The volcanic map of DEGs showed that TNFRSF4+ Tregs were more activated and immunosuppressive with high expression of Treg functional signatures such as GITR (TNFRSF18), CCR8, LAYN, TNFRSF9, CTLA4, REL, FoxP3, MAGEH1, and IL2RA (Fig. 5D), which was correlated with poor prognosis in NSCLC, colorectal cancer (CRC) and breast cancer (BC) patients [28–32]. This evidence concerns the gene LAYN and colorectal cancer.