Shi Mingsu, Zhou Rongmei, Shen Weiai, Liang Yu, Zhang Yihan, Liu Lingyun, Shao Runyi, Fang Yanxi, Zhao Chen, Wu Lianqun
Eye Institute and Department of Ophthalmology, Eye and ENT Hospital, Fudan University, Shanghai, China.
Key Laboratory of Myopia and Related Eye Diseases, National Health Commission (NHC), Shanghai, China.
Invest Ophthalmol Vis Sci. 2025 Mar 3;66(3):46. doi: 10.1167/iovs.66.3.46.
Thyroid eye disease (TED) is a visually debilitating and cosmetically disfiguring orbital disorder, characterized by the remodeling of extraocular muscles (EOMs). This study aimed to investigate the role of long non-coding RNA (lncRNA) ENST00000581911 in the EOMs of TED.
LncRNA microarray analysis was performed on EOM tissues sampled from patients with TED and patients with concomitant esotropia. LncRNA ENST00000581911 was identified and subjected to bioinformatics analysis. High-throughput RNA sequencing, CCK-8 assay, CFSE staining, and ELISA were used to investigate the regulatory function of ENST00000581911 in vitro. Furthermore, RNA pull-down, liquid chromatography-tandem mass spectrometry (LC-MS/MS), and western blot (WB) analyses were applied to identify the RNA-binding protein (RBP) interacting with ENST00000581911.
A total of 1261 lncRNAs were found to be differentially expressed in the EOMs of TED, with 648 upregulated and 613 downregulated lncRNAs. Among these, the upregulated lncRNA ENST00000581911 exhibited the highest expression level, as validated by quantitative real-time PCR (qRT-PCR). Functional analysis demonstrated that ENST00000581911 might be involved in inflammatory response, regulation of muscle contraction, and amino sugar and nucleotide sugar metabolism. RNA sequencing of ENST00000581911-overexpressing and control orbital fibroblasts (OFs) showed that ENST00000581911 might play a regulatory role in DNA replication, extracellular matrix, and cell cycle. Furthermore, KHSRP was identified as the RBP of ENST00000581911. Overexpression of ENST00000581911 promoted cell proliferation and hyaluronic acid secretion in OFs, whereas silencing KHSRP attenuated these effects.
This study provides novel insights into the role of lncRNA ENST00000581911 in the pathogenesis of EOM remodeling in TED. ENST00000581911 may serve as a potential therapeutic target of TED.
甲状腺眼病（TED）是一种导致视力受损和外貌毁损的眼眶疾病，其特征为眼外肌（EOM）重塑。本研究旨在探讨长链非编码RNA（lncRNA）ENST00000581911在TED患者眼外肌中的作用。
对TED患者和伴有内斜视患者的眼外肌组织进行lncRNA芯片分析。鉴定出lncRNA ENST00000581911并进行生物信息学分析。采用高通量RNA测序、CCK-8法、CFSE染色和ELISA法在体外研究ENST00000581911的调控功能。此外，应用RNA下拉、液相色谱-串联质谱（LC-MS/MS）和蛋白质免疫印迹（WB）分析来鉴定与ENST00000581911相互作用的RNA结合蛋白（RBP）。
共发现1261个lncRNA在TED患者的眼外肌中差异表达，其中648个lncRNA上调，613个lncRNA下调。其中，上调的lncRNA ENST00000581911表达水平最高，经定量实时PCR（qRT-PCR）验证。功能分析表明，ENST00000581911可能参与炎症反应、肌肉收缩调节以及氨基糖和核苷酸糖代谢。对过表达ENST00000581911的眼眶成纤维细胞（OF）和对照OF进行RNA测序表明，ENST00000581911可能在DNA复制、细胞外基质和细胞周期中发挥调控作用。此外，鉴定出KHSRP为ENST00000581911的RBP。ENST00000581911的过表达促进了OF的细胞增殖和透明质酸分泌，而沉默KHSRP减弱了这些作用。
本研究为lncRNA ENST00000581911在TED患者眼外肌重塑发病机制中的作用提供了新的见解。ENST00000581911可能作为TED的潜在治疗靶点。
