Eradication of extensively drug resistant Pseudomonas aeruginosa causing ventilator-associated pneumonia in acute lymphoblastic leukemia patient using phage therapy

Eradication of extensively drug resistant Pseudomonas aeruginosa causing ventilator-associated pneumonia in acute lymphoblastic leukemia patient using phage therapy | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Eradication of extensively drug resistant Pseudomonas aeruginosa causing ventilator-associated pneumonia in acute lymphoblastic leukemia patient using phage therapy Gregorio Iraola, Josefina Puig, Fabio Grill, Mateo Gómez, Ignacio Ferrés, and 6 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7802317/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted You are reading this latest preprint version Abstract Ventilator-associated pneumonia (VAP) are among the most frequent hospital-acquired infections, representing a significant morbidity and mortality risk. Increasing antibiotic resistance acquired by major bacterial pathogens associated with VAP, especially Pseudomonas aeruginosa, is rapidly narrowing therapeutic options with standard-of-care antibiotics. In response to the urgent need for alternative therapeutic approaches, phage therapy has recently reemerged as an efficacious way to combat resistant infections. Here, we report the case of a 17-year-old patient diagnosed with acute lymphoblastic leukemia who suffered from persistent VAP and sepsis caused by extensively drug-resistant P. aeruginosa for over 6 months, which originated after decompressive craniectomy due to hemorrhagic stroke. The patient received a 12-day nebulized treatment with a single phage belonging to a new Pakpunavirus species, in combination with intravenous antibiotics. This therapy was well tolerated and was associated with clinical improvement and microbiological eradication of P. aeruginosa, eventually allowing the discharge of the patient from the intensive care unit and from hospital. In vitro testing of the selected phage showed a broad-host range and anti-biofilm activity in a panel of unrelated clinical P. aeruginosa strains. Together, our results support the use of individually-tailored phage preparations and its potential scalability and transition to semi-personalized medicines using broader host-range phages, as a promising alternative to combat the antimicrobial resistance pandemic. Biological sciences/Microbiology/Bacteriophages Health sciences/Health care/Therapeutics/Biological therapy Figures Figure 1 Figure 2 Figure 3 Figure 4 Full Text Additional Declarations Yes there is potential Competing Interest. J.P. and G.I. are cofounders and shareholders of Kinzbio. M.G., I.F., E.C., M.V.G., and A.G. are current employees at Kinzbio. The remaining authors declare no competing interests. Patient Consent: Written informed consent was obtained from the parents of the patient for the publication of this clinical case report and any accompanying data. Supplementary Files SupplementaryInformation.docx Supplementary Figure S1 SupplementaryTableS21.xlsx Supplementary Table S2 SupplementaryTableS11.xlsx Supplementary Table S1 SupplementaryTableS31.xlsx Supplementary Table S3 Cite Share Download PDF Status: Under Review Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-7802317","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":530197881,"identity":"1e126dd1-3d1d-4914-9f14-85cba34c4241","order_by":0,"name":"Gregorio 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20:00:30","extension":"xlsx","order_by":9,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":9319,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementaryTableS31.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-7802317/v1/bce5f99e9403f21d4efe3444.xlsx"},{"id":93968586,"identity":"5a04ca9f-ff63-4c75-a177-bcc1166c01d4","added_by":"auto","created_at":"2025-10-20 20:00:29","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":197577,"visible":true,"origin":"","legend":"\u003cp\u003eTimeline of main clinical developments. The figure summarizes the main clinical, diagnostic, microbiologic and therapeutic findings and events from the patient’s first hospitalization until the end of phage therapy intervention.\u003c/p\u003e","description":"","filename":"Figure1.png","url":"https://assets-eu.researchsquare.com/files/rs-7802317/v1/dc3239fe2f2bc3677371d939.png"},{"id":93968590,"identity":"c4f5a07d-ee1c-41ff-b2e5-c0d4748300d3","added_by":"auto","created_at":"2025-10-20 20:00:29","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":359813,"visible":true,"origin":"","legend":"\u003cp\u003ePhage susceptibility testing of patient’s derived strains. (A) Six phages showed potential activity against the patient's isolates based on the complete clearance of P. aeruginosa lawns using spot testing. (B) The six phages were tested in liquid cultures using multiplicity of infection (MOI) of 1 and 10; phages FPs17 and FPs21 showed the best inhibitory capacity. (C) These two phages were assayed and compared with each other using additional MOIs (0.1, 1 and 10); FPs21 was selected as a therapeutic candidate showing complete bacterial suppression for up to 11 hours.\u003c/p\u003e","description":"","filename":"Figure2.png","url":"https://assets-eu.researchsquare.com/files/rs-7802317/v1/c57473c658c08958e6999954.png"},{"id":93968891,"identity":"2e34251a-4b58-4131-a55a-aca6bc1cd9da","added_by":"auto","created_at":"2025-10-20 20:08:29","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":928596,"visible":true,"origin":"","legend":"\u003cp\u003eComparative phylogenomics of patient’s derived P. aeruginosa strains. (A) Whole-genome phylogeny showing the placement of patient’s derived strains in the context of a global collection of 571 P. aeruginosa genomes. Tree tips are colored according to the presence of the ExoS/ExoU virulence determinants. (B) Zoom in patient’s strains' phylogenetic vicinity showing their genetic relatedness with the epidemic ST395 clone. The rightmost panel shows the presence/absence of antimicrobial resistance for different antibiotic families for these strains.\u003c/p\u003e","description":"","filename":"Figure3.png","url":"https://assets-eu.researchsquare.com/files/rs-7802317/v1/6f3a524af45efdbcba871aac.png"},{"id":93968897,"identity":"43d1741b-70cf-4e38-80d7-aef2694547d5","added_by":"auto","created_at":"2025-10-20 20:08:30","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":189227,"visible":true,"origin":"","legend":"\u003cp\u003eFigure 4. Biofilm disruption assays. The therapeutic phage FPs21 was tested for its capacity to disrupt biofilms formed by patient strain 387 and unrelated clinical P. aeruginosa strains. In all cases, a significant reduction was observed (A) with an average reduction of biofilm mass of about 70% (B). Statistical significance is marked by asterisks indicating p \u0026lt; 0.05 (Wilcoxon test).\u003c/p\u003e","description":"","filename":"Figure4.png","url":"https://assets-eu.researchsquare.com/files/rs-7802317/v1/00f491268a97c38915d9938c.png"},{"id":93969372,"identity":"bcc43917-c84d-471c-9ee5-3473d94d4238","added_by":"auto","created_at":"2025-10-20 20:24:34","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1121275,"visible":true,"origin":"","legend":"Article File","description":"","filename":"paperpakpunavirussubmit.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7802317/v1_covered_accebb1e-c7fc-4d34-bd59-460b0fd8897e.pdf"},{"id":93968587,"identity":"16e0e8c0-7043-4a40-a783-7f4771ecac35","added_by":"auto","created_at":"2025-10-20 20:00:29","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":835458,"visible":true,"origin":"","legend":"\u003cp\u003eSupplementary Figure S1\u003c/p\u003e","description":"","filename":"SupplementaryInformation.docx","url":"https://assets-eu.researchsquare.com/files/rs-7802317/v1/62bde92a84f0cd7938968e8b.docx"},{"id":93968890,"identity":"c283eaa1-f4c6-46ff-95e7-6cdb75478ba3","added_by":"auto","created_at":"2025-10-20 20:08:29","extension":"xlsx","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":10577,"visible":true,"origin":"","legend":"\u003cp\u003eSupplementary Table S2\u003c/p\u003e","description":"","filename":"SupplementaryTableS21.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-7802317/v1/c70c179b3f2228e0df412e21.xlsx"},{"id":93968892,"identity":"fca74365-34bc-46bd-8a7d-13bc96bf7a90","added_by":"auto","created_at":"2025-10-20 20:08:29","extension":"xlsx","order_by":3,"title":"","display":"","copyAsset":false,"role":"supplement","size":6625,"visible":true,"origin":"","legend":"\u003cp\u003eSupplementary Table S1\u003c/p\u003e","description":"","filename":"SupplementaryTableS11.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-7802317/v1/a011f42b1b773a5d4a225708.xlsx"},{"id":93968894,"identity":"b23e0037-9c19-447a-808b-e8951dc82ab4","added_by":"auto","created_at":"2025-10-20 20:08:29","extension":"xlsx","order_by":4,"title":"","display":"","copyAsset":false,"role":"supplement","size":9319,"visible":true,"origin":"","legend":"Supplementary Table S3","description":"","filename":"SupplementaryTableS31.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-7802317/v1/c30d1f77e8afe057d360660e.xlsx"}],"financialInterests":"\u003cp\u003e\u003cstrong\u003eYes\u003c/strong\u003e there is potential Competing Interest. J.P. and G.I. are cofounders and shareholders of Kinzbio. M.G., I.F., E.C., M.V.G., and A.G. are current employees at Kinzbio. The remaining authors declare no competing interests.\u003c/p\u003e\n\u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n\u003cp\u003ePatient Consent: Written informed consent was obtained from the parents of the patient for the publication of this clinical case report and any accompanying data.\u003c/p\u003e","formattedTitle":"Eradication of extensively drug resistant Pseudomonas aeruginosa causing ventilator-associated pneumonia in acute lymphoblastic leukemia patient using phage therapy","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"nature-portfolio","isNatureJournal":true,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"","title":"Nature Portfolio","twitterHandle":"","acdcEnabled":false,"dfaEnabled":false,"editorialSystem":"ejp","reportingPortfolio":"","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"","lastPublishedDoi":"10.21203/rs.3.rs-7802317/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7802317/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Ventilator-associated pneumonia (VAP) are among the most frequent hospital-acquired infections, representing a significant morbidity and mortality risk. Increasing antibiotic resistance acquired by major bacterial pathogens associated with VAP, especially Pseudomonas aeruginosa, is rapidly narrowing therapeutic options with standard-of-care antibiotics. In response to the urgent need for alternative therapeutic approaches, phage therapy has recently reemerged as an efficacious way to combat resistant infections. Here, we report the case of a 17-year-old patient diagnosed with acute lymphoblastic leukemia who suffered from persistent VAP and sepsis caused by extensively drug-resistant P. aeruginosa for over 6 months, which originated after decompressive craniectomy due to hemorrhagic stroke. The patient received a 12-day nebulized treatment with a single phage belonging to a new Pakpunavirus species, in combination with intravenous antibiotics. This therapy was well tolerated and was associated with clinical improvement and microbiological eradication of P. aeruginosa, eventually allowing the discharge of the patient from the intensive care unit and from hospital. In vitro testing of the selected phage showed a broad-host range and anti-biofilm activity in a panel of unrelated clinical P. aeruginosa strains. Together, our results support the use of individually-tailored phage preparations and its potential scalability and transition to semi-personalized medicines using broader host-range phages, as a promising alternative to combat the antimicrobial resistance pandemic.","manuscriptTitle":"Eradication of extensively drug resistant Pseudomonas aeruginosa causing ventilator-associated pneumonia in acute lymphoblastic leukemia patient using phage therapy","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-10-20 20:00:25","doi":"10.21203/rs.3.rs-7802317/v1","editorialEvents":[],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"nature-communications","isNatureJournal":true,"hasQc":false,"allowDirectSubmit":false,"externalIdentity":"NCOMMS","sideBox":"Learn more about [Nature Communications](http://www.nature.com/ncomms/)","snPcode":"","submissionUrl":"https://mts-ncomms.nature.com/","title":"Nature Communications","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"ejp","reportingPortfolio":"Nature Communications","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"5c2f0a89-41bb-4d28-8a71-dfe39d577807","owner":[],"postedDate":"October 20th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[{"id":56361771,"name":"Biological sciences/Microbiology/Bacteriophages"},{"id":56361772,"name":"Health sciences/Health care/Therapeutics/Biological therapy"}],"tags":[],"updatedAt":"2026-02-23T16:16:38+00:00","versionOfRecord":[],"versionCreatedAt":"2025-10-20 20:00:25","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7802317","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7802317","identity":"rs-7802317","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}