{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,24]],"date-time":"2026-10-24T01:33:16Z","timestamp":1792805596077,"version":"4.5.0"},"reference-count":35,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2025,4,18]],"date-time":"2025-04-18T00:00:00Z","timestamp":1744934400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Instituto de Salud Carlos III (ISCIII)","award":["RED 2024\/0007\/0035"],"award-info":[{"award-number":["RED 2024\/0007\/0035"]}],"id":[{"id":"https:\/\/ror.org\/00ca2c886","id-type":"ROR","asserted-by":"crossref"}]},{"name":"Instituto de Salud Carlos III (ISCIII)","award":["PI17\/01026"],"award-info":[{"award-number":["PI17\/01026"]}],"id":[{"id":"https:\/\/ror.org\/00ca2c886","id-type":"ROR","asserted-by":"crossref"}]},{"name":"Instituto de Salud Carlos III (ISCIII)","award":["C2-0009-2020"],"award-info":[{"award-number":["C2-0009-2020"]}],"id":[{"id":"https:\/\/ror.org\/00ca2c886","id-type":"ROR","asserted-by":"crossref"}]},{"name":"European Union","award":["RED 2024\/0007\/0035"],"award-info":[{"award-number":["RED 2024\/0007\/0035"]}],"id":[{"id":"https:\/\/ror.org\/019w4f821","id-type":"ROR","asserted-by":"crossref"}]},{"name":"European Union","award":["PI17\/01026"],"award-info":[{"award-number":["PI17\/01026"]}],"id":[{"id":"https:\/\/ror.org\/019w4f821","id-type":"ROR","asserted-by":"crossref"}]},{"name":"European Union","award":["C2-0009-2020"],"award-info":[{"award-number":["C2-0009-2020"]}],"id":[{"id":"https:\/\/ror.org\/019w4f821","id-type":"ROR","asserted-by":"crossref"}]},{"name":"Junta de Andaluc\u00eda, Programa Nicol\u00e1s Monardes","award":["RED 2024\/0007\/0035"],"award-info":[{"award-number":["RED 2024\/0007\/0035"]}],"id":[{"id":"https:\/\/ror.org\/01jem9c82","id-type":"ROR","asserted-by":"crossref"}]},{"name":"Junta de Andaluc\u00eda, Programa Nicol\u00e1s Monardes","award":["PI17\/01026"],"award-info":[{"award-number":["PI17\/01026"]}],"id":[{"id":"https:\/\/ror.org\/01jem9c82","id-type":"ROR","asserted-by":"crossref"}]},{"name":"Junta de Andaluc\u00eda, Programa Nicol\u00e1s Monardes","award":["C2-0009-2020"],"award-info":[{"award-number":["C2-0009-2020"]}],"id":[{"id":"https:\/\/ror.org\/01jem9c82","id-type":"ROR","asserted-by":"crossref"}]},{"name":"Karolinska Institutet Foundations and Funds","award":["RED 2024\/0007\/0035"],"award-info":[{"award-number":["RED 2024\/0007\/0035"]}],"id":[{"id":"https:\/\/ror.org\/056d84691","id-type":"ROR","asserted-by":"crossref"}]},{"name":"Karolinska Institutet Foundations and Funds","award":["PI17\/01026"],"award-info":[{"award-number":["PI17\/01026"]}],"id":[{"id":"https:\/\/ror.org\/056d84691","id-type":"ROR","asserted-by":"crossref"}]},{"name":"Karolinska Institutet Foundations and Funds","award":["C2-0009-2020"],"award-info":[{"award-number":["C2-0009-2020"]}],"id":[{"id":"https:\/\/ror.org\/056d84691","id-type":"ROR","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Vascular endothelial growth factor (VEGF) is a key mediator of exudative age-related macular degeneration (eAMD), yet non-invasive biomarkers for disease monitoring remain limited. This study evaluates VEGF levels in human tear fluid as a potential biomarker for eAMD and investigates the molecular dynamics of VEGF in a laser-induced choroidal neovascularization (lCNV) mouse model. Tear VEGF levels were quantified using proximity qPCR immunoassays in eAMD patients (n = 29) and healthy controls (n = 21) and correlated with optical coherence tomography (OCT) findings. Molecular analyses, including immunohistochemistry, gene expression profiling, and phosphorylation assays, were conducted on choroid\u2013retinal pigment epithelium (RPE) and lacrimal gland (LG) tissues from lCNV mice (n = 25). Tear VEGF levels were significantly elevated in eAMD patients, correlating with disease severity. Females exhibited higher VEGF levels, a pattern not replicated in the mouse model. In lCNV mice, VEGF overexpression originated from the choroid\u2013RPE, driven by hypoxic and inflammatory signaling, with no significant LG contribution. Increased VEGF, IL-6, and vimentin expression, along with NF-\u03baB and STAT3 activation, were observed. These findings suggest that tear VEGF is a promising non-invasive biomarker for eAMD, warranting further validation for clinical application in disease monitoring and treatment optimization.<\/jats:p>","DOI":"10.3390\/ijms26083855","type":"journal-article","created":{"date-parts":[[2025,4,18]],"date-time":"2025-04-18T08:24:07Z","timestamp":1744964647000},"page":"3855","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["VEGF in Tears as a Biomarker for Exudative Age-Related Macular Degeneration: Molecular Dynamics in a Mouse Model and Human Samples"],"prefix":"10.3390","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6039-4246","authenticated-orcid":false,"given":"Seyed Mohamad Mehdi","family":"Moshtaghion","sequence":"first","affiliation":[{"name":"Department of Integrative Pathophysiology and Therapies, Andalusian Molecular Biology and Regenerative Medicine Centre (CABIMER), Junta de Andaluc\u00eda, CSIC, University of Seville, University Pablo de Olavide, 41092 Seville, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-3772-8730","authenticated-orcid":false,"given":"Filippo","family":"Locri","sequence":"additional","affiliation":[{"name":"Department of Clinical Neuroscience, Division of Eye and Vision, St. Erik Eye Hospital, Karolinska Institutet, 17177 Stockholm, Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1167-6316","authenticated-orcid":false,"given":"Alvaro Plaza","family":"Reyes","sequence":"additional","affiliation":[{"name":"Department of Integrative Pathophysiology and Therapies, Andalusian Molecular Biology and Regenerative Medicine Centre (CABIMER), Junta de Andaluc\u00eda, CSIC, University of Seville, University Pablo de Olavide, 41092 Seville, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Flavia","family":"Plastino","sequence":"additional","affiliation":[{"name":"Department of Clinical Neuroscience, Division of Eye and Vision, St. Erik Eye Hospital, Karolinska Institutet, 17177 Stockholm, Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2194-7490","authenticated-orcid":false,"given":"Anders","family":"Kvanta","sequence":"additional","affiliation":[{"name":"Department of Clinical Neuroscience, Division of Eye and Vision, St. Erik Eye Hospital, Karolinska Institutet, 17177 Stockholm, Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria Jose","family":"Morillo-Sanchez","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, Virgen Macarena University Hospital, 41009 Seville, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Enrique","family":"Rodr\u00edguez-de-la-R\u00faa","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, Virgen Macarena University Hospital, 41009 Seville, Spain"},{"name":"Department of Surgery, Ophthalmology Area, University of Seville, 41009 Seville, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4851-680X","authenticated-orcid":false,"given":"Estanislao","family":"Gutierrez-Sanchez","sequence":"additional","affiliation":[{"name":"Department of Ophthalmology, Virgen Macarena University Hospital, 41009 Seville, Spain"},{"name":"Department of Surgery, Ophthalmology Area, University of Seville, 41009 Seville, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adoraci\u00f3n","family":"Montero-S\u00e1nchez","sequence":"additional","affiliation":[{"name":"Department of Integrative Pathophysiology and Therapies, Andalusian Molecular Biology and Regenerative Medicine Centre (CABIMER), Junta de Andaluc\u00eda, CSIC, University of Seville, University Pablo de Olavide, 41092 Seville, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7569-3663","authenticated-orcid":false,"given":"Helena","family":"Lucena-Padros","sequence":"additional","affiliation":[{"name":"Department of Maternofetal Medicine, Genetics and Reproduction, Institute of Biomedicine of Seville, CSIC, Virgen del Rocio University Hospital, University of Seville, 41013 Seville, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2926-2376","authenticated-orcid":false,"given":"Helder","family":"Andr\u00e9","sequence":"additional","affiliation":[{"name":"Department of Clinical Neuroscience, Division of Eye and Vision, St. Erik Eye Hospital, Karolinska Institutet, 17177 Stockholm, Sweden"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5752-0205","authenticated-orcid":false,"given":"Francisco J.","family":"D\u00edaz-Corrales","sequence":"additional","affiliation":[{"name":"Department of Integrative Pathophysiology and Therapies, Andalusian Molecular Biology and Regenerative Medicine Centre (CABIMER), Junta de Andaluc\u00eda, CSIC, University of Seville, University Pablo de Olavide, 41092 Seville, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,4,18]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"AMD160","DOI":"10.1167\/iovs.18-24882","article-title":"Soft Drusen in Age-Related Macular Degeneration: Biology and Targeting Via the Oil Spill Strategies","volume":"59","author":"Curcio","year":"2018","journal-title":"Investig. Opthalmol. Vis. Sci."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1038\/s41572-021-00265-2","article-title":"Age-related macular degeneration","volume":"7","author":"Fleckenstein","year":"2021","journal-title":"Nat. Rev. Dis. Primers"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"221","DOI":"10.2174\/138945011794182755","article-title":"Age related macular degeneration and visual disability","volume":"12","author":"Christoforidis","year":"2011","journal-title":"Curr. Drug Targets"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"246","DOI":"10.1007\/s00347-014-3217-7","article-title":"Retrospective investigation of anti-VEGF treatment reality and effectiveness in patients with neovascular age-related macular degeneration (AMD) in Germany: Treatment reality of ranibizumab for neovascular AMD in Germany","volume":"112","author":"Ziemssen","year":"2015","journal-title":"Ophthalmologe"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1144","DOI":"10.1136\/bjophthalmol-2014-305702","article-title":"Guidelines for the management of neovascular age-related macular degeneration by the European Society of Retina Specialists (EURETINA)","volume":"98","author":"Chong","year":"2014","journal-title":"Br. J. Ophthalmol."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Nguyen, C.L., Oh, L.J., Wong, E., Wei, J., and Chilov, M. (2018). Anti-vascular endothelial growth factor for neovascular age-related macular degeneration: A meta-analysis of randomized controlled trials. BMC Ophthalmol., 18.","DOI":"10.1186\/s12886-018-0785-3"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"3699","DOI":"10.1093\/hmg\/ddr270","article-title":"Common variants near FRK\/COL10A1 and VEGFA are associated with advanced age-related macular degeneration","volume":"20","author":"Yu","year":"2011","journal-title":"Hum. Mol. Genet."},{"key":"ref_8","first-page":"339","article-title":"VEGF and PEDF levels in the rat retina: Effects of aging and AMD-like retinopathy","volume":"31","author":"Kozhevnikova","year":"2018","journal-title":"Adv. Gerontol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"787","DOI":"10.1038\/eye.2013.107","article-title":"Adverse events and complications associated with intravitreal injection of anti-VEGF agents: A review of literature","volume":"27","author":"Falavarjani","year":"2013","journal-title":"Eye"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"487","DOI":"10.1016\/j.mam.2012.06.003","article-title":"Animal models of age related macular degeneration","volume":"33","author":"Pennesi","year":"2012","journal-title":"Mol. Asp. Med."},{"key":"ref_11","first-page":"1211","article-title":"Vascular endothelial growth factor A polymorphisms and age-related macular degeneration: A systematic review and meta-analysis","volume":"19","author":"Huang","year":"2013","journal-title":"Mol. Vis."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"132","DOI":"10.1016\/j.preteyeres.2017.11.004","article-title":"Towards the application of precision medicine in Age-Related Macular Degeneration","volume":"63","author":"Cascella","year":"2017","journal-title":"Prog. Retin. Eye Res."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Oca, A.I., P\u00e9rez-Sala, \u00c1., Pariente, A., Ochoa, R., Velilla, S., Pel\u00e1ez, R., and Larr\u00e1yoz, I.M. (2021). Predictive Biomarkers of Age-Related Macular Degeneration Response to Anti-VEGF Treatment. J. Pers. Med., 11.","DOI":"10.3390\/jpm11121329"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Salas, A., Badia, A., Fontrodona, L., Zapata, M., Garc\u00eda-Arum\u00ed, J., and Duarri, A. (2023). Neovascular Progression and Retinal Dysfunction in the Laser-Induced Choroidal Neovascularization Mouse Model. Biomedicines, 11.","DOI":"10.3390\/biomedicines11092445"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"155","DOI":"10.1186\/s12964-022-00973-6","article-title":"PGE2 promotes macrophage recruitment and neovascularization in murine wet-type AMD models","volume":"20","author":"Zhan","year":"2022","journal-title":"Cell Commun. Signal."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Xu, N., Sun, T., Wang, Y., Tong, X., Lu, S., Yang, F., Wang, J., Bo, Q., Sun, J., and Sun, X. (2023). Dynamic changes in macrophage morphology during the progression of choroidal neovascularization in a laser-induced choroidal neovascularization mouse model. BMC Ophthalmol., 23.","DOI":"10.1186\/s12886-023-03141-7"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"523","DOI":"10.1002\/jlb.57.4.523","article-title":"The role of PECAM-1 (CD31) in leukocyte emigration: Studies in vitro and in vivo","volume":"57","author":"Muller","year":"1995","journal-title":"J. Leukoc. Biol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1147","DOI":"10.1016\/S0140-6736(18)31550-2","article-title":"Age-related macular degeneration","volume":"392","author":"Mitchell","year":"2018","journal-title":"Lancet"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"64","DOI":"10.1016\/j.preteyeres.2016.04.003","article-title":"Risk factors and biomarkers of age-related macular degeneration","volume":"54","author":"Lambert","year":"2016","journal-title":"Prog. Retin. Eye Res."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Lukacs, R., Schneider, M., Nagy, Z.Z., Sandor, G.L., Kaan, K., Asztalos, A., Enyedi, L., Pek, G., Barcsay, G., and Szabo, A. (2023). Seven-year outcomes following intensive anti-vascular endothelial growth factor therapy in patients with exudative age-related macular degeneration. BMC Ophthalmol., 23.","DOI":"10.1186\/s12886-023-02843-2"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Corazza, P., D\u2019alterio, F.M., Kabbani, J., Alam, M.M.R., Mercuri, S., Orlans, H.O., and Younis, S. (2021). Long-term outcomes of intravitreal anti-VEGF therapies in patients affected by neovascular age-related macular degeneration: A real-life study. BMC Ophthalmol., 21.","DOI":"10.1186\/s12886-021-02055-6"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Moraru, A.D., Danielescu, C., Iorga, R.E., Moraru, R.L., Zemba, M., and Branisteanu, D.C. (2024). Review of Guideline Recommendations for Optimal Anti-VEGF Therapy in Age-Related Macular Degeneration. Life, 14.","DOI":"10.3390\/life14101220"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"309","DOI":"10.1177\/107155760000700507","article-title":"Changes in serum levels of vascular endothelial growth factor in males and females throughout life","volume":"7","author":"Tziotis","year":"2000","journal-title":"J. Soc. Gynecol. Investig."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"317","DOI":"10.1111\/ceo.14364","article-title":"Gender differences in retinal diseases: A review","volume":"52","author":"Tillmann","year":"2024","journal-title":"Clin. Exp. Ophthalmol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1016\/j.ab.2005.01.018","article-title":"Proximity ligation assays for sensitive and specific protein analyses","volume":"345","author":"Gustafsdottir","year":"2005","journal-title":"Anal. Biochem."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"e102","DOI":"10.1093\/nar\/gkr424","article-title":"Homogeneous antibody-based proximity extension assays provide sensitive and specific detection of low-abundant proteins in human blood","volume":"39","author":"Lundberg","year":"2011","journal-title":"Nucleic Acids Res."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Shahidatul-Adha, M., Zunaina, E., and Aini-Amalina, M.N. (2022). Evaluation of vascular endothelial growth factor (VEGF) level in the tears and serum of age-related macular degeneration patients. Sci. Rep., 12.","DOI":"10.1038\/s41598-022-08492-7"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"616","DOI":"10.1016\/j.ophtha.2019.11.004","article-title":"Consensus Nomenclature for Reporting Neovascular Age-Related Macular Degeneration Data: Consensus on Neovascular Age-Related Macular Degeneration Nomenclature Study Group","volume":"127","author":"Spaide","year":"2020","journal-title":"Ophthalmology"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"844","DOI":"10.1016\/j.ophtha.2012.10.036","article-title":"Clinical classification of age-related macular degeneration","volume":"120","author":"Ferris","year":"2013","journal-title":"Ophthalmology"},{"key":"ref_30","first-page":"288","article-title":"Concordance of OCT and FA-based MNV classifications in neovascular AMD and correlation with fluid volumes","volume":"64","author":"Deak","year":"2023","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"270","DOI":"10.1016\/j.jalz.2011.03.008","article-title":"The diagnosis of mild cognitive impairment due to Alzheimer\u2019s disease: Recommendations from the National Institute on Aging-Alzheimer\u2019s Association workgroups on diagnostic guidelines for Alzheimer\u2019s disease","volume":"7","author":"Albert","year":"2011","journal-title":"Alzheimer Dement."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Takei, A., Ekstr\u00f6m, M., Mammadzada, P., Aronsson, M., Yu, M., Kvanta, A., and Andr\u00e9, H. (2017). Gene Transfer of Prolyl Hydroxylase Domain 2 Inhibits Hypoxia-inducible Angiogenesis in a Model of Choroidal Neovascularization. Sci. Rep., 7.","DOI":"10.1038\/srep42546"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"6591","DOI":"10.1167\/iovs.15-16476","article-title":"Hypoxia-Inducible Factor-1\u03b1 Is Associated With Sprouting Angiogenesis in the Murine Laser-Induced Choroidal Neovascularization Model","volume":"56","author":"Tunik","year":"2015","journal-title":"Investig. Opthalmol. Vis. Sci."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Shinomiya, K., Ueta, M., and Kinoshita, S. (2018). A new dry eye mouse model produced by exorbital and intraorbital lacrimal gland excision. Sci. Rep., 8.","DOI":"10.1038\/s41598-018-19578-6"},{"key":"ref_35","first-page":"e51970","article-title":"Manipulating the Murine Lacrimal Gland","volume":"93","author":"Finley","year":"2014","journal-title":"JoVE (J. Vis. Exp.)"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/8\/3855\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T17:17:24Z","timestamp":1760030244000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/8\/3855"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,4,18]]},"references-count":35,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2025,4]]}},"alternative-id":["ijms26083855"],"URL":"https:\/\/doi.org\/10.3390\/ijms26083855","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,4,18]]}}}