Location of CD39+ T cell subpopulations within tumors predict differential outcomes in non-small cell lung cancer
Purpose An improved mechanistic understanding of immunosuppressive pathways in non-small cell lung cancer (NSCLC) is important to develop novel diagnostic and therapeutic approaches. Here, we investigate the prognostic significance of the ectonucleotidases CD39 and CD73 in NSCLC.Experimental design...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMJ Publishing Group
2023-08-01
|
| Series: | Journal for ImmunoTherapy of Cancer |
| Online Access: | https://jitc.bmj.com/content/11/8/e006770.full |
| _version_ | 1797315932148727808 |
|---|---|
| author | Richard O’Connor David A Dorward Ahsan R Akram Lilian Koppensteiner Layla Mathieson Samuel Pattle |
| author_facet | Richard O’Connor David A Dorward Ahsan R Akram Lilian Koppensteiner Layla Mathieson Samuel Pattle |
| author_sort | Richard O’Connor |
| collection | DOAJ |
| description | Purpose An improved mechanistic understanding of immunosuppressive pathways in non-small cell lung cancer (NSCLC) is important to develop novel diagnostic and therapeutic approaches. Here, we investigate the prognostic significance of the ectonucleotidases CD39 and CD73 in NSCLC.Experimental design The expression and localization of CD39, CD73 and CD103 was digitally quantified in a cohort of 162 early treatment naïve NSCLC patients using multiplex-immunofluorescence and related to patient outcome. Expression among different cell-populations was assessed via flow cytometry. Targeted RNA-Seq was performed on CD4+ and CD8+ T cells from digested NSCLC tumor tissue and single-cell RNA-Seq data was analyzed to investigate the functional significance of CD39+ T cell populations.Results We demonstrate that flow cytometry of early untreated NSCLC patients shows an upregulation of CD39 expression in the tumor tissue among natural killer (NK) cells, fibroblasts and T cells. CD73 expression is mainly found among fibroblasts and Epcam+cells in the tumor tissue. Multiplex Immunofluorescence in a cohort of 162 early untreated NSCLC patients demonstrates that CD39 expression is mainly localized in the tumor stroma while CD73 expression is equally distributed between tumor nest and stroma, and high expression of CD39 and CD73 in the tumor stroma is associated with poor recurrence-free survival (RFS) at 5 years. Additionally, we find that CD8+T cells located in the tumor nest express CD103 and the density of CD39+CD103+CD8+ T cells in the tumor nest predicts improved RFS at 5 years. Targeted RNA-Seq shows that the tumor microenvironment of NSCLC upregulates regulatory pathways in CD4+ T cells and exhaustion in CD8+ T cells, and analysis of a single cell RNA sequencing dataset shows that CD39+CD4+ cells are enriched in Treg signature gene-sets, and CD39+CD103+ cytotoxic T lymphocyte show gene signatures indicative of an exhausted cytotoxic phenotype with upregulated expression of CXCL13.Conclusions Knowledge of patterns of distribution and location are required to understand the prognostic impact of CD39+ T cell populations in NSCLC. This study provides an improved understanding of spatial and functional characteristics of CD39+ T cells and their significance to patient outcome. |
| first_indexed | 2024-03-08T03:12:00Z |
| format | Article |
| id | doaj.art-aa9f797358354e38a4f06c44b000a342 |
| institution | Directory Open Access Journal |
| issn | 2051-1426 |
| language | English |
| last_indexed | 2024-03-08T03:12:00Z |
| publishDate | 2023-08-01 |
| publisher | BMJ Publishing Group |
| record_format | Article |
| series | Journal for ImmunoTherapy of Cancer |
| spelling | doaj.art-aa9f797358354e38a4f06c44b000a3422024-02-13T05:25:09ZengBMJ Publishing GroupJournal for ImmunoTherapy of Cancer2051-14262023-08-0111810.1136/jitc-2023-006770Location of CD39+ T cell subpopulations within tumors predict differential outcomes in non-small cell lung cancerRichard O’Connor0David A Dorward1Ahsan R Akram2Lilian Koppensteiner3Layla Mathieson4Samuel Pattle5Centre for Inflammation Research, The University of Edinburgh, Edinburgh, UKDepartment of Pathology, Royal Infirmary, Edinburgh, UKCentre for Inflammation Research, The University of Edinburgh, Edinburgh, UKCentre for Inflammation Research, The University of Edinburgh, Edinburgh, UKCentre for Inflammation Research, The University of Edinburgh, Edinburgh, UKDepartment of Pathology, Royal Infirmary, Edinburgh, UKPurpose An improved mechanistic understanding of immunosuppressive pathways in non-small cell lung cancer (NSCLC) is important to develop novel diagnostic and therapeutic approaches. Here, we investigate the prognostic significance of the ectonucleotidases CD39 and CD73 in NSCLC.Experimental design The expression and localization of CD39, CD73 and CD103 was digitally quantified in a cohort of 162 early treatment naïve NSCLC patients using multiplex-immunofluorescence and related to patient outcome. Expression among different cell-populations was assessed via flow cytometry. Targeted RNA-Seq was performed on CD4+ and CD8+ T cells from digested NSCLC tumor tissue and single-cell RNA-Seq data was analyzed to investigate the functional significance of CD39+ T cell populations.Results We demonstrate that flow cytometry of early untreated NSCLC patients shows an upregulation of CD39 expression in the tumor tissue among natural killer (NK) cells, fibroblasts and T cells. CD73 expression is mainly found among fibroblasts and Epcam+cells in the tumor tissue. Multiplex Immunofluorescence in a cohort of 162 early untreated NSCLC patients demonstrates that CD39 expression is mainly localized in the tumor stroma while CD73 expression is equally distributed between tumor nest and stroma, and high expression of CD39 and CD73 in the tumor stroma is associated with poor recurrence-free survival (RFS) at 5 years. Additionally, we find that CD8+T cells located in the tumor nest express CD103 and the density of CD39+CD103+CD8+ T cells in the tumor nest predicts improved RFS at 5 years. Targeted RNA-Seq shows that the tumor microenvironment of NSCLC upregulates regulatory pathways in CD4+ T cells and exhaustion in CD8+ T cells, and analysis of a single cell RNA sequencing dataset shows that CD39+CD4+ cells are enriched in Treg signature gene-sets, and CD39+CD103+ cytotoxic T lymphocyte show gene signatures indicative of an exhausted cytotoxic phenotype with upregulated expression of CXCL13.Conclusions Knowledge of patterns of distribution and location are required to understand the prognostic impact of CD39+ T cell populations in NSCLC. This study provides an improved understanding of spatial and functional characteristics of CD39+ T cells and their significance to patient outcome.https://jitc.bmj.com/content/11/8/e006770.full |
| spellingShingle | Richard O’Connor David A Dorward Ahsan R Akram Lilian Koppensteiner Layla Mathieson Samuel Pattle Location of CD39+ T cell subpopulations within tumors predict differential outcomes in non-small cell lung cancer Journal for ImmunoTherapy of Cancer |
| title | Location of CD39+ T cell subpopulations within tumors predict differential outcomes in non-small cell lung cancer |
| title_full | Location of CD39+ T cell subpopulations within tumors predict differential outcomes in non-small cell lung cancer |
| title_fullStr | Location of CD39+ T cell subpopulations within tumors predict differential outcomes in non-small cell lung cancer |
| title_full_unstemmed | Location of CD39+ T cell subpopulations within tumors predict differential outcomes in non-small cell lung cancer |
| title_short | Location of CD39+ T cell subpopulations within tumors predict differential outcomes in non-small cell lung cancer |
| title_sort | location of cd39 t cell subpopulations within tumors predict differential outcomes in non small cell lung cancer |
| url | https://jitc.bmj.com/content/11/8/e006770.full |
| work_keys_str_mv | AT richardoconnor locationofcd39tcellsubpopulationswithintumorspredictdifferentialoutcomesinnonsmallcelllungcancer AT davidadorward locationofcd39tcellsubpopulationswithintumorspredictdifferentialoutcomesinnonsmallcelllungcancer AT ahsanrakram locationofcd39tcellsubpopulationswithintumorspredictdifferentialoutcomesinnonsmallcelllungcancer AT liliankoppensteiner locationofcd39tcellsubpopulationswithintumorspredictdifferentialoutcomesinnonsmallcelllungcancer AT laylamathieson locationofcd39tcellsubpopulationswithintumorspredictdifferentialoutcomesinnonsmallcelllungcancer AT samuelpattle locationofcd39tcellsubpopulationswithintumorspredictdifferentialoutcomesinnonsmallcelllungcancer |