SWATH-MS for discovery of early biomarkers of drug-induced cardiotoxicity using an animal doxorubicin model

SWATH-MS for discovery of early biomarkers of drug-induced cardiotoxicity using an animal doxorubicin model | 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 Research Article SWATH-MS for discovery of early biomarkers of drug-induced cardiotoxicity using an animal doxorubicin model Rohan Shotton, Amy Campbell, Ivona Baricevic-Jones, Rachel Reed, and 5 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7251224/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 10 You are reading this latest preprint version Abstract Background Clinical use of anthracyclines is limited by cumulative dose-dependent cardiotoxicity, though the mechanisms responsible for this are complex and incompletely understood. Existing biomarkers of anthracycline-related cardiotoxicity, such as troponin and NT-proBNP, lack sensitivity for the earliest stages of cardiotoxicity, therefore limiting scope to switch to alternative, safer treatment. As such, a reliable early biomarker of anthracycline-related cardiotoxicity remains a key unmet research need. Methods We investigated the rat myocardial proteome to identify novel biomarkers of anthracycline-related cardiotoxicity warranting further investigation for routine clinical application. Six groups of 8 Han-Wistar rats were dosed weekly for 1-7 doses with either doxorubicin 1.25mg/kg or saline control, followed by terminal bleeds. Sequential window acquisition of all theoretical mass spectra (SWATH-MS) was used to compare the rat myocardial proteome at serial timepoints and between doxorubicin- and saline-treated animals. A shortlist of candidate biomarkers was generated by identifying proteins with significant log 2 fold changes at early/mid timepoint dysregulation, sustained to later timepoints. Pathway analysis for human ortholog proteins was undertaken using the Uniprot database and STRING. Results Three proteins were differentially expressed at early, mid- and late timepoints (myosin light chain 3, desmin and cysteine-rich protein 2). A further 10 proteins were differentially expressed at mid- and late timepoints. Pathway analysis identified biological processes enriched within the myocardial proteome, predominantly related to muscle, metal ion, haemostasis, lipid and inflammation processes. A candidate biomarker shortlist of 34 proteins was derived for future study. Conclusion We used a rat anthracycline dosing schedule to recapitulate the development of cardiotoxicity observed in humans. We describe a shortlist of 34 proteins with promise as candidate biomarkers of the earliest stages of anthracycline-related cardiotoxicity and summarise pathway analysis of functionally enriched biological processes within the rat myocardial proteome during anthracycline treatment. Further study is underway to explore these shortlisted proteins in rat plasma, with a human study planned to validate best-performing biomarkers. Full Text Additional Declarations No competing interests reported. Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Revision requested 12 Sep, 2025 Reviews received at journal 22 Aug, 2025 Reviews received at journal 14 Aug, 2025 Reviewers agreed at journal 12 Aug, 2025 Reviewers agreed at journal 11 Aug, 2025 Reviewers agreed at journal 11 Aug, 2025 Reviewers invited by journal 11 Aug, 2025 Editor assigned by journal 07 Aug, 2025 Submission checks completed at journal 01 Aug, 2025 First submitted to journal 30 Jul, 2025 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-7251224","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":501564292,"identity":"c37a7497-2311-465d-84b8-f2691a0c2338","order_by":0,"name":"Rohan Shotton","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAtElEQVRIiWNgGAWjYFACHgjFD2MQp+VAAgODZANEiwTxWgwOEKvFvP/swccff9jkGd/uPfyBcYdNHUEtMjfykg0OJKQVm905lybBeCaNsC0SEjxmEgcSDiduu5FjxsDYdpgILfxnIFo2z8gx/sDY9p8ILQw5EC0bJHIMJBjbDhDjsBxjgzNpaYkzgA6TSGxLlmwgwmGGDypsbBL7QQ772GbHT9AWVJBAovpRMApGwSgYBTgAAB+EOfWyygnGAAAAAElFTkSuQmCC","orcid":"","institution":"University of Manchester","correspondingAuthor":true,"prefix":"","firstName":"Rohan","middleName":"","lastName":"Shotton","suffix":""},{"id":501564293,"identity":"eb4fd241-feed-4605-8812-ee7459d9ae5f","order_by":1,"name":"Amy Campbell","email":"","orcid":"","institution":"University of Manchester","correspondingAuthor":false,"prefix":"","firstName":"Amy","middleName":"","lastName":"Campbell","suffix":""},{"id":501564294,"identity":"7c39dd5a-1e99-4544-aa18-2ecb09dc4b0f","order_by":2,"name":"Ivona Baricevic-Jones","email":"","orcid":"","institution":"University of Manchester","correspondingAuthor":false,"prefix":"","firstName":"Ivona","middleName":"","lastName":"Baricevic-Jones","suffix":""},{"id":501564295,"identity":"6d4d8897-700a-4fce-8209-f85c14300303","order_by":3,"name":"Rachel Reed","email":"","orcid":"","institution":"University of Manchester","correspondingAuthor":false,"prefix":"","firstName":"Rachel","middleName":"","lastName":"Reed","suffix":""},{"id":501564296,"identity":"a07fafd5-5d9b-4fc4-9fed-27784c784dd6","order_by":4,"name":"Janet Kelsall","email":"","orcid":"","institution":"University of Manchester","correspondingAuthor":false,"prefix":"","firstName":"Janet","middleName":"","lastName":"Kelsall","suffix":""},{"id":501564297,"identity":"2def18d4-0686-4706-909a-341d3f9e53d6","order_by":5,"name":"Joanna Williams","email":"","orcid":"","institution":"University of Manchester","correspondingAuthor":false,"prefix":"","firstName":"Joanna","middleName":"","lastName":"Williams","suffix":""},{"id":501564298,"identity":"0059eb13-a319-4e49-8815-cf6d1ff40a4b","order_by":6,"name":"Richard D. 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