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Molecular studies of phages-Klebsiella pneumoniae in mucoid environment: innovative use of mucolytic agents prior to the administration of lytic phages

Mucins are important glycoproteins that form a protective layer throughout the gastrointestinal and respiratory tracts. There is scientific evidence of increase in phage-resistance in the presence of mucin for some bacterial pathogens. Manipulation in mucin composition may ultimately influence the e...

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Autores principales: Pacios, Olga, Blasco, Lucía, Ortiz Cartagena, Concha, Bleriot, Inés, Fernández-García, Laura, López, María, Barrio-Pujante, Antonio, Cuenca, Felipe Fernández, Aracil, Belén, Oteo-Iglesias, Jesús, Tomás, María
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10598653/
https://www.ncbi.nlm.nih.gov/pubmed/37886069
http://dx.doi.org/10.3389/fmicb.2023.1286046
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author Pacios, Olga
Blasco, Lucía
Ortiz Cartagena, Concha
Bleriot, Inés
Fernández-García, Laura
López, María
Barrio-Pujante, Antonio
Cuenca, Felipe Fernández
Aracil, Belén
Oteo-Iglesias, Jesús
Tomás, María
author_facet Pacios, Olga
Blasco, Lucía
Ortiz Cartagena, Concha
Bleriot, Inés
Fernández-García, Laura
López, María
Barrio-Pujante, Antonio
Cuenca, Felipe Fernández
Aracil, Belén
Oteo-Iglesias, Jesús
Tomás, María
author_sort Pacios, Olga
collection PubMed
description Mucins are important glycoproteins that form a protective layer throughout the gastrointestinal and respiratory tracts. There is scientific evidence of increase in phage-resistance in the presence of mucin for some bacterial pathogens. Manipulation in mucin composition may ultimately influence the effectiveness of phage therapy. In this work, two clinical strains of K. pneumoniae (K3574 and K3325), were exposed to the lytic bacteriophage vB_KpnS-VAC35 in the presence and absence of mucin on a long-term co-evolution assay, in an attempt to mimic in vitro the exposure to mucins that bacteria and their phages face in vivo. Enumerations of the bacterial and phage counts at regular time intervals were conducted, and extraction of the genomic DNA of co-evolved bacteria to the phage, the mucin and both was performed. We determined the frequency of phage-resistant mutants in the presence and absence of mucin and including a mucolytic agent (N-acetyl L-cysteine, NAC), and sequenced them using Nanopore. We phenotypically demonstrated that the presence of mucin induces the emergence of bacterial resistance against lytic phages, effectively decreased in the presence of NAC. In addition, the genomic analysis revealed some of the genes relevant to the development of phage resistance in long-term co-evolution, with a special focus on the mucoid environment. Genes involved in the metabolism of carbohydrates were mutated in the presence of mucin. In conclusion, the use of mucolytic agents prior to the administration of lytic phages could be an interesting therapeutic option when addressing K. pneumoniae infections in environments where mucin is overproduced.
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spelling pubmed-105986532023-10-26 Molecular studies of phages-Klebsiella pneumoniae in mucoid environment: innovative use of mucolytic agents prior to the administration of lytic phages Pacios, Olga Blasco, Lucía Ortiz Cartagena, Concha Bleriot, Inés Fernández-García, Laura López, María Barrio-Pujante, Antonio Cuenca, Felipe Fernández Aracil, Belén Oteo-Iglesias, Jesús Tomás, María Front Microbiol Microbiology Mucins are important glycoproteins that form a protective layer throughout the gastrointestinal and respiratory tracts. There is scientific evidence of increase in phage-resistance in the presence of mucin for some bacterial pathogens. Manipulation in mucin composition may ultimately influence the effectiveness of phage therapy. In this work, two clinical strains of K. pneumoniae (K3574 and K3325), were exposed to the lytic bacteriophage vB_KpnS-VAC35 in the presence and absence of mucin on a long-term co-evolution assay, in an attempt to mimic in vitro the exposure to mucins that bacteria and their phages face in vivo. Enumerations of the bacterial and phage counts at regular time intervals were conducted, and extraction of the genomic DNA of co-evolved bacteria to the phage, the mucin and both was performed. We determined the frequency of phage-resistant mutants in the presence and absence of mucin and including a mucolytic agent (N-acetyl L-cysteine, NAC), and sequenced them using Nanopore. We phenotypically demonstrated that the presence of mucin induces the emergence of bacterial resistance against lytic phages, effectively decreased in the presence of NAC. In addition, the genomic analysis revealed some of the genes relevant to the development of phage resistance in long-term co-evolution, with a special focus on the mucoid environment. Genes involved in the metabolism of carbohydrates were mutated in the presence of mucin. In conclusion, the use of mucolytic agents prior to the administration of lytic phages could be an interesting therapeutic option when addressing K. pneumoniae infections in environments where mucin is overproduced. Frontiers Media S.A. 2023-10-11 /pmc/articles/PMC10598653/ /pubmed/37886069 http://dx.doi.org/10.3389/fmicb.2023.1286046 Text en Copyright © 2023 Pacios, Blasco, Ortiz Cartagena, Bleriot, Fernández-García, López, Barrio-Pujante, Cuenca, Aracil, Oteo-Iglesias and Tomás. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Microbiology
Pacios, Olga
Blasco, Lucía
Ortiz Cartagena, Concha
Bleriot, Inés
Fernández-García, Laura
López, María
Barrio-Pujante, Antonio
Cuenca, Felipe Fernández
Aracil, Belén
Oteo-Iglesias, Jesús
Tomás, María
Molecular studies of phages-Klebsiella pneumoniae in mucoid environment: innovative use of mucolytic agents prior to the administration of lytic phages
title Molecular studies of phages-Klebsiella pneumoniae in mucoid environment: innovative use of mucolytic agents prior to the administration of lytic phages
title_full Molecular studies of phages-Klebsiella pneumoniae in mucoid environment: innovative use of mucolytic agents prior to the administration of lytic phages
title_fullStr Molecular studies of phages-Klebsiella pneumoniae in mucoid environment: innovative use of mucolytic agents prior to the administration of lytic phages
title_full_unstemmed Molecular studies of phages-Klebsiella pneumoniae in mucoid environment: innovative use of mucolytic agents prior to the administration of lytic phages
title_short Molecular studies of phages-Klebsiella pneumoniae in mucoid environment: innovative use of mucolytic agents prior to the administration of lytic phages
title_sort molecular studies of phages-klebsiella pneumoniae in mucoid environment: innovative use of mucolytic agents prior to the administration of lytic phages
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10598653/
https://www.ncbi.nlm.nih.gov/pubmed/37886069
http://dx.doi.org/10.3389/fmicb.2023.1286046
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