Determinants of tumor immunogenicity in human CRC. (A) Network of TILs and immunomodulatory molecules. Shown are only candidate genes (hexagons) which were significantly associated with survival in the TCGA cohort. The thickness of the edges is proportional to the strength of the pairwise gene correlation. The size of the circles (TILs) is inversely proportional to the log-rank P-value, with large circles representing lower P-values. Significant association with OS is indicated by a blue node border. Bad prognosis is represented by red node color, good prognosis by green node color. (B) Survival analysis for BCMA using RNA sequencing data (TCGA cohort) as well as microarray data from 11 independent cohorts. OS, overall survival; DFS, disease-free survival. A forest plot with the corresponding hazard ratios is shown on the right. Red asterisks mark the cohorts for which the results are significant. CI, confidence interval. (C) Table with the major determinants of tumor immunogenicity identified using linear regression modeling, the corresponding coefficients and the adjusted P-values. Shown are also the results of the survival analysis for the TCGA cohort, including OS hazard ratio and the P-values. (D) Expression of the chemokine CCR8 in patients with and without metastasis in the TCGA cohort calculated from the RNA sequencing data. (E) Kaplan-Meier curves for germline mutations in CCR8 for the TCGA cohort. (F) Survival analysis for CCR8 in the TCGA cohort (RNA sequencing data) and 10 independent cohorts (microarray data). Red asterisks mark the cohorts for which the results are significant. (G) Hematoxylin and eosin staining of tissue sections from an orthotopic mouse model using murine MC38 cell line in wild-type C57Bl/6 and immunodeficient RAG1-/- mice. IF, immune cell infiltration; T, tumor. Black bar: 200 μm. (H,I) Endoscopic scoring (H) and tumor growth (I) after injection of MC38 cells (104) into the submucosa of wild-type C57Bl/6 and RAG1-/- mice. Error bars represent standard error of the mean. (J) Expression of CCR8 and its ligand CCL1 in the orthotopic mouse model of CRC. * P = 0.038, ** P = 0.017. Error bars represent standard error of the mean.