Fig. 5. B.fragilis promotes tumor resistance in a VitD-dependent manner.

(A-B) Meta-analysis of metagenomic data to determine (A) common features in microbial gene products (top 20/62 features in each direction shown, 20/62) and (B) last known taxon associated with differences in VitD availability. Fecal samples were sequenced from WT or Gc^-/- mice that had been fed with VitD₃ standard (2 IU/g), deficient (0 IU/g) or high (10 IU/g) diet for 3.5 weeks. Comparison is of mice with high VitD availability [WT + VitD₃^High (n=13), Gc^-/- + VitD₃^Standard (n=20), Gc^-/- + VitD₃^High (n=13) vs. mice with normal or low VitD availability [WT + VitD₃^Standard (n=22), WT + VitD₃^Deficient (n=10), Gc^-/- + VitD₃^Deficient (n=10)]. In (A, B), count of significant features indicated in the Venn diagram and shown by color scale (top) and ranked bar plots (bottom) show common features across 3 meta-analyses as indicated. (C) Growth profile of 0.2 x 10⁶ 5555 Braf^V600E cancer cells implanted into separately housed WT C57BL/6 groups of mice (n=10 per group) fed with VitD₃ standard (left graph) or deficient diet (right graph), starting 3.5 weeks before receiving B. fragilis, P. brevis or vehicle. Mice received 10⁹ B. fragilis or P. brevis by oral gavage on days -14, -12 and -10 prior to tumor inoculation. Data in (A, B) are presented as average log₂ median fold change from three meta-analyses of data from two independent experiments. Data in (C) are presented as tumor volume (mm³) ± SEM and are representative of two independent experiments for P. brevis and 3 independent experiments for B. fragilis. In (A, B), p values were calculated using the Mann–Whitney–Wilcoxon U test on parts per million (PPM) relative abundances for that feature in samples within each group for pairwise comparisons. The combined p value (cp) for meta-analysis of within-group comparisons was calculated using Fishers P value. For each feature type, the cut-offs for the meta-analysis were: p< 0.2, cp< 0.1, false discovery rate (FDR)<0.15. Tumor growth profiles (A, C and D) were compared using Bonferroni-corrected two-way ANOVA. ****p< 0.0001; ns, not significant.