Comprehensive Analyses of the Heterogeneity and Prognostic Significance of Tumor-Infiltrating Immune Cells in Non-Small-cell Lung Cancer: Development and Validation of an Individualized Prognostic Model.

Understanding the role of tumor-infiltrating immune cells (TIICs) in non-small cell lung cancer (NSCLC) is critical to finding new prognostic biomarkers and improving prognostic evaluation. Herein, we aimed to comprehensively analyze tumor-infiltrating pattern of TIICs in NSCLC and build a TIICs-associated, risk-stratification prognostic model for clinical practice. We applied CIBERSORT and ESTIMATE computational methods to analyze RNA-seq samples of 852 NSCLC patients from The Cancer Genome Atlas (TCGA). Prognotic factors were identified by univariate and multivariate Cox regression analyses for overall survival (OS). A novel model was developed to predict the 1-, 3- and 5-year OS of NSCLC based on the TCGA cohort, validated by external validation cohorts (GSE31210, GSE37745), and then evaluated by C-indexes and calibration plots. Significant heterogeneity in the infiltrating patterns of TIICs was shown among various pathological subtypes of NSCLC and between different genders. Further analyses showed that abundances of naive B cells (NBCs), T cells and mast cells (MCs) were positively correlated with prognosis. Tumor samples with high T cells abundances tended to have higher expression levels of immune checkpoint genes (PD-1, PD-L1, CTLA-4). A new immune-gene related index (IGRI) was built by five immune-related differentially expressed genes (DEGs) including BTK, CCR2, CLEC10A, NCR3 and PRKCB, which were closely correlated with TIICs abundances and prognosis. Tumor stage, IGRI, abundances of NBCs, T cells, MCs and NK cells were significant independent prognostic factors and were included in the nomogram as predictors. The internal and external calibration plots of the nomogram were in excellent agreement. This study reveals that TIICs are significantly correlated with clinicopathological features and prognosis in NSCLC and thus can be potential prognostic biomarker or therapeutic target. The remarkable heterogeneity of TIICs suggests that specific infiltrating patterns of TIICs should also be taken into consideration when determining individualized immunotherapy strategies for NSCLC patients.