PDF(Pubmed)
Abstract：
BACKGROUND: 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.
METHODS: 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.
RESULTS: 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.
CONCLUSIONS: 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.
METHODS: 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.
RESULTS: 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.
CONCLUSIONS: 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.
摘要：
背景：玻璃体内注射血管生成抑制剂已被证明对大多数眼血管生成患者有效。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获益.tRNA衍生的小RNA（tsRNA）作为一类重要的非编码RNA分子，通过调节多个靶标来协调人类疾病进展中的关键作用。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剂。本研究试图阐明tRNA-Cys-5-0007在眼部血管生成中的精确作用。
方法:采用定量逆转录PCR(qRT-PCR)检测tRNA-Cys-5-0007表达。EdU化验，发芽测定，transwell分析，和Matrigel分析用于阐明tRNA-Cys-5-0007在内皮血管生成作用中的参与。STZ诱导的糖尿病模型,OIR模型,和激光诱导的CNV模型用于复制眼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。生物信息学分析，荧光素酶活性测定,RNA下拉法,体外研究用于阐明tRNA-Cys-5-0007的抗血管生成机制。使用外来体制剂来增强tRNA-Cys-5-0007的协同抗血管生成和抗炎功效。
结果：tRNA-Cys-5-0007表达在血管生成条件下下调。相反，tRNA-Cys-5-0007过表达在视网膜内皮细胞中表现出抗血管生成作用，扩散减少证明了这一点，发芽，迁移,和管形成能力。在糖尿病患者中，激光诱导CNV,和OIR模型，tRNA-Cys-5-0007过表达导致眼血管渗漏减少，抑制血管生成，减少眼部炎症。机械上,这些作用归因于tRNA-Cys-5-0007靶向血管内皮生长因子A(VEGFA)和TGF-β1.外泌体制剂的利用进一步增强了tRNA-Cys-5-0007的协同抗血管生成和抗炎功效。
结论：同时靶向tRNA-Cys-5-0007用于抗血管生成和抗炎治疗有望增强当前抗血管生成治疗的有效性。
方法:采用定量逆转录PCR(qRT-PCR)检测tRNA-Cys-5-0007表达。EdU化验，发芽测定，transwell分析，和Matrigel分析用于阐明tRNA-Cys-5-0007在内皮血管生成作用中的参与。STZ诱导的糖尿病模型,OIR模型,和激光诱导的CNV模型用于复制眼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。生物信息学分析，荧光素酶活性测定,RNA下拉法,体外研究用于阐明tRNA-Cys-5-0007的抗血管生成机制。使用外来体制剂来增强tRNA-Cys-5-0007的协同抗血管生成和抗炎功效。
结果：tRNA-Cys-5-0007表达在血管生成条件下下调。相反，tRNA-Cys-5-0007过表达在视网膜内皮细胞中表现出抗血管生成作用，扩散减少证明了这一点，发芽，迁移,和管形成能力。在糖尿病患者中，激光诱导CNV,和OIR模型，tRNA-Cys-5-0007过表达导致眼血管渗漏减少，抑制血管生成，减少眼部炎症。机械上,这些作用归因于tRNA-Cys-5-0007靶向血管内皮生长因子A(VEGFA)和TGF-β1.外泌体制剂的利用进一步增强了tRNA-Cys-5-0007的协同抗血管生成和抗炎功效。
结论：同时靶向tRNA-Cys-5-0007用于抗血管生成和抗炎治疗有望增强当前抗血管生成治疗的有效性。
