Department of Ophthalmology, Eye Institute, Eye & ENT Hospital, Shanghai Medical College, Fudan University, Shanghai, China; and.
Department of Ophthalmology, NHC Key Laboratory of Myopia (Fudan University), Laboratory of Myopia, Chinese Academy of Medical Sciences, Shanghai, China.
Cornea. 2022 Dec 1;41(12):1545-1552. doi: 10.1097/ICO.0000000000003065. Epub 2022 Aug 10.
Circular RNAs (circRNAs) are a novel class of endogenous noncoding RNAs that regulate gene expression through the competitive endogenous RNA (ceRNA) mechanism. CircRNA-associated-ceRNA networks are closely related to oxidative stress-related diseases. Oxidative stress-induced dysfunction of the corneal endothelium (CE) is a major pathological feature in many corneal diseases. This study was aimed to analyze circRNA-associated-ceRNA networks in oxidative stress-induced CE dysfunction.
A CE dysfunction model was established using human corneal endothelial cells (HCECs) treated with H 2 O 2 at a concentration of 250 μM for 4 hours at 37°C. High-throughput sequencing was conducted to determine the expression profiles of circRNA, miRNA, and mRNA. Bioinformatic analyses, including Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes analysis, were conducted to identify the potential biological modules and pathologic pathways of dysregulated circRNAs. CircRNA-associated-ceRNA networks were established based on the data of sequencing and bioinformatic analyses.
We obtained 108 differentially expressed circRNAs, including 77 upregulated and 31 downregulated circRNAs. GO analysis suggested that dysregulated circRNAs were mainly targeted to protein quality control for misfolded or incompletely synthesized proteins (biologic process), nuclear chromatin (cellular component), and ubiquitin protein ligase binding (molecular function). GO terms related to CE functions responding to oxidative stress were also identified. Kyoto Encyclopedia of Genes and Genomes pathway analysis indicated that dysregulated circRNAs were mostly enriched in the adherens junction pathway. Network analysis identified several potential therapeutic targets for CE dysfunction.
CircRNAs are significantly dysregulated in HCECs under oxidative stress. The circRNA-associated-ceRNA networks are closely related to HCEC functions. Targeting these networks might provide novel therapies for CE dysfunction.
环状 RNA（circRNA）是一类新型的内源性非编码 RNA，通过竞争性内源性 RNA（ceRNA）机制调节基因表达。circRNA 相关 ceRNA 网络与氧化应激相关疾病密切相关。氧化应激诱导的角膜内皮细胞（CE）功能障碍是许多角膜疾病的主要病理特征。本研究旨在分析氧化应激诱导的 CE 功能障碍中 circRNA 相关 ceRNA 网络。
用浓度为 250μM 的 H2O2 处理人角膜内皮细胞（HCEC）4 小时，在 37°C 下建立 CE 功能障碍模型。采用高通量测序技术检测 circRNA、miRNA 和 mRNA 的表达谱。进行生物信息学分析，包括基因本体论（GO）分析和京都基因与基因组百科全书分析，以鉴定失调 circRNA 的潜在生物学模块和病理途径。基于测序和生物信息学分析的数据建立 circRNA 相关 ceRNA 网络。
我们获得了 108 个差异表达的 circRNA，包括 77 个上调和 31 个下调的 circRNA。GO 分析表明，失调的 circRNA 主要靶向蛋白质质量控制，用于错误折叠或不完全合成的蛋白质（生物过程）、核染色质（细胞成分）和泛素蛋白连接酶结合（分子功能）。还确定了与 CE 对氧化应激反应功能相关的 GO 术语。京都基因与基因组百科全书通路分析表明，失调的 circRNA 主要富集在黏着连接途径中。网络分析确定了几个潜在的 CE 功能障碍治疗靶点。
氧化应激下 HCEC 中的 circRNA 显著失调。circRNA 相关 ceRNA 网络与 HCEC 功能密切相关。针对这些网络可能为 CE 功能障碍提供新的治疗方法。
