Expression of the immune checkpoint receptors PD-1, LAG3, and TIM3 in the immune context of stage II and III gastric cancer by using single and chromogenic multiplex immunohistochemistry

Abstract

We sought to determine the clinicopathological significance of PD-1, LAG3, and TIM3 in gastric cancer (GC) by examining their expression and immune context. Immunohistochemistry (IHC) for PD-1, TIM3, LAG3, and tumor-infiltrating immune cell (TIIC) markers was performed in 385 stage II/III GCs. Epstein-Barr virus (EBV) and microsatellite stability (MSI) testing were performed for molecular classification. Chromogenic multiplex IHC (mIHC) for PD1, TIM3, LAG3, CD3, CD8, FOXP3, CD68, and cytokeratin was performed in 58 of the total samples. PD-1, LAG3, and TIM3 expression in TIICs was observed in 91 (23.6%), 193 (50.1%), and 257 (66.8%) GCs by single IHC, respectively. The expression was associated with EBV + and MSI-H molecular subtypes (p ≤ 0.001). A positive expression of LAG3 in the invasive margin of the tumor was associated with better prognosis in univariate (p = .020) and multivariate (p = .026) survival analyses. The expression of different immune checkpoint receptors (ICRs) was significantly positively correlated. Dual or triple ICR expression was more frequent in high PD-1 and TIM3 density groups than in low-density groups by mIHC (all p ≤ 0.05). ICRs were mainly expressed in CD3 + CD8 + and CD3 + CD8 − T cells. Fifty-eight GCs were classified into three groups by clustering analysis based on mIHC, and the group with the highest ICR expression in TIICs showed significantly better outcomes in progression-free survival (p = .020). In GC, PD-1, LAG3, and TIM3 expression is positively correlated and associated with better prognosis.Our study provides information for the application of effective immune checkpoint inhibitors against GC.

Immune Checkpoint LAG3 and Its Ligand FGL1 in Cancer

“…LAG3 expression is correlated with that of other immune checkpoints. In gastric cancer, the expression of programmed cell death protein 1 (PD-1) is positively correlated with that of LAG3 and T-cell immunoglobulin domain and mucin domain 3 (TIM3), whereas the expression of LAG3 is associated with the expression of TIM3 ( 27 ). The co-expression of LAG3 and PD1 or PD ligand 1 (PD-L1) has been proven in breast cancer.…”

“…The single expression of LAG3 + and the dual expression of PD-1 + /LAG3 + predict better PFS, and the dual expression of TIM3 + /LAG3 + predict better OS and PFS. Furthermore, in advanced gastric cancer, a higher ratio of LAG3 + CD4 + /CD4 + T cells and LAG3 + CD8 + /CD8 + T cells is associated with better prognosis, although, at the invasive tumor margin, higher LAG3 expression is associated with better prognosis ( 27 ). In FGL1 + gastric cancer, where a higher expression of FGL1 is positively associated with gastric cancer stage and lymph node metastasis, as well as the poor OS ( 44 ).…”

“…LAG3 is expressed in various tumors, such as KIRC, gastric cancer, breast cancer, B-cell lymphoma, and lung cancer (3,(24)(25)(26). In gastric cancer, compared to CD68 + macrophages, higher expression of LAG3 can be found in CD3 + /CD8 + T cells (27). The differential expression of LAG3 in different tumor cells indicates a different patient prognosis.…”

Nature vs. Nurture: The Two Opposing Behaviors of Cytotoxic T Lymphocytes in the Tumor Microenvironment

“…A new phase I/II clinical trial (NCT03459222) was recently opened to investigate the efficacy of co-targeting LAG-3, PD-1, and CTLA-4, which, by further enhancing the efficacy of single and double targeting approaches, might become a novel combinatorial strategy for cancer treatment in the near future. In this context, it is noteworthy that PD-1, LAG-3, and TIM-3 expression is coordinately increased in gastric cancer patients with better disease prognosis [ 185 ].…”

A novel pyroptosis-related gene signature to predict outcomes in laryngeal squamous cell carcinoma

“…Nevertheless, many patients do not experience good clinical outcomes because of tumor heterogeneity. Therefore, we also compared the expression of ICI-related genes ( PD1, CTLA4, LAG3, PD-L1, and HAVCR2 ) between the high- and low-risk groups [ 57 – 59 ] and found that PD1 , CTLA4, and LAG3 were significantly increased in the low-risk group, suggesting that the low-risk patients might have a better response to ICI therapy. We confirmed this using TCIA, finding that the low-risk group had a higher immunophenoscore for PD-1 inhibitor alone or the combination of PD1 and CTLA4 inhibitor.…”