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: Pacios Santamaría, Olga, Blasco Otero, Lucía, Ortiz Cartagena, Concha, Bleriot Rial, Ines Maria, Fernández García, Laura, López, M., Barrio Pujante, Antonio, Cuenca, F.F., Aracil, B., Oteo-Iglesias, J., Tomás Carmona, María del Mar
Tipo de recurso: artículo
Fecha de publicación:2023
País:España
Institución:Servizo Galego de Saúde (SERGAS)
Repositorio:RUNA. Repositorio da Consellería de Sanidade e Sergas
OAI Identifier:oai:runa.sergas.gal:20.500.11940/21747
Acceso en línea:https://portalcientifico.sergas.gal//documentos/6546ecdf32348009d228e6e6
http://hdl.handle.net/20.500.11940/21747
Access Level:acceso abierto
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spelling Molecular studies of phages-Klebsiella pneumoniae in mucoid environment: innovative use of mucolytic agents prior to the administration of lytic phagesPacios Santamaría, OlgaBlasco Otero, LucíaOrtiz Cartagena, ConchaBleriot Rial, Ines MariaFernández García, LauraLópez, M.Barrio Pujante, AntonioCuenca, F.F.Aracil, B.Oteo-Iglesias, J.Tomás Carmona, María del MarINIBICAS A CoruñaCHUACAS A CoruñaCHUACAS A CoruñaINIBICAS A CoruñaCHUACINIBICAS A CoruñaCHUACMucins 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.The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This study has been funded by Instituto de Salud Carlos III (ISCIII) through the projects PI19/00878 and PI22/00323 and co-funded by the European Union, and by the Study Group on Mechanisms of Action and Resistance to Antimicrobials, GEMARA (SEIMC). (SEIMC, http://www.seimc.org/). This research was also supported by CIBERINFEC (CIBER21/13/00095) and by Personalized and precision medicine grant from the Instituto de Salud Carlos III (MePRAM Project, PMP22/00092). MT was financially supported by the Miguel Servet Research Programme (SERGAS and ISCIII). OP, LF-G, and ML were financially supported by the grants IN606A-2020/035, IN606B-2021/013, and IN606C-2022/002, respectively (GAIN, Xunta de Galicia). IB was financially supported by the pFIS program (ISCIII, FI20/00302). Finally, to thank to PIRASOA laboratory which is the reference laboratory for molecular typing of nosocomial pathogens and detection of mechanisms of resistance to antimicrobials of health interest in Andalusia, Virgen Macarena Hospital, Seville, to send us the clinical isolates. Thanks to Alvaro Pascual and Luis Martinez-Martinez from Virgen Macarena Hospital, Seville and Reina Sofia Hospital, Cordoba.2023info:eu-repo/semantics/articlehttps://portalcientifico.sergas.gal//documentos/6546ecdf32348009d228e6e6http://hdl.handle.net/20.500.11940/21747reponame:RUNA. Repositorio da Consellería de Sanidade e Sergasinstname:Servizo Galego de Saúde (SERGAS)Ingléshttp://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccessoai:runa.sergas.gal:20.500.11940/217472026-06-12T08:40:47Z
dc.title.none.fl_str_mv 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
spellingShingle Molecular studies of phages-Klebsiella pneumoniae in mucoid environment: innovative use of mucolytic agents prior to the administration of lytic phages
Pacios Santamaría, Olga
INIBIC
AS A Coruña
CHUAC
AS A Coruña
CHUAC
AS A Coruña
INIBIC
AS A Coruña
CHUAC
INIBIC
AS A Coruña
CHUAC
title_short 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_sort Molecular studies of phages-Klebsiella pneumoniae in mucoid environment: innovative use of mucolytic agents prior to the administration of lytic phages
dc.creator.none.fl_str_mv Pacios Santamaría, Olga
Blasco Otero, Lucía
Ortiz Cartagena, Concha
Bleriot Rial, Ines Maria
Fernández García, Laura
López, M.
Barrio Pujante, Antonio
Cuenca, F.F.
Aracil, B.
Oteo-Iglesias, J.
Tomás Carmona, María del Mar
author Pacios Santamaría, Olga
author_facet Pacios Santamaría, Olga
Blasco Otero, Lucía
Ortiz Cartagena, Concha
Bleriot Rial, Ines Maria
Fernández García, Laura
López, M.
Barrio Pujante, Antonio
Cuenca, F.F.
Aracil, B.
Oteo-Iglesias, J.
Tomás Carmona, María del Mar
author_role author
author2 Blasco Otero, Lucía
Ortiz Cartagena, Concha
Bleriot Rial, Ines Maria
Fernández García, Laura
López, M.
Barrio Pujante, Antonio
Cuenca, F.F.
Aracil, B.
Oteo-Iglesias, J.
Tomás Carmona, María del Mar
author2_role author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv INIBIC
AS A Coruña
CHUAC
AS A Coruña
CHUAC
AS A Coruña
INIBIC
AS A Coruña
CHUAC
INIBIC
AS A Coruña
CHUAC
topic INIBIC
AS A Coruña
CHUAC
AS A Coruña
CHUAC
AS A Coruña
INIBIC
AS A Coruña
CHUAC
INIBIC
AS A Coruña
CHUAC
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.
publishDate 2023
dc.date.none.fl_str_mv 2023
dc.type.none.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv https://portalcientifico.sergas.gal//documentos/6546ecdf32348009d228e6e6
http://hdl.handle.net/20.500.11940/21747
url https://portalcientifico.sergas.gal//documentos/6546ecdf32348009d228e6e6
http://hdl.handle.net/20.500.11940/21747
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.rights.none.fl_str_mv http://creativecommons.org/licenses/by/4.0/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by/4.0/
eu_rights_str_mv openAccess
dc.source.none.fl_str_mv reponame:RUNA. Repositorio da Consellería de Sanidade e Sergas
instname:Servizo Galego de Saúde (SERGAS)
instname_str Servizo Galego de Saúde (SERGAS)
reponame_str RUNA. Repositorio da Consellería de Sanidade e Sergas
collection RUNA. Repositorio da Consellería de Sanidade e Sergas
repository.name.fl_str_mv
repository.mail.fl_str_mv
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