Figure 5. NAD+ depletion induces apoptotic priming and anti-apoptotic dependencies in TNBC by dysregulation of specific metabolic pathways rather than an overall metabolic collapse.

(A) Metabolomics data of intermediates of the glycolysis, pentose phosphate pathway (PPP), tricarboxylic acid (TCA) cycle, nucleoside synthesis and the oxidative stress pathways extracted from cell lines treated for 48h with 0.01 μM FK866 or 0.1 μM GPP78. Data are expressed as fold changes compared to vehicle treated samples. Data are average of technical duplicates. Sensitive models are indicated in red, intermediately sensitive models are indicated in gray, insensitive models are indicated in blue. (B) Metabolomics data of intermediates of the glycolysis, pentose phosphate pathway (PPP), tricarboxylic acid (TCA) cycle, nucleoside synthesis and the oxidative stress pathways extracted from PDX tumors treated in vivo with 7.5 mg/kg or 15 mg/kg FK866 according to treatment schedule in Fig. 3D. Data are expressed as fold changes compared to vehicle-treated samples. Data are average of n = 2 to 4 mice per treatment arm. (C) Microscopy-based annexin V/Hoechst cell viability data of MDA-MB-468 and MDA-MB-231 cells co-treated for 72 hours with 0.01 μM FK866 and 0.1 μM BH3 mimetics in the presence or absence of different metabolites. Data are means of 3 independent experiments. (D) Percentage delta priming of TNBC cells after 48 hours exposure to 0.01 μM FK866 in the absence or presence of 25 μM adenine. Data are average of 3 independent experiments. (E) NAD⁺ levels after 48 hours NAMPT inhibition (0.01 μM FK866) in presence or absence of 25 μM adenine. Data are average and standard deviation of 3 independent experiments. Statistical analysis as done using one-way ANOVA with Tukey’s post-hoc test: ****P <0.0001; no statistical labelling indicates no statistical difference was observed.