In patients with advanced non-small cell lung cancer (NSCLC) LAG-3 is expressed on activated TILs and predicts resistance to PD-1 axis blockers

Abstract

Background: The functional role and clinical utility of measuring PD-1, LAG-3 and TIM-3 in NSCLC tumor tissue remain poorly understood even though corresponding clinical trials with blocking agents are ongoing. We analyzed the expression of these targets in association with key functional immune metrics and outcome after treatment with PD-1 axis blockers in human NSCLC. Methods: We performed CyTOF on immune cell suspensions from 20 primary human NSCLCs to map the distribution of PD-1, LAG-3 and TIM-3 and explore their function. We analyzed RNA levels of these markers in TCGA NSCLC datasets and their association with CD4/CD8 mRNA and mutational burden. Using multiplex quantitative immunofluorescence (QIF) we measured the levels of CD3, PD-1, LAG-3 and TIM-3 in 66 pre-treatment tumor samples from NSCLC patients receiving PD-1 axis blockers in 2 independent cohorts (Cohort #1, Yale, N ¼ 42 cases [Training set] and cohort #2, Cleveland Clinic/University of Navarra, N ¼ 24 [Validation set]. Results: In primary NSCLCs, PD-1 was predominantly expressed on T-and NKT cells. LAG-3 expression was higher in CD8 þ , CD4 þ CD25 þ FOXP3 þ and NKT cell subsets, but low/absent in antigen-presenting cells (APCs). TIM-3 was broadly expressed in adaptive and innate immune cells, with the highest levels in APCs. Expression of all 3 markers in T-cells was associated with lymphocyte activation (CD69/HLA-DR), effector function (Granzyme-B) and proliferation (Ki-67). LAG-3-expressing T-cells showed higher association with early activation and effector function than TILs expressing PD-1 or TIM-3. In TCGA, PD-1 and LAG-3 transcripts strongly correlated with CD8, while TIM-3 was associated with CD4 mRNA. There was limited association between the markers and tumor mutational burden. In pre-treatment specimens from both cohorts of patients treated with PD-1 blockers, elevated LAG-3 but not PD-1 or TIM-3 protein were significantly associated with shorter OS. Conclusions: PD-1, LAG-3 and TIM-3 show variable expression and are associated with T-cell activation and effector function in NSCLC. Elevated T-cell LAG-3 in baseline tumor samples predicts primary resistance to PD-1-axis blockers.

Medical Treatment of Lung Cancer: Can Immune Cells Predict the Response? A Systematic Review

“…Increasing IFN-γ response is a current goal of pharmacological development to boost ICB, notably with STING (stimulator of interferon genes) agonist ( 88 ). Finally, there are limited data for alternative checkpoints such as LAG-3 expression on T cells, associated with shorter OS with PD-1 inhibitors ( 89 ). In conclusion, an approach combining both T cell exclusion and dysfunction [as the gene expression TIDE computational method ( 85 )] may be particularly relevant to predict ICB response.…”

Differential expression of HAVCR2 gene in pan-cancer: A potential biomarker for survival and immunotherapy

“…Some studies provided evidence that PD-1 could interact with TIM-3 and Gal-9 upregulated by inflammatory cytokines to attenuate apoptosis of T cells in cancers (Yang et al, 2021). The levels of PD-1 and TIM-3 protein were significantly correlated in non-small cell lung cancer, suggesting their interplay role in cancers (Datar et al, 2019). The level of TIM-3 is also a potential biomarker for anti-angiogenesis and immunotherapy to evaluate the therapeutic effect (Pignon et al, 2019;Liu et al, 2021).…”

Chemotherapy negatively impacts the tumor immune microenvironment in NSCLC: an analysis of pre- and post-treatment biopsies in the multi-center SAKK19/09 study

“…Currently, multiple clinical trials investigate the therapeutic potential of combined CTLA4/PD-1 blockade in NSCLC patients (NCT03409614, NCT02453282, NCT03319316). Besides PD1 and CTLA4, LAG3 is another important immune checkpoint receptor that was found to be expressed on activated TILs but not NSCLC cells [ 31 ]. Preclinical studies have found that inhibition of LAG3 allowed cytotoxic T-cells to regain cytotoxic function similar to PD-1/PD-L1 inhibition [ 32 ].…”