Ma Yan, Zhang Ying, Zhang Hui-Ying, Zhao Ya, Li Xiu-Miao, Jiang Yi-Fei, Yao Mu-Di, Jiang Qin, Yan Biao
The Fourth School of Clinical Medicine, Nanjing Medical University, Nanjing, 210000, China.
The Affiliated Eye Hospital, Nanjing Medical University, Nanjing, 210000, China.
J Transl Med. 2024 Jun 12;22(1):562. doi: 10.1186/s12967-024-05338-w.
Intravitreal injections of angiogenesis inhibitors have proved efficacious in the majority of patients with ocular angiogenesis. However, one-fourth of all treated patients fail to derive benefits from intravitreal injections. tRNA-derived small RNA (tsRNA) emerges as a crucial class of non-coding RNA molecules, orchestrating key roles in the progression of human diseases by modulating multiple targets. Through our prior sequencing analyses and bioinformatics predictions, tRNA-Cys-5-0007 has shown as a potential regulator of ocular angiogenesis. This study endeavors to elucidate the precise role of tRNA-Cys-5-0007 in the context of ocular angiogenesis.
Quantitative reverse transcription PCR (qRT-PCR) assays were employed to detect tRNA-Cys-5-0007expression. EdU assays, sprouting assays, transwell assays, and Matrigel assays were conducted to elucidate the involvement of tRNA-Cys-5-0007 in endothelial angiogenic effects. STZ-induced diabetic model, OIR model, and laser-induced CNV model were utilized to replicate the pivotal features of ocular vascular diseases and evaluate the influence of tRNA-Cys-5-0007 on ocular angiogenesis and inflammatory responses. Bioinformatics analysis, luciferase activity assays, RNA pull-down assays, and in vitro studies were employed to elucidate the anti-angiogenic mechanism of tRNA-Cys-5-0007. Exosomal formulation was employed to enhance the synergistic anti-angiogenic and anti-inflammatory efficacy of tRNA-Cys-5-0007.
tRNA-Cys-5-0007 expression was down-regulated under angiogenic conditions. Conversely, tRNA-Cys-5-0007 overexpression exhibited anti-angiogenic effects in retinal endothelial cells, as evidenced by reduced proliferation, sprouting, migration, and tube formation abilities. In diabetic, laser-induced CNV, and OIR models, tRNA-Cys-5-0007 overexpression led to decreased ocular vessel leakage, inhibited angiogenesis, and reduced ocular inflammation. Mechanistically, these effects were attributed to the targeting of vascular endothelial growth factor A (VEGFA) and TGF-β1 by tRNA-Cys-5-0007. The utilization of an exosomal formulation further potentiated the synergistic anti-angiogenic and anti-inflammatory efficacy of tRNA-Cys-5-0007.
Concurrent targeting of tRNA-Cys-5-0007 for anti-angiogenic and anti-inflammatory therapy holds promise for enhancing the effectiveness of current anti-angiogenic therapy.
玻璃体内注射血管生成抑制剂已被证明对大多数眼部血管生成患者有效。然而，所有接受治疗的患者中有四分之一未能从玻璃体内注射中获益。tRNA衍生的小RNA（tsRNA）作为一类关键的非编码RNA分子出现，通过调节多个靶点在人类疾病进展中发挥关键作用。通过我们之前的测序分析和生物信息学预测，tRNA-Cys-5-0007已显示为眼部血管生成的潜在调节因子。本研究旨在阐明tRNA-Cys-5-0007在眼部血管生成中的精确作用。
采用定量逆转录PCR（qRT-PCR）检测tRNA-Cys-5-0007的表达。进行EdU检测、发芽检测、Transwell检测和基质胶检测，以阐明tRNA-Cys-5-0007在内皮细胞血管生成作用中的参与情况。利用链脲佐菌素诱导的糖尿病模型、视网膜病变模型和激光诱导的脉络膜新生血管模型来复制眼部血管疾病的关键特征，并评估tRNA-Cys-5-0007对眼部血管生成和炎症反应的影响。采用生物信息学分析、荧光素酶活性检测、RNA下拉检测和体外研究来阐明tRNA-Cys-5-0007的抗血管生成机制。采用外泌体制剂来增强tRNA-Cys-5-0007的协同抗血管生成和抗炎功效。
在血管生成条件下，tRNA-Cys-5-0007的表达下调。相反，tRNA-Cys-5-0007的过表达在视网膜内皮细胞中表现出抗血管生成作用，增殖、发芽、迁移和管形成能力降低证明了这一点。在糖尿病、激光诱导的脉络膜新生血管和视网膜病变模型中，tRNA-Cys-5-0007的过表达导致眼部血管渗漏减少、血管生成受到抑制和眼部炎症减轻。从机制上讲，这些作用归因于tRNA-Cys-5-0007对血管内皮生长因子A（VEGFA）和转化生长因子-β1的靶向作用。外泌体制剂的使用进一步增强了tRNA-Cys-5-0007的协同抗血管生成和抗炎功效。
同时靶向tRNA-Cys-5-0007进行抗血管生成和抗炎治疗有望提高当前抗血管生成治疗的有效性。
