasesofmelanomaamultiplexwholeslideimmunofluorescencestudy AT maximilianhaist roleofhypoxiaandtheadenosinesysteminimmuneevasionandprognosisofpatientswithbrainmetastasesofmelanomaamultiplexwholeslideimmunofluorescencestudy AT carmenloquai roleofhypoxiaandtheadenosinesysteminimmuneevasionandprognosisofpatientswithbrainmetastasesofmelanomaamultiplexwholeslideimmunofluorescencestudy AT stephangrabbe roleofhypoxiaandtheadenosinesysteminimmuneevasionandprognosisofpatientswithbrainmetastasesofmelanomaamultiplexwholeslideimmunofluorescencestudy AT matthiasrapp roleofhypoxiaandtheadenosinesysteminimmuneevasionandprognosisofpatientswithbrainmetastasesofmelanomaamultiplexwholeslideimmunofluorescencestudy AT wilfriedroth roleofhypoxiaandtheadenosinesysteminimmuneevasionandprognosisofpatientswithbrainmetastasesofmelanomaamultiplexwholeslideimmunofluorescencestudy AT petervaupel roleofhypoxiaandtheadenosinesysteminimmuneevasionandprognosisofpatientswithbrainmetastasesofmelanomaamultiplexwholeslideimmunofluorescencestudy AT heinzschmidberger roleofhypoxiaandtheadenosinesysteminimmuneevasionandprognosisofpatientswithbrainmetastasesofmelanomaamultiplexwholeslideimmunofluorescencestudy |