{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T09:18:07Z","timestamp":1785403087593,"version":"3.56.0"},"posted":{"date-parts":[[2026,4,16]]},"group-title":"Cell Biology","reference-count":35,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,4,16]],"date-time":"2026-04-16T00:00:00Z","timestamp":1776297600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"award":["RO1AA024082"],"award-info":[{"award-number":["RO1AA024082"]}],"id":[{"id":"https:\/\/ror.org\/001d55x84","id-type":"ROR","asserted-by":"publisher"}]},{"award":["RO1AA031225"],"award-info":[{"award-number":["RO1AA031225"]}],"id":[{"id":"https:\/\/ror.org\/001d55x84","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,4,16]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:sec>\n                  <jats:title>Background<\/jats:title>\n                  <jats:p>Alcohol consumption influences cardiovascular disease, but whether it does so by affecting endothelial plasticity is unknown. We tested whether alcohol regulates endothelial-to-mesenchymal transition (EndMT) to influence arterial pathology.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>HCAEC and HUVEC were exposed to inflammatory cytokines (TGF\u03b2 \u00b1 IL1\u03b2) or hypoxia in the presence of ethanol (0-100 mM). EndMT was assessed by changes in cell marker expression, SNAIL levels, and migration assays. In vivo, carotid ligation was performed in mice gavaged with\/without either daily moderate ethanol (2-drink equivalent\/d) or episodic binge exposure (7-drink equivalent, 2 days\/week) and myo-endothelial cell population assessed.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>Cytokines and hypoxia induced EndMT in vitro, characterized by loss of endothelial markers, increased mesenchymal markers, elevated SNAIL, and enhanced migratory capacity. Low-to-moderate dose ethanol (5\u201325 mM) attenuated these changes, preserving endothelial phenotype, whereas high dose ethanol (50\u2013100 mM) either had no effect or exacerbated EndMT. The inhibitory effect of moderate ethanol on cytokine- and hypoxia-induced changes in \u03b1SMA and Cdh5 expression was abrogated by \u03b3-secretase inhibition, consistent with involvement of Notch signaling. Carotid ligation induced neointimal formation and accumulation of myo-endothelial cells indicative of EndMT. Daily moderate ethanol significantly attenuated neointimal hyperplasia and diminished the myo-endothelial cell population, whereas in contrast, episodic binge ethanol exposure increased pathologic remodeling and myo-endothelial cell abundance.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Conclusions<\/jats:title>\n                  <jats:p>Alcohol modulates endothelial trans-differentiation in a biphasic manner. Low-to-moderate alcohol exposure suppresses EndMT and limits pathological remodeling, whereas binge-level exposure promotes these processes. These findings identify regulation of endothelial plasticity as a potential novel mechanism linking alcohol consumption patterns to vascular disease risk.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>NEW AND NOTEWORTHY<\/jats:title>\n                  <jats:p>We identify a previously unrecognized biphasic effect of alcohol on endothelial phenotypic plasticity. Low-to-moderate dose alcohol suppresses endothelial-to-mesenchymal transition (EndMT), whereas high-level (binge) exposure promotes this pro-atherogenic process. Given the central role of EndMT in vascular remodelling and atherosclerosis, these findings provide a mechanistic framework linking alcohol consumption patterns and cardiovascular disease risk \u2013 potentially explaining both the protective effect at low\/moderate levels, and the detrimental impact of heavy alcohol use.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Graphical abstract<\/jats:title>\n                  <jats:fig id=\"ufig1\" position=\"float\" fig-type=\"figure\" orientation=\"portrait\">\n                    <jats:graphic xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"718463v1_ufig1\" position=\"float\" orientation=\"portrait\"\/>\n                  <\/jats:fig>\n                  <jats:p>\n                    Injurious stimuli can trigger endothelial cells (EC) to undergo endothelial-to-mesenchymal transition (EndMT) that contributes to arterial remodeling and disease. EndMT is regulated in a biphasic manner by alcohol with low-to-moderate levels (1-3 drink equivalent) suppressing EndMT and attenuating vascular remodeling, whereas higher level\/binge exposure (7 drink equivalent) promotes these processes.\n                    <jats:italic>Graphic created using Biorender<\/jats:italic>\n                    .\n                  <\/jats:p>\n                <\/jats:sec>","DOI":"10.64898\/2026.04.14.718463","type":"posted-content","created":{"date-parts":[[2026,4,17]],"date-time":"2026-04-17T00:45:15Z","timestamp":1776386715000},"source":"Crossref","is-referenced-by-count":0,"title":["A Biphasic Effect of Alcohol on Endothelial Plasticity Through Regulation of Endothelial-to-Mesenchymal Transition"],"prefix":"10.64898","author":[{"given":"Weimin","family":"Liu","sequence":"first","affiliation":[{"name":"Vascular Biology and Therapeutics Laboratory, School of Biotechnology, Dublin City University, Dublin, Ireland and Department of Surgery, University of Rochester Medical Center, Rochester, NY, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-6186-248X","authenticated-orcid":false,"given":"Yusof","family":"Gusti","sequence":"additional","affiliation":[{"name":"Vascular Biology and Therapeutics Laboratory, School of Biotechnology, Dublin City University, Dublin, Ireland and Department of Surgery, University of Rochester Medical Center, Rochester, NY, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0608-6162","authenticated-orcid":false,"given":"Fathima","family":"Athar","sequence":"additional","affiliation":[{"name":"Vascular Biology and Therapeutics Laboratory, School of Biotechnology, Dublin City University, Dublin, Ireland and Department of Surgery, University of Rochester Medical Center, Rochester, NY, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4401-6912","authenticated-orcid":false,"given":"Naresh K","family":"Rajendran","sequence":"additional","affiliation":[{"name":"Vascular Biology and Therapeutics Laboratory, School of Biotechnology, Dublin City University, Dublin, Ireland and Department of Surgery, University of Rochester Medical Center, Rochester, NY, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5385-6502","authenticated-orcid":false,"given":"Paul A.","family":"Cahill","sequence":"additional","affiliation":[{"name":"Vascular Biology and Therapeutics Laboratory, School of Biotechnology, Dublin City University, Dublin, Ireland and Department of Surgery, University of Rochester Medical Center, Rochester, NY, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8642-4418","authenticated-orcid":false,"given":"Eileen M.","family":"Redmond","sequence":"additional","affiliation":[{"name":"Vascular Biology and Therapeutics Laboratory, School of Biotechnology, Dublin City University, Dublin, Ireland and Department of Surgery, University of Rochester Medical Center, Rochester, NY, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026042006001694000_2026.04.14.718463v1.1","doi-asserted-by":"publisher","DOI":"10.1161\/ATVBAHA.121.313788"},{"key":"2026042006001694000_2026.04.14.718463v1.2","doi-asserted-by":"crossref","unstructured":"Alvarez-Mon MA , Martinez-Urbistondo D , Barberia-Latasa M , Vazquez-Ruiz Z , Ruiz- Canela M , Bes-Rastrollo M , and Martinez-Gonzalez MA . The Unfinished Debate on Wine and Other Alcoholic Beverages: Conflicting Evidence, Public Health Messages and the Missing Trial. Nutrients 18, 2026.","DOI":"10.3390\/nu18030529"},{"key":"2026042006001694000_2026.04.14.718463v1.3","doi-asserted-by":"crossref","first-page":"695","DOI":"10.1080\/14728222.2019.1641198","article-title":"The Notch pathway: a novel therapeutic target for cardiovascular diseases?","volume":"23","year":"2019","journal-title":"Expert Opin Ther Targets"},{"key":"2026042006001694000_2026.04.14.718463v1.4","doi-asserted-by":"publisher","DOI":"10.1016\/j.atherosclerosis.2016.03.007"},{"key":"2026042006001694000_2026.04.14.718463v1.5","doi-asserted-by":"crossref","first-page":"702","DOI":"10.1016\/j.kint.2022.12.022","article-title":"Osteopontin mediation of disturbed flow-induced endothelial mesenchymal transition through CD44 is a novel mechanism of neointimal hyperplasia in arteriovenous fistulae for hemodialysis access","volume":"103","year":"2023","journal-title":"Kidney Int"},{"key":"2026042006001694000_2026.04.14.718463v1.6","doi-asserted-by":"crossref","first-page":"53","DOI":"10.3389\/fcvm.2020.00053","article-title":"Endothelial-to-Mesenchymal Transition, Vascular Inflammation, and Atherosclerosis","volume":"7","year":"2020","journal-title":"Front Cardiovasc Med"},{"key":"2026042006001694000_2026.04.14.718463v1.7","doi-asserted-by":"publisher","DOI":"10.1007\/s10654-018-0374-z"},{"key":"2026042006001694000_2026.04.14.718463v1.8","doi-asserted-by":"publisher","DOI":"10.1016\/j.jacc.2010.01.006"},{"key":"2026042006001694000_2026.04.14.718463v1.9","doi-asserted-by":"crossref","first-page":"411","DOI":"10.1093\/ajh\/hpaf027","article-title":"Endothelial Cell Phenotypic Plasticity in Cardiovascular Physiology and Disease: Mechanisms and Therapeutic Prospects","volume":"38","year":"2025","journal-title":"Am J Hypertens"},{"key":"2026042006001694000_2026.04.14.718463v1.10","doi-asserted-by":"publisher","DOI":"10.1001\/archinte.166.22.2437"},{"key":"2026042006001694000_2026.04.14.718463v1.11","doi-asserted-by":"publisher","DOI":"10.1038\/ncomms11853"},{"key":"2026042006001694000_2026.04.14.718463v1.12","doi-asserted-by":"publisher","DOI":"10.1038\/nrcardio.2015.91"},{"key":"2026042006001694000_2026.04.14.718463v1.13","doi-asserted-by":"crossref","first-page":"2051","DOI":"10.1111\/acer.13499","article-title":"Alcohol Reduces Arterial Remodeling by Inhibiting Sonic Hedgehog- Stimulated Stem Cell Antigen-1 Positive Progenitor Stem Cell Expansion","volume":"41","year":"2017","journal-title":"Alcoholism, clinical and experimental research"},{"key":"2026042006001694000_2026.04.14.718463v1.14","doi-asserted-by":"crossref","unstructured":"Gusti Y , Liu W , Athar F , Cahill PA , and Redmond EM . Endothelial Homeostasis Under the Influence of Alcohol-Relevance to Atherosclerotic Cardiovascular Disease. Nutrients 17, 2025.","DOI":"10.3390\/nu17050802"},{"key":"2026042006001694000_2026.04.14.718463v1.15","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-food-032519-051640"},{"key":"2026042006001694000_2026.04.14.718463v1.16","doi-asserted-by":"publisher","DOI":"10.1016\/j.ejcb.2018.07.005"},{"key":"2026042006001694000_2026.04.14.718463v1.17","doi-asserted-by":"publisher","DOI":"10.1016\/j.jacc.2018.09.089"},{"key":"2026042006001694000_2026.04.14.718463v1.18","doi-asserted-by":"crossref","first-page":"1408","DOI":"10.1038\/s44161-024-00569-y","article-title":"Chronic inflammation and vascular cell plasticity in atherosclerosis","volume":"3","year":"2024","journal-title":"Nat Cardiovasc Res"},{"key":"2026042006001694000_2026.04.14.718463v1.19","first-page":"6485","article-title":"Roles of notch signaling pathway and endothelial- mesenchymal transition in vascular endothelial dysfunction and atherosclerosis","volume":"22","year":"2018","journal-title":"Eur Rev Med Pharmacol Sci"},{"key":"2026042006001694000_2026.04.14.718463v1.20","doi-asserted-by":"crossref","first-page":"1734","DOI":"10.1111\/acer.14415","article-title":"Moderate Alcohol Consumption Targets S100beta(+) Vascular Stem Cells and Attenuates Injury- Induced Neointimal Hyperplasia","volume":"44","year":"2020","journal-title":"Alcoholism, clinical and experimental research"},{"key":"2026042006001694000_2026.04.14.718463v1.21","doi-asserted-by":"crossref","first-page":"448","DOI":"10.1016\/j.atherosclerosis.2011.08.034","article-title":"Differential effects of daily-moderate versus weekend-binge alcohol consumption on atherosclerotic plaque development in mice","volume":"219","year":"2011","journal-title":"Atherosclerosis"},{"key":"2026042006001694000_2026.04.14.718463v1.22","doi-asserted-by":"publisher","DOI":"10.1097\/MOH.0000000000000425"},{"key":"2026042006001694000_2026.04.14.718463v1.23","doi-asserted-by":"publisher","DOI":"10.1161\/ATVBAHA.110.215681"},{"key":"2026042006001694000_2026.04.14.718463v1.24","first-page":"e7","article-title":"Roerecke M, on behalf the American Heart Association Council on L, Cardiometabolic H, Council on C, Stroke N","volume":"152","year":"2025","journal-title":"Council on Clinical C, and Stroke C. Alcohol Use and Cardiovascular Disease: A Scientific Statement From the American Heart Association. Circulation"},{"key":"2026042006001694000_2026.04.14.718463v1.25","doi-asserted-by":"crossref","first-page":"1467","DOI":"10.1111\/acer.15138","article-title":"Alcohol and vascular endothelial function: Biphasic effect highlights the importance of dose","volume":"47","year":"2023","journal-title":"Alcohol Clin Exp Res (Hoboken"},{"key":"2026042006001694000_2026.04.14.718463v1.26","doi-asserted-by":"crossref","first-page":"e15544","DOI":"10.14814\/phy2.15544","article-title":"Caveolin-1 inhibition mediates the opposing effects of alcohol on gamma-secretase activity in arterial endothelial and smooth muscle cells","volume":"11","year":"2023","journal-title":"Physiol Rep"},{"key":"2026042006001694000_2026.04.14.718463v1.27","doi-asserted-by":"crossref","first-page":"2217","DOI":"10.1111\/acer.14706","article-title":"Moderate dose alcohol protects against serum amyloid protein A1-induced endothelial dysfunction via both notch- dependent and notch-independent pathways","volume":"45","year":"2021","journal-title":"Alcoholism, clinical and experimental research"},{"key":"2026042006001694000_2026.04.14.718463v1.28","doi-asserted-by":"publisher","DOI":"10.1136\/bmj.d671"},{"key":"2026042006001694000_2026.04.14.718463v1.29","doi-asserted-by":"publisher","DOI":"10.1093\/cvr\/cvx253"},{"key":"2026042006001694000_2026.04.14.718463v1.30","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0232565"},{"key":"2026042006001694000_2026.04.14.718463v1.31","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1016\/j.vph.2018.10.004","article-title":"Protective effect of rapamycin on endothelial-to-mesenchymal transition in HUVECs through the Notch signaling pathway","volume":"113","year":"2019","journal-title":"Vascul Pharmacol"},{"key":"2026042006001694000_2026.04.14.718463v1.32","doi-asserted-by":"crossref","first-page":"1768","DOI":"10.1093\/cvr\/cvae157","article-title":"Inhibition of endothelial-to- mesenchymal transition in a large animal preclinical arteriovenous fistula model leads to improved remodelling and reduced stenosis","volume":"120","year":"2024","journal-title":"Cardiovascular research"},{"key":"2026042006001694000_2026.04.14.718463v1.33","doi-asserted-by":"crossref","first-page":"781","DOI":"10.2174\/1871530319666191023141638","article-title":"Hepatocyte Growth Factor Attenuates the Development of TGF-beta1- Induced EndMT through Down-regulating the Notch Signaling","volume":"20","year":"2020","journal-title":"Endocr Metab Immune Disord Drug Targets"},{"key":"2026042006001694000_2026.04.14.718463v1.34","doi-asserted-by":"crossref","first-page":"419","DOI":"10.1016\/j.jare.2024.12.049","article-title":"Reconceptualizing Endothelial-to-mesenchymal transition in atherosclerosis: Signaling pathways and prospective targeting strategies","volume":"77","year":"2025","journal-title":"J Adv Res"},{"key":"2026042006001694000_2026.04.14.718463v1.35","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCRESAHA.117.312136"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.64898\/2026.04.14.718463","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,4,20]],"date-time":"2026-04-20T13:00:48Z","timestamp":1776690048000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.64898\/2026.04.14.718463"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,4,16]]},"references-count":35,"URL":"https:\/\/doi.org\/10.64898\/2026.04.14.718463","relation":{},"subject":[],"published":{"date-parts":[[2026,4,16]]},"subtype":"preprint"}}