{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,17]],"date-time":"2026-10-17T17:57:25Z","timestamp":1792259845792,"version":"4.4.0"},"reference-count":58,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2021,7,5]],"date-time":"2021-07-05T00:00:00Z","timestamp":1625443200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100008798","name":"Ministry of Science Research and Technology","doi-asserted-by":"publisher","award":["2016YFA0102100"],"award-info":[{"award-number":["2016YFA0102100"]}],"id":[{"id":"10.13039\/501100008798","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100014857","name":"National Natural Science Foundation of China-Guangdong Joint Fund","doi-asserted-by":"publisher","award":["31730041, 81422009, 81401940, 81770560, 81800533, 81870430, 31671523"],"award-info":[{"award-number":["31730041, 81422009, 81401940, 81770560, 81800533, 81870430, 31671523"]}],"id":[{"id":"10.13039\/501100014857","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell Dev. Biol."],"abstract":"<jats:p>Tissue-specific endothelial cells are more than simply a barrier lining capillaries and are proved to be capable of remarkable plasticity to become active collagen matrix-producing myofibroblasts (MFs) in solid organs with fibrosis. Liver sinusoidal endothelial cells (LSECs) also participate in the development of hepatic fibrosis, but the exact roles and underlying mechanism have been poorly understood in addition to capillarization. In this study, we demonstrate, by using single-cell RNA sequencing, lineage tracing, and colocalization analysis, that fibrotic LSECs undergo partial endothelial mesenchymal transition (EndMT) with a subset of LSECs acquiring an MF-like phenotype. These phenotypic changes make LSECs substantial producers of extracellular matrix (ECM) preferentially deposited in liver sinusoids but not septal\/portal scars as demonstrated by immunofluorescence in animal models and patients with fibrosis\/cirrhosis, likely due to their limited migration. Bioinformatic analysis verifies that LSECs undergo successive phenotypic transitions from capillarization to mesenchymal-like cells in liver fibrosis. Furthermore, blockade of LSEC capillarization by using YC-1, a selective eNOS-sGC activator, effectively attenuates liver damage and fibrogenesis as well as mesenchymal features of LSECs, suggesting that capillarization of LSECs might be upstream to their mesenchymal transition during fibrosis. In conclusion, we report that capillarized LSECs undergo a partial EndMT characterized by increased ECM production without activating cell mobility, leading to perisinusoidal ECM deposition that aggravate liver function and fibrogenesis. Targeting this transitional process may be of great value for antifibrotic treatment of liver fibrosis.<\/jats:p>","DOI":"10.3389\/fcell.2021.671081","type":"journal-article","created":{"date-parts":[[2021,7,2]],"date-time":"2021-07-02T07:14:00Z","timestamp":1625210040000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":68,"title":["Capillarized Liver Sinusoidal Endothelial Cells Undergo Partial Endothelial-Mesenchymal Transition to Actively Deposit Sinusoidal ECM in Liver Fibrosis"],"prefix":"10.3389","volume":"9","author":[{"given":"Bai","family":"Ruan","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Juan-Li","family":"Duan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hao","family":"Xu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kai-Shan","family":"Tao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hua","family":"Han","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guo-Rui","family":"Dou","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lin","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2021,7,5]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.3390\/cells8091069","article-title":"Single-cell transcriptomes reveal characteristic features of mouse hepatocytes with liver cholestatic injury.","volume":"8","author":"Chang","year":"2019","journal-title":"Cells"},{"key":"B2","doi-asserted-by":"publisher","first-page":"156","DOI":"10.1016\/j.jhep.2014.07.035","article-title":"S100A4 promotes liver fibrosis via activation of hepatic stellate cells.","volume":"62","author":"Chen","year":"2015","journal-title":"J. Hepatol."},{"key":"B3","doi-asserted-by":"publisher","first-page":"9286","DOI":"10.1074\/jbc.M111040200","article-title":"Identification of the key regions within the mouse pro-alpha 2(I) collagen gene far-upstream enhancer.","volume":"277","author":"De Val","year":"2002","journal-title":"J. Biol. Chem."},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.1038\/ncomms14361","article-title":"The molecular basis of endothelial cell plasticity.","volume":"8","author":"Dejana","year":"2017","journal-title":"Nat. Commun."},{"key":"B5","doi-asserted-by":"publisher","first-page":"1740","DOI":"10.1002\/hep.27376","article-title":"Liver sinusoidal endothelial cells in hepatic fibrosis.","volume":"61","author":"DeLeve","year":"2015","journal-title":"Hepatology"},{"key":"B6","doi-asserted-by":"publisher","first-page":"920","DOI":"10.1002\/hep.22351","article-title":"Sinusoidal endothelial cells prevent rat stellate cell activation and promote reversion to quiescence.","volume":"48","author":"Deleve","year":"2008","journal-title":"Hepatology"},{"key":"B7","doi-asserted-by":"publisher","first-page":"97","DOI":"10.1038\/nature12681","article-title":"Divergent angiocrine signals from vascular niche balance liver regeneration and fibrosis.","volume":"505","author":"Ding","year":"2014","journal-title":"Nature"},{"key":"B8","doi-asserted-by":"publisher","first-page":"1832","DOI":"10.1016\/j.celrep.2019.10.024","article-title":"Single-cell transcriptomics uncovers zonation of function in the mesenchyme during liver fibrosis.","volume":"29","author":"Dobie","year":"2019","journal-title":"Cell Rep."},{"key":"B9","doi-asserted-by":"publisher","first-page":"677","DOI":"10.1002\/hep.29834","article-title":"Endothelial Notch activation reshapes the angiocrine of sinusoidal endothelia to aggravate liver fibrosis and blunt regeneration in mice.","volume":"68","author":"Duan","year":"2018","journal-title":"Hepatology"},{"key":"B10","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-017-01169-0","article-title":"Dynamic regulation of canonical TGFbeta signalling by endothelial transcription factor ERG protects from liver fibrogenesis.","volume":"8","author":"Dufton","year":"2017","journal-title":"Nat. Commun."},{"key":"B11","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20190103","article-title":"Transforming growth factor-beta in tissue fibrosis.","volume":"217","author":"Frangogiannis","year":"2020","journal-title":"J. Exp. Med."},{"key":"B12","doi-asserted-by":"publisher","first-page":"1099","DOI":"10.1172\/JCI90086","article-title":"GATA4-dependent organ-specific endothelial differentiation controls liver development and embryonic hematopoiesis.","volume":"127","author":"Geraud","year":"2017","journal-title":"J. Clin. Invest."},{"key":"B13","doi-asserted-by":"publisher","first-page":"3124","DOI":"10.1016\/j.jacc.2019.10.036","article-title":"Transition of macrophages to fibroblast-like cells in healing myocardial infarction.","volume":"74","author":"Haider","year":"2019","journal-title":"J. Am. Coll. Cardiol."},{"key":"B14","doi-asserted-by":"publisher","first-page":"1278","DOI":"10.1016\/j.jhep.2019.02.012","article-title":"Role of liver sinusoidal endothelial cells in non-alcoholic fatty liver disease.","volume":"70","author":"Hammoutene","year":"2019","journal-title":"J. Hepatol."},{"key":"B15","doi-asserted-by":"publisher","first-page":"161","DOI":"10.1165\/rcmb.2009-0031OC","article-title":"Endothelial-mesenchymal transition in bleomycin-induced pulmonary fibrosis.","volume":"43","author":"Hashimoto","year":"2010","journal-title":"Am. J. Respir. Cell Mol. Biol."},{"key":"B16","doi-asserted-by":"publisher","first-page":"E3297","DOI":"10.1073\/pnas.1400062111","article-title":"Origin of myofibroblasts in the fibrotic liver in mice.","volume":"111","author":"Iwaisako","year":"2014","journal-title":"Proc. Natl. Acad. Sci. U. S. A."},{"key":"B17","doi-asserted-by":"publisher","first-page":"107","DOI":"10.1053\/jhep.2003.50005","article-title":"Foxf1 +\/- mice exhibit defective stellate cell activation and abnormal liver regeneration following CCl4 injury.","volume":"37","author":"Kalinichenko","year":"2003","journal-title":"Hepatology"},{"key":"B18","doi-asserted-by":"publisher","first-page":"1039","DOI":"10.1002\/hep.28948","article-title":"The origin of fibrogenic myofibroblasts in fibrotic liver.","volume":"65","author":"Kisseleva","year":"2017","journal-title":"Hepatology"},{"key":"B19","doi-asserted-by":"publisher","first-page":"429","DOI":"10.1016\/j.jhep.2006.04.014","article-title":"Bone marrow-derived fibrocytes participate in pathogenesis of liver fibrosis.","volume":"45","author":"Kisseleva","year":"2006","journal-title":"J. Hepatol."},{"key":"B20","doi-asserted-by":"publisher","first-page":"G246","DOI":"10.1152\/ajpgi.00118.2016","article-title":"Angiocrine signaling in the hepatic sinusoids in health and disease.","volume":"311","author":"Kostallari","year":"2016","journal-title":"Am. J. Physiol. Gastrointest. Liver Physiol."},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.3390\/cells8050503","article-title":"Single cell RNA sequencing identifies subsets of hepatic stellate cells and myofibroblasts in liver fibrosis.","volume":"8","author":"Krenkel","year":"2019","journal-title":"Cells"},{"key":"B22","doi-asserted-by":"publisher","first-page":"445","DOI":"10.1038\/s41569-018-0023-y","article-title":"Reassessing endothelial-to-mesenchymal transition in cardiovascular diseases.","volume":"15","author":"Li","year":"2018","journal-title":"Nat. Rev. Cardiol."},{"key":"B23","doi-asserted-by":"publisher","first-page":"2324","DOI":"10.1073\/pnas.1214136110","article-title":"Mesothelial cells give rise to hepatic stellate cells and myofibroblasts via mesothelial-mesenchymal transition in liver injury.","volume":"110","author":"Li","year":"2013","journal-title":"Proc. Natl. Acad. Sci. U. S. A."},{"key":"B24","doi-asserted-by":"publisher","first-page":"1252","DOI":"10.1038\/nmat5024","article-title":"Mechanotransduction-modulated fibrotic microniches reveal the contribution of angiogenesis in liver fibrosis.","volume":"16","author":"Liu","year":"2017","journal-title":"Nat. Mater."},{"key":"B25","doi-asserted-by":"publisher","first-page":"10832","DOI":"10.1073\/pnas.1910650117","article-title":"Matrix-transmitted paratensile signaling enables myofibroblast-fibroblast cross talk in fibrosis expansion.","volume":"117","author":"Liu","year":"2020","journal-title":"Proc. Natl. Acad. Sci. U. S. A."},{"key":"B26","doi-asserted-by":"publisher","first-page":"128","DOI":"10.1038\/s41586-018-0522-3","article-title":"Mechanosensing by beta1 integrin induces angiocrine signals for liver growth and survival.","volume":"562","author":"Lorenz","year":"2018","journal-title":"Nature"},{"key":"B27","doi-asserted-by":"publisher","first-page":"1137","DOI":"10.1016\/j.jhep.2016.01.010","article-title":"Characterization of hepatic stellate cells, portal fibroblasts, and mesothelial cells in normal and fibrotic livers.","volume":"64","author":"Lua","year":"2016","journal-title":"J. Hepatol."},{"key":"B28","doi-asserted-by":"publisher","first-page":"1641","DOI":"10.1172\/JCI114886","article-title":"Extracellular matrix gene expression increases preferentially in rat lipocytes and sinusoidal endothelial cells during hepatic fibrosis in vivo.","volume":"86","author":"Maher","year":"1990","journal-title":"J. Clin. Invest."},{"key":"B29","doi-asserted-by":"publisher","first-page":"608","DOI":"10.1016\/j.jhep.2016.04.018","article-title":"Sinusoidal communication in liver fibrosis and regeneration.","volume":"65","author":"Marrone","year":"2016","journal-title":"J. Hepatol."},{"key":"B30","doi-asserted-by":"publisher","first-page":"305","DOI":"10.1038\/nprot.2015.017","article-title":"High-yield and high-purity isolation of hepatic stellate cells from normal and fibrotic mouse livers.","volume":"10","author":"Mederacke","year":"2015","journal-title":"Nat. Protoc."},{"key":"B31","doi-asserted-by":"publisher","DOI":"10.1038\/ncomms3823","article-title":"Fate tracing reveals hepatic stellate cells as dominant contributors to liver fibrosis independent of its aetiology.","volume":"4","author":"Mederacke","year":"2013","journal-title":"Nat. Commun."},{"key":"B32","doi-asserted-by":"publisher","first-page":"2380","DOI":"10.1172\/JCI66904","article-title":"Smoothened is a master regulator of adult liver repair.","volume":"123","author":"Michelotti","year":"2013","journal-title":"J. Clin. Invest."},{"key":"B33","doi-asserted-by":"publisher","first-page":"359","DOI":"10.1016\/j.jhep.2018.05.008","article-title":"Liver regeneration requires Yap1-TGFbeta-dependent epithelial-mesenchymal transition in hepatocytes.","volume":"69","author":"Oh","year":"2018","journal-title":"J. Hepatol."},{"key":"B34","doi-asserted-by":"publisher","first-page":"814","DOI":"10.1016\/j.cmet.2020.09.006","article-title":"Generation of a p16 reporter mouse and its use to characterize and target p16 (high) cells in vivo.","volume":"32","author":"Omori","year":"2020","journal-title":"Cell Metab."},{"key":"B35","doi-asserted-by":"publisher","first-page":"1281","DOI":"10.1152\/physrev.00021.2018","article-title":"Endothelial to mesenchymal transition: role in physiology and in the pathogenesis of human diseases.","volume":"99","author":"Piera-Velazquez","year":"2019","journal-title":"Physiol. Rev."},{"key":"B36","doi-asserted-by":"publisher","first-page":"212","DOI":"10.1016\/j.jhep.2016.07.009","article-title":"Liver sinusoidal endothelial cells: physiology and role in liver diseases.","volume":"66","author":"Poisson","year":"2017","journal-title":"J. Hepatol."},{"key":"B37","doi-asserted-by":"publisher","first-page":"316","DOI":"10.1038\/nature17040","article-title":"Angiocrine functions of organ-specific endothelial cells.","volume":"529","author":"Rafii","year":"2016","journal-title":"Nature"},{"key":"B38","doi-asserted-by":"publisher","first-page":"512","DOI":"10.1038\/s41586-019-1631-3","article-title":"Resolving the fibrotic niche of human liver cirrhosis at single-cell level.","volume":"575","author":"Ramachandran","year":"2019","journal-title":"Nature"},{"key":"B39","doi-asserted-by":"publisher","first-page":"457","DOI":"10.1038\/s41575-020-0304-x","article-title":"Single-cell technologies in hepatology: new insights into liver biology and disease pathogenesis.","volume":"17","author":"Ramachandran","year":"2020","journal-title":"Nat. Rev. Gastroenterol. Hepatol."},{"key":"B40","doi-asserted-by":"publisher","first-page":"G492","DOI":"10.1152\/ajpgi.00428.2016","article-title":"A small population of liver endothelial cells undergoes endothelial-to-mesenchymal transition in response to chronic liver injury.","volume":"313","author":"Ribera","year":"2017","journal-title":"Am. J. Physiol. Gastrointest. Liver Physiol."},{"key":"B41","doi-asserted-by":"publisher","first-page":"856","DOI":"10.1002\/hep.1840070511","article-title":"Sinusoidal endothelial cells from guinea pig liver synthesize and secrete cellular fibronectin in vitro.","volume":"7","author":"Rieder","year":"1987","journal-title":"Hepatology"},{"key":"B42","doi-asserted-by":"publisher","first-page":"7594","DOI":"10.1073\/pnas.102173299","article-title":"Scavenger endothelial cells of vertebrates: a nonperipheral leukocyte system for high-capacity elimination of waste macromolecules.","volume":"99","author":"Seternes","year":"2002","journal-title":"Proc. Natl. Acad. Sci. U. S. A."},{"key":"B43","doi-asserted-by":"publisher","first-page":"1888","DOI":"10.1016\/j.cell.2019.05.031","article-title":"Comprehensive integration of single-cell data.","volume":"177","author":"Stuart","year":"2019","journal-title":"Cell"},{"key":"B44","doi-asserted-by":"publisher","first-page":"1139","DOI":"10.1016\/j.jcmgh.2020.12.007","article-title":"Single-Cell transcriptomics reveals zone-specific alterations of liver sinusoidal endothelial cells in cirrhosis.","volume":"11","author":"Su","year":"2020","journal-title":"Cell. Mol. Gastroenterol. Hepatol."},{"key":"B45","doi-asserted-by":"publisher","first-page":"1333","DOI":"10.1136\/gutjnl-2013-305962","article-title":"Myofibroblastic cells function as progenitors to regenerate murine livers after partial hepatectomy.","volume":"63","author":"Swiderska-Syn","year":"2014","journal-title":"Gut"},{"key":"B46","doi-asserted-by":"publisher","DOI":"10.3390\/jcm5010009","article-title":"Controversies over the epithelial-to-mesenchymal transition in liver fibrosis.","volume":"5","author":"Taura","year":"2016","journal-title":"J. Clin. Med."},{"key":"B47","doi-asserted-by":"publisher","first-page":"2119","DOI":"10.1002\/hep.31215","article-title":"Transcriptional dynamics of hepatic sinusoid-associated cells after liver injury.","volume":"72","author":"Terkelsen","year":"2020","journal-title":"Hepatology"},{"key":"B48","doi-asserted-by":"publisher","first-page":"381","DOI":"10.1038\/nbt.2859","article-title":"The dynamics and regulators of cell fate decisions are revealed by pseudotemporal ordering of single cells.","volume":"32","author":"Trapnell","year":"2014","journal-title":"Nat. Biotechnol."},{"key":"B49","doi-asserted-by":"publisher","first-page":"2053","DOI":"10.1681\/ASN.2016050573","article-title":"Macrophage-to-myofibroblast transition contributes to interstitial fibrosis in chronic renal allograft injury.","volume":"28","author":"Wang","year":"2017","journal-title":"J. Am. Soc. Nephrol."},{"key":"B50","doi-asserted-by":"publisher","first-page":"380","DOI":"10.1016\/j.jhep.2020.08.033","article-title":"Endothelial GATA4 controls liver fibrosis and regeneration by preventing a pathogenic switch in angiocrine signaling.","volume":"74","author":"Winkler","year":"2021","journal-title":"J. Hepatol."},{"key":"B51","doi-asserted-by":"publisher","first-page":"281","DOI":"10.1016\/j.cels.2018.11.005","article-title":"Scrublet: computational identification of cell doublets in single-cell transcriptomic data.","volume":"8","author":"Wolock","year":"2019","journal-title":"Cell Syst."},{"key":"B52","doi-asserted-by":"publisher","first-page":"918","DOI":"10.1053\/j.gastro.2011.12.017","article-title":"Role of differentiation of liver sinusoidal endothelial cells in progression and regression of hepatic fibrosis in rats.","volume":"142","author":"Xie","year":"2012","journal-title":"Gastroenterology"},{"key":"B53","doi-asserted-by":"publisher","first-page":"644","DOI":"10.1016\/j.molcel.2019.07.028","article-title":"Landscape of intercellular crosstalk in healthy and NASH liver revealed by single-cell secretome gene analysis.","volume":"75","author":"Xiong","year":"2019","journal-title":"Mol. Cell."},{"key":"B54","doi-asserted-by":"publisher","DOI":"10.3389\/fphar.2014.00167","article-title":"The types of hepatic myofibroblasts contributing to liver fibrosis of different etiologies.","volume":"5","author":"Xu","year":"2014","journal-title":"Front. Pharmacol."},{"key":"B55","doi-asserted-by":"publisher","first-page":"267","DOI":"10.1053\/jhep.2003.50067","article-title":"DNase I-hypersensitive sites enhance alpha1(I) collagen gene expression in hepatic stellate cells.","volume":"37","author":"Yata","year":"2003","journal-title":"Hepatology"},{"key":"B56","doi-asserted-by":"publisher","first-page":"2282","DOI":"10.1681\/ASN.2008050513","article-title":"Fibroblasts in kidney fibrosis emerge via endothelial-to-mesenchymal transition.","volume":"19","author":"Zeisberg","year":"2008","journal-title":"J. Am. Soc. Nephrol."},{"key":"B57","doi-asserted-by":"publisher","first-page":"952","DOI":"10.1038\/nm1613","article-title":"Endothelial-to-mesenchymal transition contributes to cardiac fibrosis.","volume":"13","author":"Zeisberg","year":"2007","journal-title":"Nat. Med."},{"key":"B58","doi-asserted-by":"publisher","first-page":"23337","DOI":"10.1074\/jbc.M700194200","article-title":"Fibroblasts derive from hepatocytes in liver fibrosis via epithelial to mesenchymal transition.","volume":"282","author":"Zeisberg","year":"2007","journal-title":"J. Biol. Chem."}],"container-title":["Frontiers in Cell and Developmental Biology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcell.2021.671081\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,7,2]],"date-time":"2021-07-02T07:14:04Z","timestamp":1625210044000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcell.2021.671081\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,7,5]]},"references-count":58,"alternative-id":["10.3389\/fcell.2021.671081"],"URL":"https:\/\/doi.org\/10.3389\/fcell.2021.671081","relation":{},"ISSN":["2296-634X"],"issn-type":[{"value":"2296-634X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,7,5]]},"article-number":"671081"}}