Extended Data Fig. 5: UPP1 mediates the liberation of uridine-derived ribose for central carbon metabolism.
From: Uridine-derived ribose fuels glucose-restricted pancreatic cancer
a. Relative intracellular uridine level in the UPP1 knockout PATU8988S cells (after 24 h) and ASPC1 cells (6 h) compared to the wild type supplemented with 1 mM uridine in medium with 10% dialyzed FBS and no glucose. n = 3 biologically independent samples per group per cell line. Statistical significance was measured using one-way ANOVA with Dunnett’s multiple comparisons test. PATU8988S (intracellular uridine) – comparison between WT and 1A, ***P = 0.0002; comparison between WT and 1B, ****P < 0.0001. ASPC1 (intracellular uridine) – comparison between WT and 1A or 1B, ****P < 0.0001. b. Relative extracellular uridine and uracil in UPP1 knockout ASPC1 cells compared to the wild type supplemented with 1 mM uridine for 6 h in medium with 10% dialyzed FBS and no glucose. n = 3 biologically independent samples per group. Statistical significance was measured using one-way ANOVA with Dunnett’s multiple comparisons test. Extracellular uridine: comparison between WT and 1A or 1B, ****P < 0.0001; extracellular uracil: comparison between WT and 1A or 1B, ****P < 0.0001. c. Heatmaps of significantly altered intracellular metabolites (P < 0.05) in PATU8988S cells after 24 h (left) and ASPC1 after 6 h (right), as measured by LC-MS. Data used for the intracellular uridine/uracil plots (a-b) were extracted from this profiling study. d–k. Mass isotopologue distribution of 1 mM 13C5-uridine ribose-derived carbon into the indicated metabolic pathways in wildtype (WT) or UPP1-KO PATU8988S and ASPC1 cells. M – mass; ‘Others’ – indicate M other than M+0 or M+5, where applicable. 1A and 1B denote UPP1-KO sgRNA clones. Data (a–b, d–k) are shown as mean ± s.d. Metabolomics experiments were done once. Statistics and reproducibility: Abbreviations – ADP, adenosine diphosphate; ATP, adenosine triphosphate; AMP, adenosine monophosphate; GSSG, oxidized glutathione; NAD+ nicotinamide adenine dinucleotide; PEP, phosphoenolpyruvate; TCA cycle, tricarboxylic acid cycle; UDP-GlcNAc, uridine diphosphate N-acetylglucosamine; UMP, uridine monophosphate; UTP, uridine triphosphate.
