TNF/IFNγ/CYLD/CASP8 expression is positively associated with responses in ICB therapy. (A) A cohort of advanced melanoma patients having received the anti–PD-1 antibody Nivolumab. Tumors from these patients before and after therapy were analyzed by RNA-seq. Patients were split into responding (R) and not responding (NR) groups according to Response Evaluation Criteria in Solid Tumors (RECIST) criteria. (B–H) Normalized TNF (B), IFNγ (C), IRF1 (D), CYLD (E), CASP8 (F), CASP7 (G), and NOS2 (H) levels before and after therapy for indicated patient populations in patient cohort from A. Wilcoxon signed-rank test was used to determine the significance: NS, ***P = 0.0004 (B); **P = 0.0078, ****P < 0.0001 (C); *P = 0.0391, ****P < 0.0001 (D); NS, **P = 0.0032 (E); NS, ***P = 0.0004 (F); NS, *P = 0.0403 (G); NS, *P = 0.0325 (H). Whiskers of the boxplots indicate 1.5× the interquartile ranges. (I) Kaplan–Meier survival curves for melanoma patient cohort from A which have received anti–PD-1 therapy with induced (n = 17) or not induced (n = 8) TNF/IFNγ signature. A log-rank test was performed: **P = 0.0072. (J) A cohort of metastatic melanoma patients who received anti–PD-1 therapy were analyzed. Patient tumors before and after therapy were analyzed by RNA-seq. Patients were split into R and NR groups according to RECIST criteria. (K–N) Normalized TNF (K), IFNγ (L), IRF1 (M), and CYLD (N) levels for indicated patient populations in the patient cohort from J. Mann–Whitney test was used to determine the significance: NS, ***P = 0.0006 (K); NS, ***P = 0.0006 (L); NS, ***P = 0.0003 (M); NS, ***P = 0.0003 (N). Whiskers of the boxplots indicate 1.5× the interquartile ranges.