Figure 1 from Human Colon Cancer–Derived Clostridioides difficile Strains Drive Colonic Tumorigenesis in Mice

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Abstract
C. difficile strains isolated from human colon cancer mucosa drive colonic tumor formation in GF ApcMin/+ mice. A and B, Variation in colonic tumorigenic potential in GF ApcMin/+ mice at 12 to 14 weeks post-inoculation (p.i.) by individual BF-positive tumor (BF+T; A) or BF-negative tumor (BF−T; B) colorectal cancer mucosal samples from six patients. n = 6–8 mice per group, from two independent experiments. C, 16S rRNA amplicon sequencing relative abundances of the top 20 species in the six patient tumor slurry inocula and 12 to 14 weeks p.i. stools from mice gavaged with the tumor slurries. D, A consortium of 30 bacterial isolates derived from GF mice gavaged with the 3728T slurry recapitulated colonic tumorigenesis of the original 3728T slurry. n = 5–11 mice per group. E and F, Colonic tumor counts (E) and representative methylene blue staining (F) of colonic tumors (arrows) at 12 weeks p.i. in a vancomycin/gentamicin model (see Methods) of chronic colonization with nontoxigenic (TcdA−TcdB−) or toxigenic (TcdA+TcdB+) C. difficile strains. n = 5–18 mice per group, from two independent experiments. G and H, Colonic tumor counts (G) and representative methylene blue staining (H) of colonic tumors (arrows) from GF ApcMin/+ mice gavaged with the 3728T isolates with or without the CIm_3728T or CIm_3752T C. difficile strains. n = 6–8 mice per group. I, Colonic tumors induced in GF ApcMin/+ mice gavaged with either the 3979T slurry or the 3979T slurry with C. difficile strain CIm_3728T at 10 weeks p.i. The cross represents a mouse death. n = 7–8 mice per group. Statistical significance was calculated via Kruskal–Wallis followed by Mann–Whitney tests. Mann–Whitney P values are shown. ns, not significant.