PDF(Pubmed)
Abstract：
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.
摘要：
原发性开角型青光眼是视觉障碍和失明的主要原因，通常使用药物或激光治疗，但可能需要手术。Tenon的眼成纤维细胞参与青光眼滤过手术后的伤口愈合，并可能通过促进纤维化而损害青光眼手术的有利结果。为了研究导致青光眼状态的基因表达和关键途径的变化，我们进行了全基因组RNA测序。从接受眼科手术的正常和青光眼人类供体中培养人Tenon的眼成纤维细胞（n=12）。提取mRNA并在Illumina平台上进行RNA-Seq。使用由FastQC组成的生物信息学管道鉴定差异表达基因，明星,FeatureCounts和edgeR。使用Enrichr确定生物学功能和途径的变化，并使用Cytoscape进行聚类。共有5817个基因在Tenon的正常和青光眼的眼成纤维细胞之间差异表达。富集分析显示787个明显不同的生物学功能和途径，分为176个簇。来自青光眼的Tenon\的眼成纤维细胞显示纤维化的迹象，成纤维细胞转分化为肌成纤维细胞，线粒体分裂相关变化，细胞外基质的重塑，扩散,未折叠的蛋白质反应，炎症和细胞凋亡可能与青光眼的发病机理或局部青光眼治疗的有害作用有关。青光眼Tenon的眼成纤维细胞中基因表达的改变可能导致青光眼滤过手术的不利结果。这项工作提出了青光眼与正常Tenon的眼成纤维细胞的全基因组转录组，该转录组可以鉴定具有治疗价值的基因或途径以改善手术结果。
