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...
| Authors: | ; ; ; ; ; ; ; ; ; ; |
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| Format: | Article |
| Publication date: | 2023 |
| Country: | España |
| Institution: | Servizo Galego de Saúde (SERGAS) |
| Repository: | RUNA. Repositorio da Consellería de Sanidade e Sergas |
| OAI Identifier: | oai:runa.sergas.gal:20.500.11940/21747 |
| Online Access: | https://portalcientifico.sergas.gal//documentos/6546ecdf32348009d228e6e6 http://hdl.handle.net/20.500.11940/21747 |
| Access Level: | Open access |
| Keyword: | INIBIC AS A Coruña CHUAC |
| _version_ | 1878741264615079936 |
|---|---|
| author | Pacios Santamaría, Olga |
| 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 |
| author_browse | Aracil, B. Barrio Pujante, Antonio Blasco Otero, Lucía Bleriot Rial, Ines Maria Cuenca, F.F. Fernández García, Laura López, M. Ortiz Cartagena, Concha Oteo-Iglesias, J. Pacios Santamaría, Olga Tomás Carmona, María del Mar |
| 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 |
| collection | RUNA. Repositorio da Consellería de Sanidade e Sergas |
| 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. |
| eu_rights_str_mv | openAccess |
| format | article |
| id | ES_e21b8e5a7d55c8fc3976fcd8a58ccc4b |
| institution | Servizo Galego de Saúde (SERGAS) |
| instname_str | Servizo Galego de Saúde (SERGAS) |
| network_acronym_str | ES |
| network_name_str | España |
| oai_identifier_str | oai:runa.sergas.gal:20.500.11940/21747 |
| publishDate | 2023 |
| publishDateSort | 2023 |
| reponame_str | RUNA. Repositorio da Consellería de Sanidade e Sergas |
| spelling | Molecular studies of phages-Klebsiella pneumoniae in mucoid environment: innovative use of mucolytic agents prior to the administration of lytic phages 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 INIBIC AS A Coruña CHUAC 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. 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. https://portalcientifico.sergas.gal//documentos/6546ecdf32348009d228e6e6 http://hdl.handle.net/20.500.11940/21747 |
| 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 |
| 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 | INIBIC AS A Coruña CHUAC |
| topic_facet | INIBIC AS A Coruña CHUAC |
| url | https://portalcientifico.sergas.gal//documentos/6546ecdf32348009d228e6e6 http://hdl.handle.net/20.500.11940/21747 |