Reuterin disrupts Clostridioides difficile metabolism and pathogenicity through reactive oxygen species generation.
Antibiotic resistance is one of the world's greatest public health challenges and adjunct probiotic therapies are strategies that could lessen this burden. Clostridioides difficile infection (CDI) is a prime example where adjunct probiotic therapies could decrease disease incidence through prevention. Human-derived Lactobacillus reuteri is a probiotic that produces the antimicrobial compound reuterin known to prevent C. difficile colonization of antibiotic-treated fecal microbial communities. However, the mechanism of inhibition is unclear. We show that reuterin inhibits C. difficile outgrowth from spores and vegetative cell growth, however, no effect on C. difficile germination or sporulation was observed. Consistent with published studies, we found that exposure to reuterin stimulated reactive oxygen species (ROS) in C. difficile, resulting in a concentration-dependent reduction in cell viability that was rescued by the antioxidant glutathione. Sublethal concentrations of reuterin enhanced the susceptibility of vegetative C. difficile to vancomycin and metronidazole treatment and reduced toxin synthesis by C. difficile. We also demonstrate that reuterin is protective against C. difficile toxin-mediated cellular damage in the human intestinal enteroid model. Overall, our results indicate that ROS are essential mediators of reuterin activity and show that reuterin production by L. reuteri is compatible as a therapeutic in a clinically relevant model.
['Engevik|Melinda A|MA|0000-0002-9742-9932', 'Danhof|Heather A|HA|0000-0002-6347-0021', 'Shrestha|Ritu|R|', 'Chang-Graham|Alexandra L|AL|0000-0002-3403-6787', 'Hyser|Joseph M|JM|', 'Haag|Anthony M|AM|', 'Mohammad|Mahmoud A|MA|', 'Britton|Robert A|RA|0000-0001-8983-9539', 'Versalovic|James|J|0000-0002-5765-4385', 'Sorg|Joseph A|JA|0000-0001-7822-2656', 'Spinler|Jennifer K|JK|0000-0002-7830-7665']
D000900:Anti-Bacterial Agents D016360:Clostridioides difficile D004357:Drug Synergism D004847:Epithelial Cells D005985:Glyceraldehyde D006801:Humans D052200:Lactobacillus reuteri D009940:Organoids D018384:Oxidative Stress D019936:Probiotics D011407:Propane D017382:Reactive Oxygen Species D013171:Spores, Bacterial
Reuterin Clostridioides difficile metabolism reactive oxygen species probiotics Lactobacillus reuteri enteroids organoids oxidative stress