{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T15:56:08Z","timestamp":1790610968297,"version":"4.1.0"},"reference-count":42,"publisher":"Wiley","issue":"10","license":[{"start":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T00:00:00Z","timestamp":1790553600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T00:00:00Z","timestamp":1790553600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["82370827"],"award-info":[{"award-number":["82370827"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["CNS Neurosci Ther"],"published-print":{"date-parts":[[2026,10]]},"abstract":"ABSTRACT<\/jats:title>\n \n Background<\/jats:title>\n Diabetic retinopathy (DR), a leading cause of blindness in diabetes, involves dysregulated neuroimmune crosstalk. While microglial activation and endothelial dysfunction are established in DR, the molecular mechanisms linking innate immunity to blood\u2010retinal barrier (BRB) breakdown remain elusive.<\/jats:p>\n <\/jats:sec>\n \n Methods<\/jats:title>\n We combined single\u2010cell RNA sequencing (scRNA\u2010seq) of diabetic mouse retinas with in\u00a0vivo models, retinal endothelial MFSD2A overexpression, and microglial Cathepsin S (CTSS) knockdown via adeno\u2010associated virus (AAV). Human retinal microvascular endothelial cells (HRMECs) and human microglial cells (HMC3) were subjected to high\u2010glucose conditions combined with in\u00a0vitro systems (transwell co\u2010cultures, holographic 3D tomography). Mechanistic studies employed siRNA knockdown, plasmid overexpression, exogenous CTSS supplementation, and transcytosis assays.<\/jats:p>\n <\/jats:sec>\n \n Results<\/jats:title>\n \n scRNA\u2010seq revealed diabetes\u2010induced CTSS upregulation in retinal microglia (\n p<\/jats:italic>\n \u2009=\u20090.0031) and MFSD2A downregulation in endothelial cells (\n p<\/jats:italic>\n \u2009=\u20090.01). Microglial CTSS promoted M1 polarization (\n p<\/jats:italic>\n \u2009<\u20090.001) and pro\u2010inflammatory cytokine secretion (IL\u20101\u03b2 and TNF\u2010\u03b1,\n p<\/jats:italic>\n \u2009<\u20090.05), while Ctss knockdown attenuated these effects (\n p<\/jats:italic>\n \u2009<\u20090.05). CTSS activated endothelial PAR2, suppressing endothelial MFSD2A and enhancing caveolin\u20101\u2010mediated transcytosis. Endothelial MFSD2A overexpression reduced vascular permeability independently of tight junction modulation (ZO\u20101, Occludin, Claudin\u20105,\n p<\/jats:italic>\n \u2009>\u20090.05). Microglia\u2010endothelial crosstalk was disrupted in diabetes, with ultrastructural alterations (cytoplasmic shrinkage, microglial amoeboid transformation) promoting cytotoxic interactions and exacerbate vascular endothelial damage.\n <\/jats:p>\n <\/jats:sec>\n \n Conclusions<\/jats:title>\n Hyperglycemia induces retinal microglial activation and CTSS upregulation, disrupting microglia\u2010endothelial crosstalk via the PAR2 pathway, suppressing endothelial MFSD2A expression, enhancing transcytosis, compromising iBRB integrity, and accelerating DR progression. Therapeutic strategies targeting microglial CTSS or endothelial MFSD2A represent promising avenues for preserving vision in diabetic patients.<\/jats:p>\n <\/jats:sec>","DOI":"10.1002\/cns.71187","type":"journal-article","created":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T15:07:17Z","timestamp":1790608037000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Hyperglycemia\u2010Induced Microglial\n CTSS<\/scp>\n Disrupts Blood\u2010Retinal Barrier Integrity via\n PAR2<\/scp>\n \u2010Dependent Downregulation of Endothelial\n MFSD2A<\/scp>"],"prefix":"10.1002","volume":"32","author":[{"given":"Chengye","family":"Xu","sequence":"first","affiliation":[{"name":"The Department of Endocrinology The First Affiliated Hospital of Harbin Medical University Harbin China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hongxue","family":"Li","sequence":"additional","affiliation":[{"name":"The Department of Endocrinology The First Affiliated Hospital of Harbin Medical University Harbin China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8326-3188","authenticated-orcid":false,"given":"Qian","family":"Xu","sequence":"additional","affiliation":[{"name":"The Department of Endocrinology The First Affiliated Hospital of Harbin Medical University Harbin China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kangqi","family":"Zhao","sequence":"additional","affiliation":[{"name":"The Department of Endocrinology The First Affiliated Hospital of Harbin Medical University Harbin China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yangwen","family":"Chen","sequence":"additional","affiliation":[{"name":"The Department of Endocrinology The First Affiliated Hospital of Harbin Medical University Harbin China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yan","family":"Wang","sequence":"additional","affiliation":[{"name":"The Department of Endocrinology The First Affiliated Hospital of Harbin Medical University Harbin China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wei","family":"Zou","sequence":"additional","affiliation":[{"name":"Acupuncture and Moxibustion Department III The First Affiliated Hospital of Heilongjiang University of Traditional Chinese Medicine Harbin China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Binglin","family":"Kuang","sequence":"additional","affiliation":[{"name":"Acupuncture and Moxibustion Department III The First Affiliated Hospital of Heilongjiang University of Traditional Chinese Medicine Harbin China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6309-9798","authenticated-orcid":false,"given":"Hongyu","family":"Kuang","sequence":"additional","affiliation":[{"name":"The Department of Endocrinology The First Affiliated Hospital of Harbin Medical University Harbin China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,9,28]]},"reference":[{"issue":"9735","key":"e_1_2_11_2_1","doi-asserted-by":"crossref","first-page":"124","DOI":"10.1016\/S0140-6736(09)62124-3","article-title":"Diabetic Retinopathy","volume":"376","author":"Cheung N.","year":"2010","journal-title":"Lancet"},{"key":"e_1_2_11_3_1","doi-asserted-by":"crossref","DOI":"10.3389\/fimmu.2022.1055087","article-title":"The Role of Inflammation in Immune System of Diabetic Retinopathy: Molecular Mechanisms, Pathogenetic Role and Therapeutic Implications","volume":"13","author":"Yue 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