BCL2L13 Influences Autophagy and Ceramide Metabolism without Affecting Temozolomide Resistance in Glioblastoma

BCL2L13 Influences Autophagy and Ceramide Metabolism without Affecting Temozolomide Resistance in Glioblastoma | 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 BCL2L13 Influences Autophagy and Ceramide Metabolism without Affecting Temozolomide Resistance in Glioblastoma Saeid Ghavami, Courtney Clark, Amir Barzegar Behrooz, Simone da Silva Rosa, and 14 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8812057/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Temozolomide (TMZ) resistance in glioblastoma (GBM) arises through metabolic rewiring that links mitochondrial function, autophagy balance, and sphingolipid metabolism. TMZ-resistant (R) U251 cells exhibited suppressed apoptosis and complete blockade of autophagy flux, evidenced by MAP1LC3/LC3-II and SQSTM1/p62 accumulation and insensitivity to bafilomycin A 1 . BCL2L13, strongly upregulated in R cells, emerged as a dual regulator of mitophagy and ceramide metabolism. BCL2L13 knockdown (KD) produced opposite effects in TMZ-sensitive (NR) and resistant cells: in NR cells, KD elevated LC3-II, reduced respiratory reserve, and triggered compensatory lipid synthesis; in R cells, KD decreased LC3-II without restoring flux or TMZ sensitivity. Lipidomic profiling revealed that BCL2L13 loss reactivated CERS6 in NR cells, increasing C16:0 and mid-chain ceramides, while relieving CERS2 inhibition in R cells, elevating very-long-chain (C22–C24) and glycosylated ceramides. These distinct sphingolipid signatures were confirmed by PLS-DA and KEGG enrichment, which highlighted steroid hormone, arachidonic, and linoleic acid metabolism in NR KD cells versus neuroactive ligand-receptor and signaling pathways in R KD cells. Together, these findings position BCL2L13 as a molecular integrator of mitochondrial respiration, autophagy flux, and CERS-dependent lipid remodeling, unveiling a context-specific metabolic mechanism that supports GBM cell survival under conditions of chemotherapeutic stress. Biological sciences/Cancer/Cancer therapy/Cancer therapeutic resistance Biological sciences/Cell biology/Cell death/Autophagy/Macroautophagy Autophagy flux BCL2L13 CERS-BCL2L13 interaction temozolomide-resistant glioblastoma cells Full Text Additional Declarations (Not answered) Supplementary Files SupplementarySMILES.docx Dataset 1. SMILE CODES WesternBlotReplicates.pptx Western Blot Original File Replicated Cite Share Download PDF Status: Posted 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. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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-8812057","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":596907228,"identity":"7b0097e5-2a3f-42ef-83bf-c30838dd0dcc","order_by":0,"name":"Saeid 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TMZ-resistant (R) U251 cells exhibited suppressed apoptosis and complete blockade of autophagy flux, evidenced by MAP1LC3/LC3-II and SQSTM1/p62 accumulation and insensitivity to bafilomycin A\u003csub\u003e1\u003c/sub\u003e. BCL2L13, strongly upregulated in R cells, emerged as a dual regulator of mitophagy and ceramide metabolism. BCL2L13 knockdown (KD) produced opposite effects in TMZ-sensitive (NR) and resistant cells: in NR cells, KD elevated LC3-II, reduced respiratory reserve, and triggered compensatory lipid synthesis; in R cells, KD decreased LC3-II without restoring flux or TMZ sensitivity. Lipidomic profiling revealed that BCL2L13 loss reactivated CERS6 in NR cells, increasing C16:0 and mid-chain ceramides, while relieving CERS2 inhibition in R cells, elevating very-long-chain (C22\u0026ndash;C24) and glycosylated ceramides. 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