Penicillin Binding Protein Substitutions Cooccur with Fluoroquinolone Resistance in Epidemic Lineages of Multidrug-Resistant Clostridioides difficile

Dingle, Kate E.; Freeman, Jane; Didelot, Xavier; Quan, T Phuong; Eyre, David W; Swann, Jeremy; Spittal, William; Clark, Emma; Jolley, Keith A.; Walker, A. Sarah; Wilcox, Mark H.; Crook, Derrick W.

doi:10.1128/mbio.00243-23

Cited by 4 publications

(5 citation statements)

References 102 publications

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Section: Discussionsupporting

confidence: 55%

“…However, this hypothesis is not supported by our data as 4→3 PG cross-links were more abundant in antibiotic-treated cells. In line with this result, biochemical analyses of the purified class A PBP PBP1 and the class B PBP PBP2 of C. difficile , which are respectively essential for cross-links synthesis during vegetative cell division and elongation, revealed that both enzymes are insensitive to most cephalosporins and carbapenems 32,33 The only other explanation to the observed structural changes is the inhibition of at least one D,D-carboxypeptidase. Activation of the L,D-transpeptidation pathway leading to β-lactam resistance in E. faecium and E. coli requires the production of a β-lactam-insensitive D,D-carboxypeptidase to generate the tetrapeptide substrate of LDTs 11,31 .…”

Section: Discussionmentioning

confidence: 54%

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The L,D-transpeptidation pathway is inhibited by antibiotics of the β-lactam class in Clostridioides difficile

Paiva,

Courtin,

Charpentier

et al. 2024

Preprint

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SummaryThe resistance ofClostridioides difficileto the β-lactam antibiotics cephalosporins, which target the peptidoglycan (PG) assembly, is a leading contributor to the development ofC. difficileinfections.C. difficilehas an original PG structure with a predominance of 3→3 cross-links generated by L,D-transpeptidases (LDTs).C. difficileforms spores and we show that the spore cortex PG contains exclusively 3→3 cross-links. PG and spore cortex ofC. difficilecells were largely unaffected by the deletion of the three predicted LDTs, revealing the implication of a new family of LDTS. The D,D-carboxypeptidases producing the essential LDT substrate were inactivated by cephalosporins, resulting in the inhibition of the L,D-transpeptidation pathway. In contrast, the participation of penicillin-binding proteins (PBPs) to PG cross-linking increased in the presence of the antibiotics. Our findings highlight that cephalosporin resistance is not primarily mediated by LDTs and illustrate the plasticity of the PG biosynthesis machinery inC. difficile.

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Section: Discussionsupporting

confidence: 55%

Section: Discussionmentioning

confidence: 54%

The L,D-transpeptidation pathway is inhibited by antibiotics of the β-lactam class in Clostridioides difficile

Paiva,

Courtin,

Charpentier

et al. 2024

Preprint

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“… 14 The emergence of epidemic C. difficile RT027 in the early 2000s was notably associated with resistance to fluoroquinolones and, more recently, high-level resistance to cephalosporins has been also associated with this outbreak. 15 , 16 …”

Section: Discussionmentioning

confidence: 99%

Antimicrobial susceptibility in Clostridioides difficile varies according to European region and isolate source

Freeman,

Viprey,

Ewin

et al. 2024

JAC-Antimicrobial Resistance

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Objectives Clostridioides difficile epidemiology is evolving with country-associated emerging and resistant ribotypes (RT). Antimicrobial susceptibility testing of C. difficile isolated from clinical and animal samples collected across Europe in 2018 was performed to provide antimicrobial resistance data and according to C. difficile RTs and source. Methods Samples were cultured for C. difficile and isolates PCR ribotyped. Metronidazole, vancomycin, fidaxomicin, moxifloxacin, clindamycin, imipenem, tigecycline, linezolid, rifampicin and meropenem minimum inhibitory concentrations (MICs) for 280 clinical and 126 animal isolates were determined by Wilkins–Chalgren agar dilution. Results Fidaxomicin was the most active antimicrobial (all isolates geometric mean MIC = 0.03 mg/L) with no evidence of reduced susceptibility. Metronidazole MICs were elevated among RT027 (1.87 mg/L) and RT181 clinical isolates (1.03 mg/L). RT027 and RT181 had elevated geometric mean moxifloxacin MICs (14.49 mg/L, 16.88 mg/L); clindamycin (7.5 mg/L, 9.1 mg/L) and rifampicin (0.6 mg/L, 21.5 mg/L). Five isolates (RT002, RT010 and RT016) were metronidazole resistant (MIC = 8 mg/L) and 10 (RT027; RT198) had intermediate resistance (4 mg/L). Metronidazole MICs were not elevated in animal isolates. Increased geometric mean vancomycin MICs were observed among RT078, mostly isolated from animals, but there was no resistance (MIC ≥ 4 mg/L). Clinical and animal isolates of multiple RTs showed resistance to moxifloxacin and clindamycin. No resistance to imipenem or meropenem was observed. Conclusion Increased antimicrobial resistance was detected in eastern Europe and mostly associated with RT027 and related emerging RT181, while clinical isolates from northern and western Europe had the lowest general levels of resistance.

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“…The molecular mechanisms by which MreE mutations increase resistance to specific classes of beta-lactam antibiotics [7] [8,9] are as yet undefined and may include reduced binding affinity of affected antibiotic classes, in addition to interaction with other genomic loci that can produce higher levels of resistance, a finding suggested by the fact that transfer of the mreE alleles alone into susceptible strains did not elevate most cephalosporin MICs to the level of the original parent strain (Fig 2C). The gene-level variants also provide targets to aid in the detection of cephalosporin resistance, whether by strain genomic analyses or with targeted mreE probes to evaluate primary patient samples for resistant strains.…”

Section: Discussionmentioning

confidence: 99%

“…Variants in the pathogen's penicillin-binding protein 2 (PBP2) have also been identified that have reduced binding-affinity to certain beta-lactam classes, including cephalosporins and carbapenems [7]. However, causative effects of these variants on phenotypic resistance in vitro and in vivo have not yet been defined, limiting our understanding of their role in patient disease [8][9][10].…”

Section: Introductionmentioning

confidence: 99%

Clostridioides difficileMreE (PBP2) variants facilitate clinical disease during cephalosporin exposures

Worley,

Benedetto,

Delaney

et al. 2023

Preprint

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Cephalosporins are the most common triggers of healthcare-associated Clostridioides difficile infections (CDI). Here, we confirm gene-level drivers of cephalosporin resistance and their roles in promoting disease. Genomic-epidemiologic analyses of 306 C. difficile isolates from a hospital surveillance program monitoring asymptomatic carriers and CDI patients identified prevalent third-generation cephalosporin resistance to ceftriaxone at >256 ug/mL in 26% of isolates. Resistance was associated with patient cephalosporin exposures 8-10 days before C. difficile detection. Genomic analyses identified variants in the mreE penicillin binding protein 2 (PBP2) associated with resistance to multiple beta-lactam classes. Transfer of variants into susceptible strain CD630 elevated resistance to first and third-generation cephalosporins. Transfer into the mouse-infective strain ATCC 43255 enabled disease when mice were exposed to 500ug/mL cefoperazone, a dose that inhibited the isogenic susceptible strain. Our findings establish roles of cephalosporins and mreE-cephalosporin-resistant variants in CDI and provide testable genetic loci for detecting resistance in patient strains.

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Penicillin Binding Protein Substitutions Cooccur with Fluoroquinolone Resistance in Epidemic Lineages of Multidrug-Resistant Clostridioides difficile

Cited by 4 publications

References 102 publications

The L,D-transpeptidation pathway is inhibited by antibiotics of the β-lactam class in Clostridioides difficile

The L,D-transpeptidation pathway is inhibited by antibiotics of the β-lactam class in Clostridioides difficile

Antimicrobial susceptibility in Clostridioides difficile varies according to European region and isolate source

Clostridioides difficileMreE (PBP2) variants facilitate clinical disease during cephalosporin exposures

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