{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,4,16]],"date-time":"2026-04-16T14:10:29Z","timestamp":1776348629638,"version":"3.51.2"},"posted":{"date-parts":[[2026,4,13]]},"group-title":"Molecular Biology","reference-count":34,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,4,13]],"date-time":"2026-04-13T00:00:00Z","timestamp":1776038400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,4,13]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:sec>\n                  <jats:title>Background<\/jats:title>\n                  <jats:p>Metabolically healthy obesity (MHO) is unstable, with up to 80% of individuals progressing to metabolically abnormal obesity (MAO), yet mechanisms underlying this transition remain unclear. African Americans bear a disproportionate burden of obesity-related cardiovascular disease. Circulating extracellular vesicles (EVs) mediate inter-organ communication and may drive MAO-related vascular dysfunction.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>Adults of African ancestry were classified as metabolically healthy lean (MHL, n=14), MHO (n=9), or MAO (n=16). Plasma-derived EVs were characterized and their microRNA cargo profiled. Human coronary artery endothelial cells were treated with EVs from each group to assess nitric oxide signaling, oxidative stress, inflammatory activation, and mitochondrial dynamics.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>MHO participants exhibited preserved insulin sensitivity and lower inflammation compared with MAO despite comparable adiposity. EVs from MHO carried a distinct microRNA signature enriched in miR-148a-5p, miR-181c-5p, and miR-1255a, linked to antioxidant and matrix regulatory pathways. MAO EVs were enriched in miR-3613-3p, miR-6842-3p, and miR-326, targeting inflammation and insulin resistance pathways. Compared with both MHL and MHO EVs, MAO EVs suppressed endothelial nitric oxide synthase phosphorylation and reduced nitric oxide bioavailability, with increased reactive oxygen species and ICAM-1 expression. MHO EVs induced an intermediate phenotype with disrupted mitochondrial morphology, supporting a graded continuum of endothelial stress.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Conclusions<\/jats:title>\n                  <jats:p>MHO represents a biologically active intermediate state. Circulating EVs from MHO individuals convey molecular signals that impair endothelial and mitochondrial function, predisposing to vascular injury and progression toward MAO. EV-associated microRNAs are mechanistic mediators and candidate biomarkers of metabolic and vascular deterioration in obesity.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>CLINICAL PERSPECTIVE<\/jats:title>\n                  <jats:sec>\n                    <jats:title>What Is New?<\/jats:title>\n                    <jats:list list-type=\"bullet\">\n                      <jats:list-item>\n                        <jats:p>This study systematically investigated extracellular vesicles derived from metabolically healthy obese individuals to define direct vesicle effects on endothelial function using integrated omics coupled to functional outputs.<\/jats:p>\n                      <\/jats:list-item>\n                      <jats:list-item>\n                        <jats:p>Extracellular vesicles from metabolically healthy obesity convey a distinct molecular and biological signature that distinguishes lean and metabolically abnormal obesity.<\/jats:p>\n                      <\/jats:list-item>\n                      <jats:list-item>\n                        <jats:p>Metabolic health status, rather than obesity alone, drives extracellular vesicle-mediated endothelial nitric oxide signaling, oxidative stress, inflammation, and mitochondrial dynamics.<\/jats:p>\n                      <\/jats:list-item>\n                    <\/jats:list>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>What Are the Clinical Implications?<\/jats:title>\n                    <jats:list list-type=\"bullet\">\n                      <jats:list-item>\n                        <jats:p>These findings explain why some individuals with obesity exhibit preserved vascular function while others develop early endothelial dysfunction.<\/jats:p>\n                      <\/jats:list-item>\n                      <jats:list-item>\n                        <jats:p>Stratifying obesity by metabolic health status improves cardiovascular risk assessment beyond body mass index alone.<\/jats:p>\n                      <\/jats:list-item>\n                      <jats:list-item>\n                        <jats:p>Targeting extracellular vesicle signaling pathways represents a novel strategy to prevent metabolically healthy individuals from progressing to metabolically abnormal obesity.<\/jats:p>\n                      <\/jats:list-item>\n                    <\/jats:list>\n                  <\/jats:sec>\n                <\/jats:sec>","DOI":"10.64898\/2026.04.09.717593","type":"posted-content","created":{"date-parts":[[2026,4,14]],"date-time":"2026-04-14T01:15:18Z","timestamp":1776129318000},"source":"Crossref","is-referenced-by-count":0,"title":["Extracellular Vesicles From Metabolically Healthy Obesity Convey Distinct Molecular Signals That Initiate Endothelial Dysfunction: A Multi-Omics Study in Adults of African Ancestry"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5484-5875","authenticated-orcid":false,"given":"Malak","family":"Abbas","sequence":"first","affiliation":[{"name":"National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Camryn","family":"Bragg","sequence":"additional","affiliation":[{"name":"National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2476-481X","authenticated-orcid":false,"given":"Ahmed","family":"Gharib","sequence":"additional","affiliation":[{"name":"National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Abdel G.","family":"Elkahloun","sequence":"additional","affiliation":[{"name":"National Human Genome Research Institute, National Institutes of Health, Bethesda, MD, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4090-0391","authenticated-orcid":false,"given":"Merry","family":"Lindsey","sequence":"additional","affiliation":[{"name":"Department of Biomedical Sciences, School of Graduate Studies, Meharry Medical College, Nashville, TN, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1180-2792","authenticated-orcid":false,"given":"Amadou","family":"Gaye","sequence":"additional","affiliation":[{"name":"Department of Integrative Genomics and Epidemiology, School of Graduate Studies, Meharry Medical College, Nashville, TN, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2026041606251154000_2026.04.09.717593v1.1","doi-asserted-by":"publisher","DOI":"10.3390\/biomedicines13020502"},{"key":"2026041606251154000_2026.04.09.717593v1.2","doi-asserted-by":"crossref","first-page":"363","DOI":"10.1016\/j.gtc.2023.03.006","article-title":"Health Complications of Obesity: 224 Obesity-Associated Comorbidities from a Mechanistic Perspective","volume":"52","year":"2023","journal-title":"Gastroenterol Clin North Am"},{"key":"2026041606251154000_2026.04.09.717593v1.3","doi-asserted-by":"crossref","unstructured":"Wei D , Gonzalez-Marrachelli V , Melgarejo JD , et al. Cardiovascular risk of metabolically healthy obesity in two European populations. Cardiovasc Diabetol. 2023;22:82.","DOI":"10.1186\/s12933-023-01815-6"},{"key":"2026041606251154000_2026.04.09.717593v1.4","doi-asserted-by":"crossref","unstructured":"Bi J , Song L , Wang L , et al. Transitions in metabolic health status over time and risk of heart failure. Diabetes Metab. 2022;48:101266.","DOI":"10.1016\/j.diabet.2021.101266"},{"key":"2026041606251154000_2026.04.09.717593v1.5","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pmed.1003351"},{"key":"2026041606251154000_2026.04.09.717593v1.6","doi-asserted-by":"crossref","first-page":"677","DOI":"10.1161\/ATVBAHA.122.317526","article-title":"Different Metabolic Phenotypes of Obesity and Risk of Coronary Artery Calcium Progression","volume":"42","year":"2022","journal-title":"Arterioscler Thromb Vasc Biol"},{"key":"2026041606251154000_2026.04.09.717593v1.7","doi-asserted-by":"publisher","DOI":"10.1016\/j.jacc.2018.02.055"},{"key":"2026041606251154000_2026.04.09.717593v1.8","doi-asserted-by":"crossref","unstructured":"Bou Matar D , Zhra M , Nassar WK , et al. Adipose tissue dysfunction disrupts metabolic homeostasis. Front Endocrinol (Lausanne). 2025;16:1592683.","DOI":"10.3389\/fendo.2025.1592683"},{"key":"2026041606251154000_2026.04.09.717593v1.9","doi-asserted-by":"crossref","unstructured":"Han Y , Ye S , Liu B . Roles of extracellular vesicles derived from healthy and obese adipose tissue in inter-organ crosstalk. Front Endocrinol (Lausanne). 2024;15:1409000.","DOI":"10.3389\/fendo.2024.1409000"},{"key":"2026041606251154000_2026.04.09.717593v1.10","doi-asserted-by":"publisher","DOI":"10.1021\/acsnano.2c08774"},{"key":"2026041606251154000_2026.04.09.717593v1.11","doi-asserted-by":"crossref","first-page":"e12404","DOI":"10.1002\/jev2.12404","article-title":"Minimal information for studies of extracellular vesicles (MISEV2023)","volume":"13","year":"2024","journal-title":"J Extracell Vesicles"},{"key":"2026041606251154000_2026.04.09.717593v1.12","doi-asserted-by":"crossref","unstructured":"Bond ST , Calkin AC , Drew BG . Adipose-Derived Extracellular Vesicles: Systemic Messengers and Metabolic Regulators. Front Physiol. 2022;13:837001.","DOI":"10.3389\/fphys.2022.837001"},{"key":"2026041606251154000_2026.04.09.717593v1.13","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1016\/j.semcancer.2025.09.003","article-title":"Extracellular vesicles and the lipid messengers","volume":"116","year":"2025","journal-title":"Semin Cancer Biol"},{"key":"2026041606251154000_2026.04.09.717593v1.14","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2017.08.035"},{"key":"2026041606251154000_2026.04.09.717593v1.15","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1808855115"},{"key":"2026041606251154000_2026.04.09.717593v1.16","doi-asserted-by":"crossref","unstructured":"Kornmueller K , Amri EZ , Scheideler M , Prassl R . Delivery of miRNAs to the adipose organ for metabolic health. Adv Drug Deliv Rev. 2022;181:114110.","DOI":"10.1016\/j.addr.2021.114110"},{"key":"2026041606251154000_2026.04.09.717593v1.17","doi-asserted-by":"crossref","unstructured":"Thery C , Witwer KW , Aikawa E , et al. Minimal information for studies of extracellular vesicles 2018 (MISEV2018). J Extracell Vesicles. 2018;7:1535750.","DOI":"10.1080\/20013078.2018.1535750"},{"key":"2026041606251154000_2026.04.09.717593v1.18","doi-asserted-by":"crossref","unstructured":"Robinson MD , Oshlack A . A scaling normalization method for differential expression analysis of RNA-seq data. Genome Biol. 2010;11:R25.","DOI":"10.1186\/gb-2010-11-3-r25"},{"key":"2026041606251154000_2026.04.09.717593v1.19","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/btp616"},{"key":"2026041606251154000_2026.04.09.717593v1.20","doi-asserted-by":"crossref","unstructured":"Rovira-Llopis S , Diaz-Rua R , Grau-Del Valle C , et al. Characterization of Differentially Expressed Circulating miRNAs in Metabolically Healthy versus Unhealthy Obesity. Biomedicines. 2021;9:321.","DOI":"10.3390\/biomedicines9030321"},{"key":"2026041606251154000_2026.04.09.717593v1.21","doi-asserted-by":"publisher","DOI":"10.1016\/j.cmet.2021.08.006"},{"key":"2026041606251154000_2026.04.09.717593v1.22","doi-asserted-by":"publisher","DOI":"10.1001\/archinte.168.15.1617"},{"key":"2026041606251154000_2026.04.09.717593v1.23","unstructured":"Huang JS , Zhang XY , Ramakrishnan R , et al. Association of Childhood Obesity Phenotypes With Cardiometabolic Outcomes in Adulthood. Obes Rev. 2025:e70006."},{"key":"2026041606251154000_2026.04.09.717593v1.24","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0096820"},{"key":"2026041606251154000_2026.04.09.717593v1.25","doi-asserted-by":"crossref","unstructured":"Mendez-Garcia A , Garcia-Mendoza MA , Zarate-Peralta CP , et al. Mitochondrial microRNAs (mitomiRs) as emerging biomarkers and therapeutic targets. Front Genet. 2025;16:1555563.","DOI":"10.3389\/fgene.2025.1555563"},{"key":"2026041606251154000_2026.04.09.717593v1.26","doi-asserted-by":"crossref","unstructured":"Tripathi A , John AA , Kumar D , et al. MiR-539-3p impairs osteogenesis by suppressing Wnt interaction with LRP-6. Front Endocrinol (Lausanne). 2022;13:977347.","DOI":"10.3389\/fendo.2022.977347"},{"key":"2026041606251154000_2026.04.09.717593v1.27","doi-asserted-by":"crossref","unstructured":"Fontaine M , Herkenne S , Ek O , et al. Extracellular Vesicles Mediate Communication between Endothelial and Vascular Smooth Muscle Cells. Int J Mol Sci. 2021;23:331.","DOI":"10.3390\/ijms23010331"},{"key":"2026041606251154000_2026.04.09.717593v1.28","doi-asserted-by":"crossref","unstructured":"Pei CZ , Liu B , Li YT , et al. MicroRNA-126 protects against vascular injury. Stem Cell Res Ther. 2020;11:28.","DOI":"10.1186\/s13287-020-1554-9"},{"key":"2026041606251154000_2026.04.09.717593v1.29","doi-asserted-by":"crossref","unstructured":"Yu B , Jiang Y , Wang X , Wang S . An integrated hypothesis for miR-126 in vascular disease. Med Res Arch. 2020;8.","DOI":"10.18103\/mra.v8i5.2133"},{"key":"2026041606251154000_2026.04.09.717593v1.30","doi-asserted-by":"crossref","unstructured":"Ye ZQ , Meng XH , Fang X , et al. MiR-126 regulates mesenchymal stem cell vascular repair on carotid atherosclerosis. World J Stem Cells. 2025;17:106520.","DOI":"10.4252\/wjsc.v17.i6.106520"},{"key":"2026041606251154000_2026.04.09.717593v1.31","doi-asserted-by":"crossref","unstructured":"Guo B , Gu J , Zhuang T , et al. MicroRNA-126: From biology to therapeutics. Biomed Pharmacother. 2025;185:117953.","DOI":"10.1016\/j.biopha.2025.117953"},{"key":"2026041606251154000_2026.04.09.717593v1.32","doi-asserted-by":"crossref","first-page":"6117","DOI":"10.3390\/ijms25116117","article-title":"Extracellular Vesicles in Diabetic Cardiomyopathy","volume":"25","year":"2024","journal-title":"Int J Mol Sci"},{"key":"2026041606251154000_2026.04.09.717593v1.33","doi-asserted-by":"crossref","first-page":"5709","DOI":"10.2147\/IJN.S511842","article-title":"Adipose Tissue-Derived Extracellular Vesicles Loaded with miR-141-3p Regulate Obesity-Induced Insulin Resistance","volume":"20","year":"2025","journal-title":"Int J Nanomedicine"},{"key":"2026041606251154000_2026.04.09.717593v1.34","doi-asserted-by":"crossref","unstructured":"Ji Y , Chen H , Pang L , et al. AGE induced macrophage-derived exosomes induce endothelial dysfunction in diabetes via miR-22-5p\/FOXP1. Cardiovasc Diabetol. 2025;24:158.","DOI":"10.1186\/s12933-025-02715-7"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.64898\/2026.04.09.717593","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,4,16]],"date-time":"2026-04-16T13:25:35Z","timestamp":1776345935000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.64898\/2026.04.09.717593"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,4,13]]},"references-count":34,"URL":"https:\/\/doi.org\/10.64898\/2026.04.09.717593","relation":{},"subject":[],"published":{"date-parts":[[2026,4,13]]},"subtype":"preprint"}}