A tumor cornification and immune-infiltration-based scheme for anti-PD-1 plus chemotherapy response in advanced squamous cell lung carcinoma

Context and significanceImmunotherapy combined with chemotherapy has achieved great success in lung cancer. However, traditional biomarkers such as PD-L1 expression have failed to predict the efficacy of anti-PD-1+chemo treatment in advanced squamous cell lung cancer. Based on RNA sequencing (RNA-seq) data of ORIENT-12, the authors found that tumor cornification greatly impacted the tumor microenvironment of squamous cell lung cancer, and a LICC scheme incorporating tumor cornification level and immune infiltration of activated CD8+ T cells and CD56bright NK cells was established for efficacy evaluation of anti-PD-1+chemo treatment in patients with advanced squamous cell lung cancer. Moreover, they also revealed that the tumor cornification signal was mainly mapped to SPRR3+ tumor cells, which might acquire its immune escape mechanisms via the CD24-SIGLEC10 (M2 macrophages) axis to maintain intratumoral immunosuppressive microenvironment.Highlights•Activated CD8+ T and CD56bright NK cells affected anti-PD-1+chemo efficacy in LUSC•Tumor cornification affected intratumoral immune cell infiltration•LICC scheme was proposed to select patients benefiting from anti-PD-1+chemo treatment•SPRR3+ tumor cells were responsible for tumor cornificationSummaryBackgroundAnti-PD-1 immunotherapy plus chemotherapy (combo) exhibits significantly prolonged survival for squamous cell lung cancer (LUSC). An exploration of predictive biomarkers is still needed.MethodsHigh-throughput RNA sequencing (RNA-seq) of 349 LUSC samples from the randomized, multi-center, phase 3 trial ORIENT-12 (ClinicalTrials.gov: NCT03629925) was conducted for biomarker discovery, followed by flow cytometry and multiplex immunohistochemistry (mIHC) in additional clinical cohorts, and in vitro experiments were performed for verification.ResultsA high abundance of activated CD8+ T and CD56bright natural killer (NK) cells benefited patients' outcomes (progression-free survival [PFS]; overall survival [OS]) with combo treatment. Tumor cornification level remarkably affected the infiltration of the two crucial immune cells. Thus, a novel scheme of LUSC immune infiltration and cornification characterization-based classification (LICC) was established for combo efficacy prediction. Patients who received combo treatment achieved significant PFS improvements in LICC1 (hazard ratio [HR] = 0.43, 95% confidence interval [CI]: 0.25–0.75, p = 0.0029) and LICC2 (HR = 0.32, 95% CI: 0.17–0.58, p = 0.0002) subtypes but not in the LICC3 subtype (HR = 0.86, 95% CI: 0.60–1.23, p = 0.4053). Via single-cell RNA-seq analysis, the tumor cornification signal was mainly mapped to SPRR3+ tumor cells, whose relationships with activated CD8+ T or CD56bright NK cells were verified using flow cytometry and mIHC. Our data suggest that SPRR3+ tumor cells might evade immune surveillance via the CD24-SIGLEC10 (M2 macrophage) axis to maintain a suppressive tumor microenvironment.ConclusionsTumor cornification greatly impacts immune infiltration, and the LICC scheme may guide clinical medication of anti-PD-1+chemo treatment in patients with LUSC.FundingThe study was funded by the National Key R&D Program of China, the National Natural Science Foundation of China, Shanghia Multidisplinary Cooperation Building Project for Diagnosis and Treatment of Major Disease, and Innovent Biologics, Inc.Graphical abstract