Chemogenomic Screen for Imipenem Resistance in Gram-Negative Bacteria

ABSTRACT Carbapenem-resistant Gram-negative bacteria are considered a major threat to global health. Imipenem (IMP) is used as a last line of treatment against these pathogens, but its efficacy is diminished by the emergence of resistance. We applied a whole-genome screen in Escherichia coli, Klebsi...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Jessica Y. El Khoury, Alexandra Maure, Hélène Gingras, Philippe Leprohon, Marc Ouellette
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2019
Materias:	chemical mutagenesis carbapenem resistance Escherichia coli Klebsiella pneumoniae Pseudomonas aeruginosa Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/a933a8c7d8e94f08abc8a8366fe32f0a
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:a933a8c7d8e94f08abc8a8366fe32f0a
record_format	dspace
spelling	oai:doaj.org-article:a933a8c7d8e94f08abc8a8366fe32f0a2021-12-02T19:47:35ZChemogenomic Screen for Imipenem Resistance in Gram-Negative Bacteria10.1128/mSystems.00465-192379-5077https://doaj.org/article/a933a8c7d8e94f08abc8a8366fe32f0a2019-12-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSystems.00465-19https://doaj.org/toc/2379-5077ABSTRACT Carbapenem-resistant Gram-negative bacteria are considered a major threat to global health. Imipenem (IMP) is used as a last line of treatment against these pathogens, but its efficacy is diminished by the emergence of resistance. We applied a whole-genome screen in Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa isolates that were submitted to chemical mutagenesis, selected for IMP resistance, and characterized by next-generation sequencing. A comparative analysis of IMP-resistant clones showed that most of the highly mutated genes shared by the three species encoded proteins involved in transcription or signal transduction. Of these, the rpoD gene was one of the most prevalent and an E. coli strain disrupted for rpoD displayed a 4-fold increase in resistance to IMP. E. coli and K. pneumoniae also specifically shared several mutated genes, most involved in membrane/cell envelope biogenesis, and the contribution in IMP susceptibility was experimentally proven for amidases, transferases, and transglycosidases. P. aeruginosa differed from the two Enterobacteriaceae isolates with two different resistance mechanisms, with one involving mutations in the oprD porin or, alternatively, in two-component systems. Our chemogenomic screen performed with the three species has highlighted shared and species-specific responses to IMP. IMPORTANCE Gram-negative carbapenem-resistant bacteria are a major threat to global health. The use of genome-wide screening approaches to probe for genes or mutations enabling resistance can lead to identification of molecular markers for diagnostics applications. We describe an approach called Mut-Seq that couples chemical mutagenesis and next-generation sequencing for studying resistance to imipenem in the Gram-negative bacteria Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa. The use of this approach highlighted shared and species-specific responses, and the role in resistance of a number of genes involved in membrane biogenesis, transcription, and signal transduction was functionally validated. Interestingly, some of the genes identified were previously considered promising therapeutic targets. Our genome-wide screen has the potential to be extended outside drug resistance studies and expanded to other organisms.Jessica Y. El KhouryAlexandra MaureHélène GingrasPhilippe LeprohonMarc OuelletteAmerican Society for Microbiologyarticlechemical mutagenesiscarbapenem resistanceEscherichia coliKlebsiella pneumoniaePseudomonas aeruginosaMicrobiologyQR1-502ENmSystems, Vol 4, Iss 6 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	chemical mutagenesis carbapenem resistance Escherichia coli Klebsiella pneumoniae Pseudomonas aeruginosa Microbiology QR1-502
spellingShingle	chemical mutagenesis carbapenem resistance Escherichia coli Klebsiella pneumoniae Pseudomonas aeruginosa Microbiology QR1-502 Jessica Y. El Khoury Alexandra Maure Hélène Gingras Philippe Leprohon Marc Ouellette Chemogenomic Screen for Imipenem Resistance in Gram-Negative Bacteria
description	ABSTRACT Carbapenem-resistant Gram-negative bacteria are considered a major threat to global health. Imipenem (IMP) is used as a last line of treatment against these pathogens, but its efficacy is diminished by the emergence of resistance. We applied a whole-genome screen in Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa isolates that were submitted to chemical mutagenesis, selected for IMP resistance, and characterized by next-generation sequencing. A comparative analysis of IMP-resistant clones showed that most of the highly mutated genes shared by the three species encoded proteins involved in transcription or signal transduction. Of these, the rpoD gene was one of the most prevalent and an E. coli strain disrupted for rpoD displayed a 4-fold increase in resistance to IMP. E. coli and K. pneumoniae also specifically shared several mutated genes, most involved in membrane/cell envelope biogenesis, and the contribution in IMP susceptibility was experimentally proven for amidases, transferases, and transglycosidases. P. aeruginosa differed from the two Enterobacteriaceae isolates with two different resistance mechanisms, with one involving mutations in the oprD porin or, alternatively, in two-component systems. Our chemogenomic screen performed with the three species has highlighted shared and species-specific responses to IMP. IMPORTANCE Gram-negative carbapenem-resistant bacteria are a major threat to global health. The use of genome-wide screening approaches to probe for genes or mutations enabling resistance can lead to identification of molecular markers for diagnostics applications. We describe an approach called Mut-Seq that couples chemical mutagenesis and next-generation sequencing for studying resistance to imipenem in the Gram-negative bacteria Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa. The use of this approach highlighted shared and species-specific responses, and the role in resistance of a number of genes involved in membrane biogenesis, transcription, and signal transduction was functionally validated. Interestingly, some of the genes identified were previously considered promising therapeutic targets. Our genome-wide screen has the potential to be extended outside drug resistance studies and expanded to other organisms.
format	article
author	Jessica Y. El Khoury Alexandra Maure Hélène Gingras Philippe Leprohon Marc Ouellette
author_facet	Jessica Y. El Khoury Alexandra Maure Hélène Gingras Philippe Leprohon Marc Ouellette
author_sort	Jessica Y. El Khoury
title	Chemogenomic Screen for Imipenem Resistance in Gram-Negative Bacteria
title_short	Chemogenomic Screen for Imipenem Resistance in Gram-Negative Bacteria
title_full	Chemogenomic Screen for Imipenem Resistance in Gram-Negative Bacteria
title_fullStr	Chemogenomic Screen for Imipenem Resistance in Gram-Negative Bacteria
title_full_unstemmed	Chemogenomic Screen for Imipenem Resistance in Gram-Negative Bacteria
title_sort	chemogenomic screen for imipenem resistance in gram-negative bacteria
publisher	American Society for Microbiology
publishDate	2019
url	https://doaj.org/article/a933a8c7d8e94f08abc8a8366fe32f0a
work_keys_str_mv	AT jessicayelkhoury chemogenomicscreenforimipenemresistanceingramnegativebacteria AT alexandramaure chemogenomicscreenforimipenemresistanceingramnegativebacteria AT helenegingras chemogenomicscreenforimipenemresistanceingramnegativebacteria AT philippeleprohon chemogenomicscreenforimipenemresistanceingramnegativebacteria AT marcouellette chemogenomicscreenforimipenemresistanceingramnegativebacteria
_version_	1718375982335787008

Chemogenomic Screen for Imipenem Resistance in Gram-Negative Bacteria

Ejemplares similares