Fig. 1: Defining mechanisms of short- and long-term adaptation to DHODH inhibition in PDAC by an integrated multi-omics approach.
From: De novo pyrimidine biosynthesis inhibition synergizes with BCL-XL targeting in pancreatic cancer
a Multi-omic workflow used to define adaptation to DHODH inhibition, see Methods and text for additional details (Santana-Codina, N. (2025) https://BioRender.com/a62aees). b Fold change of metabolites in the pyrimidine synthesis pathway (left: 24 h, right: 7 days). Asp: aspartate; N-Carb-L-Asp: N-Carbamoyl-L-Aspartate; DHO: dihydroorotate; UMP/UDP/UTP, uridine mono/di/triphosphate; CTP, cytidine triphosphate; dTTP, deoxythymidine triphosphate. Data are shown as mean values with error bars representing s.d. of n = 3 independent plates. Significance determined with t-test (unpaired, two-tailed) for cells treated with BQ vs. vehicle (*p < 0.05, **p < 0.01, ***p < 0.001). c Ratio of reduced-to-oxidized glutathione (GSH/GSSG) in PaTu-8988T cells. Ratios normalized to each DMSO condition and represented as fold change of 24 h and 7 day BQ-treated cells (0.5 and 5 μM), n = 3 independent plates. Significance determined with t-test (unpaired, two-tailed) for BQ vs. vehicle (*p < 0.05, **p < 0.01). d, e Volcano plot of protein abundance in PaTu-8988T cells treated with 5 μM BQ at 24 h (d) and 7 days (e). Plots display −log10 (FDR) versus log2 relative protein abundance of mean BQ to DMSO-treated samples. Red circles: log2 fold change≥1, FDR < 0.01; blue circles: log2 fold change ≤ −1, FDR < 0.01; data from 3 DMSO or 3 BQ-treated independent plates. f Enrichment map of gene set enrichment analysis (GSEA) of BQ-proteome (PaTu-8988T, 7 days, 0.5 μM). FDR < 0.01, Jaccard coefficient>0.25, node size related to number of components within a gene set, line width proportional to overlap between related gene sets. GSEA terms associated with upregulated (red), and downregulated (blue) proteins colored accordingly and grouped into nodes with associated terms. g BQ-anchored whole-genome in vitro screen in PaTu-8988T cells lentivirally transduced with the Brunello library (19,114 genes/76,441 sgRNAs), treated with BQ (0.5 or 5 μM, 2 weeks) followed by next-generation sequencing (Santana-Codina, N. (2025) https://BioRender.com/4lblqps). h Manhattan plot for depleted hits in PaTu-8988T (5 μM BQ) including significant genes (−log10(P)≥3) in BQ vs. DMSO (blue dots), apoptosis genes (orange), nucleoside salvage pathway genes (green). All source data including p values are provided as Source Data file.
