Rationale: Angiogenesis expedites tissue impairment in many diseases, including age-related macular degeneration (AMD), a leading cause of irreversible blindness in elderly. A substantial proportion of neovascular AMD patients, characterized by aberrant choroidal neovascularization (CNV), exhibit poor responses or adverse reactions to anti-VEGF therapy. Herein, we aimed to unveil the function of newly identified transfer RNA-derived small RNA, tRF-Glu-CTC, in the pathology of CNV and determine its potential in inhibiting angiogenesis./nMethods: Small non-coding RNA sequencing and quantitative polymerase chain reaction were conducted to detect expression pattern of tRF-Glu-CTC in CNV development. Immunofluorescence staining, fundus fluorescein angiography and ex vivo choroidal sprouting assays were employed for the evaluation of tRF-Glu-CTC's function in CNV development. The role of tRF-Glu-CTC in endothelial cells were determined by in vitro endothelial cell proliferation, migration and tube formation assays. Transcriptome sequencing, dual-luciferase reporter assay and in vitro experiments were conducted to investigate downstream mechanism of tRF-Glu-CTC mediated pathology./nResults: tRF-Glu-CTC exhibited substantial up-regulation in AMD patients, laser-induced CNV model, and endothelial cells under hypoxia condition, which is a hallmark of CNV. Inhibiting tRF-Glu-CTC reduced angiogenesis and hypoxia stress in the neovascular region without neuroretina toxicity in laser-induced CNV model, showing an anti-angiogenic effect comparable to bevacizumab, while overexpression of tRF-Glu-CTC significantly augmented CNV. Mechanically, under hypoxia condition, angiogenin was involved in the production of tRF-Glu-CTC, which in turn triggered endothelial cell tubulogenesis, migration and promoted the secretion of inflammatory factors via the suppression of vasohibin 1 (VASH1). When downregulating VASH1 expression, the inhibition of tRF-Glu-CTC showed minimal suppression on angiogenesis./nConclusions: This study demonstrated the important role of tRF-Glu-CTC in the progression of angiogenesis. Targeting of tRF-Glu-CTC may be an alternative to current anti-VEGF therapy for CNV in AMD and other conditions with angiogenesis.

中文翻译：

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转移 RNA 衍生片段 tRF-Glu-CTC 加剧新生血管性年龄相关性黄斑变性的发展

理由：血管生成会加速许多疾病的组织损伤，包括年龄相关性黄斑变性 (AMD)，这是老年人不可逆失明的主要原因。相当一部分以异常脉络膜新生血管（CNV）为特征的新生血管性 AMD 患者对抗 VEGF 治疗表现出较差的反应或不良反应。在此，我们的目的是揭示新鉴定的转移 RNA 衍生的小 RNA tRF-Glu-CTC 在 CNV 病理学中的功能，并确定其抑制血管生成的潜力。/n 方法：小非编码 RNA 测序和定量聚合酶进行链式反应检测tRF-Glu-CTC在CNV发育中的表达模式。采用免疫荧光染色、眼底荧光素血管造影和离体脉络膜出芽测定来评估 tRF-Glu-CTC 在 CNV 发育中的功能。通过体外内皮细胞增殖、迁移和管形成测定来确定 tRF-Glu-CTC 在内皮细胞中的作用。通过转录组测序、双荧光素酶报告基因测定和体外实验来研究 tRF-Glu-CTC 介导的病理学下游机制。/n结果： tRF-Glu-CTC 在 AMD 患者、激光诱导的 CNV 模型中表现出显着上调和缺氧条件下的内皮细胞，这是 CNV 的标志。在激光诱导的 CNV 模型中，抑制 tRF-Glu-CTC 可以减少新生血管区域的血管生成和缺氧应激，而没有神经视网膜毒性，显示出与贝伐珠单抗相当的抗血管生成作用，而 tRF-Glu-CTC 的过表达则显着增强了 CNV。机械上，在缺氧条件下，血管生成素参与tRF-Glu-CTC的产生，进而触发内皮细胞的管状形成、迁移，并通过抑制血管抑制素1（VASH1）促进炎症因子的分泌。当下调 VASH1 表达时，tRF-Glu-CTC 的抑制对血管生成显示出最小的抑制。/n结论：本研究证明了 tRF-Glu-CTC 在血管生成进展中的重要作用。tRF-Glu-CTC 靶向可能是当前针对 AMD 和其他血管生成疾病中 CNV 的抗 VEGF 疗法的替代疗法。