Eye Institute, Eye & ENT Hospital, Shanghai Medical College, Fudan University, Shanghai, China.
The Fourth School of Clinical Medicine, Nanjing Medical University, Nanjing, China.
Theranostics. 2020 Feb 10;10(7):3293-3307. doi: 10.7150/thno.39488. eCollection 2020.
: Choroidal neovascularization (CNV) is a major cause of severe vision loss and occurs in many ocular diseases, especially neovascular age-related macular degeneration (nAMD). Circular RNAs (circRNAs) are emerging as a new class of endogenous noncoding RNAs, which have been implicated in the regulation of endothelial cell dysfunction in diabetes mellitus and cancer. In this study, we aimed to determine the role of circRNA-ZBTB44 (cZBTB44) in the pathogenesis of CNV. : Quantitative polymerase chain reaction was conducted to detect cZBTB44 expression pattern during CNV development. Isolectin B4 staining, hematoxylin and eosin (HE) staining, and choroidal sprouting assay were conducted to evaluate the role of cZBTB44 in the development of CNV. Endothelial cell proliferation, migration and tube formation assays were conducted to determine the role of cZBTB44 in angiogenic effect . Bioinformatics analysis, RNA immunoprecipitation assay, luciferase assay, and studies were conducted to investigate the mechanism of cZBTB44-mediated CNV development. : cZBTB44 expression was significantly up-regulated in a laser-induced CNV mouse model and in endothelial cells upon hypoxia stress . cZBTB44 silencing retarded CNV development, while overexpression of cZBTB44 showed the opposite effects. The role of cZBTB44 in CNV development was confirmed in choroidal sprouting assay . cZBTB44 silencing reduced endothelial cell viability, proliferation, migration and tube formation . cZBTB44 acted as miR-578 sponge to sequester and inhibit miR-578 activity, which led to increased expression of vascular endothelial growth factor A (VEGFA) and vascular cell adhesion molecule-1 (VCAM1). Overexpression of miR-578 mimicked cZBTB44 silencing-mediated anti-angiogenic effects and . Furthermore, dysregulated cZBTB44 expression was detected in the clinical samples of nAMD patients. : This study provided novel insights into the molecular pathogenesis of CNV. The cZBTB44-miR-578-VEGFA/VCAM1 axis might be a potential source of novel therapeutic targets for neovascularization-related diseases.
脉络膜新生血管（CNV）是严重视力丧失的主要原因，发生在许多眼部疾病中，特别是新生血管性年龄相关性黄斑变性（nAMD）。环状 RNA（circRNA）作为一类新的内源性非编码 RNA 逐渐受到关注，其被认为与糖尿病和癌症中的内皮细胞功能障碍的调节有关。在这项研究中，我们旨在确定环状 RNA-ZBTB44（cZBTB44）在 CNV 发病机制中的作用。
定量聚合酶链反应检测 CNV 发展过程中 cZBTB44 的表达模式。异硫氰酸荧光素 B4 染色、苏木精和伊红（HE）染色和脉络膜发芽试验评估 cZBTB44 在 CNV 发展中的作用。内皮细胞增殖、迁移和管形成试验确定 cZBTB44 在血管生成作用中的作用。生物信息学分析、RNA 免疫沉淀试验、荧光素酶测定和功能研究用于研究 cZBTB44 介导的 CNV 发展的机制。
cZBTB44 在激光诱导的 CNV 小鼠模型和缺氧应激下的内皮细胞中表达显著上调。cZBTB44 沉默延迟了 CNV 的发展，而 cZBTB44 的过表达则产生了相反的效果。在脉络膜发芽试验中证实了 cZBTB44 在 CNV 发展中的作用。cZBTB44 沉默降低了内皮细胞的活力、增殖、迁移和管形成。cZBTB44 作为 miR-578 的海绵，可结合并抑制 miR-578 的活性，从而导致血管内皮生长因子 A（VEGFA）和血管细胞黏附分子 1（VCAM1）的表达增加。过表达 miR-578 模拟了 cZBTB44 沉默介导的抗血管生成作用。此外，在 nAMD 患者的临床样本中检测到 cZBTB44 表达失调。
这项研究为 CNV 的分子发病机制提供了新的见解。cZBTB44-miR-578-VEGFA/VCAM1 轴可能成为与新生血管相关疾病的新治疗靶点的潜在来源。
