{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,8,12]],"date-time":"2026-08-12T04:27:32Z","timestamp":1786508852155,"version":"build-2736575974"},"posted":{"date-parts":[[2026,8,10]]},"group-title":"Cell Biology","reference-count":45,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,8,10]],"date-time":"2026-08-10T00:00:00Z","timestamp":1786320000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"funder":[{"DOI":"10.13039\/100000053","name":"National Eye Institute","doi-asserted-by":"crossref","award":["1R01EY037135-01"],"award-info":[{"award-number":["1R01EY037135-01"]}],"id":[{"id":"10.13039\/100000053","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000053","name":"National Eye Institute","doi-asserted-by":"crossref","award":["1R21EY034198-01"],"award-info":[{"award-number":["1R21EY034198-01"]}],"id":[{"id":"10.13039\/100000053","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Mr. and Mrs. Robert and Joell Brightfelt"},{"name":"Mr. and Mrs. Lloyd and Betty Schermer"},{"name":"Ms. Mary Dawson"},{"name":"The M.D. Wagoner & M.A. Greiner Cornea Excellence Fund"},{"name":"The Cornea Transplant Research Fund"},{"name":"The Beulah and Florence Usher Chair in Cornea\/External Disease and Refractive Surgery"},{"name":"The Seidler Educational Support Fund in Cornea Research"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,8,10]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:sec>\n                  <jats:title>Purpose<\/jats:title>\n                  <jats:p>Fuchs endothelial corneal dystrophy (FECD) is a progressive disease, causing premature death of corneal endothelial cells (CECs). Iron-dependent lipid peroxidation and ferroptosis mediate cell death in FECD. We aimed to determine whether FECD progression is mediated by derangements in ferritinophagy \u2013 a form of autophagy that degrades ferritin to release labile ferrous iron \u2013 and whether ultraviolet A (UVA) exposure drives FECD progression by activating ferritinophagy.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>Endothelium-Descemet membrane (EDM) tissues were collected from patients with end-stage FECD undergoing endothelial keratoplasty and from healthy age-matched donor corneas. Separately, immortalized FECD and healthy control CEC lines were cultured. Cellular levels of NCOA4 production and LC3 activation, both markers of ferritinophagy, were quantified using western blotting and PCR. UVA-exposed immortalized cells were plated on coverslips, stained for immunohistochemistry (IHC), and analyzed using confocal microscopy. Corneal endothelial peels were stained and analyzed using laser ablation\u2013inductively coupled plasma\u2013mass spectrometry (LA-ICP-MS).<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>Surgically explanted FECD CECs showed significantly increased levels of NCOA4 compared to healthy controls. LC3 activation was increased in FECD immortalized CECs; UV exposure further increased LC3 activation. Additionally, UVA exposure showed trends of increased expression of NCOA4 in immortalized FECD and healthy CECs. On IHC of FECD surgical explant tissue, ferritin was decreased markedly, NCOA4 localized in a dramatic punctate pattern, and both ferritin and LC3 localized within cell nuclei. Spectrometry images showed higher iron levels correlating with areas of higher FECD disease burden.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Conclusions<\/jats:title>\n                  <jats:p>Our results demonstrate ferritinophagy in FECD indicated by the increase of NCOA4 and LC3 ferritinophagy markers in FECD patient and cell culture models. Our finding that UVA activates ferritinophagy implicates this mechanism in UVA-mediated FECD progression. Altogether, aberrant iron dysregulation associated with FECD and ferroptosis may be mediated by ferritinophagy, providing a biomarker to assess disease severity as well as a potential target for future medical therapeutics.<\/jats:p>\n                <\/jats:sec>","DOI":"10.64898\/2026.08.08.743691","type":"posted-content","created":{"date-parts":[[2026,8,10]],"date-time":"2026-08-10T15:25:13Z","timestamp":1786375513000},"source":"Crossref","is-referenced-by-count":0,"title":["Ferritinophagy Contributes to Iron Accumulation and Ferroptosis in Fuchs Endothelial Corneal Dystrophy"],"prefix":"10.64898","author":[{"given":"Zachary","family":"Shepard","sequence":"first","affiliation":[{"name":"University of Iowa Carver College of Medicine, Department of Ophthalmology and Visual Sciences, Iowa City, IA 52242"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jessica M.","family":"Skeie","sequence":"additional","affiliation":[{"name":"University of Iowa Carver College of Medicine, Department of Ophthalmology and Visual Sciences, Iowa City, IA 52242"},{"name":"Iowa Lions Eye Bank, Coralville, IA 52241"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hanna","family":"Shevalye","sequence":"additional","affiliation":[{"name":"University of Iowa Carver College of Medicine, Department of Ophthalmology and Visual Sciences, Iowa City, IA 52242"},{"name":"Iowa Lions Eye Bank, Coralville, IA 52241"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Timothy","family":"Eggleston","sequence":"additional","affiliation":[{"name":"University of Iowa Carver College of Medicine, Department of Ophthalmology and Visual Sciences, Iowa City, IA 52242"},{"name":"Iowa Lions Eye Bank, Coralville, IA 52241"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Linhan","family":"Li","sequence":"additional","affiliation":[{"name":"University of Iowa Carver College of Medicine, Department of Ophthalmology and Visual Sciences, Iowa City, IA 52242"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matthew G.","family":"Field","sequence":"additional","affiliation":[{"name":"Minnesota Eye Consultants, Minneapolis, MN 55305"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gregory A.","family":"Schmidt","sequence":"additional","affiliation":[{"name":"Iowa Lions Eye Bank, Coralville, IA 52241"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pornpoj","family":"Phruttiwanichakun","sequence":"additional","affiliation":[{"name":"University of Iowa College of Pharmacy, Department of Pharmaceutical Sciences and Experimental Therapeutics, Iowa City, IA 52242"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christopher S.","family":"Sales","sequence":"additional","affiliation":[{"name":"University of Iowa Carver College of Medicine, Department of Ophthalmology and Visual Sciences, Iowa City, IA 52242"},{"name":"Iowa Lions Eye Bank, Coralville, IA 52241"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aliasger K.","family":"Salem","sequence":"additional","affiliation":[{"name":"University of Iowa College of Pharmacy, Department of Pharmaceutical Sciences and Experimental Therapeutics, Iowa City, IA 52242"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mark A.","family":"Greiner","sequence":"additional","affiliation":[{"name":"University of Iowa Carver College of Medicine, Department of Ophthalmology and Visual Sciences, Iowa City, IA 52242"},{"name":"Iowa Lions Eye Bank, Coralville, IA 52241"},{"name":"University of Iowa College of Pharmacy, Department of Pharmaceutical Sciences and Experimental Therapeutics, Iowa City, IA 52242"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"issue":"9","key":"2026081120352225000_2026.08.08.743691v1.1","first-page":"2501","article-title":"Corneal endothelial cell apoptosis in patients with Fuchs\u2019 dystrophy","volume":"41","year":"2000","journal-title":"Invest Ophthalmol Vis Sci"},{"key":"2026081120352225000_2026.08.08.743691v1.2","first-page":"640234","article-title":"Fuchs Endothelial Corneal Dystrophy: Strong Association with rs613872 Not Paralleled by Changes in Corneal Endothelial TCF4 mRNA Level","volume":"2015","year":"2015","journal-title":"Biomed Res Int"},{"key":"2026081120352225000_2026.08.08.743691v1.3","doi-asserted-by":"publisher","DOI":"10.1016\/S1542-0124(12)70232-X"},{"key":"2026081120352225000_2026.08.08.743691v1.4","doi-asserted-by":"publisher","DOI":"10.2353\/ajpath.2010.100279"},{"key":"2026081120352225000_2026.08.08.743691v1.5","doi-asserted-by":"crossref","first-page":"103348","DOI":"10.1016\/j.redox.2024.103348","article-title":"TCF4 trinucleotide repeat expansions and UV irradiation increase susceptibility to ferroptosis in Fuchs endothelial corneal dystrophy","volume":"77","year":"2024","journal-title":"Redox Biol"},{"key":"2026081120352225000_2026.08.08.743691v1.6","doi-asserted-by":"crossref","unstructured":"Benischke, A.-S. , et al., Activation of mitophagy leads to decline in Mfn2 and loss of mitochondrial mass in Fuchs endothelial corneal dystrophy. Scientific Reports, 2017. 7(1).","DOI":"10.1038\/s41598-017-06523-2"},{"key":"2026081120352225000_2026.08.08.743691v1.7","doi-asserted-by":"publisher","DOI":"10.1016\/j.ajpath.2019.06.012"},{"key":"2026081120352225000_2026.08.08.743691v1.8","doi-asserted-by":"crossref","unstructured":"Iriondo, M.N. , et al., LC3 subfamily in cardiolipin-mediated mitophagy: a comparison of the LC3A, LC3B and LC3C homologs. Autophagy, 2022: p. 1\u201319.","DOI":"10.1080\/15548627.2022.2062111"},{"key":"2026081120352225000_2026.08.08.743691v1.9","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1912546116"},{"issue":"4","key":"2026081120352225000_2026.08.08.743691v1.10","doi-asserted-by":"crossref","first-page":"2319","DOI":"10.1167\/iovs.16-19097","article-title":"UV-A Irradiation Activates Nrf2-Regulated Antioxidant Defense and Induces p53\/Caspase3-Dependent Apoptosis in Corneal Endothelial Cells","volume":"57","year":"2016","journal-title":"Invest Ophthalmol Vis Sci"},{"issue":"8","key":"2026081120352225000_2026.08.08.743691v1.11","doi-asserted-by":"crossref","first-page":"1893","DOI":"10.1016\/j.bbagen.2017.05.019","article-title":"Ferroptosis: Role of lipid peroxidation, iron and ferritinophagy","volume":"1861","year":"2017","journal-title":"Biochimica et Biophysica Acta (BBA) - General Subjects"},{"key":"2026081120352225000_2026.08.08.743691v1.12","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2012.03.042"},{"key":"2026081120352225000_2026.08.08.743691v1.13","doi-asserted-by":"crossref","unstructured":"Kim, S.W. , et al., Ferroptosis-Related Genes in Neurodevelopment and Central Nervous System. Biology (Basel), 2021. 10(1).","DOI":"10.3390\/biology10010035"},{"key":"2026081120352225000_2026.08.08.743691v1.14","doi-asserted-by":"publisher","DOI":"10.1038\/s41418-020-00728-1"},{"key":"2026081120352225000_2026.08.08.743691v1.15","doi-asserted-by":"crossref","unstructured":"Santana-Codina, N. and J.D. Mancias , The Role of NCOA4-Mediated Ferritinophagy in Health and Disease. Pharmaceuticals (Basel), 2018. 11(4).","DOI":"10.3390\/ph11040114"},{"key":"2026081120352225000_2026.08.08.743691v1.16","doi-asserted-by":"publisher","DOI":"10.1016\/j.tcb.2020.02.009"},{"key":"2026081120352225000_2026.08.08.743691v1.17","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2022.884362"},{"issue":"1","key":"2026081120352225000_2026.08.08.743691v1.18","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1186\/s40478-023-01617-7","article-title":"Ferroptosis induces detrimental effects in chronic EAE and its implications for progressive MS","volume":"11","year":"2023","journal-title":"Acta Neuropathol Commun"},{"key":"2026081120352225000_2026.08.08.743691v1.19","doi-asserted-by":"publisher","DOI":"10.1038\/cr.2016.95"},{"key":"2026081120352225000_2026.08.08.743691v1.20","doi-asserted-by":"publisher","DOI":"10.1093\/hmg\/ddy018"},{"issue":"2","key":"2026081120352225000_2026.08.08.743691v1.21","doi-asserted-by":"crossref","first-page":"813","DOI":"10.3892\/ijo.2015.3276","article-title":"Antimycin A induces death of the human pulmonary fibroblast cells via ROS increase and GSH depletion","volume":"48","year":"2016","journal-title":"International Journal of Oncology"},{"key":"2026081120352225000_2026.08.08.743691v1.22","doi-asserted-by":"publisher","DOI":"10.1155\/2019\/1583656"},{"key":"2026081120352225000_2026.08.08.743691v1.23","doi-asserted-by":"publisher","DOI":"10.1007\/s11120-016-0217-2"},{"key":"2026081120352225000_2026.08.08.743691v1.24","doi-asserted-by":"crossref","first-page":"322","DOI":"10.3389\/fonc.2020.00322","article-title":"Autophagy Regulation by the Translation Machinery and Its Implications in Cancer","volume":"10","year":"2020","journal-title":"Front Oncol"},{"issue":"19","key":"2026081120352225000_2026.08.08.743691v1.25","doi-asserted-by":"crossref","first-page":"11769","DOI":"10.1021\/acs.chemrev.0c01219","article-title":"Laser Ablation-Inductively Coupled Plasma-Mass Spectrometry Imaging in Biology","volume":"121","year":"2021","journal-title":"Chem Rev"},{"key":"2026081120352225000_2026.08.08.743691v1.26","doi-asserted-by":"publisher","DOI":"10.1039\/c1ja10172b"},{"key":"2026081120352225000_2026.08.08.743691v1.27","doi-asserted-by":"publisher","DOI":"10.1039\/c2ja10383d"},{"key":"2026081120352225000_2026.08.08.743691v1.28","doi-asserted-by":"publisher","DOI":"10.1038\/onc.2011.613"},{"key":"2026081120352225000_2026.08.08.743691v1.29","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2014.04.031"},{"issue":"4","key":"2026081120352225000_2026.08.08.743691v1.30","doi-asserted-by":"crossref","first-page":"5344","DOI":"10.18632\/oncotarget.23747","article-title":"Expression and function of nuclear receptor coactivator 4 isoforms in transformed endometriotic and malignant ovarian cells","volume":"9","year":"2018","journal-title":"Oncotarget"},{"key":"2026081120352225000_2026.08.08.743691v1.31","doi-asserted-by":"publisher","DOI":"10.1038\/ncb3053"},{"key":"2026081120352225000_2026.08.08.743691v1.32","doi-asserted-by":"publisher","DOI":"10.1038\/nature13148"},{"key":"2026081120352225000_2026.08.08.743691v1.33","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1603244113"},{"key":"2026081120352225000_2026.08.08.743691v1.34","doi-asserted-by":"crossref","first-page":"119911","DOI":"10.1016\/j.biomaterials.2020.119911","article-title":"Fenton reaction-independent ferroptosis therapy via glutathione and iron redox couple sequentially triggered lipid peroxide generator","volume":"241","year":"2020","journal-title":"Biomaterials"},{"issue":"3","key":"2026081120352225000_2026.08.08.743691v1.35","doi-asserted-by":"crossref","first-page":"387","DOI":"10.1021\/acscentsci.7b00589","article-title":"Resolving the Role of Lipoxygenases in the Initiation and Execution of Ferroptosis","volume":"4","year":"2018","journal-title":"ACS Cent Sci"},{"key":"2026081120352225000_2026.08.08.743691v1.36","doi-asserted-by":"publisher","DOI":"10.1080\/15548627.2016.1187366"},{"key":"2026081120352225000_2026.08.08.743691v1.37","doi-asserted-by":"crossref","first-page":"118","DOI":"10.1016\/j.freeradbiomed.2017.03.022","article-title":"Ultraviolet A-induced oxidation in cornea: Characterization of the early oxidation-related events","volume":"108","year":"2017","journal-title":"Free Radic Biol Med"},{"key":"2026081120352225000_2026.08.08.743691v1.38","doi-asserted-by":"publisher","DOI":"10.1080\/09553000902740150"},{"key":"2026081120352225000_2026.08.08.743691v1.39","doi-asserted-by":"publisher","DOI":"10.1016\/j.bpj.2012.02.023"},{"key":"2026081120352225000_2026.08.08.743691v1.40","doi-asserted-by":"publisher","DOI":"10.3390\/cells9102315"},{"key":"2026081120352225000_2026.08.08.743691v1.41","doi-asserted-by":"publisher","DOI":"10.1016\/j.tiv.2010.03.009"},{"key":"2026081120352225000_2026.08.08.743691v1.42","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbamcr.2020.118913"},{"key":"2026081120352225000_2026.08.08.743691v1.43","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2014.12.013"},{"key":"2026081120352225000_2026.08.08.743691v1.44","doi-asserted-by":"crossref","unstructured":"Linsenmayer, T.F. , et al., Corneal Epithelial Nuclear Ferritin and Its Transporter Ferritoid Afford Unique Protection to DNA from UV Light and Reactive Oxygen Species. 2015, Springer New York. p. 39\u201366.","DOI":"10.1007\/978-1-4939-1935-2_3"},{"key":"2026081120352225000_2026.08.08.743691v1.45","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M210050200"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.64898\/2026.08.08.743691","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,12]],"date-time":"2026-08-12T03:35:36Z","timestamp":1786505736000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.64898\/2026.08.08.743691"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,10]]},"references-count":45,"URL":"https:\/\/doi.org\/10.64898\/2026.08.08.743691","relation":{},"subject":[],"published":{"date-parts":[[2026,8,10]]},"subtype":"preprint"}}