Fig. 4: Gut microbiome diversity and metabolic profile associated with durable responses to immunotherapy.

a Top 20 differentially abundant microbes (adj. p. value < 0.05, |LogFC|>1.5) between Baseline_DCB and Baseline_LCB displayed in a heatmap. Similar to tumor transcriptome, heatmaps demonstrate that across all microbes, patients with durable clinical benefit express higher microbe levels, and this expression is increased with treatment in DCB (On.TX_DCB, red), as compared to baseline DCB (Baseline_DCB, orange), baseline LCB (Baseline_LCB, light blue), and on treatment LCB (On.TX_LCB, dark blue) patient populations. b Violin plots quantify OTU abundance in all four groups for select bacterial species, all of which demonstrate higher levels of the bacterial species at baseline in DCB (Baseline_DCB, orange) as compared to LCB (Baseline_LCB, light blue), and more so with treatment in the DCB group (On.TX_DCB, dark red). *p < 0.05, **p < 0.01; derived from linear regression of log-normalized abundances, by clinical group, with multiple test corrections. c Volcano plots demonstrate variety of significant (|logFC|>1.5, adjusted p < 0.1) differentially abundant metabolites between Baseline LCB (Baseline_LCB, light blue), Baseline DCB (Baseline_DCB, orange), On-Treatment LCB (On.TX_LCB, dark blue), and On-Treatment DCB (On.TX_DCB, red) patient populations. d Expression levels of the 20 most highly differentially expressed metabolites separate Baseline_DCB and Baseline_LCB fecal samples to a high degree based on Euclidean distances, across all 4 groups. Some metabolites were decreased with treatment in DCB and LCB (indole), some were solely increased in DCB with treatment (C3-OH, TMAO, Glu, and Ser), and some lipids were increased more so in LCB, regardless of timepoint (TGs and DGs). *p < 0.05, derived from linear regression of log-normalized abundances, by clinical group. Exact p-values can be found in Data Tables 2 and 3.