Key Laboratory of Smart Drug Delivery of the Ministry of Education & Department of Pharmaceutics, School of Pharmacy, Fudan University, Shanghai 201203, China; State Key Laboratory of Medical Neurobiology and the Collaborative Innovation Center for Brain Science, Fudan University, Shanghai 200032, China.
State Key Laboratory of Oncogenes and Related Genes, Shanghai Cancer Institute, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200030, China.
J Control Release. 2018 Jun 10;279:306-315. doi: 10.1016/j.jconrel.2018.04.035. Epub 2018 Apr 19.
The receptor associated protein (RAP) is a 39 kDa chaperone protein, binding tightly to low-density lipoprotein receptor-related protein-1 (LRP1) that is overexpressed in glioma, tumor neovasculature, vasculogenic mimicry (VM), the blood-brain barrier (BBB) and the blood-brain tumor barrier (BBTB). Herein, we miniaturized the RAP protein into a short peptide RAP12 (EAKIEKHNHYQK) aiding by computer-aided peptide design technique. RAP12 contained the essential lysines at the positions 253 and 256. The binding affinity of RAP12 to LRP1 was theoretically and experimentally evaluated. In cellular level, RAP12 could effectively internalize into U87, HUVEC and bEnd.3 cells. When modified on the surface of PEG-PLA micelles (RAP12-PEG-PLA), RAP12 could effectively facilitate the penetration of micelles through the BBB/BBTB in vitro/vivo. Paclitaxel-loaded RAP12-PEG-PLA could remarkably inhibit the growth of glioma cells and the formation of tumor neovasculature and VM, significantly prolong the median survival time of nude mice bearing intracranial glioma in comparison to model mice treated with plain micelles or Taxol. These results suggested that the RAP12 held the potential for multifunctional glioma-targeted drug delivery.
受体相关蛋白 (RAP) 是一种 39kDa 的伴侣蛋白，与在神经胶质瘤、肿瘤新生血管、血管生成拟态 (VM)、血脑屏障 (BBB) 和血脑肿瘤屏障 (BBTB) 中过度表达的低密度脂蛋白受体相关蛋白-1 (LRP1) 紧密结合。在此，我们通过计算机辅助肽设计技术将 RAP 蛋白小型化为短肽 RAP12（EAKIEKHNHYQK）。RAP12 包含位于位置 253 和 256 的必需赖氨酸。通过理论和实验评估了 RAP12 与 LRP1 的结合亲和力。在细胞水平上，RAP12 可以有效地内化到 U87、HUVEC 和 bEnd.3 细胞中。当修饰在 PEG-PLA 胶束的表面上时（RAP12-PEG-PLA），RAP12 可以有效地促进胶束穿过 BBB/BBTB 在体外/体内的渗透。载紫杉醇的 RAP12-PEG-PLA 可以显著抑制神经胶质瘤细胞的生长以及肿瘤新生血管和 VM 的形成，与用普通胶束或紫杉醇治疗的模型小鼠相比，显著延长了携带颅内神经胶质瘤的裸鼠的中位生存时间。这些结果表明，RAP12 具有用于多功能神经胶质瘤靶向药物递送的潜力。
