Irene Gullo1,2,3,4, Patrícia Oliveira3,4, Maria Athelogou5, Gilza Gonçalves2,3,4,6, Marta L Pinto4,7,8, Joana Carvalho3,4, Ana Valente3,4, Hugo Pinheiro3,4,9, Sara Andrade3,4,10, Gabriela M Almeida3,4, Ralf Huss5, Kakoli Das11, Patrick Tan11,12,13, José C Machado2,3,4, Carla Oliveira14,15,16, Fátima Carneiro17,18,19,20. 1. Department of Pathology, Centro Hospitalar de São João, Al. Prof. Hernâni Monteiro, 4200-319, Porto, Portugal. 2. Department of Pathology, Faculty of Medicine of the University of Porto (FMUP), Al. Prof. Hernâni Monteiro, 4200-319, Porto, Portugal. 3. Institute of Molecular Pathology and Immunology at the University of Porto (Ipatimup), Rua Júlio Amaral de Carvalho 45, 4200-135, Porto, Portugal. 4. Instituto de Investigação e Inovação em Saúde (i3S), University of Porto, Rua Alfredo Allen 208, 4200-135, Porto, Portugal. 5. Definiens AG, Bernhard-Wicki Str 5, 80636, Munich, Germany. 6. Department of Biomedical Sciences and Medicine, University of Algarve, Campus De Gambelas, 8005-139, Faro, Portugal. 7. INEB-Institute of Biomedical Engineering, University of Porto, Rua Alfredo Allen 208, 4200-135, Porto, Portugal. 8. ICBAS-Institute of Biomedical Sciences Abel Salazar, University of Porto, Rua Jorge De Viterbo Ferreira 228, 4050-343, Porto, Portugal. 9. Hospital Senhora da Oliveira, Rua Dos Cutileiros 114, 4835-044, Guimarães, Portugal. 10. Department of Biomedicine, Faculty of Medicine of the University of Porto (FMUP), Al. Prof. Hernâni Monteiro, 4200-319, Porto, Portugal. 11. Cancer and Stem Cell Biology Program, Duke-NUS Medical School, 8 College Road, Singapore, 169857, Singapore. 12. Genome Institute of Singapore, Biopolis, 60 Biopolis St, Singapore, 138672, Singapore. 13. Cancer Science Institute of Singapore, National University of Singapore, 14 Medical Dr, Singapore, 117599, Singapore. 14. Department of Pathology, Faculty of Medicine of the University of Porto (FMUP), Al. Prof. Hernâni Monteiro, 4200-319, Porto, Portugal. carlaol@ipatimup.pt. 15. Institute of Molecular Pathology and Immunology at the University of Porto (Ipatimup), Rua Júlio Amaral de Carvalho 45, 4200-135, Porto, Portugal. carlaol@ipatimup.pt. 16. Instituto de Investigação e Inovação em Saúde (i3S), University of Porto, Rua Alfredo Allen 208, 4200-135, Porto, Portugal. carlaol@ipatimup.pt. 17. Department of Pathology, Centro Hospitalar de São João, Al. Prof. Hernâni Monteiro, 4200-319, Porto, Portugal. fcarneiro@ipatimup.pt. 18. Department of Pathology, Faculty of Medicine of the University of Porto (FMUP), Al. Prof. Hernâni Monteiro, 4200-319, Porto, Portugal. fcarneiro@ipatimup.pt. 19. Institute of Molecular Pathology and Immunology at the University of Porto (Ipatimup), Rua Júlio Amaral de Carvalho 45, 4200-135, Porto, Portugal. fcarneiro@ipatimup.pt. 20. Instituto de Investigação e Inovação em Saúde (i3S), University of Porto, Rua Alfredo Allen 208, 4200-135, Porto, Portugal. fcarneiro@ipatimup.pt.
Abstract
BACKGROUND: Gastric cancer with lymphoid stroma (GCLS) is characterized by prominent stromal infiltration of T-lymphocytes. The aim of this study was to investigate GCLS biology through analysis of clinicopathological features, EBV infection, microsatellite instability (MSI), immune gene-expression profiling and PD-L1 status in neoplastic cells and tumor immune microenvironment. METHODS: Twenty-four GCLSs were analyzed by RNA in situ hybridization for EBV (EBER), PCR/fragment analysis for MSI, immunohistochemistry (PD-L1, cytokeratin, CD3, CD8), co-immunofluorescence (CK/PD-L1, CD68/PD-L1), NanoString gene-expression assay for immune-related genes and PD-L1 copy number alterations. CD3+ and CD8+ T-cell densities were calculated by digital analysis. Fifty-four non-GCLSs were used as control group. RESULTS: GCLSs displayed distinctive clinicopathological features, such as lower pTNM stage (p = 0.02) and better overall survival (p = 0.01). EBV+ or MSI-high phenotype was found in 66.7 and 16.7% cases, respectively. GCLSs harbored a cytotoxic T-cell-inflamed profile, particularly at the invasive front of tumors (p < 0.01) and in EBV+ cases (p = 0.01). EBV+ GCLSs, when compared to EBV- GCLSs, showed higher mRNA expression of genes related to Th1/cytotoxic and immunosuppressive biomarkers. PD-L1 protein expression, observed in neoplastic and immune stromal cells (33.3 and 91.7%, respectively), and PD-L1 amplification (18.8%) were restricted to EBV+/MSI-high tumors and correlated with high values of PD-L1 mRNA expression. CONCLUSIONS: This study shows that GCLS has a distinctive clinico-pathological and molecular profile. Furthermore, through an in-depth study of tumor immune microenvironment-by digital analysis and mRNA expression profiling-it highlights the role of EBV infection in promoting an inflamed tumor microenvironment, with putative therapeutic implications.
BACKGROUND:Gastric cancer with lymphoid stroma (GCLS) is characterized by prominent stromal infiltration of T-lymphocytes. The aim of this study was to investigate GCLS biology through analysis of clinicopathological features, EBV infection, microsatellite instability (MSI), immune gene-expression profiling and PD-L1 status in neoplastic cells and tumor immune microenvironment. METHODS: Twenty-four GCLSs were analyzed by RNA in situ hybridization for EBV (EBER), PCR/fragment analysis for MSI, immunohistochemistry (PD-L1, cytokeratin, CD3, CD8), co-immunofluorescence (CK/PD-L1, CD68/PD-L1), NanoString gene-expression assay for immune-related genes and PD-L1 copy number alterations. CD3+ and CD8+ T-cell densities were calculated by digital analysis. Fifty-four non-GCLSs were used as control group. RESULTS: GCLSs displayed distinctive clinicopathological features, such as lower pTNM stage (p = 0.02) and better overall survival (p = 0.01). EBV+ or MSI-high phenotype was found in 66.7 and 16.7% cases, respectively. GCLSs harbored a cytotoxic T-cell-inflamed profile, particularly at the invasive front of tumors (p < 0.01) and in EBV+ cases (p = 0.01). EBV+ GCLSs, when compared to EBV- GCLSs, showed higher mRNA expression of genes related to Th1/cytotoxic and immunosuppressive biomarkers. PD-L1 protein expression, observed in neoplastic and immune stromal cells (33.3 and 91.7%, respectively), and PD-L1 amplification (18.8%) were restricted to EBV+/MSI-high tumors and correlated with high values of PD-L1 mRNA expression. CONCLUSIONS: This study shows that GCLS has a distinctive clinico-pathological and molecular profile. Furthermore, through an in-depth study of tumor immune microenvironment-by digital analysis and mRNA expression profiling-it highlights the role of EBV infection in promoting an inflamed tumor microenvironment, with putative therapeutic implications.
Authors: Christy Ralph; Eyad Elkord; Deborah J Burt; Jackie F O'Dwyer; Eric B Austin; Peter L Stern; Robert E Hawkins; Fiona C Thistlethwaite Journal: Clin Cancer Res Date: 2010-02-23 Impact factor: 12.531
Authors: A M Chiaravalli; M Cornaggia; D Furlan; C Capella; R Fiocca; G Tagliabue; C Klersy; E Solcia Journal: Virchows Arch Date: 2001-08 Impact factor: 4.064
Authors: Irene Gullo; Catarina Costa; Susana L Silva; Cristina Ferreira; Adriana Motta; Sara P Silva; Rúben Duarte Ferreira; Pedro Rosmaninho; Emília Faria; José Torres da Costa; Rita Câmara; Gilza Gonçalves; João Santos-Antunes; Carla Oliveira; José C Machado; Fátima Carneiro; Ana E Sousa Journal: Cells Date: 2020-06-19 Impact factor: 6.600