[HTML][HTML] RAS–mitogen-activated protein kinase signal is required for enhanced PD-L1 expression in human lung cancers

H Sumimoto, A Takano, K Teramoto, Y Daigo - PloS one, 2016 - journals.plos.org
H Sumimoto, A Takano, K Teramoto, Y Daigo
PloS one, 2016journals.plos.org
Ectopic programmed cell death ligand 1 (PD-L1) expression in non-small cell lung cancers
(NSCLCs) is related to immune evasion by cancer, and it is a molecular target of immune
checkpoint therapies. Although some altered signals in NSCLCs are responsible for ectopic
PD-L1 expression, the precise mechanisms remain obscure. Because we found a higher
frequency of EGFR/KRAS mutations in NSCLC cell lines with high PD-L1 expression (p<
0.001), we evaluated the relationships between downstream signals and PD-L1 expression …
Ectopic programmed cell death ligand 1 (PD-L1) expression in non-small cell lung cancers (NSCLCs) is related to immune evasion by cancer, and it is a molecular target of immune checkpoint therapies. Although some altered signals in NSCLCs are responsible for ectopic PD-L1 expression, the precise mechanisms remain obscure. Because we found a higher frequency of EGFR/KRAS mutations in NSCLC cell lines with high PD-L1 expression (p < 0.001), we evaluated the relationships between downstream signals and PD-L1 expression, particularly in three KRAS-mutant adenocarcinoma cell lines. The MEK inhibitor U0126 (20 μM) significantly decreased the surface PD-L1 levels by 50–60% compared with dimethyl sulfoxide (p < 0.0001). Phorbol 12-myristate 13-acetate stimulation (100 nM, 15 min) increased (p < 0.05) and two ERK2 siRNAs as well as KRAS siRNAs decreased (p < 0.05) PD-L1 expression. The transcriptional activity of the potential AP-1 site (+4785 to +5056 from the transcription start site) in the PD-L1 gene was demonstrated by luciferase assays, which was inhibited by U0126. The chromatin immunoprecipitation assay demonstrated the binding of cJUN to the AP-1 site. Two STAT3 siRNAs decreased PD-L1 expression by 10–32% in two of the three KRAS-mutant lung adenocarcinoma cell lines (p < 0.05), while the PI3K inhibitor LY294002 (40 μM) did not change the expression level. Supervised cluster analysis and gene set enrichment analysis between the PD-L1-high and -low NSCLCs revealed a correlation between PD-L1 expression and genes/pathways related to cell motility/adhesion. These results indicate that MAPK signaling is the dominant downstream signal responsible for ectopic PD-L1 expression, in which STAT3 is also involved to some extent. Furthermore, MAPK signaling may control the expression of PD-L1 and several genes related to enhanced cell motility. Our findings suggest that MAPK, along with STAT3, is important for determining PD-L1 expression, which could be useful for targeted therapies against lung cancers.
PLOS