{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,7]],"date-time":"2026-07-07T17:26:55Z","timestamp":1783445215768,"version":"3.54.6"},"reference-count":38,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2023,9,7]],"date-time":"2023-09-07T00:00:00Z","timestamp":1694044800000},"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. Cardiovasc. Med."],"abstract":"<jats:p>Peripheral arterial disease (PAD) is an age-related medical condition affecting mostly muscular arteries of the limb. It is the 3<jats:sup>rd<\/jats:sup> leading cause of atherosclerotic morbidity. The mechanical environment of endothelial cells (ECs) in PAD is characterized by disturbed blood flow (d-flow) and stiff extracellular matrices. In PAD, the stiffness of arteries is due to decreased elastin function and increased collagen content. These flow and stiffness parameters are largely missing from current models of PAD. It has been previously proven that ECs exposed to d-flow or stiff substrates lead to proatherogenic pathways, but the effect of both, d-flow and stiffness, on EC phenotype has not been fully investigated. In this study, we sought to explore the effect of sex on proatherogenic pathways that could result from exposing endothelial cells to a d-flow and stiff environment. We utilized the scRNA-seq tool to analyze the gene expression of ECs exposed to the different mechanical conditions both <jats:italic>in vitro<\/jats:italic> and <jats:italic>in vivo<\/jats:italic>. We found that male ECs exposed to different mechanical stimuli presented higher expression of genes related to fibrosis and d-flow <jats:italic>in vitro<\/jats:italic>. We validated our findings <jats:italic>in vivo<\/jats:italic> by exposing murine carotid arteries to d-flow via partial carotid artery ligation. Since women have delayed onset of arterial stiffening and subsequent PAD, this work may provide a framework for some of the pathways in which biological sex interacts with sex-based differences in PAD.<\/jats:p>","DOI":"10.3389\/fcvm.2023.1251141","type":"journal-article","created":{"date-parts":[[2023,9,7]],"date-time":"2023-09-07T11:11:25Z","timestamp":1694085085000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":9,"title":["Single-Cell RNA sequencing investigation of female-male differences under PAD conditions"],"prefix":"10.3389","volume":"10","author":[{"given":"Gloriani","family":"S\u00e1nchez Marrero","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicolas","family":"Villa-Roel","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Feifei","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christian","family":"Park","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dong-Won","family":"Kang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katherine E.","family":"Hekman","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hanjoong","family":"Jo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luke P.","family":"Brewster","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2023,9,7]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"884","DOI":"10.1001\/archinte.163.8.884","article-title":"Critical issues in peripheral arterial disease detection and management: a call to action","volume":"163","author":"Belch","year":"2003","journal-title":"Arch Intern Med"},{"key":"B2","doi-asserted-by":"publisher","first-page":"327","DOI":"10.1146\/annurev.pu.16.050195.001551","article-title":"The status of health assessment 1994","volume":"16","author":"Ware","year":"1995","journal-title":"Annu Rev Public Health"},{"key":"B3","doi-asserted-by":"publisher","first-page":"1509","DOI":"10.1161\/CIRCRESAHA.116.303849","article-title":"Epidemiology of peripheral artery disease","volume":"116","author":"Criqui","year":"2015","journal-title":"Circ Res"},{"key":"B4","doi-asserted-by":"publisher","first-page":"465","DOI":"10.1016\/0301-5629(86)90218-8","article-title":"Age-related changes in carotid artery wall properties in men","volume":"12","author":"Reneman","year":"1986","journal-title":"Ultrasound Med Biol"},{"key":"B5","doi-asserted-by":"publisher","first-page":"4208","DOI":"10.1111\/bph.14624","article-title":"Sex differences in mechanisms of arterial stiffness","volume":"176","author":"DuPont","year":"2019","journal-title":"Br J Pharmacol"},{"key":"B6","doi-asserted-by":"publisher","first-page":"496","DOI":"10.1161\/CIRCRESAHA.121.320702","article-title":"Sex differences in peripheral artery disease","volume":"130","author":"Pabon","year":"2022","journal-title":"Circ Res"},{"key":"B7","doi-asserted-by":"publisher","first-page":"H1535","DOI":"10.1152\/ajpheart.00510.2009","article-title":"Partial carotid ligation is a model of acutely induced disturbed flow, leading to rapid endothelial dysfunction and atherosclerosis","volume":"297","author":"Nam","year":"2009","journal-title":"Am J Physiol-Heart Circ Physiol"},{"key":"B8","doi-asserted-by":"publisher","first-page":"588","DOI":"10.1016\/j.ejvs.2005.06.011","article-title":"Are your arteries older than your age?","volume":"30","author":"Labropoulos","year":"2005","journal-title":"Eur J Vasc Endovasc Surg"},{"key":"B9","doi-asserted-by":"publisher","first-page":"1217","DOI":"10.1161\/CIRCULATIONAHA.116.026361","article-title":"Disturbed flow promotes arterial stiffening through thrombospondin-1","volume":"136","author":"Kim","year":"2017","journal-title":"Circulation"},{"key":"B10","doi-asserted-by":"publisher","first-page":"1620","DOI":"10.3390\/ma3031620","article-title":"Biophysical cueing and vascular endothelial cell behavior","volume":"3","author":"Wood","year":"2010","journal-title":"Materials (Basel)"},{"key":"B11","doi-asserted-by":"publisher","first-page":"108491","DOI":"10.1016\/j.celrep.2020.108491","article-title":"Endothelial reprogramming by disturbed flow revealed by single-cell RNA and chromatin accessibility study","volume":"33","author":"Andueza","year":"2020","journal-title":"Cell Rep"},{"key":"B12","doi-asserted-by":"publisher","first-page":"203","DOI":"10.1177\/1358863X11400935","article-title":"Arterial dysfunction and functional performance in patients with peripheral artery disease: a review","volume":"16","author":"Coutinho","year":"2011","journal-title":"Vasc Med"},{"key":"B13","doi-asserted-by":"publisher","first-page":"551","DOI":"10.1161\/ATVBAHA.110.220624","article-title":"Thrombospondin-1 is a plasmatic marker of peripheral arterial disease that modulates endothelial progenitor cell angiogenic properties","volume":"31","author":"Smadja","year":"2011","journal-title":"Arterioscler Thromb Vasc Biol"},{"key":"B14","doi-asserted-by":"publisher","first-page":"32","DOI":"10.1161\/CIRCRESAHA.115.303883","article-title":"Hemodynamic disturbed flow induces differential DNA methylation of endothelial kruppel-like factor 4 promoter in vitro and in vivo","volume":"115","author":"Jiang","year":"2014","journal-title":"Circ Res"},{"key":"B15","doi-asserted-by":"publisher","first-page":"3187","DOI":"10.1172\/JCI74792","article-title":"Flow-dependent epigenetic DNA methylation regulates endothelial gene expression and atherosclerosis","volume":"124","author":"Dunn","year":"2014","journal-title":"J Clin Invest"},{"key":"B16","doi-asserted-by":"publisher","first-page":"2203069","DOI":"10.1002\/adfm.202203069","article-title":"Single-cell transcriptomic census of endothelial changes induced by matrix stiffness and the association with atherosclerosis","volume":"32","author":"Zamani","year":"2022","journal-title":"Adv Funct Mater"},{"key":"B17","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1186\/2042-6410-2-11","article-title":"Reporting of sex as a variable in cardiovascular studies using cultured cells","volume":"2","author":"Taylor","year":"2011","journal-title":"Biol Sex Differ"},{"key":"B18","doi-asserted-by":"publisher","first-page":"1388","DOI":"10.1016\/j.jacc.2021.10.050","article-title":"Sex as a biological variable in cardiovascular diseases: JACC focus seminar 1\/7","volume":"79","author":"Clayton","year":"2022","journal-title":"J Am Coll Cardiol"},{"key":"B19","doi-asserted-by":"publisher","first-page":"2100735","DOI":"10.1002\/adhm.202100735","article-title":"Sex-specific response to combinations of shear stress and substrate stiffness by endothelial cells in vitro","volume":"10","author":"James","year":"2021","journal-title":"Adv Healthcare Mater"},{"key":"B20","doi-asserted-by":"publisher","first-page":"784","DOI":"10.1016\/j.jbiomech.2014.01.018","article-title":"Compressive mechanical properties of atherosclerotic plaques\u2014indentation test to characterise the local anisotropic behaviour","volume":"47","author":"Chai","year":"2014","journal-title":"J Biomech"},{"key":"B21","doi-asserted-by":"publisher","first-page":"1028","DOI":"10.1002\/jbm.a.32321","article-title":"Nanomechanical properties of calcification, fibrous tissue, and hematoma from atherosclerotic plaques","volume":"91","author":"Ebenstein","year":"2009","journal-title":"J Biomed Mater Res Part A"},{"key":"B22","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1186\/s13036-018-0107-6","article-title":"A novel method for segmenting growth of cells in sheared endothelial culture reveals the secretion of an anti-inflammatory mediator","volume":"12","author":"Ghim","year":"2018","journal-title":"J Biol Eng"},{"key":"B23","doi-asserted-by":"publisher","DOI":"10.3791\/61817","article-title":"Segmenting growth of endothelial cells in 6-well plates on an orbital shaker for mechanobiological studies","volume":"172","author":"Pang","year":"2021","journal-title":"J Vis Exp"},{"key":"B24","doi-asserted-by":"publisher","first-page":"111","DOI":"10.1016\/j.abb.2015.11.005","article-title":"The role of endothelial mechanosensitive genes in atherosclerosis and omics approaches","volume":"591","author":"Simmons","year":"2016","journal-title":"Arch Biochem Biophys"},{"key":"B25","doi-asserted-by":"publisher","first-page":"e1861","DOI":"10.3791\/1861","article-title":"A model of disturbed flow-induced atherosclerosis in mouse carotid artery by partial ligation and a simple method of RNA isolation from carotid endothelium","volume":"40","author":"Nam","year":"2010","journal-title":"J Vis Exp"},{"key":"B26","doi-asserted-by":"publisher","first-page":"513","DOI":"10.1161\/CIRCULATIONAHA.121.054108","article-title":"Genes that escape X chromosome inactivation modulate sex differences in valve myofibroblasts","volume":"145","author":"Aguado","year":"2022","journal-title":"Circulation"},{"key":"B27","doi-asserted-by":"publisher","first-page":"244","DOI":"10.1038\/nature24265","article-title":"Landscape of X chromosome inactivation across human tissues","volume":"550","author":"Tukiainen","year":"2017","journal-title":"Nature"},{"key":"B28","doi-asserted-by":"publisher","first-page":"1280","DOI":"10.1038\/s41591-019-0512-5","article-title":"Atheroprotective roles of smooth muscle cell phenotypic modulation and the TCF21 disease gene as revealed by single-cell analysis","volume":"25","author":"Wirka","year":"2019","journal-title":"Nat Med"},{"key":"B29","author":"Dong","year":""},{"key":"B30","doi-asserted-by":"publisher","first-page":"1041","DOI":"10.1007\/s10237-014-0553-0","article-title":"Differential mechanical response and microstructural organization between non-human primate femoral and carotid arteries","volume":"13","author":"Wang","year":"2014","journal-title":"Biomech Model Mechanobiol"},{"key":"B31","doi-asserted-by":"publisher","first-page":"011010","DOI":"10.1115\/1.4035262","article-title":"A novel approach to assess the in situ versus ex vivo mechanical behaviors of the coronary artery","volume":"139","author":"Wang","year":"2017","journal-title":"J Biomech Eng"},{"key":"B32","doi-asserted-by":"publisher","first-page":"660","DOI":"10.1152\/physiolgenomics.00025.2016","article-title":"Deep transcriptomic profiling reveals the similarity between endothelial cells cultured under static and oscillatory shear stress conditions","volume":"48","author":"Qiao","year":"2016","journal-title":"Physiol Genomics"},{"key":"B33","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.vph.2015.11.083","article-title":"Arterial stiffness, atherosclerosis and cardiovascular risk: pathophysiologic mechanisms and emerging clinical indications","volume":"77","author":"Palombo","year":"2016","journal-title":"Vasc Pharmacol"},{"key":"B34","doi-asserted-by":"publisher","first-page":"226","DOI":"10.1016\/j.atherosclerosis.2016.04.014","article-title":"Is increased arterial stiffness a cause or consequence of atherosclerosis?","volume":"249","author":"Hansen","year":"2016","journal-title":"Atherosclerosis"},{"key":"B35","doi-asserted-by":"publisher","first-page":"912","DOI":"10.1038\/s42255-019-0102-3","article-title":"Endothelial TGF-\u03b2 signalling drives vascular inflammation and atherosclerosis","volume":"1","author":"Chen","year":"2019","journal-title":"Nat Metabol"},{"key":"B36","doi-asserted-by":"publisher","first-page":"582","DOI":"10.1016\/j.ajpath.2022.01.005","article-title":"The impact of sex chromosomes in the sexual dimorphism of pulmonary arterial hypertension","volume":"192","author":"Predescu","year":"2022","journal-title":"Am J Pathol"},{"key":"B37","doi-asserted-by":"publisher","first-page":"375","DOI":"10.1093\/cvr\/cvu064","article-title":"The number of X chromosomes influences protection from cardiac ischaemia\/reperfusion injury in mice: one X is better than two","volume":"102","author":"Li","year":"2014","journal-title":"Cardiovasc Res"},{"key":"B38","doi-asserted-by":"publisher","first-page":"1275","DOI":"10.1161\/HYPERTENSIONAHA.109.144949","article-title":"Sex chromosome effects unmasked in angiotensin II\u2013induced hypertension","volume":"55","author":"Ji","year":"2010","journal-title":"Hypertension"}],"container-title":["Frontiers in Cardiovascular Medicine"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcvm.2023.1251141\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,9,7]],"date-time":"2023-09-07T11:11:27Z","timestamp":1694085087000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcvm.2023.1251141\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,9,7]]},"references-count":38,"alternative-id":["10.3389\/fcvm.2023.1251141"],"URL":"https:\/\/doi.org\/10.3389\/fcvm.2023.1251141","relation":{},"ISSN":["2297-055X"],"issn-type":[{"value":"2297-055X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,9,7]]},"article-number":"1251141"}}