Investigative ophthalmology & visual science最新文献

{"title":"The Spatial Transcriptomic Atlas of Human Limbus and Vital Niche Microenvironment Regulating the Fate of Limbal Epithelial Stem Cells.","authors":"Shiding Li, Hao Sun, Fei Fang, Siyi Zhang, Junzhao Chen, Chunyi Shao, Yao Fu, Liangbo Chen","doi":"10.1167/iovs.66.3.52","DOIUrl":"10.1167/iovs.66.3.52","url":null,"abstract":"<p><strong>Purpose: </strong>This study aimed to generate the spatial atlas of the human limbus using spatial transcriptomic technology and reveal the deep interaction among the niche microenvironment.</p><p><strong>Methods: </strong>The spatial transcriptomic atlas of human limbus was performed using 10× Genomics Space Ranger software platform. Single-cell RNA sequencing data of human limbal epithelial stem cells (LESCs) were downloaded for integrating analysis.</p><p><strong>Results: </strong>We profiled more than 400 spots within each sample and spatially located major cell types within the limbus area. LESCs were localized mainly in the basement membrane, and limbal niche cells were situated predominantly within the stromal area. Next, the limbus was divided into four regions based on histological structure, and the differential expressed genes among the four regions were analyzed. Notably, GPHB5 was highly expressed in the epithelium of the middle region and co-staining with deltaNp63 suggested it might be a novel potential biomarker of LESCs. Subsequently, limbal mesenchymal stem cells were found to exhibit the greatest amounts of ligands associated with LESCs. The widespread activity of COL6A2/CD44 signaling among limbal mesenchymal stem cells, melanocytes, immune cells, and LESCs indicate its essential role in mediating bidirectional communication via the collagen pathway.</p><p><strong>Conclusions: </strong>This research mapped the spatial positioning of key cells within the limbal niche and detailed interactions between major cell types. These findings provide a foundation for further LESC research and enhance our understanding of corneal biology.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 3","pages":"52"},"PeriodicalIF":5.0,"publicationDate":"2025-03-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11951063/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143700325","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":"Three-Dimensional Choroidal Vessels Assessment in Diabetic Retinopathy.","authors":"Elham Sadeghi, Katherine Du, Oluwaseyi Ajayi, Elli Davis, Nicola Valsecchi, Mohammed Nasar Ibrahim, Sandeep Chandra Bollepalli, Kiran Kumar Vupparaboina, Jose Alain Sahel, Jay Chhablani","doi":"10.1167/iovs.66.3.50","DOIUrl":"10.1167/iovs.66.3.50","url":null,"abstract":"<p><strong>Purpose: </strong>To evaluate choroidal vasculature in eyes with diabetic retinopathy (DR) using a novel three-dimensional algorithm.</p><p><strong>Methods: </strong>Patients with DR and healthy controls underwent clinical examinations and swept-source optical coherence tomography (PlexElite-9000). The choroidal layer was segmented using the ResUNet model. Phansalkar thresholding was used to binarize the choroidal vasculature. The macular area was divided into 5 sectors by a custom grid, and the 15 largest vessels in each sector were measured for mean choroidal vessel diameter (MChVD). Volumetric choroidal thickness (ChT) and the choroidal vascularity index (CVI) were calculated. A linear mixed model was used for analysis.</p><p><strong>Results: </strong>This retrospective cross-sectional study analyzed 73 eyes of 45 patients with DR (36 proliferative vs. 37 nonproliferative DR, and 42 with diabetic macular edema [DME] vs. 31 without DME), and 27 eyes of 21 age-match controls. The average MChVD was decreased in DR compared with healthy (200.472 ± 28.246 µm vs. 240.264 ± 22.350 µm; P < 0.001), as well as lower sectoral MChVD (P < 0.001); however, there was no difference in average ChT between the groups (P > 0.05). The global CVI was reduced in DR, especially in temporal and central sectors (P < 0.05). Compared with nonproliferative, proliferative DR exhibited decreased ChT (temporal, P < 0.05; other sectors, P > 0.05), CVI (P > 0.05), and MChVD (P > 0.05). DME eyes demonstrated lower but not statistically significant MChVD (196.449 ± 27.221 µm vs. 205.922 ± 29.134 µm; P > 0.05) and significantly reduced average CVI (0.365 ± 0.032 vs. 0.389 ± 0.040; P = 0.008) compared with non-DME eyes.</p><p><strong>Conclusions: </strong>DR and DME eyes showed reduced MChVD and CVI, likely owing to microvascular changes leading to ischemia. These findings highlight the need for new choroidal biomarkers to better understand DR's pathogenic mechanisms.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 3","pages":"50"},"PeriodicalIF":5.0,"publicationDate":"2025-03-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11951060/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143700335","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":"Orbital Clinicopathological Differences in Thyroid Eye Disease: An Analysis of Cytokines With Histopathological and Clinical Correlation.","authors":"Yue Li, Jiaqi Tang, Gaojing Jing, Yueyue Li, Rui Ma, Xin Kang, Liyuan Rong, Wenlu Liu, Lan Yao, Xiaohui Lv, Aijun Deng, Wei Wu, Xinji Yang","doi":"10.1167/iovs.66.3.33","DOIUrl":"10.1167/iovs.66.3.33","url":null,"abstract":"<p><strong>Purpose: </strong>To explore the pathological differences in orbital adipose/connective tissue between active and inactive thyroid eye disease (TED) subjects and their correlations with clinical characteristics.</p><p><strong>Methods: </strong>Orbital adipose/connective tissue samples from 42 TED subjects (20 active, 22 inactive) were collected during decompression surgery. We analyzed cytokine expression, inflammatory cell infiltration, inherent cell populations, and interstitial changes by Luminex and histopathology. Correlations were analyzed using Pearson and Spearman correlation analyses.</p><p><strong>Results: </strong>Among the 108 cytokines detected, active TED exhibited elevated platelet endothelial cell adhesion molecule 1 (PECAM-1), interleukin-23 (IL-23), a proliferation-inducing ligand (APRIL), IL-6, C-C motif chemokine ligand 2 (CCL2), β-nerve growth factor (NGF), and lower CCL21 and CCL5. The extent of infiltration by helper T (Th) cells and monocytes was significantly greater in the active group than in the inactive group. Adipocyte density was significantly elevated in active TED, whereas fibrosis was more prominent in inactive TED. Fifteen cytokines were significantly associated with inflammatory cell infiltration, with IL-16 showing the strongest correlations with T cells. Ten cytokines showed significant positive correlations with fibrosis. Four cytokines (IL-6, PECAM-1, IL-23 and transforming growth factor β1), Th cell infiltration and adipocyte density were significantly positively correlated with clinical activity score (CAS). Sixteen cytokines, along with adipocyte density, were significantly positively correlated with disease severity of TED.</p><p><strong>Conclusions: </strong>The orbital adipose/connective tissues of active and inactive TED subjects showed significant differences in terms of cytokines, inflammatory cells infiltration, inherent cells and interstitium. These pathological changes were correlated with clinical characteristics of TED.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 3","pages":"33"},"PeriodicalIF":5.0,"publicationDate":"2025-03-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11927299/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143656965","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":"An Approach to Predict Intraocular Diseases by Machine Learning Based on Vitreous Humor Immune Mediator Profile.","authors":"Risa Sugawara, Yoshihiko Usui, Akira Saito, Naoya Nezu, Hiroyuki Komatsu, Kinya Tsubota, Masaki Asakage, Naoyuki Yamakawa, Yoshihiro Wakabayashi, Masahiro Sugimoto, Masahiko Kuroda, Hiroshi Goto","doi":"10.1167/iovs.66.3.38","DOIUrl":"10.1167/iovs.66.3.38","url":null,"abstract":"<p><strong>Purpose: </strong>This study aimed to elucidate whether machine learning algorithms applied to vitreous levels of immune mediators predict the diagnosis of 12 representative intraocular diseases, and identify immune mediators driving the predictive power of machine learning model.</p><p><strong>Methods: </strong>Vitreous samples in 522 eyes diagnosed with 12 intraocular diseases were collected, and 28 immune mediators were measured using a cytometric bead array. The significance of each immune mediator was determined by employing five machine learning algorithms. Stratified k-fold cross-validation was performed to divide the dataset into training and test sets. The algorithms were assessed by analyzing precision, recall, accuracy, F-score, area under the receiver operating characteristics curve, area under the precision-recall curve, and feature importance.</p><p><strong>Results: </strong>Of the five machine learning models, random forest attained the maximum accuracy in the classification of 12 intraocular diseases in a multi-class setting. The random forest prediction models for vitreoretinal lymphoma, endophthalmitis, uveal melanoma, rhegmatogenous retinal detachment, and acute retinal necrosis demonstrated superior classification accuracy, precision, and recall. The top three important immune mediators for predicting vitreoretinal lymphoma were IL-10, granzyme A, and IL-6; those for endophthalmitis were IL-6, G-CSF, and IL-8; and those for uveal melanoma were RANTES, IL-8 and bFGF.</p><p><strong>Conclusions: </strong>The random forest algorithm effectively classified 28 immune mediators in the vitreous to accurately predict the diagnosis of vitreoretinal lymphoma, endophthalmitis, and uveal melanoma among 12 representative intraocular diseases. In summary, the results of this study enhance our understanding of potential new biomarkers that may contribute to elucidating the pathophysiology of intraocular diseases in the future.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 3","pages":"38"},"PeriodicalIF":5.0,"publicationDate":"2025-03-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11932427/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143656926","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":"Comparison Between MAIA and MP-3 In Healthy Subjects and Patients With Diabetes, Diabetic Retinopathy, and Age-Related Macular Degeneration.","authors":"Anna Marmalidou, Haleema Siddiqui, Muhammad Usman Jamil, Kwang Min Woo, Antonio Yaghy, A Yasin Alibhai, Hiroyuki Takahashi, Stephanie Kaiser, Keith Effert, Peter Y Zhao, Shilpa J Desai, Christopher C Robinson, Jay S Duker, Nadia K Waheed","doi":"10.1167/iovs.66.3.59","DOIUrl":"10.1167/iovs.66.3.59","url":null,"abstract":"<p><strong>Purpose: </strong>The purpose of this study was to assess the comparability of mean sensitivity (MS) values obtained using the Macular Integrity Assessment (MAIA; CenterVue S.p.A. [iCare], Padova, Italy) and Microperimeter-3 (MP-3; NIDEK CO., Ltd., Gamagori, Japan) microperimetry (MP) devices.</p><p><strong>Methods: </strong>This cross-sectional study includes subjects with healthy eyes, eyes with diabetes mellitus without diabetic retinopathy (DM no DR), diabetic retinopathy (DR), and non-exudative age-related macular degeneration (AMD). Patients underwent testing on both MAIA and MP-3 MP devices, using a 10-2 macular grid with 68 stimuli and identical parameters. A diagnosis-adjusted linear regression model and mixed modeling mapped MP-3 MS to MAIA MS and Bland-Altman analysis were used to assess the agreement.</p><p><strong>Results: </strong>One hundred eleven eyes (43 healthy eyes, 14 eyes with DM no DR, 32 eyes with DR, and 22 eyes with AMD) from 80 subjects (age = 57.2 ± 20.3 years) were enrolled. MAIA consistently showed higher MS than MP-3. The MP-3 device detected absolute scotomatous points in more eyes than MAIA (6 eyes [MAIA] vs. 10 eyes [MP-3]). Healthy eyes exhibited stronger agreements than those with DR (P < 0.001) or AMD (P = 0.015). Converting MP-3 to MAIA MS improved agreement and reduced coefficients of repeatability (CoRs) but did not fully account for inter-device variability. MP-3 classified more eyes as having relatively unstable or unstable fixation than MAIA (P = 0.014).</p><p><strong>Conclusions: </strong>Both devices effectively detect retinal functional changes and scotomas. The conversion methods developed in this study can aid cross-device comparisons, but retinal pathologies (DM and AMD) introduce additional inter-device variability. Future studies incorporating multiple devices should account for this variability in their study design.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 3","pages":"59"},"PeriodicalIF":5.0,"publicationDate":"2025-03-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11954536/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143730093","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":"Targeted siRNA Delivery Against RUNX1 Via tFNA: Inhibiting Retinal Neovascularization and Restoring Vessels Through Dll4/Notch1 Signaling.","authors":"Xiaodi Zhou, Xiaoxiao Xu, Qiong Wang, Yanting Lai, Linyan Zhang, Yunfeng Lin, Xiaoyan Ding, Limei Sun","doi":"10.1167/iovs.66.3.39","DOIUrl":"10.1167/iovs.66.3.39","url":null,"abstract":"<p><strong>Purpose: </strong>To assess the efficacy of tetrahedral framework nucleic acids (tFNAs) as a delivery system for small interfering RNA (siRNA) targeting RUNX1 (siRUNX1) in inhibiting retinal neovascularization (RNV) and restoring vascular integrity via the Dll4/Notch1 signaling pathway.</p><p><strong>Methods: </strong>tFNAs and tFNAs-siRUNX1 were synthesized using annealing of single-stranded DNAs and characterized by PAGE and high-performance capillary electrophoresis. Human umbilical vein endothelial cells were treated under hypoxic conditions with tFNAs-siRUNX1, and cellular uptake was evaluated using fluorescence microscopy and flow cytometry. Angiogenesis was assessed through EdU proliferation, tube formation, and wound-healing assays. In vivo experiments used oxygen-induced retinopathy (OIR) and laser-induced choroidal neovascularization (CNV) models in mice, with subsequent imaging by optical coherence tomography (OCT) and fundus fluorescence angiography. Gene and protein expression were analyzed by RT-PCR and Western blotting, focusing on the Dll4/Notch1 pathway and apoptosis markers.</p><p><strong>Results: </strong>tFNAs-siRUNX1 effectively inhibited endothelial cell proliferation, migration, and tube formation in vitro. In OIR and CNV models, it reduced neovascularization, nonperfusion areas, and vascular leakage. The mechanism involved modulation of the Dll4/Notch1 pathway, with decreased Dll4, Notch1, and Hes1 and increased Nts expression. tFNAs-siRUNX1 also reduced endothelial cell apoptosis via the Bcl-2/Bax pathway.</p><p><strong>Conclusions: </strong>tFNAs-siRUNX1 is a promising delivery system for targeting RNV, inhibiting neovascularization, and restoring retinal vascular integrity, providing a potential therapeutic alternative to anti-VEGF treatments.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 3","pages":"39"},"PeriodicalIF":5.0,"publicationDate":"2025-03-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11932424/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143656994","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":"Ocular Surface Involvements in the Development of Sjögren's Syndrome-Associated Dry Eye in the IL14α Transgenic Mouse.","authors":"Minjie Zhang, Yichen Liang, Han Wu, Rongrong Zong, Xiaobo Zhang, Hui He, Peter Sol Reinach, Zuguo Liu, Long Shen, Wei Li","doi":"10.1167/iovs.66.3.2","DOIUrl":"10.1167/iovs.66.3.2","url":null,"abstract":"<p><strong>Purpose: </strong>To investigate the ocular surface changes during progress of the Sjögren's Syndrome (SS), using a previously described IL14α transgenic mice (IL14α TG) SS model.</p><p><strong>Methods: </strong>The ocular surface of IL14α TG and C57BL/6 wild-type (WT) female mice were evaluated at the age of six, nine, 12, 15, and 18 months. Slit lamp microscopy observation, Oregon green dextran staining, Schirmer test, and periodic-acid-Schiff staining were assessed. Immunohistochemistry, immunofluorescence, and associated gene expression analysis by qPCR and ELISA were performed in cornea, conjunctiva, and lacrimal grand at different ages of the mice. Masson's trichome staining was conducted on lacrimal gland cryosections.</p><p><strong>Results: </strong>Compared with C57BL/6 WT mice, IL14α TG mice showed corneal barrier function damage and losses in conjunctival goblet cell density starting at nine months, whereas decreases in tear secretion started at 18 months of age. Significant increases in CD4+ T cell infiltration in the conjunctiva of IL14α TG mice was first observed at 6 months. Higher expression levels of inflammatory cytokines IL-17A, IFN-γ, IL-1β, and TNF-α in the conjunctiva, whereas MUC5AC and MUC5B had lower expression levels at nine months in the IL14α TG mice. However, lacrimal gland function-associated gene expression levels mostly decreased in IL14α TG mice at 12 months of age.</p><p><strong>Conclusions: </strong>Ocular surface tissue changes were involved in SS-like dry eye in a time-dependent manner in IL14α TG mice, and conjunctival T-cell infiltration may contribute to ocular surface pathological changes in an early stage of SS-related dry eye.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 3","pages":"2"},"PeriodicalIF":5.0,"publicationDate":"2025-03-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11887930/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143541806","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":"Ameboid Microglia as a Scavenger Role in Phagocytosis of Photoreceptor Outer Segment in an Experimental Retinal Detachment Model.","authors":"Manjing Cao, Yahan Zhang, Yan Li, Xian Zhang, Mingming Ma","doi":"10.1167/iovs.66.3.4","DOIUrl":"10.1167/iovs.66.3.4","url":null,"abstract":"<p><strong>Purpose: </strong>Photoreceptor (PR) death is the ultimate cause of irreversible vision loss in retinal detachment (RD). Previous studies have shown that microglia may have a dual role in RD. Nevertheless, the potential protective effects of microglia on PR are largely unknown. We aimed to uncover the phagocytic role of microglia in RD and propose a new concept to regulate PR survival.</p><p><strong>Methods: </strong>An RD model was conducted by injecting sodium hyaluronate into the subretinal space (SRS) of C57BL/6J wild type mice. Bioinformatics analysis was used to evaluate the highly enriched pathways and terms relating to phagocytosis in human datasets and mouse transcriptomes of RD. The observation of microglial morphology was performed by immunofluorescence through cryosection and flat mount. PLX 3397 was used for microglial ablation. Phagocytosis of the outer segment (OS) by microglia was confirmed by immunofluorescence and hematoxylin and eosin staining. Expression of phagocytic markers in microglia was detected by immunofluorescence of cryosection. The PR survival was measured by TUNEL assay and hematoxylin and eosin staining. The optical coherence tomography (OCT) images through the center of the fovea in twelve patients were obtained to observe the clinic features of IS/OS dynamics after RD.</p><p><strong>Results: </strong>The results showed that OS went through an accumulation-clearance process after RD. Ameboid microglia accumulated in the SRS and engulfed OS. Upregulation of phagocytic markers was observed in subretinal microglia. Depletion of microglia led to failure of OS clearance and retinal ruffling, which had the same characteristics as outer retinal undulation (ORU) in some patients with RD. PR did not benefit from microglial depletion, as no morphology and thickness recovery of PR was observed in the long term.</p><p><strong>Conclusions: </strong>These results elucidate that microglial phagocytosis of OS is a critical process after RD. Insufficient phagocytosis leads to the accumulation of OS in the SRS and PR abnormalities. Appropriate regulation of microglial phagocytosis to remove OS may be a new concept to regulate photoreceptor survival.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 3","pages":"4"},"PeriodicalIF":5.0,"publicationDate":"2025-03-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11892526/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143541293","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":"Single-Cell Multiomics Analysis of Early Wound Response Programs in the Mouse Corneal Epithelium.","authors":"Zhao-Jing Lu, Jin-Guo Ye, Jing-Ni Li, Jiang-Bo Liang, Ming Zhou, Qiu-Ling Hu, Qi-Kai Zhang, Yu-Heng Lin, Ying-Feng Zheng","doi":"10.1167/iovs.66.3.9","DOIUrl":"10.1167/iovs.66.3.9","url":null,"abstract":"<p><strong>Purpose: </strong>Wound healing is crucial for restoring homeostasis in living organisms. Although wound response mechanisms have been studied extensively, the gene regulatory programs involved remain to be elucidated. Here, we used single-cell RNA sequencing (RNA-seq) and ATAC sequencing (ATAC-seq) analysis to profile the regulatory landscape of mouse corneal epithelium in early wound response.</p><p><strong>Methods: </strong>We used our previously published single-cell data sets of homeostatic adult mouse corneal epithelium as the unwounded group. The wounded group data sets were obtained by sequencing the epithelium after an annular epithelial wound. Following the integration of the relevant data sets, the Seurat and ArchR packages were employed for single-cell RNA-seq and single-cell ATAC-seq data processing and downstream analysis, respectively. The Monocle 2 was used for pseudo-time analysis, CellChat for intercellular communication analysis, and pySCENIC for analyzing transcription factors. The expression of key genes was validated via immunofluorescence staining and quantitative real-time PCR.</p><p><strong>Results: </strong>Our data show that the number of cell type-specific genes decreases and the number of common transcriptional responses increases in early wound response. Concurrently, we find that the chromatin accessibility landscape undergoes significant changes across all epithelial cell types and that the wound-induced open regions are similarly distributed across the genome. Motif enrichment analysis shows that Fosl1/AP-1 binding site is highly enriched among the opened regions. However, by assessing the correlation between changes in chromatin accessibility and gene expression, we observe that only a small subset of wound-induced genes shows a high correlation with the accessibility of nearby chromatin.</p><p><strong>Conclusions: </strong>Our study provides a detailed single-cell landscape for transcriptomic and epigenetic changes in mouse corneal epithelium during early wound response, which improved our understanding of the mechanisms of wound healing.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 3","pages":"9"},"PeriodicalIF":5.0,"publicationDate":"2025-03-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11892537/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143556832","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":"STING Deficiency Promotes Th17-Like Tfh to Aggravate the Experimental Autoimmune Uveitis.","authors":"Zhuang Li, Xiuxing Liu, Zuoyi Li, Zhiqiang Xiao, Guanyu Chen, Yangyang Li, Jun Huang, Yunwei Hu, Haixiang Huang, Wenjie Zhu, Yuxun Shi, Minzhen Wang, Yanyan Xie, Wenru Su, Xiaoqing Chen, Dan Liang","doi":"10.1167/iovs.66.3.8","DOIUrl":"10.1167/iovs.66.3.8","url":null,"abstract":"<p><strong>Purpose: </strong>The purpose of this study was to explore the underlying mechanism that Th17-like T follicular helper cells (Tfh) orchestrated by STING signaling have a pathogenic role in experimental autoimmune uveitis (EAU).</p><p><strong>Methods: </strong>The differences of transcriptome and gene ontology (GO) pathway of Tfh between EAU and control mice were analyzed by single-cell RNA sequence (scRNA-seq) and bulk RNA sequence. Additionally, draining lymph nodes (DLNs) were extracted to verify the expression of IL-17A and IFN-γ in Tfh from EAU and control mice by flow cytometry. Then, the scRNA-seq and flow cytometry were used to explore the different proportion of Tfh between STING deficiency (Sting-/-) mice and wild type (WT) mice. In vitro, naïve CD4+ T cells were isolated from Sting-/- mice and WT mice to induce the Tfh under the induction condition. In addition, flow cytometry was used to detect the different induction ratio and the IL-17A expression between 2 groups of naïve CD4+ T cells.</p><p><strong>Results: </strong>Compared with control mice, marked increase of Tfh was observed in EAU, accompanied by elevated levels of Th1 and Th17 cells. Moreover, Th17-related genes, such as Rorc, Il22, Il23r, Il17a, and Il17f, and the corresponding GO pathways were upregulated in Tfh from EAU. The scRNA-seq showed that a higher proportion of Tfh was observed in the DLNs from Sting-/- mice than WT mice, which was verified by flow cytometry. When STING was knocked out, the Tfh was characterized with upregulated Th17-related phenotype in vivo, and there was a higher induction ratio of Tfh whose IL-17A expression was significantly increased in vitro. Notably, the STING expression of CD4+ T cells was downregulated in the EAU. STING-deficient EAU mice displayed more severe retinal inflammation, characterized by massive infiltration of CD4+ T cells, including Th1 and Th17 subsets. Importantly, treatment with a STING agonist alleviated inflammation of EAU.</p><p><strong>Conclusions: </strong>Th17-like Tfh cells play a pathogenic role in the EAU. STING deficiency promotes the differentiation and phenotypic transformation of Th17-like Tfh cells, exacerbating the inflammatory response in EAU. These findings highlight the potential of targeting STING to modulate Tfh cells as a therapeutic strategy for uveitis.</p>","PeriodicalId":14620,"journal":{"name":"Investigative ophthalmology & visual science","volume":"66 3","pages":"8"},"PeriodicalIF":5.0,"publicationDate":"2025-03-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11892529/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143556834","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}