Induction of immunological genes in an RB-dependent manner in PDAC
models treated with MEK and CDK4/6 inhibitors. (A) Common genes significantly
induced or repressed in palbociclib and trametinib-treated cell lines were
determined using an average log 2 fold-change greater than 1 and a
false-discovery rate less than 5% were considered to be differentially
expressed. The blue symbols denote repressed cell-cycle genes and the red
symbols denote induced genes associated with the immune system. (right panel)
Heatmap comparing the log 2 fold change of genes related to immune function
(red-colour bar) and cell cycle (blue-colour bar) in
palbociclib+trametinib-treated cell PDAC cell line models. (B) Top gene ontology
terms related to the immune system and cell cycle ranked based on p-value. (C)
Relative mRNA expression of palbociclib (green) and palbociclib+trametinib
(orange) treated cells for a subset of genes related to antigen presentation and
Interferon signalling (*p≤0.05,**p≤0.01,
***p≤0.001,****p≤0.0001, determined by Student t-test two-sided,
the average and SD are shown). (D) qRT-PCR for the immune-related genes were
performed in 3226 cell line with CRISPR-mediated deletion of RB treated with
palbociclib+trametinib (*p<=0.05,**p<=0.01,***p<=0.001,
determined by Student t-test two-sided, the average and SD are shown). (E)
Heatmap showing the log fold change of representative MEK target genes (red
colour-bar), cell cycle (blue colour-bar) and immune function (green colour-bar)
with the indicated treatments across cell line models. (F) Heatmap showing the
log fold change of representative MEK target genes (red colour-bar), cell cycle
(blue colour-bar) and immune function (green colour-bar) with the indicated
treatments across PDX models. (G) Heatmap showing K-means clustering of TCGA
pancreatic cancer cohort based on significant genes altered with
palbociclib+trametinib treatment related to immune function (red colour-bar) or
cell cycle (blue colour-bar). Pearson correlation between cluster 1 and cluster
2 with a negative correlation of −0.48 and significance
p<2.2e−16. (H) Kaplan-Meier analysis for overall survival
comparing cluster 1 and cluster 2 from the heatmap. PDAC, pancreatic ductal
adenocarcinoma.