{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T06:40:44Z","timestamp":1792737644662,"version":"4.5.0"},"reference-count":57,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,6,27]],"date-time":"2025-06-27T00:00:00Z","timestamp":1750982400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell. Neurosci."],"abstract":"<jats:p>Exosomes are small, membrane-bound vesicles secreted by most cell types into the extracellular environment. They play a crucial role in intercellular communication by transporting bioactive molecules, including proteins, lipids, and RNAs, thereby influencing the phenotype and potentially the genotype in recipient cells. In recent years, exosomes have gained increasing attention in the study of pathophysiological conditions and numerous diseases, including multiple sclerosis (MS), an autoimmune disorder with myelin sheath and neuroaxonal damage in the central nervous system. In this study, we isolated and purified serum-derived exosomes from patients with relapsing remitting MS (RR-MS) and characterized their lipid profiles using matrix-assisted laser desorption ionization-time-of-flight\/mass spectrometry (MALDI-TOF\/MS). Lipid analysis was performed in both negative and positive ion modes on intact exosomes, bypassing lipid extraction steps and significantly reducing sample-processing time. The lipid profiles of RR-MS exosomes were compared to those of exosomes isolated from the serum of healthy subjects (HS), and statistical analysis was applied to mass spectra to identify potential lipid biomarkers. The specific phospholipid marker of exosomal membranes, bis(monoacylglycero)phosphate (BMP), was clearly detected in both MALDI lipid profiles, with no significant differences in its content between the two sample groups. However, RR-MS exosomes exhibited significantly lower levels of phosphatidic acid (PA) compared to HS exosomes, despite PA being a key structural component of extracellular vesicles. Notably, comparative analysis revealed an enrichment of several lysophosphatidylcholine (LPC) species in RR-MS exosome membranes, aligning with their known proinflammatory role in MS pathology. Our most significant finding was a markedly lower phosphatidylcholine (PC) to LPC ratio in the pathological group indicating potential alterations in membrane lipid homeostasis. To the best of our knowledge, this study is the first to report a distinct lipid signature in serum-derived exosomes from RR-MS patients using direct MALDI-TOF\/MS analysis.<\/jats:p>","DOI":"10.3389\/fncel.2025.1613618","type":"journal-article","created":{"date-parts":[[2025,6,27]],"date-time":"2025-06-27T05:33:00Z","timestamp":1751002380000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":6,"title":["Circulating exosomes with unique lipid signature in relapsing remitting multiple sclerosis"],"prefix":"10.3389","volume":"19","author":[{"given":"Claudia","family":"Palazzo","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ilaria","family":"Asci","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Silvia","family":"Russo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cinzia","family":"Buccoliero","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vincenzo","family":"Mangialardi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pasqua","family":"Abbrescia","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Onofrio","family":"Valente","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maddalena","family":"Ruggieri","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Damiano","family":"Paolicelli","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Simona","family":"Lobasso","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Antonio","family":"Frigeri","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2025,6,27]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1038\/protex.2016.057","article-title":"Improved exosome isolation by sucrose gradient fractionation of ultracentrifuged crude exosome pellets.","volume":"2016","author":"Alexander","year":"2016","journal-title":"Protoc. Exch."},{"key":"B2","doi-asserted-by":"publisher","first-page":"56","DOI":"10.1016\/j.cca.2022.01.017","article-title":"Kappa free light chains: Diagnostic performance in multiple sclerosis and utility in a clinical laboratory.","volume":"528","author":"Alves Martins","year":"2022","journal-title":"Clin. Chim. Acta"},{"key":"B3","doi-asserted-by":"publisher","first-page":"327","DOI":"10.1016\/0024-3205(69)90138-6","article-title":"Plasma and platelet phospholipids in multiple sclerosis patients.","volume":"8","author":"Andreoli","year":"1969","journal-title":"Life Sci."},{"key":"B4","doi-asserted-by":"publisher","first-page":"661","DOI":"10.1136\/jnnp.36.4.661","article-title":"Significance of plasma lysolecithin in patients with multiple sclerosis: A longitudinal study.","volume":"36","author":"Andreoli","year":"1973","journal-title":"J. Neurol. Neurosurg. Psychiatry"},{"key":"B5","doi-asserted-by":"publisher","first-page":"1417","DOI":"10.1194\/jlr.D026203","article-title":"Lipidomics of intact mitochondria by MALDI-TOF\/MS.","volume":"53","author":"Angelini","year":"2012","journal-title":"J. Lipid Res."},{"key":"B6","doi-asserted-by":"publisher","first-page":"169","DOI":"10.1016\/j.chemphyslip.2014.07.001","article-title":"A fast method for the determination of the PC\/LPC ratio in intact serum by MALDI-TOF MS: An easy-to-follow lipid biomarker of inflammation.","volume":"183","author":"Angelini","year":"2014","journal-title":"Chem. Phys. Lipids"},{"key":"B7","doi-asserted-by":"publisher","first-page":"4411","DOI":"10.1002\/alz.13823","article-title":"Assessment of brain-derived extracellular vesicle enrichment for blood biomarker analysis in age-related neurodegenerative diseases: An international overview.","volume":"20","author":"Badhwar","year":"2024","journal-title":"Alzheimers Dement."},{"key":"B8","doi-asserted-by":"publisher","first-page":"411","DOI":"10.1016\/j.humimm.2014.02.021","article-title":"Elevated serum levels of lysophosphatidic acid in patients with multiple sclerosis.","volume":"75","author":"Balood","year":"2014","journal-title":"Hum. Immunol."},{"key":"B9","doi-asserted-by":"publisher","first-page":"166967","DOI":"10.1016\/j.bbadis.2023.166967","article-title":"Blood-brain barrier transporters: An overview of function, dysfunction in Alzheimer\u2019s disease and strategies for treatment.","volume":"1870","author":"Chaves","year":"2024","journal-title":"Biochim. Biophys. Acta"},{"key":"B10","doi-asserted-by":"publisher","first-page":"188","DOI":"10.1186\/s12974-019-1586-1","article-title":"Dynamic response of microglia\/macrophage polarization following demyelination in mice.","volume":"16","author":"Chu","year":"2019","journal-title":"J. Neuroinflamm."},{"key":"B11","doi-asserted-by":"publisher","first-page":"790","DOI":"10.1002\/glia.23126","article-title":"Translational readthrough generates new astrocyte AQP4 isoforms that modulate supramolecular clustering, glial endfeet localization, and water transport.","volume":"65","author":"De Bellis","year":"2017","journal-title":"Glia"},{"key":"B12","doi-asserted-by":"publisher","first-page":"2826","DOI":"10.1016\/j.jprot.2011.06.023","article-title":"Lipidomic investigations for the characterization of circulating serum lipids in multiple sclerosis.","volume":"74","author":"Del Boccio","year":"2011","journal-title":"J. Proteomics"},{"key":"B13","doi-asserted-by":"publisher","first-page":"545","DOI":"10.1038\/nri3871","article-title":"Immunopathology of multiple sclerosis.","volume":"15","author":"Dendrou","year":"2015","journal-title":"Nat. Rev. Immunol."},{"key":"B14","doi-asserted-by":"publisher","first-page":"43","DOI":"10.1038\/s41572-018-0041-4","article-title":"Multiple sclerosis.","volume":"4","author":"Filippi","year":"2018","journal-title":"Nat. Rev. Dis. Prim."},{"key":"B15","doi-asserted-by":"publisher","first-page":"925","DOI":"10.1016\/j.clinbiochem.2005.06.006","article-title":"The phosphatidylcholine\/lysophosphatidylcholine ratio in human plasma is an indicator of the severity of rheumatoid arthritis: Investigations by 31P NMR and MALDI-TOF MS.","volume":"38","author":"Fuchs","year":"2005","journal-title":"Clin. Biochem."},{"key":"B16","doi-asserted-by":"publisher","first-page":"1198044","DOI":"10.3389\/fmolb.2023.1198044","article-title":"The lipid composition of extracellular vesicles: Applications in diagnostics and therapeutic delivery.","volume":"10","author":"Ghadami","year":"2023","journal-title":"Front. Mol. Biosci."},{"key":"B17","doi-asserted-by":"publisher","first-page":"5346","DOI":"10.3390\/ijms22105346","article-title":"Exosome traceability and cell source dependence on composition and cell-cell cross talk.","volume":"22","author":"Hamzah","year":"2021","journal-title":"Int. J. Mol. Sci."},{"key":"B18","doi-asserted-by":"publisher","first-page":"255","DOI":"10.1016\/j.ijms.2014.08.026","article-title":"Ion mobility and tandem mass spectrometry of phosphatidylglycerol and bis(monoacylglycerol)phosphate (BMP).","volume":"378","author":"Hankin","year":"2015","journal-title":"Int. J. Mass Spectrom."},{"key":"B19","doi-asserted-by":"publisher","first-page":"122668","DOI":"10.1016\/j.lfs.2024.122668","article-title":"Potential therapeutic and diagnostic approaches of exosomes in multiple sclerosis pathophysiology.","volume":"347","author":"Hasaniani","year":"2024","journal-title":"Life Sci."},{"key":"B20","doi-asserted-by":"publisher","first-page":"330","DOI":"10.1016\/0005-2760(92)90063-2","article-title":"Bis(monoacylglycero)phosphate from PC12 cells, a phospholipid that can comigrate with phosphatidic acid: Molecular species analysis by fast atom bombardment mass spectrometry.","volume":"1125","author":"Holbrook","year":"1992","journal-title":"Biochim. Biophys. Acta"},{"key":"B21","doi-asserted-by":"publisher","first-page":"51","DOI":"10.1016\/j.clim.2015.06.015","article-title":"Biomarkers in multiple sclerosis.","volume":"161","author":"Housley","year":"2015","journal-title":"Clin. Immunol."},{"key":"B22","doi-asserted-by":"publisher","first-page":"77","DOI":"10.1016\/j.biochi.2022.09.016","article-title":"Exosomal lipids from membrane organization to biomarkers: Focus on an endolysosomal-specific lipid.","volume":"203","author":"Hullin-Matsuda","year":"2022","journal-title":"Biochimie"},{"key":"B23","doi-asserted-by":"publisher","first-page":"138","DOI":"10.1038\/nm1344","article-title":"Lipid microarrays identify key mediators of autoimmune brain inflammation.","volume":"12","author":"Kanter","year":"2006","journal-title":"Nat. Med."},{"key":"B24","doi-asserted-by":"publisher","first-page":"707","DOI":"10.1515\/hsz-2015-0109","article-title":"Secretory sphingomyelinase in health and disease.","volume":"396","author":"Kornhuber","year":"2015","journal-title":"Biol. Chem."},{"key":"B25","doi-asserted-by":"publisher","first-page":"2483","DOI":"10.3390\/ijms25052483","article-title":"Plasma lipidomic profiling using mass spectrometry for multiple sclerosis diagnosis and disease activity stratification (LipidMS).","volume":"25","author":"Lattau","year":"2024","journal-title":"Int. J. Mol. Sci."},{"key":"B26","doi-asserted-by":"publisher","first-page":"696","DOI":"10.1038\/s41590-018-0135-x","article-title":"Reassessing B cell contributions in multiple sclerosis.","volume":"19","author":"Li","year":"2018","journal-title":"Nat. Immunol."},{"key":"B27","doi-asserted-by":"publisher","first-page":"117443","DOI":"10.1016\/j.lfs.2020.117443","article-title":"The mechanisms of lysophosphatidylcholine in the development of diseases.","volume":"247","author":"Liu","year":"2020","journal-title":"Life Sci."},{"key":"B28","doi-asserted-by":"publisher","first-page":"748895","DOI":"10.3389\/fphys.2021.748895","article-title":"A lipidomic approach to identify potential biomarkers in exosomes from melanoma cells with different metastatic potential.","volume":"12","author":"Lobasso","year":"2021","journal-title":"Front. Physiol."},{"key":"B29","doi-asserted-by":"publisher","first-page":"3540","DOI":"10.1002\/jcp.25815","article-title":"Lipid profiling of parkin-mutant human skin fibroblasts.","volume":"232","author":"Lobasso","year":"2017","journal-title":"J. Cell. Physiol."},{"key":"B30","doi-asserted-by":"publisher","first-page":"27031","DOI":"10.3402\/jev.v4.27031","article-title":"Optimized exosome isolation protocol for cell culture supernatant and human plasma.","volume":"4","author":"Lobb","year":"2015","journal-title":"J. Extracell. Vesicles"},{"key":"B31","doi-asserted-by":"publisher","first-page":"1344","DOI":"10.3389\/fphys.2019.01344","article-title":"Alteration of cholesterol sulfate\/seminolipid ratio in semen lipid profile of men with oligoasthenozoospermia.","volume":"10","author":"Lopalco","year":"2019","journal-title":"Front. Physiol."},{"key":"B32","doi-asserted-by":"publisher","DOI":"10.1002\/jev2.12385","article-title":"MIBlood-EV: Minimal information to enhance the quality and reproducibility of blood extracellular vesicle research","author":"Lucien","year":"2023","journal-title":"J. Extracell Vesicles"},{"key":"B33","doi-asserted-by":"publisher","first-page":"204","DOI":"10.1093\/cei\/uxae032","article-title":"Dysregulated lipid metabolism networks modulate T-cell function in people with relapsing-remitting multiple sclerosis.","volume":"217","author":"Martin-Gutierrez","year":"2024","journal-title":"Clin. Exp. Immunol."},{"key":"B34","doi-asserted-by":"publisher","first-page":"5404","DOI":"10.1007\/s12035-023-03862-2","article-title":"Exosome content-mediated signaling pathways in multiple sclerosis.","volume":"61","author":"Mohammadinasr","year":"2024","journal-title":"Mol. Neurobiol."},{"key":"B35","doi-asserted-by":"publisher","first-page":"674","DOI":"10.1038\/nrm3188","article-title":"Insights into autotaxin: How to produce and present a lipid mediator.","volume":"12","author":"Moolenaar","year":"2011","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"B36","doi-asserted-by":"publisher","first-page":"949","DOI":"10.1007\/s00702-003-0007-9","article-title":"Decreased lysophosphatidylcholine\/phosphatidylcholine ratio in cerebrospinal fluid in Alzheimer? disease.","volume":"110","author":"Mulder","year":"2003","journal-title":"J. Neural Trans."},{"key":"B37","doi-asserted-by":"publisher","first-page":"1531","DOI":"10.3390\/ijms21041531","article-title":"Tissue distribution of the readthrough isoform of AQP4 reveals a dual role of AQP4ex limited to CNS.","volume":"21","author":"Palazzo","year":"2020","journal-title":"Int. J. Mol. Sci."},{"key":"B38","doi-asserted-by":"publisher","first-page":"1595","DOI":"10.3390\/brainsci12121595","article-title":"Real-life experience of the effects of cladribine tablets on lymphocyte subsets and serum neurofilament light chain levels in relapsing multiple sclerosis patients.","volume":"12","author":"Paolicelli","year":"2022","journal-title":"Brain Sci."},{"key":"B39","doi-asserted-by":"publisher","first-page":"1426700","DOI":"10.3389\/fnins.2024.1426700","article-title":"CNS cell-derived exosome signatures as blood-based biomarkers of neurodegenerative diseases.","volume":"18","author":"Park","year":"2024","journal-title":"Front. Neurosci."},{"key":"B40","doi-asserted-by":"publisher","first-page":"3071","DOI":"10.1038\/s41598-018-21497-5","article-title":"Enhanced release of acid sphingomyelinase-enriched exosomes generates a lipidomics signature in CSF of Multiple Sclerosis patients.","volume":"8","author":"Pieragostino","year":"2018","journal-title":"Sci. Rep."},{"key":"B41","doi-asserted-by":"publisher","first-page":"1563","DOI":"10.1039\/C4MB00700J","article-title":"An integrated metabolomics approach for the research of new cerebrospinal fluid biomarkers of multiple sclerosis.","volume":"11","author":"Pieragostino","year":"2015","journal-title":"Mol. BioSyst."},{"key":"B42","doi-asserted-by":"publisher","first-page":"18889","DOI":"10.1073\/pnas.0806310105","article-title":"Antigen microarrays identify unique serum autoantibody signatures in clinical and pathologic subtypes of multiple sclerosis.","volume":"105","author":"Quintana","year":"2008","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"B43","doi-asserted-by":"publisher","first-page":"108","DOI":"10.1016\/j.bbalip.2013.10.004","article-title":"Exosomes as new vesicular lipid transporters involved in cell-cell communication and various pathophysiologies.","volume":"1841","author":"Record","year":"2014","journal-title":"Biochim. Biophys. Acta"},{"key":"B44","doi-asserted-by":"publisher","first-page":"169","DOI":"10.1056\/NEJMra1401483","article-title":"Multiple sclerosis.","volume":"378","author":"Reich","year":"2018","journal-title":"N. Engl. J. Med."},{"key":"B45","doi-asserted-by":"publisher","first-page":"19847","DOI":"10.1038\/s41598-022-24231-4","article-title":"Beneficial effects of SS-31 peptide on cardiac mitochondrial dysfunction in tafazzin knockdown mice.","volume":"12","author":"Russo","year":"2022","journal-title":"Sci. Rep."},{"key":"B46","doi-asserted-by":"publisher","first-page":"620","DOI":"10.1038\/339620a0","article-title":"Vesicular removal by oligodendrocytes of membrane attack complexes formed by activated complement.","volume":"339","author":"Scolding","year":"1989","journal-title":"Nature"},{"key":"B47","doi-asserted-by":"publisher","first-page":"9","DOI":"10.1194\/jlr.R084343","article-title":"Exosomal lipid composition and the role of ether lipids and phosphoinositides in exosome biology.","volume":"60","author":"Skotland","year":"2019","journal-title":"J. Lipid Res."},{"key":"B48","doi-asserted-by":"publisher","first-page":"2","DOI":"10.3389\/fncel.2019.00002","article-title":"Phosphatidic acid: From pleiotropic functions to neuronal pathology.","volume":"13","author":"Tanguy","year":"2019","journal-title":"Front. Cell. Neurosci."},{"key":"B49","doi-asserted-by":"publisher","first-page":"53","DOI":"10.3389\/fnmol.2020.00053","article-title":"Circulating exosomal miRNA as diagnostic biomarkers of neurodegenerative diseases.","volume":"13","author":"Wang","year":"2020","journal-title":"Front. Mol. Neurosci."},{"key":"B50","doi-asserted-by":"publisher","first-page":"e12404","DOI":"10.1002\/jev2.12404","article-title":"Minimal information for studies of extracellular vesicles (MISEV2023): From basic to advanced approaches.","volume":"13","author":"Welsh","year":"2024","journal-title":"J. Extracell. Vesicles"},{"key":"B51","doi-asserted-by":"publisher","first-page":"3092","DOI":"10.1093\/brain\/awn190","article-title":"A defect of sphingolipid metabolism modifies the properties of normal appearing white matter in multiple sclerosis.","volume":"131","author":"Wheeler","year":"2008","journal-title":"Brain"},{"key":"B52","doi-asserted-by":"crossref","first-page":"925","DOI":"10.1001\/jama.1997.03540350075038","article-title":"Recommendations guiding physicians in biomedical research involving human subjects","volume":"277","year":"1997","journal-title":"JAMA"},{"key":"B53","doi-asserted-by":"publisher","first-page":"3556","DOI":"10.1007\/s12035-024-04477-x","article-title":"Dysregulation of serum exosomal lipid metabolism in schizophrenia: A biomarker perspective.","volume":"62","author":"Xu","year":"2025","journal-title":"Mol. Neurobiol."},{"key":"B54","doi-asserted-by":"publisher","first-page":"2301706","DOI":"10.1002\/adhm.202301706","article-title":"Assessing extracellular vesicles in human biofluids using flow-based analyzers.","volume":"12","author":"Yim","year":"2023","journal-title":"Adv. Healthc. Mater."},{"key":"B55","doi-asserted-by":"publisher","first-page":"120","DOI":"10.1016\/j.jneuroim.2014.06.006","article-title":"Increased autotaxin activity in multiple sclerosis.","volume":"273","author":"Zahednasab","year":"2014","journal-title":"J. Neuroimmunol."},{"key":"B56","doi-asserted-by":"publisher","DOI":"10.1101\/2024.01.04.574253","article-title":"Blood-based targeted metabolipidomics reveals altered omega fatty acid-derived lipid mediators in relapsing-remitting multiple sclerosis patients.","author":"Zahoor","year":"2024","journal-title":"bioRxiv"},{"key":"B57","doi-asserted-by":"publisher","first-page":"724","DOI":"10.1093\/labmed\/lmae039","article-title":"Lipidomic analysis of serum exosomes identifies a novel diagnostic marker for type 2 diabetes mellitus.","volume":"55","author":"Zhang","year":"2024","journal-title":"Lab. Med."}],"container-title":["Frontiers in Cellular Neuroscience"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fncel.2025.1613618\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,6,27]],"date-time":"2025-06-27T05:33:01Z","timestamp":1751002381000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fncel.2025.1613618\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,6,27]]},"references-count":57,"alternative-id":["10.3389\/fncel.2025.1613618"],"URL":"https:\/\/doi.org\/10.3389\/fncel.2025.1613618","relation":{},"ISSN":["1662-5102"],"issn-type":[{"value":"1662-5102","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,6,27]]},"article-number":"1613618"}}