Cephamycins inhibit pathogen sporulation and effectively treat recurrent Clostridioides difficile infection

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Abstract

Spore-forming bacteria encompass a diverse range of genera and species, including important human and animal pathogens, and food contaminants. Clostridioides difficile is one such bacterium and is a global health threat because it is the leading cause of antibiotic-associated diarrhoea in hospitals. A crucial mediator of C. difficile disease initiation, dissemination and re-infection is the formation of spores that are resistant to current therapeutics, which do not target sporulation. Here, we show that cephamycin antibiotics inhibit C. difficile sporulation by targeting spore-specific penicillin-binding proteins. Using a mouse disease model, we show that combined treatment with the current standard-of-care antibiotic, vancomycin, and a cephamycin prevents disease recurrence. Cephamycins were found to have broad applicability as an anti-sporulation strategy, as they inhibited sporulation in other spore-forming pathogens, including the food contaminant Bacillus cereus. This study could directly and immediately affect treatment of C. difficile infection and advance drug development to control other important spore-forming bacteria that are problematic in the food industry (B. cereus), are potential bioterrorism agents (Bacillus anthracis) and cause other animal and human infections.

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Fig. 1: Visualization of C. difficile sporulation morphology of untreated and cefoxitin-treated M7404 cells and the in vitro effect of cephamycins on C. difficile M7404 sporulation.
Fig. 2: The in vitro effect of cephamycins on B. cereus, B. subtilis and P. sordellii sporulation.
Fig. 3: Identification of the sporulation-specific PBP cephamycin targets, their role in sporulation and interaction with the cephamycins.
Fig. 4: Co-administration of cefotetan and vancomycin prevents recurrent CDI.

Data availability

The data that support the findings of this study are available from the corresponding author on reasonable request.

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Acknowledgements

This work was supported by the Australian National Health and Medical Research Council grant APP1145760 and the Australian Research Council Future Fellowship FT12010077 awarded to D.L.

Author information

Y.N.S. co-designed in vitro work, performed the majority of the experimental work and co-wrote the manuscript. M.L.H. co-designed and led in vivo experiments. M.M.A. performed toxin assays and made initial TEM observations of cefoxitin effects on spores. J.S. made initial observations of spore reduction with cefoxitin. N.D. co-designed and performed binding assays with compounds and targets. S.L.D. performed sporulation assays with other clades. B.A.C. performed cephalosporin sporulation experiments. S.M. co-designed binding assays and co-wrote the manuscript, and D.L. co-designed all experiments and co-wrote the manuscript.

Correspondence to Dena Lyras.

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Supplementary Figs. 1–6, Supplementary Tables 1–3 and Supplementary References.

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Srikhanta, Y.N., Hutton, M.L., Awad, M.M. et al. Cephamycins inhibit pathogen sporulation and effectively treat recurrent Clostridioides difficile infection. Nat Microbiol (2019) doi:10.1038/s41564-019-0519-1

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