Abstract
Nanostring nCounter targeted RNA Seq. data of CD4+ and CD8+ T cells isolated from human non-cancerous lung tissue and non-small cell lung cancer tumour tissue as shown in Koppensteiner et al. (in submission), "Location of CD39+ T cell sub-populations within tumours predict differential outcomes in non-small cell lung cancer".
An improved mechanistic understanding of immunosuppressive pathways in NSCLC is important to develop novel diagnostic and therapeutic approaches. Here, we investigate the prognostic significance of the ectonucleotidases CD39 and CD73 in NSCLC. The expression and localisation 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 amongst different cell-populations was assessed via flow cytometry. Targeted RNA-Seq was performed on CD4+ and CD8+ T cells from digested NSCLC tumour tissue and single-cell RNA-Seq data was analysed to investigate the functional significance of CD39+ T cell populations. We demonstrate that flow cytometry of early untreated NSCLC patients shows an upregulation of CD39 expression in the tumour tissue amongst NK cells, fibroblasts and T cells. CD73 expression is mainly found amongst fibroblasts and Epcam+ cells in the tumour tissue. Multiplex Immunofluorescence in a cohort of 162 early untreated NSCLC patients demonstrates that CD39 expression is mainly localised in the tumour stroma while CD73 expression is equally distributed between tumour nest and stroma, and high expression of CD39 and CD73 in the tumour stroma is associated with poor RFS at 5 years. Additionally, we find that CD8+ T cells located in the tumour nest express CD103 and the density of CD39+CD103+CD8+ T cells in the tumour nest predicts improved RFS at 5 years. Targeted RNA-Seq shows that the TME 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+ CTL show gene signatures indicative of an exhausted cytotoxic phenotype with upregulated expression of CXCL13. 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.
An improved mechanistic understanding of immunosuppressive pathways in NSCLC is important to develop novel diagnostic and therapeutic approaches. Here, we investigate the prognostic significance of the ectonucleotidases CD39 and CD73 in NSCLC. The expression and localisation 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 amongst different cell-populations was assessed via flow cytometry. Targeted RNA-Seq was performed on CD4+ and CD8+ T cells from digested NSCLC tumour tissue and single-cell RNA-Seq data was analysed to investigate the functional significance of CD39+ T cell populations. We demonstrate that flow cytometry of early untreated NSCLC patients shows an upregulation of CD39 expression in the tumour tissue amongst NK cells, fibroblasts and T cells. CD73 expression is mainly found amongst fibroblasts and Epcam+ cells in the tumour tissue. Multiplex Immunofluorescence in a cohort of 162 early untreated NSCLC patients demonstrates that CD39 expression is mainly localised in the tumour stroma while CD73 expression is equally distributed between tumour nest and stroma, and high expression of CD39 and CD73 in the tumour stroma is associated with poor RFS at 5 years. Additionally, we find that CD8+ T cells located in the tumour nest express CD103 and the density of CD39+CD103+CD8+ T cells in the tumour nest predicts improved RFS at 5 years. Targeted RNA-Seq shows that the TME 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+ CTL show gene signatures indicative of an exhausted cytotoxic phenotype with upregulated expression of CXCL13. 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.
| Date made available | 18 May 2023 |
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| Publisher | Edinburgh DataShare |
| Geographical coverage | UK,UNITED KINGDOM |