{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T04:08:18Z","timestamp":1790654898684,"version":"4.1.0"},"reference-count":35,"publisher":"Public Library of Science (PLoS)","issue":"2","license":[{"start":{"date-parts":[[2024,2,16]],"date-time":"2024-02-16T00:00:00Z","timestamp":1708041600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100007305","name":"Maternal and Child Health Bureau","doi-asserted-by":"publisher","award":["5 H30 MC00008-20-00"],"award-info":[{"award-number":["5 H30 MC00008-20-00"]}],"id":[{"id":"10.13039\/100007305","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000050","name":"National Heart, Lung, and Blood Institute","doi-asserted-by":"publisher","award":["P01 HL144457"],"award-info":[{"award-number":["P01 HL144457"]}],"id":[{"id":"10.13039\/100000050","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000019","name":"National Hemophilia Foundation","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100000019","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Children's Hospital Colorado Foundation","award":["Tanabe Bobrow Fund"],"award-info":[{"award-number":["Tanabe Bobrow Fund"]}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>Alterations in von Willebrand factor (VWF) have an important role in human health and disease. Deficiency of VWF is associated with symptoms of bleeding and excesses of VWF are associated with thrombotic outcomes. Understanding the mechanisms that drive VWF regulation can lead to a better understanding of modulation of VWF levels in humans. We identified clusterin (<jats:italic>CLU<\/jats:italic>) as a potential candidate regulator of VWF based on a single cell RNA sequencing (scRNA-seq) analysis in control endothelial cells (ECs) and von Willebrand disease (VWD) endothelial colony-forming-cells (ECFCs). We found that patients with deficiencies of VWF (von Willebrand disease, VWD) had decreased <jats:italic>CLU<\/jats:italic> expression and ECs with low <jats:italic>VWF<\/jats:italic> expression also had low <jats:italic>CLU<\/jats:italic> expression. Based on these findings, we sought to evaluate the role of <jats:italic>CLU<\/jats:italic> in the regulation of VWF, specifically as it relates to VWD. As <jats:italic>CLU<\/jats:italic> is primarily thought to be a golgi protein involved in protein chaperoning, we hypothesized that knockdown of <jats:italic>CLU<\/jats:italic> would lead to decreases in VWF and alterations in Weibel-Palade bodies (WPBs). We used both siRNA- and CRISPR-Cas9-based approaches to modulate <jats:italic>CLU<\/jats:italic> in human umbilical vein endothelial cells (HUVECs) and evaluated VWF protein levels, <jats:italic>VWF<\/jats:italic> mRNA copy number, and WPB quantity and size. We demonstrated that siRNA-based knockdown of <jats:italic>CLU<\/jats:italic> resulted in decreases in VWF content in cellular lysates and supernatants, but no significant change in WPB quantity or size. A CRISPR-Cas9-based knockdown of <jats:italic>CLU<\/jats:italic> demonstrated similar findings of decreases in intracellular VWF content but no significant change in WPB quantity or size. Our data suggests that <jats:italic>CLU<\/jats:italic> knockdown is associated with decreases in cellular VWF content but does not affect <jats:italic>VWF<\/jats:italic> mRNA levels or WPB quantity or size.<\/jats:p>","DOI":"10.1371\/journal.pone.0298133","type":"journal-article","created":{"date-parts":[[2024,2,16]],"date-time":"2024-02-16T18:24:20Z","timestamp":1708107860000},"page":"e0298133","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":6,"title":["Clusterin knockdown has effects on intracellular and secreted von Willebrand factor in human umbilical vein endothelial cells"],"prefix":"10.1371","volume":"19","author":[{"given":"Allaura A.","family":"Cox","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alice","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8056-6116","authenticated-orcid":true,"given":"Christopher J.","family":"Ng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2024,2,16]]},"reference":[{"key":"pone.0298133.ref001","doi-asserted-by":"crossref","first-page":"3697","DOI":"10.1182\/blood-2014-02-554501","article-title":"Murine coagulation factor VIII is synthesized in endothelial cells","volume":"123","author":"LA Everett","year":"2014","journal-title":"Blood"},{"key":"pone.0298133.ref002","doi-asserted-by":"crossref","DOI":"10.1111\/jth.14634","article-title":"Human endothelial cells size\u2010select their secretory granules for exocytosis to modulate their 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