Endoplasmic Reticulum-Mitochondria Contacts are apical hotspots of lipid peroxidation driving ferroptosis

Endoplasmic Reticulum-Mitochondria Contacts are apical hotspots of lipid peroxidation driving ferroptosis | 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 Endoplasmic Reticulum-Mitochondria Contacts are apical hotspots of lipid peroxidation driving ferroptosis Maria Livia Sassano, Yulia (Y) Tyurina, Antigoni Diometzidou, and 10 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4311835/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 13 Jun, 2025 Read the published version in Nature Cell Biology → Version 1 posted You are reading this latest preprint version Abstract Peroxidation of membrane phospholipids (PLs) is a hallmark of ferroptosis. ER and mitochondria have been implicated in ferroptosis, but whether intracellular PL-peroxidation ensues at their contact sites (EMCSs) is unknown. Using super-resolution live imaging we charted the immediate spatiotemporal events triggered by ferroptosis at the inter-organelle level. EMCSs expand minutes after the formation of localized and activated lipid-peroxides that secondary spread to mitochondria, promoting mitochondrial ROS accumulation and fission. Oxidative lipidomics unravels that EMCSs host distinct pro-ferroptotic polyunsaturated (PUFA)-PLs, including doubly PUFA-acylated PLs, demonstrating the persuasive propensity of EMCS to PL-peroxidation. Genetic disruption of EMCSs blunts PL-peroxidation and ferroptosis, while EMCS stabilization enhances them. Analysis of human triple-negative breast cancer (TNBC) data shows that expression of EMCShigh-gene signature associates with the ferroptosis-susceptible TNBC subtype harboring heightened levels of oxidized-PLs. Our data reveal EMCSs as apical orchestrators of lethal lipid-peroxidation production and suggest that empowering EMCSs can promote ferroptosis in vulnerable cancer cells. Biological sciences/Cell biology/Cell death Biological sciences/Cell biology/Organelles/Mitochondria Biological sciences/Cell biology/Membrane trafficking/Endoplasmic reticulum Membrane contact sites ferroptosis lipid peroxidation endoplasmic reticulum mitochondria Full Text Additional Declarations There is NO Competing Interest. Table 1 is available in the Supplementary Files section. Supplementary Files TableWORD.pdf Table1 Cite Share Download PDF Status: Published Journal Publication published 13 Jun, 2025 Read the published version in Nature Cell Biology → 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. 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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-4311835","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":294636638,"identity":"ac422dd9-b746-4786-9fbe-16395ea3fc76","order_by":0,"name":"Maria Livia Sassano","email":"","orcid":"https://orcid.org/0000-0002-2262-6915","institution":"KU, Leuven","correspondingAuthor":false,"prefix":"","firstName":"Maria","middleName":"Livia","lastName":"Sassano","suffix":""},{"id":294636639,"identity":"72f68130-2f01-49fc-aa6c-bd0462e47fa5","order_by":1,"name":"Yulia (Y) Tyurina","email":"","orcid":"","institution":"University of 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