Alpha diversity and composition of bacterial communities (beta diversity). Shannon diversity index was calculated at phylum, class, genus and species taxonomy level according to long vs short PFS (cutoff 6 months), PD-L1 expression (≥50% high, <50% low) and front-line chemotherapy (CHT)-treatment prior to ICI. There was no significant difference in Shannon diversity index regarding PFS (A), PD-L1 expression (B) and CHT-treatment (C) using either taxonomic level. Diversity (Shannon and Simpson) indices and p-values for all alpha-diversity comparisons are listed in STable 3. Ordination plot using UMAP was generated from normalized, CLR-transformed bacterial abundances in the same comparisons. Permanova analysis was used to assess significant differences between the composition of bacterial communities. There was a significant difference between long- vs short PFS patients (F = 3.379, p = 0.0006, D) and between CHT-treated vs CHT-naive patients (F = 2.139, p = 0.0156, F), whereas no significant difference was detected between PD-L1 high vs PD-L1 low patients (F = 0.916, p = 0.0156, D) regarding the composition of bacterial species. Metric data are shown as mean and corresponding standard deviation (SD). Statistical significance *P < 0.05; **P < 0.01, ***P<.001. N/A: data not available.