Uncovering novel susceptibility targets to enhance the efficacy of third-generation cephalosporins against ESBL-producing uropathogenic Escherichia coli.

Phan, M-D; Bottomley, AL; Peters, KM; Harry, EJ; Schembri, MA

Uncovering novel susceptibility targets to enhance the efficacy of third-generation cephalosporins against ESBL-producing uropathogenic Escherichia coli.

Phan, M-D Bottomley, AL Peters, KM Harry, EJ Schembri, MA

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Publisher:: Oxford University Press (OUP)
Publication Type:: Journal Article
Citation:: The Journal of antimicrobial chemotherapy, 2020, 75, (6), pp. 1415-1423
Issue Date:: 2020-06

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Field	Value	Language
dc.contributor.author	Phan, M-D
dc.contributor.author	Bottomley, AL
dc.contributor.author	Peters, KM
dc.contributor.author	Harry, EJ
dc.contributor.author	Schembri, MA
dc.date.accessioned	2020-08-13T05:25:01Z
dc.date.available	2020-01-13
dc.date.available	2020-08-13T05:25:01Z
dc.date.issued	2020-06
dc.identifier.citation	The Journal of antimicrobial chemotherapy, 2020, 75, (6), pp. 1415-1423
dc.identifier.issn	0305-7453
dc.identifier.issn	1460-2091
dc.identifier.uri	http://hdl.handle.net/10453/142095
dc.description.abstract	BACKGROUND:Uropathogenic Escherichia coli (UPEC) are a major cause of urinary tract infection (UTI), one of the most common infectious diseases in humans. UPEC are increasingly associated with resistance to multiple antibiotics. This includes resistance to third-generation cephalosporins, a common class of antibiotics frequently used to treat UTI. METHODS:We employed a high-throughput genome-wide screen using saturated transposon mutagenesis and transposon directed insertion-site sequencing (TraDIS) together with phenotypic resistance assessment to identify key genes required for survival of the MDR UPEC ST131 strain EC958 in the presence of the third-generation cephalosporin cefotaxime. RESULTS:We showed that blaCMY-23 is the major ESBL gene in EC958 responsible for mediating resistance to cefotaxime. Our screen also revealed that mutation of genes involved in cell division and the twin-arginine translocation pathway sensitized EC958 to cefotaxime. The role of these cell-division and protein-secretion genes in cefotaxime resistance was confirmed through the construction of mutants and phenotypic testing. Mutation of these genes also sensitized EC958 to other cephalosporins. CONCLUSIONS:This work provides an exemplar for the application of TraDIS to define molecular mechanisms of resistance to antibiotics. The identification of mutants that sensitize UPEC to cefotaxime, despite the presence of a cephalosporinase, provides a framework for the development of new approaches to treat infections caused by MDR pathogens.
dc.format	Print
dc.language	eng
dc.publisher	Oxford University Press (OUP)
dc.relation	http://purl.org/au-research/grants/arc/DP150102062
dc.relation.ispartof	The Journal of antimicrobial chemotherapy
dc.relation.isbasedon	10.1093/jac/dkaa023
dc.rights	info:eu-repo/semantics/restrictedAccess
dc.subject	0605 Microbiology, 1108 Medical Microbiology, 1115 Pharmacology and Pharmaceutical Sciences
dc.subject.classification	Microbiology
dc.title	Uncovering novel susceptibility targets to enhance the efficacy of third-generation cephalosporins against ESBL-producing uropathogenic Escherichia coli.
dc.type	Journal Article
utslib.citation.volume	75
utslib.location.activity	England
utslib.for	0605 Microbiology
utslib.for	1108 Medical Microbiology
utslib.for	1115 Pharmacology and Pharmaceutical Sciences
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Strength - ithree - Institute of Infection, Immunity and Innovation
pubs.organisational-group	/University of Technology Sydney
utslib.copyright.status	closed_access	*
dc.date.updated	2020-08-13T05:24:55Z
pubs.issue	6
pubs.publication-status	Published
pubs.volume	75
utslib.citation.issue	6

Abstract:

BACKGROUND:Uropathogenic Escherichia coli (UPEC) are a major cause of urinary tract infection (UTI), one of the most common infectious diseases in humans. UPEC are increasingly associated with resistance to multiple antibiotics. This includes resistance to third-generation cephalosporins, a common class of antibiotics frequently used to treat UTI. METHODS:We employed a high-throughput genome-wide screen using saturated transposon mutagenesis and transposon directed insertion-site sequencing (TraDIS) together with phenotypic resistance assessment to identify key genes required for survival of the MDR UPEC ST131 strain EC958 in the presence of the third-generation cephalosporin cefotaxime. RESULTS:We showed that blaCMY-23 is the major ESBL gene in EC958 responsible for mediating resistance to cefotaxime. Our screen also revealed that mutation of genes involved in cell division and the twin-arginine translocation pathway sensitized EC958 to cefotaxime. The role of these cell-division and protein-secretion genes in cefotaxime resistance was confirmed through the construction of mutants and phenotypic testing. Mutation of these genes also sensitized EC958 to other cephalosporins. CONCLUSIONS:This work provides an exemplar for the application of TraDIS to define molecular mechanisms of resistance to antibiotics. The identification of mutants that sensitize UPEC to cefotaxime, despite the presence of a cephalosporinase, provides a framework for the development of new approaches to treat infections caused by MDR pathogens.

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http://hdl.handle.net/10453/142095