{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T20:09:00Z","timestamp":1792786140804,"version":"4.5.0"},"reference-count":25,"publisher":"Oxford University Press (OUP)","issue":"10","license":[{"start":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T00:00:00Z","timestamp":1790035200000},"content-version":"vor","delay-in-days":4,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"name":"National Institute of Diabetes and Digestive and Kidney Diseases of the National Institutes of Health","award":["R01DK126477"],"award-info":[{"award-number":["R01DK126477"]}],"id":[{"id":"https:\/\/ror.org\/00adh9b73","id-type":"ROR","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,9,18]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Urinary plasminogen has been implicated in the progression of proteinuric kidney diseases, yet its direct role in mediating injury along the nephron remains incompletely understood. Experimental models commonly used to study plasminogen-mediated damage are limited by incomplete phenotypic fidelity or interspecies differences, hindering translational relevance. Here, we used kidney organoids generated from H9 human embryonic stem cells to investigate the effects of plasminogen exposure and to evaluate urokinase-type plasminogen activator (uPA) inhibition with amiloride as a therapeutic strategy. Kidney organoids were exposed to plasminogen in the presence or absence of amiloride, and uPA activity was quantified using a fluorescence-based assay. Compartment-specific injury was assessed by confocal microscopy, automated image analysis, and AI-assisted nuclear segmentation, while targeted transcriptomic changes were evaluated using bulk quantitative PCR. Plasminogen exposure induced glomerular injury, characterized by reduced podocalyxin, nephrin, podocin, and WT1 expression, decreased glomerular cell density, and increased apoptosis. Proximal tubules exhibited reduced LTL and megalin expression, with increased KIM-1, whereas distal tubules were relatively preserved. Amiloride effectively inhibited uPA activity and partially restored podocyte and proximal tubular marker expression, but did not prevent apoptosis, indicating incomplete protection against plasminogen-induced injury. These findings support preferential plasminogen-induced injury along the glomerular\u2013proximal tubular axis and highlight the utility of human kidney organoids for modeling this process, while underscoring the need for further validation and broader molecular profiling to strengthen their application in mechanistic studies and therapeutic discovery.<\/jats:p>","DOI":"10.1093\/stcltm\/szag078","type":"journal-article","created":{"date-parts":[[2026,8,14]],"date-time":"2026-08-14T11:32:46Z","timestamp":1786707166000},"source":"Crossref","is-referenced-by-count":0,"title":["Kidney organoids as a platform to study plasminogen-induced injury and uPA-based therapies"],"prefix":"10.1093","volume":"15","author":[{"given":"Marina","family":"de Cos","sequence":"first","affiliation":[{"name":"Dr. Barbara T. Murphy Division of Nephrology, Icahn School of Medicine at Mount Sinai , New York, NY,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Samuel Mon-Wei","family":"Yu","sequence":"additional","affiliation":[{"name":"Dr. Barbara T. Murphy Division of Nephrology, Icahn School of Medicine at Mount Sinai , New York, NY,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jacob","family":"Wright","sequence":"additional","affiliation":[{"name":"Dr. Barbara T. Murphy Division of Nephrology, Icahn School of Medicine at Mount Sinai , New York, NY,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nanditha","family":"Anandakrishnan","sequence":"additional","affiliation":[{"name":"Dr. Barbara T. Murphy Division of Nephrology, Icahn School of Medicine at Mount Sinai , New York, NY,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kristin","family":"Meliambro","sequence":"additional","affiliation":[{"name":"Dr. Barbara T. Murphy Division of Nephrology, Icahn School of Medicine at Mount Sinai , New York, NY,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jenny S","family":"Wong","sequence":"additional","affiliation":[{"name":"Division of Renal-Electrolyte and Hypertension, University of Pennsylvania Perelman School of Medicine , Philadelphia, PA,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Evren","family":"Azeloglu","sequence":"additional","affiliation":[{"name":"Dr. Barbara T. Murphy Division of Nephrology, Icahn School of Medicine at Mount Sinai , New York, NY,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3855-8651","authenticated-orcid":false,"given":"Kirk N","family":"Campbell","sequence":"additional","affiliation":[{"name":"Division of Renal-Electrolyte and Hypertension, University of Pennsylvania Perelman School of Medicine , Philadelphia, PA,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2026,9,21]]},"reference":[{"key":"2026092201310131600_szag078-B1","doi-asserted-by":"publisher","first-page":"809","DOI":"10.1046\/j.1523-1755.2003.00840.x","article-title":"Pathophysiology of proteinuria","volume":"63","author":"D\u2019Amico","year":"2003","journal-title":"Kidney Int"},{"key":"2026092201310131600_szag078-B2","doi-asserted-by":"publisher","first-page":"2881","DOI":"10.1182\/blood.2020008951","article-title":"Plasminogen: an enigmatic zymogen","volume":"137","author":"Keragala","year":"2021","journal-title":"Blood"},{"key":"2026092201310131600_szag078-B3","doi-asserted-by":"publisher","first-page":"F753","DOI":"10.1152\/ajprenal.00257.2025","article-title":"Therapeutic potential of tubular serine protease inhibitors in proteinuria","volume":"329","author":"Boes","year":"2025","journal-title":"Am J Physiol Renal Physiol"},{"key":"2026092201310131600_szag078-B4","doi-asserted-by":"publisher","first-page":"16191","DOI":"10.1096\/fj.202000413R","article-title":"Plasminogenuria is associated with podocyte injury, edema, and kidney dysfunction in incident glomerular disease","volume":"34","author":"Egerman","year":"2020","journal-title":"Faseb J"},{"key":"2026092201310131600_szag078-B5","doi-asserted-by":"publisher","first-page":"F1308","DOI":"10.1152\/ajprenal.00162.2016","article-title":"Podocyte injury: the role of proteinuria, urinary plasminogen, and oxidative stress","volume":"311","author":"Raij","year":"2016","journal-title":"Am J Physiol Renal Physiol"},{"key":"2026092201310131600_szag078-B6","doi-asserted-by":"publisher","first-page":"187","DOI":"10.1016\/0014-5793(87)80039-x","article-title":"Amiloride selectively inhibits the urokinase-type plasminogen activator","volume":"214","author":"Vassalli","year":"1987","journal-title":"FEBS Lett"},{"key":"2026092201310131600_szag078-B7","doi-asserted-by":"publisher","first-page":"1746","DOI":"10.1093\/ndt\/gfr612","article-title":"Amiloride off-target effect inhibits podocyte urokinase receptor expression and reduces proteinuria","volume":"27","author":"Zhang","year":"2012","journal-title":"Nephrol Dial Transplant"},{"key":"2026092201310131600_szag078-B8","doi-asserted-by":"publisher","first-page":"410","DOI":"10.1681\/ASN.0000000000000312","article-title":"Amiloride reduces urokinase\/plasminogen-driven intratubular complement activation in glomerular proteinuria","volume":"35","author":"Isaksson","year":"2024","journal-title":"J Am Soc Nephrol"},{"key":"2026092201310131600_szag078-B9","doi-asserted-by":"publisher","first-page":"205","DOI":"10.1053\/j.ajkd.2024.01.520","article-title":"Urinary plasminogen as a marker of disease progression in human glomerular disease","volume":"84","author":"de Cos","year":"2024","journal-title":"Am J Kidney Dis"},{"key":"2026092201310131600_szag078-B10","doi-asserted-by":"publisher","first-page":"97","DOI":"10.1016\/j.jash.2017.12.003","article-title":"Urine albumin is a superior predictor of preeclampsia compared to urine plasminogen in type I diabetes patients","volume":"12","author":"Nielsen","year":"2018","journal-title":"J Am Soc Hypertens"},{"key":"2026092201310131600_szag078-B11","doi-asserted-by":"publisher","first-page":"451","DOI":"10.1007\/s00431-025-06278-3","article-title":"Urinary plasminogen as an early marker of diabetic kidney disease in children with type 1 diabetes mellitus: a cross-sectional study","volume":"184","author":"\u0130n\u00f6z\u00fc","year":"2025","journal-title":"Eur J Pediatr"},{"key":"2026092201310131600_szag078-B12","doi-asserted-by":"publisher","first-page":"26","DOI":"10.1038\/sj.ki.5002291","article-title":"Podocytes in culture: past, present, and future","volume":"72","author":"Shankland","year":"2007","journal-title":"Kidney Int"},{"key":"2026092201310131600_szag078-B13","doi-asserted-by":"publisher","first-page":"10982","DOI":"10.1074\/jbc.M109.093492","article-title":"Structure-based engineering of species selectivity in the interaction between urokinase and its receptor: implication for preclinical cancer therapy","volume":"285","author":"Lin","year":"2010","journal-title":"J Biol Chem"},{"key":"2026092201310131600_szag078-B14","doi-asserted-by":"publisher","first-page":"383","DOI":"10.1146\/annurev-bioeng-072623-044010","article-title":"Kidney disease modeling with organoids and organs-on-chips","volume":"26","author":"Musah","year":"2024","journal-title":"Annu Rev Biomed Eng"},{"key":"2026092201310131600_szag078-B15","doi-asserted-by":"publisher","DOI":"10.1242\/dev.200198","article-title":"Human pluripotent stem cell-derived kidney organoids for personalized congenital and idiopathic nephrotic syndrome modeling","volume":"149","author":"Jansen","year":"2022","journal-title":"Development"},{"key":"2026092201310131600_szag078-B16","doi-asserted-by":"publisher","first-page":"195","DOI":"10.1038\/nprot.2016.170","article-title":"Generation of nephron progenitor cells and kidney organoids from human pluripotent stem cells","volume":"12","author":"Morizane","year":"2017","journal-title":"Nat Protoc"},{"key":"2026092201310131600_szag078-B17","doi-asserted-by":"publisher","first-page":"100","DOI":"10.1038\/s41592-020-01018-x","article-title":"Cellpose: a generalist algorithm for cellular segmentation","volume":"18","author":"Stringer","year":"2021","journal-title":"Nat Methods"},{"key":"2026092201310131600_szag078-B18","doi-asserted-by":"publisher","author":"Pachitariu","DOI":"10.1101\/2025.04.28.651001"},{"key":"2026092201310131600_szag078-B19","doi-asserted-by":"publisher","first-page":"50","DOI":"10.1038\/s41440-022-01064-2","article-title":"The serine protease plasmin plays detrimental roles in epithelial sodium channel activation and podocyte injury in dahl salt-sensitive rats","volume":"46","author":"Deng","year":"2023","journal-title":"Hypertens Res"},{"key":"2026092201310131600_szag078-B20","doi-asserted-by":"publisher","DOI":"10.3390\/ijms242417563","article-title":"Proteolytic activation of the epithelial sodium channel (ENaC): its mechanisms and implications","volume":"24","author":"Aufy","year":"2023","journal-title":"Int J Mol Sci"},{"key":"2026092201310131600_szag078-B21","doi-asserted-by":"publisher","first-page":"299","DOI":"10.1681\/ASN.2008040364","article-title":"Plasmin in nephrotic urine activates the epithelial sodium channel","volume":"20","author":"Svenningsen","year":"2009","journal-title":"J Am Soc Nephrol"},{"key":"2026092201310131600_szag078-B22","doi-asserted-by":"crossref","first-page":"e2404767","DOI":"10.1002\/adhm.202404767","article-title":"Engineered 3D kidney glomerular microtissues to model podocyte-centric diseases for the validation of new drug targets","volume":"14","author":"Buttomicron","year":"2025","journal-title":"Adv Healthc Mater"},{"key":"2026092201310131600_szag078-B23","doi-asserted-by":"publisher","first-page":"689083","DOI":"10.3389\/fphys.2021.689083","article-title":"Modeling the glomerular filtration barrier and intercellular crosstalk","volume":"12","author":"Ebefors","year":"2021","journal-title":"Front Physiol"},{"key":"2026092201310131600_szag078-B24","doi-asserted-by":"publisher","first-page":"457","DOI":"10.1007\/s00441-021-03499-4","article-title":"Kidney organoid systems for studies of immune-mediated kidney diseases: challenges and opportunities","volume":"385","author":"Stein","year":"2021","journal-title":"Cell Tissue Res"},{"key":"2026092201310131600_szag078-B25","doi-asserted-by":"publisher","first-page":"191","DOI":"10.1007\/s00467-018-4177-y","article-title":"Kidney and organoid single-cell transcriptomics: the end of the beginning","volume":"35","author":"Wilson","year":"2020","journal-title":"Pediatr Nephrol"}],"container-title":["Stem Cells Translational Medicine"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/academic.oup.com\/stcltm\/article-pdf\/15\/10\/szag078\/71299784\/szag078.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/stcltm\/article-pdf\/15\/10\/szag078\/71299784\/szag078.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T05:31:07Z","timestamp":1790055067000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/stcltm\/article\/doi\/10.1093\/stcltm\/szag078\/8826370"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,18]]},"references-count":25,"journal-issue":{"issue":"10","published-print":{"date-parts":[[2026,9,18]]}},"URL":"https:\/\/doi.org\/10.1093\/stcltm\/szag078","relation":{},"ISSN":["2157-6564","2157-6580"],"issn-type":[{"value":"2157-6564","type":"print"},{"value":"2157-6580","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2026,10]]},"published":{"date-parts":[[2026,9,18]]},"article-number":"szag078"}}