Seyed Behrad Piran

Background: Programmed cell death protein-1 (PD-1) and its ligands, PD-L1 and PD-L2, constitute a central immune checkpoint pathway that regulates T-cell activity and tumor immune escape, while their relationship with STAT signaling remains incompletely understood in ccRCC. Methods: We analyzed 27 paired ccRCC and adjacent non-tumorous human kidney tissue samples. mRNA levels of PD-1, PD-L1, PD-L2, STAT1, and STAT3 were quantified by RT-qPCR. In addition, representative human ccRCC cell lines (CAKI-2 and A-498) were treated with IFN-γ to assess the time-dependent modulation of immune checkpoint molecules and STAT pathway activation. Results: PD-L1 and PD-L2 were significantly upregulated in tumor tissues compared with adjacent normal kidney tissue. Exploratory observation suggests grade dependent increase. Whereas PD-1 was predominantly downregulated, IFN-γ treatment induced a rapid transcriptional upregulation of PD-L1 and PD-L2 in RCC cell lines, with maximal protein accumulation observed at 72 h. STAT1, but not STAT3, exhibited dynamic induction following IFN-γ stimulation, showing temporal association with PD-L1 and PD-L2 upregulation, indicating cell-line-specific regulatory effects. Correlation analyses confirmed a strong correlation between PD-1 and its ligands, whereas STAT1 and STAT3 expression showed no direct association with PD-L1 or PD-L2 levels in cancer samples. Conclusions: Our findings demonstrate that PD-L1 and PD-L2 are frequently upregulated in ccRCC and dynamically regulated by IFN-γ/STAT1-dependent signaling. Our results provide additional insight into the mechanisms of immune escape and underscore the potential of integrated profiling of PD-1 ligands and STAT signaling to guide personalized immunotherapeutic strategies in ccRCC.