Department of Ophthalmology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Department of Preventative Ophthalmology, Shanghai Eye Disease Prevention and Treatment Center/Shanghai Eye Hospital, Shanghai, China.
Invest Ophthalmol Vis Sci. 2021 Jan 4;62(1):16. doi: 10.1167/iovs.62.1.16.
Abundant evidence has shown benefits of antivascular endothelial growth factor (anti-VEGF) therapies in neovascular eye diseases. However, the high cost, side effects, and inconvenience of frequent injections demand alternative novel drug candidates. This study aimed to analyze antiangiogenic effects of peptide H-KI20 and illustrated signaling mechanisms.
Live cell culture and tracing, wound healing assay, and tube formation were performed in human retinal microvascular endothelial cells (HRECs). The chick embryo chorioallantoic membrane and mouse oxygen-induced ischemic retinopathy model were applied to examine the effects of H-KI20 in vivo. The intracellular signaling pathways were examined. Molecular docking and surface plasmon resonance assay were used to validate the direct interaction of H-KI20 and c-Jun N-terminal kinase 2 (JNK2).
H-KI20 had high penetration ability in vitro and in vivo. It inhibited motility, migration, and tube formation of HRECs, without cytotoxicity, and inhibited angiogenesis in vivo. Furthermore, H-KI20 treatment reduced the phosphorylation level of activating transcription factor 2 (ATF2) stimulated by VEGF via downregulating p-JNK. H-KI20 bound to JNK2 directly with a dissociation constant value of 83.68 µM. The knockdown of ATF2 attenuated VEGF-induced tube formation and decreased the movement speed of HRECs.
H-KI20 inhibited angiogenesis both in vitro and in vivo. The ratios of p-ATF2/ATF2 and p-JNK/JNK stimulated by VEGF were decreased by H-KI20, and H-KI20 targeted JNK2 directly. In addition, the pivotal role of ATF2 in VEGF-induced retinal neovascularization was elucidated for the first time. Taken together, H-KI20 displays potential for pathological retinal angiogenesis as a sustained and low-toxic peptide.
大量证据表明抗血管内皮生长因子（anti-VEGF）疗法在新生血管性眼病中的益处。然而，高成本、副作用和频繁注射的不便性要求替代新型候选药物。本研究旨在分析肽 H-KI20 的抗血管生成作用，并阐明其信号机制。
在人视网膜微血管内皮细胞（HRECs）中进行活细胞培养和示踪、划痕愈合试验和管形成试验。应用鸡胚绒毛尿囊膜和小鼠氧诱导缺血性视网膜病变模型来检测 H-KI20 在体内的作用。检测细胞内信号通路。采用分子对接和表面等离子体共振试验来验证 H-KI20 与 c-Jun N 端激酶 2（JNK2）的直接相互作用。
H-KI20 在体外和体内均具有高穿透能力。它抑制 HRECs 的运动、迁移和管形成，无细胞毒性，并抑制体内血管生成。此外，H-KI20 处理通过下调 p-JNK 来降低 VEGF 刺激的激活转录因子 2（ATF2）的磷酸化水平。H-KI20 与 JNK2 直接结合，解离常数为 83.68 µM。ATF2 的敲低减弱了 VEGF 诱导的管形成，并降低了 HRECs 的运动速度。
H-KI20 抑制了体内外的血管生成。H-KI20 降低了 VEGF 刺激的 p-ATF2/ATF2 和 p-JNK/JNK 的比值，并且 H-KI20 直接靶向 JNK2。此外，首次阐明了 ATF2 在 VEGF 诱导的视网膜新生血管形成中的关键作用。总之，H-KI20 作为一种持续且低毒的肽，具有病理性视网膜血管生成的潜力。
