通过上调 tRNA 衍生片段 tRF-22 的表达靶向脉络膜血管病变，以控制近视的进展。
Targeting choroidal vasculopathy via up-regulation of tRNA-derived fragment tRF-22 expression for controlling progression of myopia.
机构信息
Eye Institute and Department of Ophthalmology, Eye & ENT Hospital, Fudan University, Shanghai, 200031, China.
NHC Key Laboratory of Myopia (Fudan University), Key Laboratory of Myopia, Chinese Academy of Medical Sciences, Shanghai, 200031, China.
出版信息
J Transl Med. 2023 Jun 24;21(1):412. doi: 10.1186/s12967-023-04274-5.
BACKGROUND
Myopia has emerged as a major public health concern globally, which is tightly associated with scleral extracellular matrix (ECM) remodeling and choroidal vasculopathy. Choroidal vasculopathy has gradually been recognized as a critical trigger of myopic pathology. However, the precise mechanism controlling choroidal vasculopathy remains unclear. Transfer RNA-derived fragments (tRFs) are known as a novel class of small non-coding RNAs that plays important roles in several biological and pathological processes. In this study, we investigated the role of tRF-22-8BWS72092 (tRF-22) in choroidal vasculopathy and myopia progression.
METHODS
The tRF-22 expression pattern under myopia-related stresses was detected by qRT-PCR. MTT assays, EdU incorporation assays, Transwell migration assays, and Matrigel assays were conducted to detect the role of tRF-22 in choroidal endothelial cell function in vitro. Isolectin B4 staining and choroidal sprouting assay ex vivo were conducted to detect the role of tRF-22 in choroidal vascular dysfunction in vivo. Immunofluorescent staining, western blot assays and ocular biometric parameters measurement were performed to examine whether altering tRF-22 expression in choroid affects scleral hypoxia and ECM remodeling and myopia progression in vivo. Bioinformatics analysis and luciferase activity assays were conducted to identify the downstream targets of tRF-22. RNA-sequencing combined with m6A-qPCR assays were used to identify the m6A modified targets of METTL3. Gain-of-function and Loss-of-function analysis were performed to reveal the mechanism of tRF-22/METTL3-mediated choroidal vascular dysfunction.
RESULTS
The results revealed that tRF-22 expression was significantly down-regulated in myopic choroid. tRF-22 overexpression alleviated choroidal vasculopathy and retarded the progression of myopia in vivo. tRF-22 regulated choroidal endothelial cell viability, proliferation, migration, and tube formation ability in vitro. Mechanistically, tRF-22 interacted with METTL3 and blocked mA methylation of Axin1 and Arid1b mRNA transcripts, which led to increased expression of Axin1 and Arid1b.
CONCLUSIONS
Our study reveals that the intervention of choroidal vasculopathy via tRF-22-METTL3- Axin1/Arid1b axis is a promising strategy for the treatment of patients with myopic pathology.
背景
近视已成为全球主要的公共卫生问题，它与巩膜细胞外基质（ECM）重塑和脉络膜血管病变密切相关。脉络膜血管病变已逐渐被认为是近视病理的关键触发因素。然而，控制脉络膜血管病变的确切机制尚不清楚。转移 RNA 衍生片段（tRFs）是一类新的小非编码 RNA，在多种生物学和病理过程中发挥重要作用。在这项研究中，我们研究了 tRF-22 在脉络膜血管病变和近视进展中的作用。
方法
通过 qRT-PCR 检测与近视相关应激下 tRF-22 的表达模式。MTT 测定、EdU 掺入测定、Transwell 迁移测定和 Matrigel 测定用于检测 tRF-22 在体外脉络膜内皮细胞功能中的作用。异硫氰酸荧光素 B4 染色和脉络膜发芽测定在体内检测 tRF-22 在脉络膜血管功能障碍中的作用。免疫荧光染色、Western blot 测定和眼生物参数测量用于检查改变脉络膜中 tRF-22 的表达是否影响巩膜缺氧和 ECM 重塑以及体内近视进展。生物信息学分析和荧光素酶活性测定用于鉴定 tRF-22 的下游靶标。RNA 测序结合 m6A-qPCR 测定用于鉴定 METTL3 的 m6A 修饰靶标。功能获得和功能丧失分析用于揭示 tRF-22/METTL3 介导的脉络膜血管功能障碍的机制。
结果
结果显示，近视脉络膜中 tRF-22 的表达明显下调。tRF-22 过表达可减轻脉络膜血管病变并延缓体内近视进展。tRF-22 调节体外脉络膜内皮细胞活力、增殖、迁移和管状形成能力。机制上，tRF-22 与 METTL3 相互作用并阻断 Axin1 和 Arid1b mRNA 转录物的 mA 甲基化，导致 Axin1 和 Arid1b 的表达增加。
结论
我们的研究表明，通过 tRF-22-METTL3-Axin1/Arid1b 轴干预脉络膜血管病变是治疗近视患者的一种有前途的策略。
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