TGFβ-induced myofibroblastic C2-CAF reprograms oral-cancer cells to acquire embryonic-like transcriptome state.(A) Feature plot showing expressions of epithelial and CAF marker modules on UMAP projection from three conditions of co-cultures as indicated. Circled clusters are annotated as CAF clusters with high CAF marker module scores and negativity for epithelial markers module scores. (B) UMAP plot shows re-clustering of CAF clusters from all the three conditions merged, revealing 13 clusters with a total of 11,391 cells. A split view of major clusters in a sample specific manner is provided on side panel. (C) (i) An UMAP plot visualizing sample wise grouped CAF clusters. (ii) Monocle3 pseudotime -time analysis showing CAF dynamic transition along the trajectory. (D) Violin plot showing enrichment of TGFβ and Tie2 signaling AUC scores generated by R tool ‘AUCell’, upon TGFβ treatment of CAF, which was significantly decreased followed by Tie2-inhibition. (E) AUC scoring of CAF from classified patient groups (High BMP4 (C1-like)/High ITGA3 (C2-like)) from Puram et al. and Quah et al. HNSCC datasets shows significant enrichment of C1-CAF DEGs in High BMP4 group, and C2-CAF DEGs and Tie2 signaling in High ITGA3 group. (F) Subset of 32,354 epithelial cells from all three conditions were merged and re-clustered, identified 16 clusters, projected on UMAP plot. (G) Pseudotime analysis exploring transition trajectory of cancer cells. (H) Bubble plot showing GO biological process analysis of gene-set among single cell and bulk RNA sequencing of cancer cells co cultured with TGF-CAF. Size of bubble represents numbers of associated genes and colour corresponds to given p value
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