{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,6,24]],"date-time":"2026-06-24T05:59:04Z","timestamp":1782280744177,"version":"3.54.5"},"posted":{"date-parts":[[2025,9,16]]},"group-title":"Microbiology","reference-count":45,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,9,16]],"date-time":"2025-09-16T00:00:00Z","timestamp":1757980800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,9,16]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  <jats:italic>Pseudomonas aeruginosa<\/jats:italic>\n                  is an opportunistic human pathogen and a frequent cause of multidrug-resistant infections. This organism continues to evade antimicrobial therapy despite the clinical introduction of new anti-pseudomonal antibiotics over the past several years. One of these agents is cefiderocol (FDC), a novel siderophore-cephalosporin conjugate antibiotic that was designed to overcome both intrinsic and acquired \u03b2-lactam resistance mechanisms in\n                  <jats:italic>P. aeruginosa<\/jats:italic>\n                  . However, studies have demonstrated that inactivation of TonB-dependent receptors, most notably the catechol siderophore receptor\n                  <jats:italic>piuA<\/jats:italic>\n                  can substantially curtail the drug\u2019s ability to permeate the bacterial outer membrane, leading to rapid development of resistance. In this study, we examined the FDC resistance mechanisms of the laboratory strain PA14. We demonstrated that inactivation of the ferripyochelin receptor FptA was a first-step mutation towards FDC resistance. Through transposon mutagenesis, we identified several resistance pathways following\n                  <jats:italic>fptA<\/jats:italic>\n                  inactivation, such as the loss of an additional FDC import porin and overexpression of the MuxABC-OpmB multidrug efflux system. Introduction of clinically-identified mutations analogous to these transposon insertions in the absence of\n                  <jats:italic>fptA<\/jats:italic>\n                  conferred full FDC non-susceptibility while preserving the activity of other antipseudomonal \u03b2-lactam antibiotics. We also demonstrated that inactivation of\n                  <jats:italic>fptA<\/jats:italic>\n                  in a pyoverdine biosynthetic mutant disrupted bacterial iron homeostasis and conferred a fitness disadvantage. These FDC resistance mechanisms identified in PA14 highlight the long-term challenges of using FDC treatment for drug-resistant\n                  <jats:italic>P. aeruginosa<\/jats:italic>\n                  infections.\n                <\/jats:p>","DOI":"10.1101\/2025.09.16.676464","type":"posted-content","created":{"date-parts":[[2025,9,17]],"date-time":"2025-09-17T04:15:15Z","timestamp":1758082515000},"source":"Crossref","is-referenced-by-count":0,"title":["Loss of the Ferripyochelin Receptor FptA Drives Reduced Cefiderocol Susceptibility and Impairs Fitness in\n                  <i>Pseudomonas aeruginosa<\/i>\n                  PA14"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5314-0961","authenticated-orcid":false,"given":"Donghoon","family":"Kang","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1806-8260","authenticated-orcid":false,"given":"Rodrigo P.","family":"Baptista","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9054-9774","authenticated-orcid":false,"given":"Cesar A.","family":"Arias","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1518-2973","authenticated-orcid":false,"given":"William R.","family":"Miller","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2025091916001042000_2025.09.16.676464v1.1","first-page":"1","article-title":"Antimicrobial-resistant pathogens associated with adult healthcare-associated infections: Summary of data reported to the National Healthcare Safety Network, 2015-2017","volume":"41","year":"2020","journal-title":"Infect Control Hosp Epidemiol"},{"key":"2025091916001042000_2025.09.16.676464v1.2","doi-asserted-by":"publisher","DOI":"10.1093\/cid\/ciy378"},{"key":"2025091916001042000_2025.09.16.676464v1.3","doi-asserted-by":"publisher","DOI":"10.1007\/s10096-021-04308-0"},{"key":"2025091916001042000_2025.09.16.676464v1.4","doi-asserted-by":"crossref","first-page":"165","DOI":"10.1186\/s12866-023-02864-3","article-title":"Relebactam restores susceptibility of resistant Pseudomonas aeruginosa and Enterobacterales and enhances imipenem activity against chromosomal AmpC-producing species: analysis of global SMART 2018-2020","volume":"23","year":"2023","journal-title":"BMC Microbiol"},{"key":"2025091916001042000_2025.09.16.676464v1.5","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.01405-16"},{"key":"2025091916001042000_2025.09.16.676464v1.6","doi-asserted-by":"crossref","first-page":"e0100923","DOI":"10.1128\/aac.01009-23","article-title":"The emergence of cefiderocol resistance in Pseudomonas aeruginosa from a heteroresistant isolate during prolonged therapy","volume":"68","year":"2024","journal-title":"Antimicrob Agents Chemother"},{"key":"2025091916001042000_2025.09.16.676464v1.7","doi-asserted-by":"crossref","first-page":"dlaf129","DOI":"10.1093\/jacamr\/dlaf129","article-title":"Cefepime-zidebactam therapy for extensively drug-resistant Pseudomonas aeruginosa and Klebsiella pneumoniae infection as a bridge to liver transplantation","volume":"7","year":"2025","journal-title":"JAC Antimicrob Resist"},{"key":"2025091916001042000_2025.09.16.676464v1.8","doi-asserted-by":"crossref","first-page":"178","DOI":"10.1016\/j.cmi.2023.08.029","article-title":"Global prevalence of cefiderocol non-susceptibility in Enterobacterales, Pseudomonas aeruginosa, Acinetobacter baumannii, and Stenotrophomonas maltophilia: a systematic review and meta-analysis","volume":"30","year":"2024","journal-title":"Clin Microbiol Infect"},{"key":"2025091916001042000_2025.09.16.676464v1.9","doi-asserted-by":"crossref","unstructured":"Luscher A , Moynie L , Auguste PS , Bumann D , Mazza L , Pletzer D , Naismith JH , Kohler T . 2018. TonB-Dependent Receptor Repertoire of Pseudomonas aeruginosa for Uptake of Siderophore-Drug Conjugates. Antimicrob Agents Chemother 62.","DOI":"10.1128\/AAC.00097-18"},{"key":"2025091916001042000_2025.09.16.676464v1.10","doi-asserted-by":"crossref","first-page":"e0149822","DOI":"10.1128\/mbio.01498-22","article-title":"Plant-Derived Catechols Are Substrates of TonB-Dependent Transporters and Sensitize Pseudomonas aeruginosa to Siderophore-Drug Conjugates","volume":"13","year":"2022","journal-title":"mBio"},{"key":"2025091916001042000_2025.09.16.676464v1.11","doi-asserted-by":"publisher","DOI":"10.1128\/aac.00127-24"},{"key":"2025091916001042000_2025.09.16.676464v1.12","doi-asserted-by":"crossref","first-page":"e0092123","DOI":"10.1128\/aac.00921-23","article-title":"Identification of cpxS mutational resistome in Pseudomonas aeruginosa","volume":"67","year":"2023","journal-title":"Antimicrob Agents Chemother"},{"key":"2025091916001042000_2025.09.16.676464v1.13","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1005932"},{"key":"2025091916001042000_2025.09.16.676464v1.14","doi-asserted-by":"crossref","first-page":"631","DOI":"10.1038\/s41564-024-01601-4","article-title":"Siderophores promote cooperative interspecies and intraspecies cross-protection against antibiotics in vitro","volume":"9","year":"2024","journal-title":"Nat Microbiol"},{"key":"2025091916001042000_2025.09.16.676464v1.15","doi-asserted-by":"crossref","first-page":"1785","DOI":"10.1093\/jac\/dkad172","article-title":"In vitro dynamics and mechanisms of cefiderocol resistance development in wild-type, mutator and XDR Pseudomonas aeruginosa","volume":"78","year":"2023","journal-title":"J Antimicrob Chemother"},{"key":"2025091916001042000_2025.09.16.676464v1.16","doi-asserted-by":"crossref","first-page":"2591","DOI":"10.1093\/jac\/dkae263","article-title":"Impact of chromosomally encoded resistance mechanisms and transferable beta-lactamases on the activity of cefiderocol and innovative beta-lactam\/beta-lactamase inhibitor combinations against Pseudomonas aeruginosa","volume":"79","year":"2024","journal-title":"J Antimicrob Chemother"},{"key":"2025091916001042000_2025.09.16.676464v1.17","doi-asserted-by":"crossref","first-page":"dlad107","DOI":"10.1093\/jacamr\/dlad107","article-title":"In vitro activity of cefiderocol against Pseudomonas aeruginosa demonstrating evolved resistance to novel beta-lactam\/beta-lactamase inhibitors","volume":"5","year":"2023","journal-title":"JAC Antimicrob Resist"},{"key":"2025091916001042000_2025.09.16.676464v1.18","doi-asserted-by":"publisher","DOI":"10.1093\/ofid\/ofab311"},{"key":"2025091916001042000_2025.09.16.676464v1.19","doi-asserted-by":"publisher","DOI":"10.1021\/ic00104a059"},{"key":"2025091916001042000_2025.09.16.676464v1.20","doi-asserted-by":"publisher","DOI":"10.1099\/mic.0.2006\/002915-0"},{"key":"2025091916001042000_2025.09.16.676464v1.21","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00010-09"},{"key":"2025091916001042000_2025.09.16.676464v1.22","doi-asserted-by":"publisher","DOI":"10.1016\/0003-2697(87)90612-9"},{"key":"2025091916001042000_2025.09.16.676464v1.23","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0511100103"},{"key":"2025091916001042000_2025.09.16.676464v1.24","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1016\/j.jgar.2023.06.003","article-title":"Rapid development of cefiderocol resistance in a carbapenem-resistant Pseudomonas aeruginosa isolate associated with mutations in the pyoverdine biosynthesis pathway","volume":"34","year":"2023","journal-title":"J Glob Antimicrob Resist"},{"key":"2025091916001042000_2025.09.16.676464v1.25","doi-asserted-by":"crossref","unstructured":"Karakonstantis S , Rousaki M , Kritsotakis EI . 2022. Cefiderocol: Systematic Review of Mechanisms of Resistance, Heteroresistance and In Vivo Emergence of Resistance. 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Performance Standards for Antimicrobial Susceptibility Testing, 35th Edition. M-100., 34 ed. 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