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| Status |
Public on Mar 24, 2022 |
| Title |
The licorice metabolite enoxolone attenuates Clostridioides difficile pathophysiology by corrupting its metabolic and toxin production networks |
| Organism |
Clostridioides difficile |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Toxins TcdA and TcdB are the main virulence factors of Clostridioides difficile, a leading cause of hospital-acquired diarrhea. We investigated the therapeutic potential of inhibiting the biosynthesis of TcdA and TcdB. Accordingly, screening of structurally diverse phytochemicals with medicinal properties identified 18b-glycyrrhetinic acid (enoxolone) as an inhibitor of TcdA and TcdB biosynthesis. Enoxolone also inhibited sporulation. In a CDI colitis model, enoxolone when combined with vancomycin protected mice from becoming moribund and the combination was more effective than vancomycin alone, a standard of care antibiotic for CDI. While enoxolone alone reduced the in vivo load of toxins, the monotherapy did not protect mice from CDI. Affinity based proteomics identified ATP synthase subunit alpha (AtpA) and adenine deaminase (Ade) as possible molecular targets for enoxolone. Silencing of mRNA for Ade and AtpA also reduced toxin biosynthesis, while molecular interaction analysis showed that enoxolone directly bound to Ade. Ade converts adenine to hypoxanthine as an early step in the purine salvage pathway. Metabolomics revealed enoxolone caused cells to accumulate adenosine and deplete hypoxanthine and ATP. Accordingly, supplementation with hypoxanthine partly restored toxin production. Enoxolone also impacted phosphate metabolism by reducing the amounts of cellular phosphate. Thus, supplementation with triethyl phosphate as a source of phosphate also partly restored toxin production. When hypoxanthine and triethyl phosphate were combined, toxin production was fully restored in the presence of enoxolone. Taken together, studies with enoxolone revealed metabolic pathways that affect C. difficile toxin production and could represent potential anti-virulence drug targets.
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| Overall design |
mRNA profiles of C. difficile R20291 upon exposure to enoxolone (16 µM) or DMSO for 30 minutes.
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| Contributor(s) |
Hurdle JG, Marreddy RK, Lee RE, Phelps GA |
| Citation missing |
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| Submission date |
Mar 21, 2022 |
| Last update date |
Mar 24, 2022 |
| Contact name |
Richard Lee |
| E-mail(s) |
richard.lee@stjude.org
|
| Phone |
9015956617
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| Organization name |
St. Jude Children's Research Hospital
|
| Department |
Department of Chemical Biology and Therapeutics
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| Lab |
Lee
|
| Street address |
262 Danny Thomas Place
|
| City |
Memphis |
| State/province |
TN |
| ZIP/Postal code |
38105 |
| Country |
USA |
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| Platforms (1) |
| GPL30071 |
Illumina NovaSeq 6000 (Clostridioides difficile) |
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| Samples (6)
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| Relations |
| BioProject |
PRJNA818416 |
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