Extended Data Fig. 8: Fructose is transformed into polar metabolites in a KHK-dependent manner.
From: Dietary fructose enhances tumour growth indirectly via interorgan lipid transfer
(a) Schematic for the generation of hepatocyte conditioned media (CM) and CaSki conditioned media (CM). Image created with BioRender.com. (b) Heatmap of labelled metabolites measured from hepatocyte CM (n = 3). White indicates no isotope labelling. Dark green indicates extensive isotope labelling. Except for fructose, which was the tracer, these metabolites were excreted into the media by primary mouse hepatocytes incubated for 24 h with [U-13C] fructose. (c) Heatmap of log2 fold changes based on the ratio of metabolite intensity from CaSki CM relative to metabolite intensity from hepatocyte CM (n = 3). See (a) for design schematic. Dark blue indicates a low ratio (i.e., metabolite is high in hepatocyte CM and low in CaSki CM). Red indicates a high ratio (i.e., metabolite is low in hepatocyte CM and high in CaSki CM). Only metabolites from (b) that were at least 1% labelled were analysed. (d) Concentration of labelled fructose in the serum of nude mice 30 min after administration of 1.5 g/kg [U-13C] fructose and 75 min after the indicated dose of PF-06835919 (n = 3-4, per dose). (e) Isotopologue distribution for serum glucose from nude mice 30 min after administration of [U-13C] fructose and 75 min after the indicated dose of PF-06835919 (n = 3-4, per dose). (f) Concentration of PF-06835919 in nude mouse serum after i.p. injection at a dose of 50 mg/kg. Measurements were made at 1 h, 4 h, 8 h, and 25 h post-injection (n = 4 mice per data point). Nonsignificant differences are not indicated. (g) CaSki tumour outgrowth in mice on a control diet administered a daily i.p. injection of vehicle (n = 4; Control Diet + No drug), a control diet administered 50 mg/kg PF-06835919 (n = 4; Control Diet + KHK Drug), or a high-fructose diet administered 50 mg/kg PF-06835919 (n = 3 Fructose Diet + KHK Drug). Data from Fig. 1g (Fructose Diet + No Drug, n = 7) provided a benchmark to show the impact of PF-06835919 on fructose-mediated CaSki tumour growth. All experiments from this figure were performed once. P values determined by ordinary one-way ANOVA with Tukey’s multiple comparisons test (d, e) or ordinary one-way ANOVA with Tukey’s multiple comparisons test of AUC (g). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, and n.s. is not significant. (g) shows mean ± standard error. The rest of data shown as mean ± standard deviation.
