{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,6,20]],"date-time":"2026-06-20T04:50:55Z","timestamp":1781931055668,"version":"3.54.5"},"posted":{"date-parts":[[2025,11,19]]},"group-title":"Pathology","reference-count":52,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,11,19]],"date-time":"2025-11-19T00:00:00Z","timestamp":1763510400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"crossref","award":["R35HL155649"],"award-info":[{"award-number":["R35HL155649"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"crossref","award":["TL1TR001997"],"award-info":[{"award-number":["TL1TR001997"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"crossref","award":["R01HL078871"],"award-info":[{"award-number":["R01HL078871"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"crossref","award":["R01HL163870"],"award-info":[{"award-number":["R01HL163870"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000968","name":"American Heart Association","doi-asserted-by":"crossref","award":["23MERIT1036341"],"award-info":[{"award-number":["23MERIT1036341"]}],"id":[{"id":"10.13039\/100000968","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,6,17]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:sec>\n                  <jats:title>Aims<\/jats:title>\n                  <jats:p>Plasminogen activator inhibitor-1 (PAI-1) regulates plasmin-mediated proteolysis, thereby influencing vascular stability and tissue remodeling. Angiotensin II (AngII) induces an increase in PAI-1 during the development of ascending thoracic aortic aneurysm (ATAA). The initial purpose of this study was to determine whether deletion of PAI-1 influenced development of ATAA. Subsequently, this study aimed to define the early pathological events preceding cardiac fibrosis in PAI-1 deficiency and the structural domain responsible for its protective effect.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Methods and results<\/jats:title>\n                  <jats:p>\n                    AngII was infused for 4 weeks in whole-body PAI-1 deficient (PAI-1\u2212\/\u2212) mice and their wild-type littermates (PAI-1+\/+) to examine the role of PAI-1 in ATAA. PAI-1 deficiency did not alter AngII-induced aortopathy but revealed a striking cardiac phenotype characterized by replacement fibrosis predominantly within the epicardium and posterior septum. Ferric iron, indicative of prior hemorrhage, was coincident with fibrosis. Similar phenotypes were observed in PAI-1\u2212\/\u2212 mice infused with norepinephrine for 4 weeks. To define the pathological events preceding cardiac fibrosis, either AngII or norepinephrine was infused for 1 week in PAI-1+\/+ or \u2212\/\u2212 mice. Both infusions induced extensive epicardial hemorrhage and posterior septal fibrosis in PAI-1\u2212\/\u2212 mice. To explore the initiation of cardiac pathology, AngII was infused for approximately 1 day. PAI-1\u2212\/\u2212 mice developed diffuse hemorrhage and cardiomyocyte injury localized to the posterior septum, pathologic changes that preceded overt fibrosis. Finally, to determine the protective domain of PAI-1, saline or AngII was administered to mice harboring loss-of-function point mutations in the protease inhibitory (PAI-1\n                    <jats:sup>Ala\/Ala<\/jats:sup>\n                    ) or somatomedin B-binding domains (PAI-1\n                    <jats:sup>AK\/AK<\/jats:sup>\n                    ). Compared to saline infusion, 1 week of AngII induced hemorrhage and heterogeneous fibrosis in PAI-1\n                    <jats:sup>Ala\/Ala<\/jats:sup>\n                    , but not PAI-1\n                    <jats:sup>AK\/AK<\/jats:sup>\n                    mice.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Conclusions<\/jats:title>\n                  <jats:p>These findings support that, under hemodynamic stress, PAI-1 deficiency promotes early cardiac hemorrhage and cardiomyocyte injury that lead to fibrosis. Mutational studies implicate dysregulated plasmin generation as an initiator of cardiac injury and fibrosis.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>TRANSLATIONAL PERSPECTIVE<\/jats:title>\n                  <jats:p>Cardiac fibrosis has been reported in a human population with PAI-1 deficiency and currently lacks targeted therapy. Our findings demonstrate that in animal models, PAI-1 deficiency confers susceptibility to cardiac injury in response to hemodynamic stress, which may accelerate fibrotic remodeling. Mutational disruption of the protease-inhibitory domain of PAI-1 induced similar pathology, supporting a protective role for this function. These observations suggest that interventions aimed at controlling hypertension, promoting endothelial integrity, or regulating plasmin activation could reduce fibrotic remodeling in this population.<\/jats:p>\n                <\/jats:sec>","DOI":"10.1101\/2025.11.19.689269","type":"posted-content","created":{"date-parts":[[2025,11,20]],"date-time":"2025-11-20T01:05:39Z","timestamp":1763600739000},"source":"Crossref","is-referenced-by-count":2,"title":["Cardiac Hemorrhage Precedes Hypertension-induced Fibrosis in Plasminogen Activator Inhibitor-1 Deficient Mice"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6433-2464","authenticated-orcid":false,"given":"Alex C.","family":"Pettey","sequence":"first","affiliation":[{"name":"Department of Physiology"},{"name":"Saha Cardiovascular Research Center"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1374-2946","authenticated-orcid":false,"given":"Sohei","family":"Ito","sequence":"additional","affiliation":[{"name":"Saha Cardiovascular Research Center"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael K.","family":"Franklin","sequence":"additional","affiliation":[{"name":"Saha Cardiovascular Research Center"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Deborah A.","family":"Howatt","sequence":"additional","affiliation":[{"name":"Saha Cardiovascular Research Center"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jessica J.","family":"Moorleghen","sequence":"additional","affiliation":[{"name":"Saha Cardiovascular Research Center"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bryana M.","family":"Levitan","sequence":"additional","affiliation":[{"name":"Department of Physiology"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David B.","family":"Graf","sequence":"additional","affiliation":[{"name":"Saha Cardiovascular Research Center"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Valerie Z.","family":"Guzm\u00e1n","sequence":"additional","affiliation":[{"name":"Saha Cardiovascular Research Center"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nancy","family":"Zhang","sequence":"additional","affiliation":[{"name":"Saha Cardiovascular Research Center"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3126-1935","authenticated-orcid":false,"given":"Daniel A.","family":"Lawrence","sequence":"additional","affiliation":[{"name":"Division of Cardiovascular Medicine, Department of Internal Medicine"},{"name":"Department of Molecular and Integrative Physiology"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-9920-9328","authenticated-orcid":false,"given":"Thomas H.","family":"Sisson","sequence":"additional","affiliation":[{"name":"Division of Pulmonary and Critical Care Medicine, Department of Internal Medicine University of Michigan, Ann Arbor, MI"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0017-6236","authenticated-orcid":false,"given":"Hisashi","family":"Sawada","sequence":"additional","affiliation":[{"name":"Department of Physiology"},{"name":"Saha Cardiovascular Research Center"},{"name":"Saha Aortic Center University of Kentucky, Lexington, KY"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jeffrey E.","family":"Saffitz","sequence":"additional","affiliation":[{"name":"Department of Pathology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0577-2558","authenticated-orcid":false,"given":"Hong S.","family":"Lu","sequence":"additional","affiliation":[{"name":"Department of Physiology"},{"name":"Saha Cardiovascular Research Center"},{"name":"Saha Aortic Center University of Kentucky, Lexington, KY"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2093-3775","authenticated-orcid":false,"given":"Alan","family":"Daugherty","sequence":"additional","affiliation":[{"name":"Department of Physiology"},{"name":"Saha Cardiovascular Research Center"},{"name":"Saha Aortic Center University of Kentucky, Lexington, KY"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026061921101063000_2025.11.19.689269v2.1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jacc.2020.11.010"},{"key":"2026061921101063000_2025.11.19.689269v2.2","doi-asserted-by":"publisher","DOI":"10.1161\/ATVBAHA.118.310956"},{"key":"2026061921101063000_2025.11.19.689269v2.3","doi-asserted-by":"crossref","first-page":"226","DOI":"10.31083\/j.rcm2408226","article-title":"A review of therapeutic strategies against cardiac fibrosis: from classical pharmacology to novel molecular, epigenetic, and biotechnological approaches","volume":"24","year":"2023","journal-title":"Rev Cardiovasc Med"},{"key":"2026061921101063000_2025.11.19.689269v2.4","doi-asserted-by":"publisher","DOI":"10.2174\/138945007781662337"},{"key":"2026061921101063000_2025.11.19.689269v2.5","doi-asserted-by":"crossref","first-page":"622473","DOI":"10.3389\/fcvm.2020.622473","article-title":"Targeting PAI-1 in cardiovascular disease: Structural insights into PAI-1 functionality and inhibition","volume":"7","year":"2020","journal-title":"Front Cardiovasc Med"},{"key":"2026061921101063000_2025.11.19.689269v2.6","doi-asserted-by":"publisher","DOI":"10.1002\/jcp.22783"},{"key":"2026061921101063000_2025.11.19.689269v2.7","doi-asserted-by":"publisher","DOI":"10.1161\/01.RES.0000141427.61023.f4"},{"key":"2026061921101063000_2025.11.19.689269v2.8","doi-asserted-by":"crossref","first-page":"365","DOI":"10.1161\/01.ATV.0000152356.85791.52","article-title":"Pharmacological inhibition and genetic deficiency of plasminogen activator inhibitor-1 attenuates angiotensin II\/salt-induced aortic remodeling","volume":"25","year":"2005","journal-title":"Arterioscler Thromb Vasc Biol"},{"key":"2026061921101063000_2025.11.19.689269v2.9","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2009-09-244962"},{"key":"2026061921101063000_2025.11.19.689269v2.10","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCULATIONAHA.110.955245"},{"key":"2026061921101063000_2025.11.19.689269v2.11","doi-asserted-by":"publisher","DOI":"10.1097\/HJH.0b013e32834840e8"},{"key":"2026061921101063000_2025.11.19.689269v2.12","doi-asserted-by":"crossref","first-page":"365","DOI":"10.1038\/s41598-017-00418-y","article-title":"Plasminogen activator inhibitor-1 protects mice against cardiac fibrosis by inhibiting urokinase-type plasminogen activator-mediated plasminogen activation","volume":"7","year":"2017","journal-title":"Scientific Reports"},{"key":"2026061921101063000_2025.11.19.689269v2.13","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCULATIONAHA.117.028145"},{"key":"2026061921101063000_2025.11.19.689269v2.14","doi-asserted-by":"publisher","DOI":"10.1161\/01.CIR.98.3.249"},{"key":"2026061921101063000_2025.11.19.689269v2.15","doi-asserted-by":"publisher","DOI":"10.1038\/sj.gt.3303069"},{"key":"2026061921101063000_2025.11.19.689269v2.16","doi-asserted-by":"publisher","DOI":"10.1152\/ajpheart.00620.2011"},{"key":"2026061921101063000_2025.11.19.689269v2.17","doi-asserted-by":"crossref","first-page":"e70562","DOI":"10.1096\/fj.202403133RR","article-title":"Plasminogen activator inhibitor 1 controls abdominal aortic aneurysm formation via the modulation of TGF-beta\/Smad2\/3 signaling in mice","volume":"39","year":"2025","journal-title":"FASEB J"},{"key":"2026061921101063000_2025.11.19.689269v2.18","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2010-12-324574"},{"key":"2026061921101063000_2025.11.19.689269v2.19","doi-asserted-by":"crossref","first-page":"e186131","DOI":"10.1172\/jci.insight.186131","article-title":"PAI-1 interaction with sortilin-related receptor 1 is required for lung fibrosis","volume":"10","year":"2025","journal-title":"JCI Insight"},{"key":"2026061921101063000_2025.11.19.689269v2.20","doi-asserted-by":"publisher","DOI":"10.3390\/ijms22052721"},{"key":"2026061921101063000_2025.11.19.689269v2.21","doi-asserted-by":"crossref","unstructured":"Liu Y , Saunders TL , Sisson T , Blackburn R , Ginsberg DS , Day D. Optimizing CRISPR\/Cas9 system to precisely model plasminogen activator inhibitor-1 point mutations in mice. bioRxiv 2018:254805.","DOI":"10.1101\/254805"},{"key":"2026061921101063000_2025.11.19.689269v2.22","doi-asserted-by":"publisher","DOI":"10.1113\/JP280389"},{"key":"2026061921101063000_2025.11.19.689269v2.23","doi-asserted-by":"publisher","DOI":"10.3791\/59013"},{"key":"2026061921101063000_2025.11.19.689269v2.24","doi-asserted-by":"publisher","DOI":"10.1093\/cvr\/cvaa110"},{"key":"2026061921101063000_2025.11.19.689269v2.25","doi-asserted-by":"publisher","DOI":"10.3390\/biom12020339"},{"key":"2026061921101063000_2025.11.19.689269v2.26","doi-asserted-by":"publisher","DOI":"10.1161\/ATVBAHA.121.315983"},{"key":"2026061921101063000_2025.11.19.689269v2.27","unstructured":"Team RC. R: A language and environment for statistical computing. R Foundation for Statistical Computing 2021: https:\/\/www.R-project.org\/."},{"key":"2026061921101063000_2025.11.19.689269v2.28","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/btp616"},{"key":"2026061921101063000_2025.11.19.689269v2.29","unstructured":"Korotkevich G , Sukhov V , Budin N , Shpak B , Artyomov MN , Sergushichev A. Fast gene set enrichment analysis. bioRxiv 2019."},{"key":"2026061921101063000_2025.11.19.689269v2.30","doi-asserted-by":"crossref","unstructured":"Dolgalev I. msigdbr: MSigDB gene sets for multiple organisms in a tidy data format. R package version 2022;7.","DOI":"10.32614\/CRAN.package.babelgene"},{"key":"2026061921101063000_2025.11.19.689269v2.31","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0294236"},{"key":"2026061921101063000_2025.11.19.689269v2.32","doi-asserted-by":"publisher","DOI":"10.1016\/j.ajpath.2014.05.014"},{"key":"2026061921101063000_2025.11.19.689269v2.33","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCULATIONAHA.121.058173"},{"key":"2026061921101063000_2025.11.19.689269v2.34","doi-asserted-by":"publisher","DOI":"10.1097\/01.ASN.0000035087.11758.ED"},{"key":"2026061921101063000_2025.11.19.689269v2.35","doi-asserted-by":"publisher","DOI":"10.1161\/CIRCRESAHA.109.212837"},{"key":"2026061921101063000_2025.11.19.689269v2.36","doi-asserted-by":"publisher","DOI":"10.3389\/fphys.2014.00396"},{"key":"2026061921101063000_2025.11.19.689269v2.37","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.270.43.25309"},{"key":"2026061921101063000_2025.11.19.689269v2.38","doi-asserted-by":"publisher","DOI":"10.1161\/hc3301.092803"},{"key":"2026061921101063000_2025.11.19.689269v2.39","doi-asserted-by":"publisher","DOI":"10.1006\/jmcc.2002.2001"},{"key":"2026061921101063000_2025.11.19.689269v2.40","doi-asserted-by":"publisher","DOI":"10.1111\/bph.15650"},{"key":"2026061921101063000_2025.11.19.689269v2.41","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2008-07-169003"},{"key":"2026061921101063000_2025.11.19.689269v2.42","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.242501899"},{"key":"2026061921101063000_2025.11.19.689269v2.43","doi-asserted-by":"publisher","DOI":"10.1002\/path.2454"},{"key":"2026061921101063000_2025.11.19.689269v2.44","doi-asserted-by":"publisher","DOI":"10.1111\/j.1538-7836.2007.02649.x"},{"key":"2026061921101063000_2025.11.19.689269v2.45","doi-asserted-by":"publisher","DOI":"10.1111\/j.1538-7836.2007.02849.x"},{"key":"2026061921101063000_2025.11.19.689269v2.46","doi-asserted-by":"crossref","first-page":"110555","DOI":"10.1016\/j.cellsig.2022.110555","article-title":"Cardiomyocyte PAI-1 influences the cardiac transcriptome and limits the extent of cardiac fibrosis in response to left ventricular pressure overload","volume":"104","year":"2023","journal-title":"Cell Signal"},{"key":"2026061921101063000_2025.11.19.689269v2.47","doi-asserted-by":"publisher","DOI":"10.1155\/2011\/562481"},{"key":"2026061921101063000_2025.11.19.689269v2.48","doi-asserted-by":"publisher","DOI":"10.1161\/ATVBAHA.114.304554"},{"key":"2026061921101063000_2025.11.19.689269v2.49","doi-asserted-by":"crossref","first-page":"633","DOI":"10.1038\/labinvest.2014.51","article-title":"Vitronectin-binding PAI-1 protects against the development of cardiac fibrosis through interaction with fibroblasts","volume":"94","year":"2014","journal-title":"Lab Invest"},{"key":"2026061921101063000_2025.11.19.689269v2.50","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M105980200"},{"key":"2026061921101063000_2025.11.19.689269v2.51","doi-asserted-by":"publisher","DOI":"10.1096\/fj.01-0552com"},{"key":"2026061921101063000_2025.11.19.689269v2.52","doi-asserted-by":"crossref","first-page":"841","DOI":"10.1001\/jamacardio.2020.6909","article-title":"Identification of cardiac fibrosis in young adults with a homozygous frameshift variant in SERPINE1","volume":"6","year":"2021","journal-title":"JAMA Cardiology"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1101\/2025.11.19.689269","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,6,20]],"date-time":"2026-06-20T04:10:24Z","timestamp":1781928624000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.1101\/2025.11.19.689269"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,11,19]]},"references-count":52,"URL":"https:\/\/doi.org\/10.1101\/2025.11.19.689269","relation":{},"subject":[],"published":{"date-parts":[[2025,11,19]]},"subtype":"preprint"}}