Phage Resistance in Multidrug-Resistant <named-content content-type="genus-species">Klebsiella pneumoniae</named-content> ST258 Evolves via Diverse Mutations That Culminate in Impaired Adsorption

ABSTRACT The evolution of phage resistance poses an inevitable threat to the efficacy of phage therapy. The strategic selection of phage combinations that impose high genetic barriers to resistance and/or high compensatory fitness costs may mitigate this threat. However, for such a strategy to be ef...

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Autores principales:	Shayla Hesse, Manoj Rajaure, Erin Wall, Joy Johnson, Valery Bliskovsky, Susan Gottesman, Sankar Adhya
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	bacteriophage cocktails bacteriophage resistance bacteriophage therapy Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/4e041000c950464486454e9b28544844
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spelling	oai:doaj.org-article:4e041000c950464486454e9b285448442021-11-15T15:56:58ZPhage Resistance in Multidrug-Resistant <named-content content-type="genus-species">Klebsiella pneumoniae</named-content> ST258 Evolves via Diverse Mutations That Culminate in Impaired Adsorption10.1128/mBio.02530-192150-7511https://doaj.org/article/4e041000c950464486454e9b285448442020-02-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.02530-19https://doaj.org/toc/2150-7511ABSTRACT The evolution of phage resistance poses an inevitable threat to the efficacy of phage therapy. The strategic selection of phage combinations that impose high genetic barriers to resistance and/or high compensatory fitness costs may mitigate this threat. However, for such a strategy to be effective, the evolution of phage resistance must be sufficiently constrained to be consistent. In this study, we isolated lytic phages capable of infecting a modified Klebsiella pneumoniae clinical isolate and characterized a total of 57 phage-resistant mutants that evolved from their prolonged coculture in vitro. Single- and double-phage-resistant mutants were isolated from independently evolved replicate cocultures grown in broth or on plates. Among resistant isolates evolved against the same phage under the same conditions, mutations conferring resistance occurred in different genes, yet in each case, the putative functions of these genes clustered around the synthesis or assembly of specific cell surface structures. All resistant mutants demonstrated impaired phage adsorption, providing a strong indication that these cell surface structures functioned as phage receptors. Combinations of phages targeting different host receptors reduced the incidence of resistance, while, conversely, one three-phage cocktail containing two phages targeting the same receptor increased the incidence of resistance (relative to its two-phage, nonredundant receptor-targeting counterpart). Together, these data suggest that laboratory characterization of phage-resistant mutants is a useful tool to help optimize therapeutic phage selection and cocktail design. IMPORTANCE The therapeutic use of bacteriophage (phage) is garnering renewed interest in the setting of difficult-to-treat infections. Phage resistance is one major limitation of phage therapy; therefore, developing effective strategies to avert or lessen its impact is critical. Characterization of in vitro phage resistance may be an important first step in evaluating the relative likelihood with which phage-resistant populations emerge, the most likely phenotypes of resistant mutants, and the effect of certain phage cocktail combinations in increasing or decreasing the genetic barrier to resistance. If this information confers predictive power in vivo, then routine studies of phage-resistant mutants and their in vitro evolution should be a valuable means for improving the safety and efficacy of phage therapy in humans.Shayla HesseManoj RajaureErin WallJoy JohnsonValery BliskovskySusan GottesmanSankar AdhyaAmerican Society for Microbiologyarticlebacteriophage cocktailsbacteriophage resistancebacteriophage therapyMicrobiologyQR1-502ENmBio, Vol 11, Iss 1 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	bacteriophage cocktails bacteriophage resistance bacteriophage therapy Microbiology QR1-502
spellingShingle	bacteriophage cocktails bacteriophage resistance bacteriophage therapy Microbiology QR1-502 Shayla Hesse Manoj Rajaure Erin Wall Joy Johnson Valery Bliskovsky Susan Gottesman Sankar Adhya Phage Resistance in Multidrug-Resistant <named-content content-type="genus-species">Klebsiella pneumoniae</named-content> ST258 Evolves via Diverse Mutations That Culminate in Impaired Adsorption
description	ABSTRACT The evolution of phage resistance poses an inevitable threat to the efficacy of phage therapy. The strategic selection of phage combinations that impose high genetic barriers to resistance and/or high compensatory fitness costs may mitigate this threat. However, for such a strategy to be effective, the evolution of phage resistance must be sufficiently constrained to be consistent. In this study, we isolated lytic phages capable of infecting a modified Klebsiella pneumoniae clinical isolate and characterized a total of 57 phage-resistant mutants that evolved from their prolonged coculture in vitro. Single- and double-phage-resistant mutants were isolated from independently evolved replicate cocultures grown in broth or on plates. Among resistant isolates evolved against the same phage under the same conditions, mutations conferring resistance occurred in different genes, yet in each case, the putative functions of these genes clustered around the synthesis or assembly of specific cell surface structures. All resistant mutants demonstrated impaired phage adsorption, providing a strong indication that these cell surface structures functioned as phage receptors. Combinations of phages targeting different host receptors reduced the incidence of resistance, while, conversely, one three-phage cocktail containing two phages targeting the same receptor increased the incidence of resistance (relative to its two-phage, nonredundant receptor-targeting counterpart). Together, these data suggest that laboratory characterization of phage-resistant mutants is a useful tool to help optimize therapeutic phage selection and cocktail design. IMPORTANCE The therapeutic use of bacteriophage (phage) is garnering renewed interest in the setting of difficult-to-treat infections. Phage resistance is one major limitation of phage therapy; therefore, developing effective strategies to avert or lessen its impact is critical. Characterization of in vitro phage resistance may be an important first step in evaluating the relative likelihood with which phage-resistant populations emerge, the most likely phenotypes of resistant mutants, and the effect of certain phage cocktail combinations in increasing or decreasing the genetic barrier to resistance. If this information confers predictive power in vivo, then routine studies of phage-resistant mutants and their in vitro evolution should be a valuable means for improving the safety and efficacy of phage therapy in humans.
format	article
author	Shayla Hesse Manoj Rajaure Erin Wall Joy Johnson Valery Bliskovsky Susan Gottesman Sankar Adhya
author_facet	Shayla Hesse Manoj Rajaure Erin Wall Joy Johnson Valery Bliskovsky Susan Gottesman Sankar Adhya
author_sort	Shayla Hesse
title	Phage Resistance in Multidrug-Resistant <named-content content-type="genus-species">Klebsiella pneumoniae</named-content> ST258 Evolves via Diverse Mutations That Culminate in Impaired Adsorption
title_short	Phage Resistance in Multidrug-Resistant <named-content content-type="genus-species">Klebsiella pneumoniae</named-content> ST258 Evolves via Diverse Mutations That Culminate in Impaired Adsorption
title_full	Phage Resistance in Multidrug-Resistant <named-content content-type="genus-species">Klebsiella pneumoniae</named-content> ST258 Evolves via Diverse Mutations That Culminate in Impaired Adsorption
title_fullStr	Phage Resistance in Multidrug-Resistant <named-content content-type="genus-species">Klebsiella pneumoniae</named-content> ST258 Evolves via Diverse Mutations That Culminate in Impaired Adsorption
title_full_unstemmed	Phage Resistance in Multidrug-Resistant <named-content content-type="genus-species">Klebsiella pneumoniae</named-content> ST258 Evolves via Diverse Mutations That Culminate in Impaired Adsorption
title_sort	phage resistance in multidrug-resistant <named-content content-type="genus-species">klebsiella pneumoniae</named-content> st258 evolves via diverse mutations that culminate in impaired adsorption
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/4e041000c950464486454e9b28544844
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Phage Resistance in Multidrug-Resistant <named-content content-type="genus-species">Klebsiella pneumoniae</named-content> ST258 Evolves via Diverse Mutations That Culminate in Impaired Adsorption

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