Galectin-9 in non-small cell lung cancer.

Lung cancer has the highest incidence and fatality among all cancers. Our research analyzed the expression of galectin-9 on non-small cell lung cancer (NSCLC) tumor cells and tumor infiltrating lymphocytes (TILs). We analyzed the expression level of galectin-9 protein in 136 NSCLC primary tumor samples by immunohistochemistry (IHC). We tested 136 surgical resected primary NSCLC tumor tissues for galectin-9 from Medical University of Gdansk, Poland. We found that on tumor cells, galectin-9 level only had correlation with T cell immunoglobulin and mucin-domain containing-3 (TIM-3) level (Correlation Coefficient = 0.360, p < 0.001). On TILs, galectin-9 level had broad connections with other checkpoints including programmed cell death protein-1 (PD-1) (Correlation Coefficient = 0.332, p < 0.001), programmed cell death-ligand 1 (PD-L1) (Correlation Coefficient = 0.247, p = 0.004) and TIM-3 (Correlation Coefficient = 0.350, p < 0.001). Interestingly, galectin-9 level on TILs also had positive relation with PD-L1 level on tumor cells (Correlation Coefficient = 0.278, p = 0.001), galectin-9 level on tumor cells (Correlation Coefficient = 0.181, p = 0.035) and TIL percentage (Correlation Coefficient = 0.236, p = 0.006). High level of galectin-9 on TILs indicated shorter RFS (recurrence-free survival) (RFS 1.82 years, 95% CI 0.795-2.845 vs. 0.67 years, 95% CI 0.086-1.254, P = 0.033). Patients with galectin-9 positive tumor cells display longer overall survival (OS) (1.76 years, 95% CI 0.222-3.298 vs. 3.10 years, 95% CI 2.662-3.538, P = 0.039) CONCLUSIONS: We found galectin-9 expression on both NSCLC tumor cells and TILs. Galectin-9 expression was found in all NSCLC pathological type. Galectin-9 level on TILs had correlation with TIM-3, PD-1 and PD-L1 level. On tumor cells, galectin-9 level had correlation with TIM-3 level. Patients with low galectin-9 level on tumor cells or high galectin-9 level on TILs were more likely to have poor prognosis.

He Y ，Jia K ，Dziadziuszko R ，Zhao S ，Zhang X ，Deng J ，Wang H ，Hirsch FR ，Zhou C ... -  《-》

PD-1, PD-L1 Protein Expression in Non-Small Cell Lung Cancer and Their Relationship with Tumor-Infiltrating Lymphocytes.

He Y ，Rozeboom L ，Rivard CJ ，Ellison K ，Dziadziuszko R ，Yu H ，Zhou C ，Hirsch FR ... -  《MEDICAL SCIENCE MONITOR》

T cell immunoglobulin and mucin-domain containing-3 in non-small cell lung cancer.

Immunotherapy has shown promising effect for non-small cell lung cancer (NSCLC) patients. Yet the biomarkers for predicting immunotherapy efficiency are still lacking. We tested 139 surgical resected NSCLC primary tumor samples from Medical University of Gdansk, Poland, for T cell immunoglobulin and mucin-domain containing-3 (TIM-3) level by immunohistochemistry (IHC), analyzed the expression of TIM-3 protein on NSCLC tumor cells and tumor infiltrating lymphocytes (TILs). TIM-3 on TILs was correlated with programmed cell death protein-1 (PD-1) on TILs (correlation coefficient =0.346, P<0.001) and programmed cell death protein-ligand 1 (PD-L1) on TILs (correlation coefficient =0.313, P<0.001), PD-L1 level on tumor cells (correlation coefficient =0.255, P=0.002), TIM-3 level on tumor cells (correlation coefficient =0.262, P=0.002) and TIL percentage (correlation coefficient =0.172, P=0.043). TIM-3 level on tumor cells only had correlation with PD-L1 level (correlation coefficient =0.170, P=0.045). High level of TIM-3 on TILs indicated shorter recurrence-free survival (RFS) and overall survival (OS) (RFS 1.800 years, 95% CI, 1.230-2.370 vs. 0.870 years, 95% CI, 0.212-1.528, P=0.048) (OS 2.960 years, 95% CI, 2.268-3.652 vs. 1.080 years, 95% CI, 0.228-1.932, P=0.034). TIM-3 is expressed on NSCLC tumor cells and TILs in all NSCLC pathological type. TIM-3 level on TILs had correlation with PD-1 and PD-L1 level. NSCLC patients with high TIM-3 level on TILs were more likely to have poor prognosis.

Jia K ，He Y ，Dziadziuszko R ，Zhao S ，Zhang X ，Deng J ，Wang H ，Hirsch FR ，Zhou C ，Yu H ，Zhang L ... -  《-》

LAG-3 Protein Expression in Non-Small Cell Lung Cancer and Its Relationship with PD-1/PD-L1 and Tumor-Infiltrating Lymphocytes.

Immunotherapy targeting the programmed death 1 (PD-1)/programmed death ligand 1 (PD-L1) checkpoint has shown promising efficacy in patients with NSCLC. Lymphocyte activating 3 gene (LAG-3) is another important checkpoint, and its role in NSCLC is still not clear. In this study we investigated lymphocyte activing 3 (LAG-3) protein expression; its correlation with PD-1, PD-L1, and tumor-infiltrating lymphocytes (TILs); and its association with survival in NSCLC. The expression of LAG-3 (EPR4392 [Abcam, Cambridge, MA]) protein was assessed in 55 NSCLC cell lines by immunohistochemistry. LAG-3, PD-1 (NAT 105 [Cell Marque, Rocklin, CA]), and PD-L1 (22C3 [Dako, Carpenteria, CA]) protein expression was evaluated by immunohistochemistry, and TIL abundance was scored in 139 surgically resected specimens from patients with NSCLC. We also verified results in samples from 62 patients with untreated NSCLC and detected a correlation between LAG-3 expression and EGFR and KRAS mutation and echinoderm microtubule associated protein like 4 gene (EML4)-anaplastic lymphoma receptor tyrosine kinase gene (ALK) rearrangement. LAG-3 was not expressed on any of the 55 NSCLC cell lines. However, LAG-3 was expressed on the TILs in 36 patients with NSCLC (25.9%). Sixty patient samples (43.2%) were positive for PD-1 on the TILs, and 25 (18.0%) were positive for PD-L1 on tumor cells. Neither LAG-3 nor PD-1 was expressed on the tumor cells. LAG-3 was overexpressed on the TILs in nonadenocarcinoma compared with in adenocarcinoma (p = 0.031). LAG-3 expression on TILs was significantly correlated with that of PD-1 on TILs (p < 0.001) and PD-L1 on tumor cells (p = 0.041) but not with TIL percentage (p = 0.244). With the logistic regression model, the ORs for LAG-3 were 0.320 (95% confidence interval [CI]: 0.110-0.929) and 4.364 (95% CI: 1.898-10.031) when nonadenocarcinoma was compared with adenocarcinoma and TILs that were negative for PD-1 were compared with those positive for PD-1. Recurrence-free survival was significantly different in patients whose TILs were LAG-3-negative as opposed to LAG-3-positive (1.91 years [95% CI: 0.76-3.06] versus 0.87 years [95% CI: 0.27-1.47] [p = 0.025]). Likewise, LAG-3 status of TILs (negative versus positive) did significantly affect overall survival (OS) (3.04 years [95% CI: 2.76-3.32] versus 1.08 years [95% CI: 0.42-1.74] [p = 0.039]). Using Kaplan-Meier analysis, we found that patients with both PD-L1-negative tumor cells and LAG-3-negative TILs have longer recurrence-free survival than patients who are either PD-L1- or LAG-3-positive or both PD-L1- and LAG-3-positive (2.09 years [95% CI: 0.90-3.28] versus 1.42 years [95% CI: 0.46-2.34] versus 0.67 years [95% CI: 0.00-1.45] [p = 0.007]). In the verification stage, high expression of LAG-3 was also significantly correlated with higher expression of PD-1 on TILs (p = 0.016) and PD-L1 on tumor cells (p = 0.014). There was no correlation between LAG-3 expression and EGFR (p = 0.325) and KRAS mutation (p = 1.000) and ALK fusion (p = 0.562). LAG-3 is expressed on TILs in tumor tissues of some patients with NSCLC. Its expression was higher in nonadenocarcinoma and correlated with PD-1/PD-L1 expression. LAG-3 positivity or both LAG-3 and PD-L1 positivity was correlated with early postoperative recurrence. LAG-3 was related to poor prognosis.

He Y ，Yu H ，Rozeboom L ，Rivard CJ ，Ellison K ，Dziadziuszko R ，Suda K ，Ren S ，Wu C ，Hou L ，Zhou C ，Hirsch FR ... -  《-》

Dynamic changes in PD-L1 expression and CD8(+) T cell infiltration in non-small cell lung cancer following chemoradiation therapy.

The treatment for stage III non-small cell lung cancer (NSCLC) is quite variable because stage III NSCLC is a heterogenous disease. Programmed death ligand 1 (PD-L1) and CD8+ tumor infiltrating lymphocytes (TILs) are thought to be related to treatment outcome in many tumors. To improve treatment outcome in stage III NSCLC, it is necessary to obtain data on PD-L1 expression and CD8+ TIL counts following CCRT and their relationship to treatment outcome. We retrospectively enrolled 43 patients with stage III NSCLC treated with neoadjuvant CCRT followed by surgery at Yonsei Cancer Center Severance Hospital in Korea between June 2008 and October 2010. PD-L1 level and CD8+ TIL numbers in tumors following CCRT were analyzed by immunohistochemistry, and their association with patient survival was evaluated with Kaplan-Meier method. More than half patients (52%) showed up- or downregulation of PD-L1 expression, and most patients (81%) showed change in CD8+ TIL counts by CCRT. Patients with PD-L1 expression following CCRT tended to have shorter recurrence free survival (RFS) (P = 0.182) or overall survival (OS) (P = 0.215) compared to the ones without PD-L1 expression. In the survival analysis with pre-CCRT specimens, neither RFS nor OS showed statistically significant differences. Patients with increased CD8+ TIL counts following CCRT regardless of pathological response strongly showed longer OS (median: not reached vs. 14.2 months for others; P = 0.017). CCRT dynamically alters PD-L1 expression and CD8+ TIL numbers in stage III NSCLC. Our data provide a rationale for combining CCRT and immunotherapy for the treatment of potentially resectable NSCLC.

Choe EA ，Cha YJ ，Kim JH ，Pyo KH ，Hong MH ，Park SY ，Shim HS ，Jung I ，Lee CY ，Cho BC ，Kim HR ... -  《-》