Fig. 2: Dysregulated expression of SERPINE1 (PAI-1) promotes chemoresistance and metastasis in oxaliplatin-treated HCT116 cells.

A Significantly enriched hallmarks in HCT116-LD and MD using Gene Set Enrichment Analysis (GSEA) on differentially expressed genes obtained from bulk transcriptomic data of HCT116 resistant models. Red and blue indicates positively and negatively regulated genes, respectively, with the values of normalized enrichment score (NES) as indicated. Pathways with annotated with * denotes hallmarks commonly associated with drug-resistance and metastasis. Pathways and genes explored in this manuscript are highlighted black boxes. B Venn diagram of overlapping and exclusive genes from the leading edge of the coagulation hallmark in HCT116-LD and MD (top). Overlapping genes shared between HCT116-LD and MD from the leading edge of the coagulation hallmark (bottom). C Immunoblotting of PAI-1 in HCT116 PAR, LD, MD, and HD. D Fold change of PAI-1 protein in cell culture supernatant of HCT116 LD, MD, and HD relative to PAR determined using ELISA. E UMAP projection and sample type annotation of scRNA-seq data from the four scRNA-seq libraries: HCT116-PAR/LD/MD/HD. F UMAP projection and Louvain-based clustering of the four samples identifies 5 distinct cell states (cl 0-5) defining the parental and resistant conditions. G Dot plot showing differentially expressed genes (DEGs) of the 5 identified clusters (cl 0-5). RESIST-M gene signature, wherein RESIST-M1 genes denote SERPINE1, SMARCD3, and RESIST-M2 genes denote SC5D, FDPS, MVD, HMGCS1, HMGCR, CYP51A1, ACAT2 can distinguish the cell states defined by clusters 1, 2, 4. H UMAP projection and annotation of the 5 identified cell states based on DEGs. I Proportions of the different annotated clusters across the sample type shows loss and gain in clusters with high RESIST-M2 and high RESIST-M1 signature, respectively. J Dot plot showing differential expression of RESIST-M genes as they increase and decrease, respectively, depending on the dose of oxaliplatin and type of metastatic phenotype acquired. K Violin plot showing Ucell score for RESIST-M1 and M2 genes, increasing and decreasing, respectively, as the selection pressure of oxaliplatin is increased. Statistical significance was determined using Ordinary one-way ANOVA followed by Dunnett’s multiple comparisons test for (D).