Fig. 3: Multi-omics analysis of the effects of disulfiram in rats.

Proteomic analysis of liver proteins (a–g) and untargeted serum metabolomics (h–j) in rats exposed to disulfiram. a Heat map visualization of protein abundance in livers of rats fed laboratory SD (control) either supplemented or not with a low or high dose of DSF. Upregulation (red font), downregulation (blue font). n = 8 per group. b Volcano plots of liver protein abundance changes after diet supplementation with low or high dose of DSF were plotted with the y-axis showing the Benjamini–Hochberg corrected −log10 (P) and the x-axis showing the log2 fold change of protein abundance (DSF/control SD) calculated from the median LFQ intensity values. The blue and red symbols denote significant changes with low and high DSF, respectively, and the black symbols denote nonsignificant changes. Significance is defined as >40% median fold changes in either direction. c Two-way Venn diagram depicting the distribution of unique and common proteins whose expression was impacted by DSF (low or high dose) vs. control SD. Upregulation (red font), downregulation (blue font). Significance is defined as >40% median fold changes in either direction. d Abundance of a select group of proteins significantly impacted by DSF treatment. Normalized LFQ intensity values are represented in box and whisker plot format (n = 8 per group). Statistics for the effects of DSF intervention represent the p-value from a one-way ANOVA with Dunnett’s post hoc tests. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001. e Clustering of liver proteins significantly impacted by DSF treatment as provided by String protein–protein interaction database. High confidence interaction score was selected (0.7). For reasons of clarity, some of the proteins present in the redox & drug metabolism cluster were labeled ‘a’ to ‘m’. These are: a, Aldh1a1; b, Ugdh; c, Cyp2b1; d, Cyp2e1; e, Cyp4a1; f, Cyp3a1; g, Ugt1a1; h, Ugt2b1; i, Cyp2c6v1; j, Cyp2c13; k, Cyp1a2; l, Cyp2c12; m, Gsta2. f Top enriched ten pathways generated from these experimental and predicted interactions map for “metabolic pathways,” “Redox and drug metabolism,” Tryptophan metabolism,” and “fatty acid (FA) and branched-chain amino acid (BCAA) catabolism.” g Gene-set enrichment analysis (GSEA) depicting a set of proteins involved in “metabolism of xenobiotics by CYP450” whose expression was significantly impacted by DSF supplementation. h PLS-DA of untargeted serum metabolomics (n = 8 in each group). i Heat map illustrates the relative average of each metabolite contributing to the group separation between control and DSF (low and high). j Relative abundance of a select group of metabolites. The data are represented in box and whisker plot format. *P < 0.05; **P < 0.01; ****P < 0.0001 vs. control group. Related to Supplementary Fig. 3 and Supplementary Tables 4 and 5.