Tang Hanfeng, Lin Yi, Hu Jianzhang
Department of Ophthalmology, Fujian Medical University Union Hospital, Fu Zhou, China.
Invest Ophthalmol Vis Sci. 2025 Mar 3;66(3):27. doi: 10.1167/iovs.66.3.27.
Fungal keratitis (FK) is a challenging and sight-threatening corneal disease caused by fungal infections. Although long noncoding RNAs (lncRNAs) have been explored in various infectious diseases, their specific roles in FK remain largely unexplored.
A mouse model of FK was created by infecting corneal stromal cells with Fusarium solani. High-throughput lncRNA expression profiling was conducted on FK-affected corneal tissues to identify differentially expressed lncRNAs. Reverse transcription quantitative PCR (RT-qPCR) was used to validate the results. A competing endogenous RNA (ceRNA) network was constructed. Additionally, a specific antisense oligonucleotide (ASO) targeting lncRNA ENSMUST00000226838/Osr2 (Osr2) was developed for therapeutic evaluation. Inflammatory markers IL-1β, IL-6, and TNF-α were measured, and corneal inflammation was assessed through histological analysis and slit-lamp examination. Fluorescent in situ hybridization (FISH) was used to confirm Osr2 localization, whereas a dual-luciferase reporter assay verified interactions between Osr2 and miR-30a-3p.
We identified 1143 differentially expressed lncRNAs in FK, with 701 upregulated and 442 downregulated. The ceRNA network analysis indicated that lncRNA Osr2 regulates Xcr1 expression through miR-30a-3p. Treatment with ASO-Osr2 significantly reduced corneal inflammation, and FISH confirmed lncRNA Osr2 distribution in both the nucleus and cytoplasm. Dual-luciferase assays demonstrated the interaction between Osr2 and miR-30a-3p, highlighting their potential roles in the progression of FK.
This study outlined the lncRNA expression profile in FK and established a ceRNA regulatory network, identifying lncRNA Osr2 as a crucial modulator of FK pathogenesis through its interaction with miR-30a-3p. These findings highlighted lncRNA Osr2 as a promising therapeutic target for the treatment of FK.
真菌性角膜炎（FK）是一种由真菌感染引起的具有挑战性且威胁视力的角膜疾病。尽管长链非编码RNA（lncRNAs）已在多种传染病中得到研究，但其在FK中的具体作用仍 largely未被探索。
通过用茄病镰刀菌感染角膜基质细胞建立FK小鼠模型。对受FK影响的角膜组织进行高通量lncRNA表达谱分析，以鉴定差异表达的lncRNAs。采用逆转录定量PCR（RT-qPCR）验证结果。构建竞争性内源RNA（ceRNA）网络。此外，开发了一种靶向lncRNA ENSMUST00000226838/Osr2（Osr2）的特异性反义寡核苷酸（ASO）用于治疗评估。检测炎症标志物白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）和肿瘤坏死因子-α（TNF-α），并通过组织学分析和裂隙灯检查评估角膜炎症。采用荧光原位杂交（FISH）确认Osr2的定位，而双荧光素酶报告基因测定验证Osr2与miR-30a-3p之间的相互作用。
我们在FK中鉴定出1143个差异表达的lncRNAs，其中701个上调，442个下调。ceRNA网络分析表明，lncRNA Osr2通过miR-30a-3p调节Xcr1表达。用ASO-Osr2治疗可显著减轻角膜炎症，FISH证实lncRNA Osr2在细胞核和细胞质中均有分布。双荧光素酶测定证明了Osr2与miR-30a-3p之间的相互作用，突出了它们在FK进展中的潜在作用。
本研究概述了FK中的lncRNA表达谱并建立了ceRNA调控网络，通过lncRNA Osr2与miR-30a-3p的相互作用确定其为FK发病机制的关键调节因子。这些发现突出了lncRNA Osr2作为治疗FK的有前景的治疗靶点。
