Fig. 4: Low stromal FAK expression enhance metabolic pathways in malignant cells.

a Phosphoproteomics analysis defines a significant enrichment of several metabolic processes in mouse malignant cells exposed to CM from FAK-depleted CAFs compared with malignant cells exposed to WT-CAF CM. b Table: the expression levels of several glycolysis enzymes are significantly upregulated (red) in proteomics analysis of mouse malignant cells exposed to FAK-depleted CAF CM when compared with malignant cells exposed to WT-CAF CM. Box and whisker plots indicate significant transcriptional upregulation of genes encoding glycolytic enzymes in the cancer cells of patient tumours with low stromal FAK compared with high stromal FAK. c Box and whisker plots, elevated IDH, SDH and FH gene transcription in the epithelial compartment of patient tumours with low stromal FAK. Table: Mouse proteomics data indicate a significant enrichment of IDH peptides in mouse malignant cells exposed to CM from FAK-depleted CAFs compared with malignant cells exposed to WT-CAF CM. d Box and Whisker plots show elevated transcription of genes encoding enzymes of fatty acid metabolism in the epithelial gene set of human breast cancer with low stromal FAK. Table: HADHA upregulation at the protein level in mouse malignant cells exposed to CM from FAK-depleted CAFs. For all human data, n = 39 breast cancer patients with high stromal FAK, n = 9 breast cancer patients with low stromal FAK (from Finak et al. dataset); for all mouse data n = 3 mouse malignant cell preparations exposed to WT-CAF CM and n = 3 mouse malignant cell preparations exposed to FAK-depleted CAF CM. Box and whisker plots—box denotes the interquartile range (IQR, Q1 25th percentile—Q3 75th percentile), and whisker denotes the maximum (Q3 + 1.5IQR) and minimum (Q1–1.5IQR). The median value is shown by the horizontal line within the box. *P < 0.05,**P < 0.01,***P < 0.001,****P < 0.0001. nsd, no significant difference. Statistical analysis, two-sided Student’s t-test. e The transcriptome of cancer cells in breast cancer patients with low vs high stromal FAK were compared using Reactome database. GSEA shows that many pathways regulating cellular metabolism are upregulated in the cancer cells in patient tumours with low stromal FAK expression levels. Disc sizes reflect the number of differentially expressed hits from each pathway. Colour bar indicates FDR q-value significance.