Investigative ophthalmology & visual science最新文献

{"title":"Fatty Acid Desaturase 1 Knockdown Promotes Wound Healing and Functional Recovery of the Corneal Epithelium in Diabetes.","authors":"Yangqi Zhao, Yi Dong, Qingqing Zheng, Yue Zhao, Yingjie Ni, Peijin Qiu, Chuannan Chen, Mengyue Xu, Chaoyang Hong, Ting Shen","doi":"10.1167/iovs.66.6.6","DOIUrl":"10.1167/iovs.66.6.6","url":null,"abstract":"<p><strong>Purpose: </strong>Fatty acid desaturase 1 (FADS1) is significantly and specifically upregulated following diabetic corneal injury. However, its role in diabetic keratopathy remains unclear. This study aimed to investigate the impact of FADS1 on wound healing and functional recovery of the diabetic corneal epithelium and explore its potential mechanisms.</p><p><strong>Methods: </strong>Using high-glucose-induced corneal epithelial cells and a streptozotocin-induced type 1 diabetic mouse model, FADS1 expression was suppressed via FADS1 small interfering RNA (siRNA). Cell migration was assessed using scratch and transwell assays. Wound healing and functional recovery of the corneal epithelium were evaluated using sodium fluorescein staining, anterior segment optical coherence tomography, hematoxylin and eosin staining, and immunofluorescence staining.</p><p><strong>Results: </strong>FADS1 knockdown promoted wound healing and functional recovery of the diabetic corneal epithelium both in vivo and in vitro. Suppression of FADS1 enhanced high-glucose-induced corneal epithelial cell migration, which was dependent on elevated levels of the upstream metabolite γ-linolenic acid. This effect was mediated through the activation of the mitogen-activated protein kinase signaling pathway and the accumulation of autophagosomes.</p><p><strong>Conclusions: </strong>After diabetic corneal epithelial injury, FADS1 expression is specifically upregulated. Knockdown of FADS1 promotes wound healing and functional recovery, suggesting a novel therapeutic strategy for diabetic keratopathy.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 6","pages":"6"},"PeriodicalIF":5.0,"publicationDate":"2025-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12136103/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144208525","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Choroidal Hyperreflective Foci Represent a Common Finding Across Different Types of Macular Atrophy.","authors":"Lorena Ulla, Claudio Foti, Alessandro Arrigo, Maurizio Battaglia Parodi, Maria Vittoria Cicinelli, Paola Marolo, Elisabetta Miserocchi, Mario Peronetti, Francesco Bandello, Enrico Borrelli, Michele Reibaldi","doi":"10.1167/iovs.66.6.42","DOIUrl":"10.1167/iovs.66.6.42","url":null,"abstract":"<p><strong>Purpose: </strong>To evaluate the presence, distribution, and potential implications of choroidal hyperreflective foci (HRFs) across different types of macular atrophy and their relationship with the size of atrophic regions.</p><p><strong>Methods: </strong>This retrospective case series analyzed 95 eyes from 87 patients with macular atrophy caused by various conditions, including geographic atrophy due to age-related macular degeneration, pachychoroid disease, punctate inner choroidopathy, angioid streaks, and Stargardt disease. Structural optical coherence tomography (OCT) images were evaluated to identify HRFs in different choroidal layers. HRF counts were correlated with the size of macular atrophy and underlying conditions using multiple regression analysis.</p><p><strong>Results: </strong>HRFs were observed in all cases, with their number significantly associated with the size of macular atrophy (P < 0.001). The highest HRF counts were found in Stargardt disease, while the lowest were in pachychoroid disease. However, no association was identified between HRF quantity and specific disease type (P = 0.2). HRFs were primarily located in the choriocapillaris, Sattler's layer, and near Bruch's membrane but were absent within choroidal vessels.</p><p><strong>Conclusions: </strong>HRFs represent a common OCT finding in various disorders associated with macular atrophy. Their quantity correlates with atrophy size rather than disease type, suggesting they are likely normal choroidal components rendered visible by RPE loss. Future studies are warranted to elucidate their origin and potential implications for disease progression.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 6","pages":"42"},"PeriodicalIF":5.0,"publicationDate":"2025-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12173085/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144284390","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Plin2 Coordinates Immune and Metabolic Reprogramming in Lacrimal Gland Aging.","authors":"Xiaoting Pei, Shuting Xuan, Jingwen Yang, Mengru Ba, Tingting Yang, Duliurui Huang, Di Qi, Dingli Lu, Shenzhen Huang, Zhijie Li","doi":"10.1167/iovs.66.6.79","DOIUrl":"10.1167/iovs.66.6.79","url":null,"abstract":"<p><strong>Purpose: </strong>Aging impairs lacrimal gland function, contributing to dry eye syndrome and reduced quality of life. This study aimed to identify core molecular regulators of lacrimal gland aging and delineate their roles in immune imbalance and metabolic dysfunction.</p><p><strong>Methods: </strong>Bulk transcriptomic profiling and single-cell RNA sequencing (scRNA-seq) were performed on lacrimal glands from young and aged C57BL/6 mice. Differentially expressed genes (DEGs) were screened, followed by machine learning-based feature selection using least absolute shrinkage and selection operator regression and support vector machine-recursive feature elimination. Immune cell composition was inferred using the CIBERSORT algorithm, and functional enrichment (gene ontology, Kyoto Encyclopedia of Genes and Genomes, and gene set enrichment analysis) and cell-type-resolved spatial analyses were conducted to elucidate aging-associated pathways.</p><p><strong>Results: </strong>Among 659 DEGs, Perilipin-2 (Plin2) emerged as the sole hub gene identified by both algorithms, with perfect classification performance (area under the curve = 1.00). Aging was associated with increased infiltration of pro-inflammatory cells (naïve B cells, M1 macrophages) and depletion of anti-inflammatory subsets (plasma cells, M2 macrophages). Plin2 expression exhibited a significant inverse association with pro-senescent immune populations and a positive correlation with regulatory immune cells. Functional analyses linked Plin2 to T/B cell receptor signaling, cytokine interaction, and ribosomal biosynthesis. scRNA-seq revealed Plin2 downregulation in fibroblasts and Mono/Mφ/DC subsets in aged glands (P < 0.001), indicating stromal-immune dysfunction.</p><p><strong>Conclusions: </strong>Our study identifies Plin2 as a master regulator of immune-metabolic aging in the lacrimal gland. Its decline accelerates inflammaging and metabolic reprogramming, contributing to tissue atrophy and tear deficiency. Plin2 represents a promising candidate biomarker and potential therapeutic target for age-related ocular diseases.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 6","pages":"79"},"PeriodicalIF":5.0,"publicationDate":"2025-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12205569/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144496678","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"FGF10 Protects the Corneal Epithelium From Dry Eye-Induced Oxidative and ER Stress Via Upregulation of SLC7A11.","authors":"Zhiwei Zha, Juan Li, Shuang Xie, Xunjie Shang, Zixin Xu, Haiyuan Chang, Kaisheng Wang, Yang Liu, Wei Chen","doi":"10.1167/iovs.66.6.72","DOIUrl":"10.1167/iovs.66.6.72","url":null,"abstract":"<p><strong>Purpose: </strong>To investigate the protective effect of fibroblast growth factor 10 (FGF10) on the corneal epithelium in dry eye disease (DED) and reveal the underlying mechanism.</p><p><strong>Methods: </strong>DED mouse model was induced via scopolamine injections and low-humidity airflow to evaluate the therapeutic effects of FGF10. Mice received topical FGF10 (5, 25, or 125 µg/mL) or vehicle for seven days. Corneal fluorescein staining, oxidative stress (ROS levels), endoplasmic reticulum (ER) stress, and apoptosis were evaluated. To investigate protective mechanisms on corneal epithelium cells, hyperosmolar-stressed HCE-2 cells were treated with 100 ng/mL FGF10, and RNA sequencing was performed. Transcriptomic analysis identified SLC7A11, a key regulator of cellular antioxidant defense, as significantly upregulated by FGF10. SLC7A11's functional importance was validated through siRNA-mediated silencing in HCE-2 cells and AAV-mediated overexpression in mouse model.</p><p><strong>Results: </strong>FGF10 treatment significantly improved corneal epithelial integrity in dry eye mice, reducing fluorescein staining, ROS level, and ER stress markers, while increasing Bcl-2 and decreasing BAX. RNA sequencing revealed that FGF10 stimulated antioxidant signaling pathways and upregulated SLC7A11 expression. FGF10 also increased SLC7A11 protein levels in HCE-2 cells and dry eye corneas. Silencing of SLC7A11 in vitro prevented FGF10-induced reductions in ROS, ER stress, and apoptosis. Furthermore, AAV-mediated overexpression of SLC7A11 in dry eye mice recapitulated the protective effects observed with FGF10 treatment.</p><p><strong>Conclusions: </strong>FGF10 protects mouse corneal epithelium and HCE-2 cells from oxidative stress, ER stress, and apoptosis, partially through SLC7A11 upregulation. The FGF10-SLC7A11 pathway represents a promising therapeutic target in dry eye.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 6","pages":"72"},"PeriodicalIF":5.0,"publicationDate":"2025-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12204230/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144475177","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Impact of Optic Nerve Tortuosity, Globe Proptosis, and Size on Retinal Ganglion Cell Thickness Across General, Glaucoma, and Myopic Populations.","authors":"Charis Y N Chiang, Xiaofei Wang, Stuart K Gardiner, Martin Buist, Michaël J A Girard","doi":"10.1167/iovs.66.6.4","DOIUrl":"10.1167/iovs.66.6.4","url":null,"abstract":"<p><strong>Purpose: </strong>The purpose of this study was to investigate the impact of optic nerve tortuosity (ONT), and the interaction of globe proptosis and size on retinal ganglion cell (RGC) thickness, using retinal nerve fiber layer (RNFL) thickness, across general, glaucoma, and myopic populations.</p><p><strong>Methods: </strong>This study analyzed 17,940 eyes from the UKBiobank cohort (ID 76442), including 72 glaucomatous and 2475 myopic eyes. Artificial intelligence models were developed to derive RNFL thickness corrected for ocular magnification from 3D optical coherence tomography scans and orbit features from 3D magnetic resonance images, including ONT, globe proptosis, axial length, and a novel feature: the interzygomatic line-to-posterior pole (ILPP) distance - a composite marker of globe proptosis and size. Generalized estimating equation (GEE) models evaluated associations between orbital and retinal features.</p><p><strong>Results: </strong>RNFL thickness was positively correlated with ONT and ILPP distance (r = 0.065, P < 0.001 and r = 0.206, P < 0.001, respectively) in the general population. The same was true for glaucoma (r = 0.040, P = 0.74 and r = 0.224, P = 0.059), and for myopia (r = 0.069, P < 0.001 and r = 0.100, P < 0.001). GEE models revealed that straighter optic nerves and shorter ILPP distance were predictive of thinner RNFL in all populations.</p><p><strong>Conclusions: </strong>Straighter optic nerves and decreased ILPP distance could cause RNFL thinning, possibly due to greater traction forces. ILPP distance emerged as a potential biomarker of axonal health. These findings underscore the importance of orbit structures in RGC axonal health and warrant further research into orbit biomechanics.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 6","pages":"4"},"PeriodicalIF":5.0,"publicationDate":"2025-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12136101/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144199150","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Age-Related Dysregulation of α5β1 and αvβ3 Integrin Activity Alters Contractile Properties of Trabecular Meshwork Cells.","authors":"Kassidy L Johns, Jennifer A Faralli, Mark S Filla, Nandini S Shah, Ying Ying Sun, Kate E Keller, Donna M Peters","doi":"10.1167/iovs.66.6.31","DOIUrl":"10.1167/iovs.66.6.31","url":null,"abstract":"<p><strong>Purpose: </strong>Age and elevated intraocular pressure are major risk factors for primary open-angle glaucoma (POAG) which is caused by a restriction in aqueous humor outflow from the anterior chamber. In this study, we investigated whether age-related changes in integrin subunit expression and activity signifies an early event in initiating fibrotic-like changes in the TM that could restrict outflow.</p><p><strong>Methods: </strong>Human trabecular meshwork (TM) cells from young (<40 years) and old (>50 years) donor eyes were used. Flow cytometry, RT-qPCR, and immunofluorescence microscopy were used to evaluate levels of integrin and αSMA expression. On-cell westerns were used to determine fibronectin levels. Collagen gel contraction assays were used to determine contractile properties of cells and shRNA was used to knockdown α5β1 integrin levels.</p><p><strong>Results: </strong>Studies revealed a significant decrease in α5 integrin expression in TM cells from older individuals. This loss was accompanied by an increase in activated but not total αvβ3 integrin levels. TM cells from older donors expressed higher levels of αSMA mRNA, assembled αSMA-containing stress fibers, and contracted collagen gels significantly more than young TM cells. TM cells from old donors also assembled higher levels of insoluble fibronectin fibrils and contained higher levels of EDB+ fibronectin in their extracellular matrix. shRNA knockdown of α5 integrin subunits showed that the increase in αvβ3 integrin activity was due to lower levels of α5 integrin expression.</p><p><strong>Conclusions: </strong>These studies suggest that age-related dysregulation of α5β1 and αvβ3 integrin signaling may represent an important early molecular event in inducing fibrogenic pathways associated with POAG.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 6","pages":"31"},"PeriodicalIF":5.0,"publicationDate":"2025-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12161370/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144247913","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"HIF-1α Promotes Inflammatory Responses in Aspergillus Fumigatus Keratitis by Activating Pyroptosis Through Caspase-8/GSDMD Pathway.","authors":"Hua Yang, Mengzhu Liu, Shiqi Song, Qiang Xu, Jieun Lee, Jintao Sun, Shasha Xue, Xiaoyan Sun, Chengye Che","doi":"10.1167/iovs.66.6.32","DOIUrl":"10.1167/iovs.66.6.32","url":null,"abstract":"<p><strong>Purpose: </strong>This research was designed to explore the expression patterns and functional significance of hypoxia-inducible factor-1α (HIF-1α) in the inflammatory response associated with Aspergillus fumigatus (A. fumigatus) keratitis.</p><p><strong>Methods: </strong>Mouse models of A. fumigatus keratitis were created by scraping the corneal epithelium and applying A. fumigatus on the corneal surface. In the in vitro experiments, human corneal epithelial cells (HCECs) and THP-1 macrophages stimulated by A. fumigatus were used to investigate the cellular responses. HIF-1α was inhibited using LW6. Western blot, immunofluorescence, and quantitative reverse transcription polymerase chain reaction (qRT-PCR) were performed to assess the expression levels of HIF-1α in A. fumigatus keratitis. The inflammatory response was evaluated using clinical scoring, corneal thickness measurements, hematoxylin and eosin (H&E) staining, corneal fluorescein sodium staining, and a cell scratch test. The polarization of macrophages was determined using flow cytometry. The molecular mechanisms of HIF-1α were assessed by qRT-PCR and Western blot.</p><p><strong>Results: </strong>In A. fumigatus keratitis, the expression of HIF-1α was significantly increased at both the mRNA and protein levels. Compared with the controls, HIF-1α inhibitor accelerated corneal epithelial repair, reduced the infiltration of macrophages, induced shift in macrophage polarization, and attenuated the inflammatory response. HIF-1α exerts a pro-inflammatory effect in A. fumigatus keratitis by modulating the expression of inflammatory mediators and engaging in pyroptosis via the caspase-8/GSDMD signaling pathway.</p><p><strong>Conclusions: </strong>In conclusion, HIF-1α promotes A. fumigatus keratitis by inhibiting corneal epithelial repair and promoting inflammation, leading to increased severity of the disease.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 6","pages":"32"},"PeriodicalIF":5.0,"publicationDate":"2025-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12165259/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144258026","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Relationship of Choroidal Thickness With Age-Related Macular Degeneration in Asians: An Asian Eye Epidemiology Consortium Meta-Analysis.","authors":"Mariko Sasaki, Kai Xiong Cheong, Crystal Chun Yuen Chong, Marco Yu, Akiko Hanyuda, Kenya Yuki, Kazuno Negishi, Sawako Hashimoto, Kohta Fujiwara, Kohei Sonoda, Ya Xing Wang, Fei Gao, Anyarak Amornpetchsathaporn, Methaphon Chainakul, Ramyaa Srinivasan, Rehana Khan, Rajiv Raman, Paisan Ruamviboonsuk, Sung Ho Kim, Su Jeong Song, Mohammad Hassan Emamian, Hassan Hashemi, Akbar Fotouhi, Juping Liu, Xiaorong Li, Jost B Jonas, Chui Ming Gemmy Cheung, Tien Yin Wong, Ching-Yu Cheng, Yih Chung Tham, Yasuo Yanagi, Anna Cheng Sim Tan","doi":"10.1167/iovs.66.6.36","DOIUrl":"10.1167/iovs.66.6.36","url":null,"abstract":"<p><strong>Purpose: </strong>To compare the choroidal thickness (CT) of participants with various stages of age-related macular degeneration vs. normal controls through a meta-analysis of studies conducted within the Asian Eye Epidemiology Consortium.</p><p><strong>Methods: </strong>Eight population-based studies from China, Iran, Japan, and Singapore were included. Axial length and spherical equivalent measurements and imaging with color fundus photography and spectral-domain optical coherence tomography were performed. Random-effects meta-analysis was performed to examine the association between AMD and its stages (early AMD, intermediate AMD [iAMD], neovascular AMD [nAMD], and geographic atrophy [GA]) with CT, while adjusting for age, sex, current smoking status, and axial length/spherical equivalent.</p><p><strong>Results: </strong>Of 17,916 participants, 13,116 participants (mean age, 62.15 ± 9.66 years) were included into the study. The mean unadjusted CT was 245.01 ± 84.04 µm (mean CT, 255.4 µm [no AMD], 263.59 µm [early AMD], 270.64 µm [iAMD], 273.32 µm [nAMD], and 156.50 µm [GA]). The presence of AMD was associated with a thicker choroid (β = 11.51; 95% confidence interval [CI], 4.10-18.92). AMD severity was also positively associated with CT. Early AMD (β = 8.75; 95% CI, 0.03-17.47), iAMD (β = 19.68; 95% CI, 13.20-26.16), and nAMD (β = 34.15; 95% CI, 6.84-61.46) were each positively associated with a thicker CT after adjusting for age, sex, smoking, and spherical equivalent. GA was not significantly associated with CT.</p><p><strong>Conclusions: </strong>In a large Asian cohort, AMD is associated with a thicker choroid in early AMD, iAMD, and nAMD, but not in GA. Studying the CT will help to better characterize Asian AMD phenotypes, which may show differences compared with AMD phenotypes in Western populations.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 6","pages":"36"},"PeriodicalIF":5.0,"publicationDate":"2025-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12165263/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144258030","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"NR4A1 Alleviates Subretinal Fibrosis by Inhibiting Macrophage to Myofibroblast Transition.","authors":"Rufei Yang, Tingting Zong, Ning Wang, Feng Wang, Ying Su","doi":"10.1167/iovs.66.6.47","DOIUrl":"10.1167/iovs.66.6.47","url":null,"abstract":"<p><strong>Purpose: </strong>Subretinal fibrosis (SF) secondary to neovascular age-related macular degeneration (nAMD) is the most predominant cause of central visual impairment in all patients with AMD. NR4A1 is a member of the nuclear orphan receptor superfamily, and has shown inhibitory effects on fibrosis in tissues such as the dermis, intestines, and heart. Cytosporone B (Csn-B) is a natural agonist of NR4A1. This study aims to explore whether NR4A1 plays a role in SF associated with nAMD.</p><p><strong>Methods: </strong>Mice RPE-choroid-sclera flat mounts were prepared for serial observation of changes in macrophage infiltration, as well as macrophage to myofibroblast transformation (MMT). The morphology of MMT cells and differences in extracellular matrix (ECM) expression were further observed in TGF-β1-induced THP-1 cells. The role of NR4A1 in MMT was confirmed by small interfering RNA (siRNA) after changes in NR4A1 were observed. To determine whether NR4A1 could be a target for SF treatment, we intervened with the Csn-B and observed the MMT and SF changes.</p><p><strong>Results: </strong>Macrophages were rapidly recruited in the early stage and gradually decreased after the second week. MMT was observed in the lesions and the maximum number of MMT cells was observed at the third week. NR4A1 was transiently upregulated with induction, followed by a gradual decrease and a continuous phosphorylation. The knockdown of NR4A1 promoted MMT and ECM expression, whereas treatment with Csn-B had an inhibitory effect. P-NR4A1 expression was significantly suppressed in Csn-B-treated MMT cells. Finally, MK-2206 was found to inhibit sustained TGF-β1-induced NR4A1 phosphorylation and also ECM expression.</p><p><strong>Conclusions: </strong>NR4A1 inhibits MMT and reduces ECM deposition in SF. Its agonist Csn-B inhibits MMT by inhibiting AKT-induced NR4A1 phosphorylation, which then attenuates SF.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 6","pages":"47"},"PeriodicalIF":5.0,"publicationDate":"2025-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12178436/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144302083","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"RIPK3-Mediated Necroptosis Drives Macrophage Infiltration and Corneal Neovascularization After Alkali Burn.","authors":"Yue Li, Boyu Yang, Qi Chen, Yuchen Zhou, Wen Hu, Xiaojia Huang, Guangyi Huang, Ningning Tang, Fen Tang, Hui Huang, Qianqian Lan, Wenjing He, Fan Xu, Yiming Ye, Li Jiang","doi":"10.1167/iovs.66.6.54","DOIUrl":"10.1167/iovs.66.6.54","url":null,"abstract":"<p><strong>Purpose: </strong>To reveal the role of receptor-interacting protein kinase 3 (RIPK3) in regulating macrophage inflammation and corneal neovascularization (CoNV) induced by alkali burn.</p><p><strong>Methods: </strong>A corneal alkali burn (AB) model was established in C57BL/6J (wild-type) and RIPK3fl/flCx3cr1+/cre (RIPK3-/-, RIPK3 knockout [KO]) mice using sodium hydroxide. Anterior segment optical coherence tomography and hematoxylin and eosin staining were used to evaluate the impact of RIPK3 on corneal edema and morphology. CoNV was detected by slit-lamp microscopy and whole-mount immunofluorescence staining of cornea. Corneal macrophage and necroptotic cell death was analyzed through immunofluorescence staining and propidium iodide (PI) staining. Activation of the necroptosis pathway was examined after corneal AB by western blot.</p><p><strong>Results: </strong>Necroptosis-related proteins RIPK1, RIPK3, and mixed lineage kinase domain-like (MLKL) were upregulated and activated following corneal AB. Among these, RIPK3 demonstrated the most pronounced increase. Notably, the elevated level of RIPK3 was prominently colocalized with the infiltrating F4/80+ macrophages. RIPK3 KO significantly alleviated corneal edema and morphology defects. Additionally, as the corneal morphological defects progressed, macrophages became activated, and CoNV and lymphangiogenesis (LyG) were enhanced. RIPK3 KO markedly reduced AB-induced macrophage accumulation, as well as CoNV and LyG. RIPK3 KO mice also showed a meaningful decrease in PI+ necroptotic cells. Mechanistically, AB-induced necroptosis stimulated the expression of MLKL and fibroblast growth factor 2 (FGF2), whereas RIPK3 deficiency decreased their expression.</p><p><strong>Conclusions: </strong>This study revealed that RIPK3-mediated necroptosis drives macrophage inflammation and CoNV. Targeting RIPK3 could effectively suppress these responses by inhibiting the MLKL/FGF2 pathway, making it a promising therapeutic strategy for corneal AB.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 6","pages":"54"},"PeriodicalIF":5.0,"publicationDate":"2025-06-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12178437/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144316932","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}