Key Laboratory of Chemical Biology (Ministry of Education), Department of Pharmacology, School of Pharmaceutical Sciences, Shandong University, Jinan 250012, China.
Department of General Surgery, Shandong University Affiliated Shandong Provincial Hospital, Jinan 250021, China.
Life Sci. 2021 Apr 1;270:119113. doi: 10.1016/j.lfs.2021.119113. Epub 2021 Jan 27.
This study aimed to design and screen a dual functional fusion peptide that could penetrate the blood-brain barrier and target neuropilin 1 (NRP1) overexpressed in vascular endothelial cells for the anti-angiogenesis of glioma treatment.
At the cellular level, the in vitro anti-angiogenic activity of six NRP1 targeting peptides was screened by testing the ability to inhibit the proliferation and tube formation of HUVECs. Then, the in vitro anti-angiogenic activity of two fusion peptides containing different linkers was screened by testing the ability to inhibit HUVECs proliferation, tube formation and migration. The effect of fusion peptide on VEGFR2 related signal pathway was confirmed by Western-blotting. Surface plasmon resonance technology was used to detect the affinity of the fusion peptide to NRP1. The ability of FITC-labeled peptides to penetrate cells was confirmed by cell uptake assay. By establishing an orthotopic glioma model, we evaluated the ability of FITC-labeled peptides to penetrate the blood-brain barrier and their anti-glioma growth activity in vivo.
We found that NRP1 targeting peptide RP7 and linker cysteine were the most suitable key components in the fusion peptide. We also found that the fusion peptide Tat-C-RP7 we constructed had the strongest ability to penetrate the blood-brain barrier and anti-angiogenic activity in vitro and in vivo.
At present, NRP1 targeting peptide as a drug delivery tool and molecular probe seems to have received more attention. We constructed a fusion peptide Tat-C-RP7 with strong anti-angiogenic activity for the treatment of glioma.
本研究旨在设计和筛选一种双重功能融合肽，该融合肽能够穿透血脑屏障，并靶向血管内皮细胞中过表达的神经纤毛蛋白 1（NRP1），用于治疗胶质瘤的抗血管生成。
在细胞水平上，通过检测抑制 HUVEC 增殖和管腔形成的能力，筛选六种靶向 NRP1 的肽的体外抗血管生成活性。然后，通过检测抑制 HUVEC 增殖、管腔形成和迁移的能力，筛选两种含有不同连接子的融合肽的体外抗血管生成活性。通过 Western-blotting 验证融合肽对 VEGFR2 相关信号通路的影响。表面等离子体共振技术用于检测融合肽与 NRP1 的亲和力。通过细胞摄取实验证实 FITC 标记肽穿透细胞的能力。通过建立原位脑胶质瘤模型，评估 FITC 标记肽穿透血脑屏障的能力及其体内抗脑胶质瘤生长活性。
我们发现 NRP1 靶向肽 RP7 和半胱氨酸连接子是融合肽中最合适的关键成分。我们还发现，我们构建的融合肽 Tat-C-RP7 在体外和体内具有最强的穿透血脑屏障和抗血管生成活性。
目前，NRP1 靶向肽作为药物递送工具和分子探针似乎受到了更多的关注。我们构建了一种具有强抗血管生成活性的融合肽 Tat-C-RP7，用于治疗脑胶质瘤。
