In vivo evolution of resistance to novel β-lactam/β-lactamase inhibitor combinations through overproduction of the horizontally acquired extended-spectrum AmpC β-lactamase FOX-14 and porin disruption in Serratia marcescens
| UDC.coleccion | Investigación | |
| UDC.departamento | Fisioterapia, Medicina e Ciencias Biomédicas | |
| UDC.endPage | 640 | |
| UDC.grupoInv | Investigación en Microbiología (INIBIC) | |
| UDC.institutoCentro | INIBIC - Instituto de Investigacións Biomédicas de A Coruña | |
| UDC.issue | 3 | |
| UDC.journalTitle | European Journal of Clinical Microbiology and Infectious Diseases | |
| UDC.startPage | 631 | |
| UDC.volume | 45 | |
| dc.contributor.author | Rodríguez-Pallares, Salud | |
| dc.contributor.author | González-Pinto, Lucía | |
| dc.contributor.author | Tarriño-León, María | |
| dc.contributor.author | Aja-Macaya, Pablo | |
| dc.contributor.author | Sánchez-Peña, Lucía | |
| dc.contributor.author | Moscoso, Miriam | |
| dc.contributor.author | Pérez-Rodríguez, Gloria | |
| dc.contributor.author | Blanco Martín, Tania | |
| dc.contributor.author | Beceiro Casas, Alejandro | |
| dc.contributor.author | Bou, Germán | |
| dc.contributor.author | Arca-Suárez, Jorge | |
| dc.date.accessioned | 2026-03-17T12:44:39Z | |
| dc.date.available | 2026-03-17T12:44:39Z | |
| dc.date.issued | 2025-11-11 | |
| dc.description.abstract | [Abstract] We investigated the genetic and biochemical determinants that reduce the activity of several β-lactam/β-lactamase inhibitor combinations against Serratia marcescens clinical isolates after carbapenem exposure. We analyzed paired clinical S. marcescens isolates recovered before and after treatment of a prolonged infection with ertapenem. We tested the susceptibility of the isolates to ceftazidime/avibactam, aztreonam/avibactam, imipenem/relebactam and meropenem/vaborbactam. Mechanisms of resistance were investigated using whole-genome sequencing and RNA expression analysis. The blaFOX-14 gene was cloned into Escherichia coli, to assess the phenotype in wild-type and low-permeability backgrounds. FOX-14 was purified to determine steady-state kinetics and avibactam IC50. The baseline isolate was susceptible to ceftazidime/avibactam, aztreonam/avibactam, imipenem/relebactam and meropenem/vaborbactam, whereas the post-therapy isolate displayed high-level resistance. The post-therapy isolate harboured a premature stop in ompF, loss of an additional putative porin, and overexpressed of blaFOX-14 due to triplication of the blaFOX-14 surrounding region. Cloning of blaFOX-14 conferred an extended-spectrum AmpC phenotype in E. coli, which was further accentuated under reduced permeability. Complementation of the post-therapy resistant clinical isolate with functional OmpF restored susceptibility to new β-lactam/β-lactamase inhibitor combinations. Purified FOX-14 displayed marked cephalosporinase activity against ceftazidime and cefepime, with low Km values, while negligible turnover for aztreonam and ertapenem and a low avibactam IC50. In S. marcescens, in vivo resistance in clinical isolates emerged from FOX-14 overproduction combined with reduced outer-membrane permeability, predominantly due to OmpF disruption. These findings elucidate resistance to newer β-lactam/β-lactamase inhibitor combinations and warrant close monitoring of antimicrobial activity during carbapenem-based therapy for S. marcescens infections. | |
| dc.description.sponsorship | This work was supported by the Instituto de Salud Carlos III (ISCIII projects: PI24/00920, PI23/00851, PI22/01212, PI21/00704 and PI20/01212) and co-funded by the European Union. The research was also funded by Centro de Investigación Biomédica en Red de Enfermedades Infecciosas (CIBERINFEC; CB21/13/00055), integrated in the National Plan for Scientific Research, Development and Technological Innovation 2013–2016, and funded by the ISCIII-General Subdirection of Assessment and Promotion of the Research-European Regional Development Fund (FEDER) “A way of making Europe.” The study was also funded by Axencia Galega de Innovacion (GAIN), Conselleria de Innovacion, Conselleria de Emprego e Industria through “Proxectos de excelencia” (IN607D 2021/12 to A.B. and IN607D 2024/008 to J.A.-S.), and ‘’Axudas para a consolidación de grupos de investigación que pola súa produción científica e a súa actividade de I + D constitúan unha referencia no Sistema galego de I + D + I’’ (IN607A 2024/09 to G.B.). S.R.-P. was financially supported by the Río Hortega Program (ISCIII, CM23/00104). L.G.-P. was financially supported by the PFIS Program (ISCIII, FI23/00074). L.S.-P. was financially supported by Fundación Publica Galega de investigación Biomédica (INIBIC) and Xunta de Galicia (IN606A 2024/022). M.-M. was financially supported by CIBERINFEC. G.P.-R. was financially supported by the ISCIII Project PI22/01212. T.B.-M. was financially supported by the Río Hortega Program (ISCIII, CM23/00095). J.A.-S. was financially supported by the Juan Rodés Program (ISCIII, JR21/00026). | |
| dc.identifier.citation | Rodríguez-Pallares, S., González-Pinto, L., Tarriño-León, M., Aja-Macaya, P., Sánchez-Peña, L., Moscoso, M., Pérez-Rodríguez, G., Blanco-Martín, T., Beceiro, A., Bou, G., & Arca-Suárez, J. (2026). In vivo evolution of resistance to novel β-lactam/β-lactamase inhibitor combinations through overproduction of the horizontally acquired extended-spectrum AmpC β-lactamase FOX-14 and porin disruption in Serratia marcescens. European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology, 45(3), 631–640. | |
| dc.identifier.doi | 10.1007/S10096-025-05352-W | |
| dc.identifier.issn | 1435-4373 | |
| dc.identifier.uri | https://hdl.handle.net/2183/47742 | |
| dc.language.iso | eng | |
| dc.publisher | Springer Nature | |
| dc.relation.projectID | info:eu-repo/grantAgreement/ISCIII/Plan Estatal de Investigación Científica, Técnica y de Innovación 2021-2023/PI22%2F01212/ES/Inhibidores de carbapenemasas: actividad frente a Enterobacterales productores de carbapenemasas, mecanismos e impacto en la evolución de la resistencia antimicrobiana (PROTECT)/ | |
| dc.relation.projectID | Xunta de Galicia; IN607D 2021/12 | |
| dc.relation.projectID | Xunta de Galicia; IN607D 2024/008 | |
| dc.relation.uri | https://doi.org/10.1007/S10096-025-05352-W | |
| dc.rights | This version of the article has been accepted for publication, after peer review and is subject to Springer Nature’s AM terms of use, but is not the Version of Record and does not reflect post-acceptance improvements, or any corrections. The Version of Record is available online at Springer Link. | |
| dc.rights.accessRights | embargoed access | |
| dc.subject | Anti-Bacterial Agents | |
| dc.subject | Bacterial Proteins | |
| dc.subject | Porins | |
| dc.subject | Serratia marcescens | |
| dc.subject | beta-Lactamase Inhibitors | |
| dc.subject | beta-Lactams | |
| dc.title | In vivo evolution of resistance to novel β-lactam/β-lactamase inhibitor combinations through overproduction of the horizontally acquired extended-spectrum AmpC β-lactamase FOX-14 and porin disruption in Serratia marcescens | |
| dc.type | journal article | |
| dc.type.hasVersion | AM | |
| dspace.entity.type | Publication | |
| relation.isAuthorOfPublication | 909e08d1-6ed1-4b99-9e9e-c64eb72e7dea | |
| relation.isAuthorOfPublication.latestForDiscovery | 909e08d1-6ed1-4b99-9e9e-c64eb72e7dea |