Transcriptomic Modulation by Exosomes Derived from Human Adipose Stem Cells in Neuronal Cells: Insights from mRNA Sequencing Analysis

Transcriptomic Modulation by Exosomes Derived from Human Adipose Stem Cells in Neuronal Cells: Insights from mRNA Sequencing Analysis | 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 Transcriptomic Modulation by Exosomes Derived from Human Adipose Stem Cells in Neuronal Cells: Insights from mRNA Sequencing Analysis Ekkaphot Khongkla, Kornkanok Promtap, Jitrawadee Meerasri, Pornphawit Mo-Mai, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7778530/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 04 Jan, 2026 Read the published version in Scientific Reports → Version 1 posted 13 You are reading this latest preprint version Abstract Brain aging, which influences neurological function across cellular and molecular domains, is a critical concern in the elderly population. Therapeutic strategies for mitigating age-related neurodegeneration should target molecular pathways that are primarily involved in neuroinflammation. Exosomes derived from human adipose tissue mesenchymal stem cells (hASCs) have demonstrated anti-inflammatory and rejuvenating properties, making them promising agents for neurochemical intervention. However, their transcriptomic impact on neuronal cells remains largely unexplored. To address this research question, we applied high-throughput mRNA sequencing and downstream bioinformatic analysis. As an in vitro model for aging and neurodegeneration, CNS mouse-derived CAD cells were exposed to D-galactose (DG) to trigger molecular responses and were used to evaluate the efficacy of the isolated exosomes. The hASC-exosomes were isolated via ultrafiltration and subsequently characterized via nanoparticle tracking analysis, cryo-EM microscopy, and immunoassays. The internalization of PHK26-tagged hASC exosomes in the cytosol of the neuronal cells was monitored. Illumina-based mRNA sequencing has allowed expression profiling of more than 27,000 genes. Comparative transcriptomic profiling revealed 3,951 differentially expressed genes (DEGs) associated with DG-induced cells and 3,091 DEGs modulated by hASC-exosome treatment. In DG-treated cells, many genes were upregulated in response to cellular stress. The DEGs whose expression was upregulated in response to DG play roles in the DNA damage response, cellular senescence, and apoptosis. In the presence of hASC-derived exosomes, many DEGs (1,948) were downregulated, suggesting that the exosomes suppressed stress-induced gene expression. The functional pathway analysis indicated that hASC-exosomes significantly downregulated processes related largely to translation, neuroinflammation, cellular senescence, apoptosis, and other age-associated molecular pathways. A set of genes involved in the inflammatory response and regulated by hASC-exosomes was identified. Our study provides transcriptomic evidence supporting the regulatory role of hASC-derived exosomes in attenuating the expression of inflammatory and neurodegenerative markers, positioning them as potential candidates for antiaging neurotherapeutics. Biological sciences/Cell biology Biological sciences/Molecular biology Biological sciences/Neuroscience Biological sciences/Stem cells Human adipose tissue stem cells Exosomes mRNA sequencing Neuroinflammation CAD cells D-galactose Full Text Additional Declarations No competing interests reported. Supplementary Files SupplementaryFile13Dvideo.zip SupplementaryFile2Expressionresult.zip SupplementaryFile3DEGanalysis.zip SupplementaryFile4Uncroppedwesternblotimages.zip Supplementaryinformation.pdf Cite Share Download PDF Status: Published Journal Publication published 04 Jan, 2026 Read the published version in Scientific Reports → Version 1 posted Editorial decision: Revision requested 26 Nov, 2025 Reviews received at journal 24 Nov, 2025 Reviewers agreed at journal 13 Nov, 2025 Reviewers agreed at journal 13 Nov, 2025 Reviewers agreed at journal 12 Nov, 2025 Reviews received at journal 31 Oct, 2025 Reviewers agreed at journal 23 Oct, 2025 Reviewers agreed at journal 23 Oct, 2025 Reviewers invited by journal 22 Oct, 2025 Editor assigned by journal 22 Oct, 2025 Editor invited by journal 22 Oct, 2025 Submission checks completed at journal 18 Oct, 2025 First submitted to journal 18 Oct, 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. 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-7778530","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":538465180,"identity":"e8df02cd-886a-4bea-90dd-2d7a375caa9e","order_by":0,"name":"Ekkaphot 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cells, Exosomes, mRNA sequencing, Neuroinflammation, CAD cells, D-galactose","lastPublishedDoi":"10.21203/rs.3.rs-7778530/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7778530/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eBrain aging, which influences neurological function across cellular and molecular domains, is a critical concern in the elderly population. Therapeutic strategies for mitigating age-related neurodegeneration should target molecular pathways that are primarily involved in neuroinflammation. Exosomes derived from human adipose tissue mesenchymal stem cells (hASCs) have demonstrated anti-inflammatory and rejuvenating properties, making them promising agents for neurochemical intervention. However, their transcriptomic impact on neuronal cells remains largely unexplored. To address this research question, we applied high-throughput mRNA sequencing and downstream bioinformatic analysis. As an in vitro model for aging and neurodegeneration, CNS mouse-derived CAD cells were exposed to D-galactose (DG) to trigger molecular responses and were used to evaluate the efficacy of the isolated exosomes. The hASC-exosomes were isolated via ultrafiltration and subsequently characterized via nanoparticle tracking analysis, cryo-EM microscopy, and immunoassays. The internalization of PHK26-tagged hASC exosomes in the cytosol of the neuronal cells was monitored. Illumina-based mRNA sequencing has allowed expression profiling of more than 27,000 genes. Comparative transcriptomic profiling revealed 3,951 differentially expressed genes (DEGs) associated with DG-induced cells and 3,091 DEGs modulated by hASC-exosome treatment. In DG-treated cells, many genes were upregulated in response to cellular stress. The DEGs whose expression was upregulated in response to DG play roles in the DNA damage response, cellular senescence, and apoptosis. In the presence of hASC-derived exosomes, many DEGs (1,948) were downregulated, suggesting that the exosomes suppressed stress-induced gene expression. The functional pathway analysis indicated that hASC-exosomes significantly downregulated processes related largely to translation, neuroinflammation, cellular senescence, apoptosis, and other age-associated molecular pathways. A set of genes involved in the inflammatory response and regulated by hASC-exosomes was identified. Our study provides transcriptomic evidence supporting the regulatory role of hASC-derived exosomes in attenuating the expression of inflammatory and neurodegenerative markers, positioning them as potential candidates for antiaging neurotherapeutics.\u003c/p\u003e","manuscriptTitle":"Transcriptomic Modulation by Exosomes Derived from Human Adipose Stem Cells in Neuronal Cells: Insights from mRNA Sequencing Analysis","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-11-03 18:25:45","doi":"10.21203/rs.3.rs-7778530/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-11-26T07:39:59+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-11-25T04:44:05+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"92188140183032858616793592939897812765","date":"2025-11-13T23:57:06+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"200575797898199115714987095681340323319","date":"2025-11-13T14:09:17+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"82972224233961122047216300740668662611","date":"2025-11-12T18:36:15+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-10-31T20:03:29+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"88211290954738190170663399666343742919","date":"2025-10-23T21:09:48+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"221702946467451605821425694754198703857","date":"2025-10-23T10:59:09+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-10-22T13:40:04+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-10-22T13:34:23+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2025-10-22T12:49:21+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-10-18T10:32:48+00:00","index":"","fulltext":""},{"type":"submitted","content":"Scientific Reports","date":"2025-10-18T10:28:42+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"f50c6f25-ac3c-40cd-a56d-6be26273ce92","owner":[],"postedDate":"November 3rd, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":57273598,"name":"Biological sciences/Cell biology"},{"id":57273599,"name":"Biological sciences/Molecular biology"},{"id":57273600,"name":"Biological sciences/Neuroscience"},{"id":57273601,"name":"Biological sciences/Stem cells"}],"tags":[],"updatedAt":"2026-01-05T16:00:15+00:00","versionOfRecord":{"articleIdentity":"rs-7778530","link":"https://doi.org/10.1038/s41598-025-34661-5","journal":{"identity":"scientific-reports","isVorOnly":false,"title":"Scientific Reports"},"publishedOn":"2026-01-04 15:57:40","publishedOnDateReadable":"January 4th, 2026"},"versionCreatedAt":"2025-11-03 18:25:45","video":"","vorDoi":"10.1038/s41598-025-34661-5","vorDoiUrl":"https://doi.org/10.1038/s41598-025-34661-5","workflowStages":[]},"version":"v1","identity":"rs-7778530","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7778530","identity":"rs-7778530","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}