Centre for Genomic Medicine, Biomedical Sciences Research Institute, Ulster University, Coleraine, Northern Ireland, United Kingdom.
Department of Eye and Vision Science, Insitute of Life Course and Medical Sciences, University of Liverpool, Liverpool, United Kingdom.
PLoS One. 2024 Jul 11;19(7):e0307227. doi: 10.1371/journal.pone.0307227. eCollection 2024.
Primary open angle glaucoma is a leading cause of visual impairment and blindness which is commonly treated with drugs or laser but may require surgery. Tenon's ocular fibroblasts are involved in wound-healing after glaucoma filtration surgery and may compromise a favourable outcome of glaucoma surgery by contributing to fibrosis. To investigate changes in gene expression and key pathways contributing to the glaucomatous state we performed genome-wide RNA sequencing. Human Tenon's ocular fibroblasts were cultured from normal and glaucomatous human donors undergoing eye surgery (n = 12). mRNA was extracted and RNA-Seq performed on the Illumina platform. Differentially expressed genes were identified using a bioinformatics pipeline consisting of FastQC, STAR, FeatureCounts and edgeR. Changes in biological functions and pathways were determined using Enrichr and clustered using Cytoscape. A total of 5817 genes were differentially expressed between Tenon's ocular fibroblasts from normal versus glaucomatous eyes. Enrichment analysis showed 787 significantly different biological functions and pathways which were clustered into 176 clusters. Tenon's ocular fibroblasts from glaucomatous eyes showed signs of fibrosis with fibroblast to myofibroblast transdifferentiation and associated changes in mitochondrial fission, remodeling of the extracellular matrix, proliferation, unfolded protein response, inflammation and apoptosis which may relate to the pathogenesis of glaucoma or the detrimental effects of topical glaucoma therapies. Altered gene expression in glaucomatous Tenon's ocular fibroblasts may contribute to an unfavourable outcome of glaucoma filtration surgery. This work presents a genome-wide transcriptome of glaucomatous versus normal Tenon's ocular fibroblasts which may identify genes or pathways of therapeutic value to improve surgical outcomes.
原发性开角型青光眼是导致视力损害和失明的主要原因，通常采用药物或激光治疗，但可能需要手术。Tenon 氏眼纤维母细胞参与青光眼滤过手术后的伤口愈合过程，通过促进纤维化，可能会影响青光眼手术的良好效果。为了研究导致青光眼状态的基因表达变化和关键途径，我们进行了全基因组 RNA 测序。从接受眼部手术的正常和青光眼患者（n = 12）中培养了人 Tenon 氏眼纤维母细胞。提取 mRNA 并在 Illumina 平台上进行 RNA-Seq。使用由 FastQC、STAR、FeatureCounts 和 edgeR 组成的生物信息学管道识别差异表达基因。使用 Enrichr 确定生物学功能和途径的变化，并使用 Cytoscape 进行聚类。正常眼与青光眼眼中的 Tenon 氏眼纤维母细胞之间有 5817 个基因差异表达。富集分析显示有 787 个显著不同的生物学功能和途径，它们被聚类为 176 个簇。青光眼眼中的 Tenon 氏眼纤维母细胞显示出纤维化的迹象，伴有成纤维细胞向肌成纤维细胞的转分化，以及线粒体裂变、细胞外基质重塑、增殖、未折叠蛋白反应、炎症和细胞凋亡的相关变化，这些可能与青光眼的发病机制或局部青光眼治疗的有害影响有关。青光眼 Tenon 氏眼纤维母细胞中的基因表达改变可能导致青光眼滤过手术的不良结果。这项工作展示了青光眼与正常 Tenon 氏眼纤维母细胞的全基因组转录组，这可能会识别出具有治疗价值的基因或途径，以改善手术结果。
