Repurposing clofarabine against enteric pathogens via metabolic starvation
- 1University of Sheffield
Abstract
Enteropathogenic and enterohaemorrhagic Escherichia coli (EPEC/EHEC) cause severe global diarrhoeal disease, exacerbated by rising antimicrobial resistance. Pathogenesis relies on "attaching and effacing" lesions, where bacteria generate actin pedestals to adhere, replicate, and destroy intestinal microvilli. To identify novel anti-virulence candidates targeting this process, we conducted a high-throughput phenotypic screen quantifying pedestal formation via high-content fluorescence microscopy. We identified the anticancer nucleotide analogue clofarabine as a potent inhibitor of EPEC growth and pathogenesis. Distinct from its oncology mechanism - inducing DNA chain termination and apoptosis - clofarabine competes with bacterial ATP-dependent processes to impede gene transcription, protein synthesis, and Type III Secretion System (T3SS) function. Remarkably, EPEC counteracts this inhibition by cleaving clofarabine, enabling restored protein synthesis and resumed growth. Furthermore, we report for the first time that clofarabine suppresses protein translation in human monocytes, suggesting an uncharacterised translational impact on mammalian cells. These findings reveal host and bacterial translation as interconnected vulnerabilities to purine antimetabolites, offering a novel strategy to disarm enteric pathogens while uncovering a bacterial enzymatic mechanism for neutralizing host-targeted therapeutics.
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BibTeX
@misc{deans2026repurposing,
title = {{Repurposing clofarabine against enteric pathogens via metabolic starvation}},
author = {N. D. Deans and K. L. Naylor and S. Srour and B. Mallick and T. K. Pham and J. W. Sheffield and O. F. Ansell and C. D. Edington-Siddon and E. A. Burt and G. Stafford and M. Collins and D. Humphreys},
howpublished = {bioRxiv},
year = {2026},
month = oct,
doi = {10.64898/2026.10.09.757903},
url = {https://www.biorxiv.org/content/10.64898/2026.10.09.757903v1},
note = {CC BY},
}
RIS
TY - UNPB
TI - Repurposing clofarabine against enteric pathogens via metabolic starvation
AU - N. D. Deans
AU - K. L. Naylor
AU - S. Srour
AU - B. Mallick
AU - T. K. Pham
AU - J. W. Sheffield
AU - O. F. Ansell
AU - C. D. Edington-Siddon
AU - E. A. Burt
AU - G. Stafford
AU - M. Collins
AU - D. Humphreys
PY - 2026
DA - 2026/10/09
PB - bioRxiv
DO - 10.64898/2026.10.09.757903
UR - https://www.biorxiv.org/content/10.64898/2026.10.09.757903v1
AB - Enteropathogenic and enterohaemorrhagic Escherichia coli (EPEC/EHEC) cause severe global diarrhoeal disease, exacerbated by rising antimicrobial resistance. Pathogenesis relies on "attaching and effacing" lesions, where bacteria generate actin pedestals to adhere, replicate, and destroy intestinal microvilli. To identify novel anti-virulence candidates targeting this process, we conducted a high-throughput phenotypic screen quantifying pedestal formation via high-content fluorescence microscopy. We identified the anticancer nucleotide analogue clofarabine as a potent inhibitor of EPEC growth and pathogenesis. Distinct from its oncology mechanism - inducing DNA chain termination and apoptosis - clofarabine competes with bacterial ATP-dependent processes to impede gene transcription, protein synthesis, and Type III Secretion System (T3SS) function. Remarkably, EPEC counteracts this inhibition by cleaving clofarabine, enabling restored protein synthesis and resumed growth. Furthermore, we report for the first time that clofarabine suppresses protein translation in human monocytes, suggesting an uncharacterised translational impact on mammalian cells. These findings reveal host and bacterial translation as interconnected vulnerabilities to purine antimetabolites, offering a novel strategy to disarm enteric pathogens while uncovering a bacterial enzymatic mechanism for neutralizing host-targeted therapeutics.
ER -