Fig. 6
From: miR200-regulated CXCL12β promotes fibroblast heterogeneity and immunosuppression in ovarian cancers
CXCL12β mRNA is targeted by miR-141/200a in CAF-S4. a Schema (from https://genome.ucsc.edu) of CXCL12 human genomic locus. Two predicted miR141/200a sites are shown, site 1 specific of CXCL12β and site 2 also detected in CXCL12ε. b Normalized luciferase activity of CXCL12β-3’UTR-luciferase reporter construct after co-transfection with miR200s. Values are fold changes of Firefly/Renilla activity ratio (normalized to control) ± SEM. n = 2. P values from Student’s t-test. c CXCL12α and CXCL12β mRNA levels in miR-200-overexpressing CAF-S1. Data are mean of fold change ± SEM. n = 3. P values from Student’s t-test. d Correlation plots between CXCL12β mRNA and miR-141 or miR-200a levels. P values from Spearman’s test. e miR-141/miR-200a levels in non-mesenchymal (N = 38) versus mesenchymal (N = 45) HGSOC (Institut Curie cohort). Normalized cycle thresholds are centered to the mean (ΔΔCt). P value from Student’s t-test. f Reduced (GSH) and oxidized (GSSG) glutathione levels evaluated by mass spectrometry in non-mesenchymal (N = 19) and mesenchymal (N = 25) HGSOC. P value from Student’s t-test (non-mesenchymal/mesenchymal) and paired ttest (GSH/GSSG). g Significant enrichment of electron transport chain (ETC) gene signature in CAF-S4. P refers to false discovery rate q-value. h, i PTPN6 (h) and ZEB1, MAPK14, CTNNB1 (miR-141/200a-target genes7,35,36) (i) mRNA levels in CAF-S1 and CAF-S4. Data are mean ± SEM. P values from Student’s t-test. j Model, mesenchymal HGSOC7,13 accumulate a dense stroma enriched in CAF-S1 fibroblasts (Right). CAF-S1, characterized by expression of FAP, SMA and PDGFRβ, promotes attraction of regulatory T cells through CXCL12β and CXCL12α. CAF-S4 fibroblasts accumulate in non-mesenchymal HGSOC (Left), characterized by genes correlated with miR-2007,13, involved in oxidative stress response. Accordingly, non-mesenchymal HGSOC suffer from a chronic oxidative stress. CAF-S4 fibroblasts exhibit lower levels of CXCL12β mRNA than CAF-S1, and thus show reduced attraction of regulatory T lymphocytes. In contrast to CXCL12α, CXCL12β is targeted by miR-141/200a that accumulate in CAF-S4-enriched non-mesenchymal HGSOC
