{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T17:14:36Z","timestamp":1788455676094,"version":"build-2803163510"},"reference-count":48,"publisher":"Wiley","issue":"18","license":[{"start":{"date-parts":[[2025,5,29]],"date-time":"2025-05-29T00:00:00Z","timestamp":1748476800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001646","name":"Robert Bosch Stiftung","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100001646","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["bpspubs.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["British J Pharmacology"],"published-print":{"date-parts":[[2025,9]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background and purpose<\/jats:title><jats:p>Thiopurines are used in paediatric inflammatory bowel disease (IBD), but some patients do not respond. Because the gut microbiota influences drug efficacy and IBD\u2010patient microbiota presents increased bacterial abundance, we investigated the impact of candidate Enterobacteriaceae on drug cytotoxicity, metabolism and efficacy.<\/jats:p><\/jats:sec><jats:sec><jats:title>Experimental approach<\/jats:title><jats:p>Thiopurines were exposed in vitro to bacteria for 4\u2009h at 37\u00b0C and drug concentrations measured by UV spectrophotometry. Cytotoxic effects and drug metabolite concentrations on NALM6 and JURKAT cells were determined after treatment with thiopurines exposed or not to bacteria. Drugs were measured in <jats:styled-content style=\"fixed-case\"><jats:italic>Klebsiella pneumoniae<\/jats:italic><\/jats:styled-content> lysates and bacterial conditioned media were used for metabolomic analyses. Shotgun metagenomic sequencing was performed on eight IBD\u2010patient faecal stools.<\/jats:p><\/jats:sec><jats:sec><jats:title>Key results<\/jats:title><jats:p>Incubation of thiopurines with <jats:styled-content style=\"fixed-case\"><jats:italic>K. pneumoniae<\/jats:italic><\/jats:styled-content>, but not <jats:italic>Escherichia <jats:styled-content style=\"fixed-case\">coli<\/jats:styled-content><\/jats:italic> and <jats:styled-content style=\"fixed-case\"><jats:italic>Salmonella enterica<\/jats:italic><\/jats:styled-content>, reduced thiopurine concentrations and cytotoxicity on NALM6 and JURKAT cells. Thiopurine metabolites were lower in cells treated with drugs previously exposed to <jats:styled-content style=\"fixed-case\"><jats:italic>K. pneumoniae<\/jats:italic><\/jats:styled-content>. Internalisation of drugs was demonstrated by their detection in lysates after bacterial incubation. Untargeted metabolomics revealed biotransformation of thiopurines by <jats:italic><jats:styled-content style=\"fixed-case\">K. pneumoniae<\/jats:styled-content>,<\/jats:italic> as reactions of deconjugation, reduction, glycosylation, acetylation or conjugation with propionic acid. Incubation with thiopurines led to changes in the secretion of endogenous bacterial metabolites. <jats:styled-content style=\"fixed-case\"><jats:italic>K. pneumoniae<\/jats:italic><\/jats:styled-content> faecal abundance was associated with lower thiopurine metabolite concentrations in erythrocytes of paediatric IBD\u2010patients.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions and Implications<\/jats:title><jats:p><jats:styled-content style=\"fixed-case\"><jats:italic>K. pneumoniae<\/jats:italic><\/jats:styled-content> decreases the cytotoxicity of thiopurines through internalisation of MP and TG. We revealed potential bacterial drug biotransformation, as well as negative correlations between bacterial abundance and drug metabolites.<\/jats:p><\/jats:sec>","DOI":"10.1111\/bph.70089","type":"journal-article","created":{"date-parts":[[2025,5,29]],"date-time":"2025-05-29T03:49:03Z","timestamp":1748490543000},"page":"4281-4298","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["<scp><i>Klebsiella pneumoniae<\/i><\/scp> contributes to altered cytotoxicity of thiopurines in vitro: Possible implications of biotransformation and bacterial metabolism"],"prefix":"10.1111","volume":"182","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1304-6190","authenticated-orcid":false,"given":"Martina","family":"Franzin","sequence":"first","affiliation":[{"name":"Department of Advanced Translational Diagnostics Institute for Maternal and Child Health, IRCCS \u2018Burlo Garofolo\u2019  Trieste 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