Abstract

The blood–brain barrier (BBB) and complex tumour immunosuppressive micro-environment posed austere challenges for combatting brain tumours such as the glioblastoma. In this study, we have developed a novel dual functional dendrimer drug delivery system (DDS) by the PAMAM and loaded with siLSINCT5 (NP- siRNA) for efficiently across the BBB to inhibit glioblastoma. To achieve the goal of BBB crossing, on the surface of NP-siRNA was decorated with the cell penetrating peptides tLyp-1 (tLypNP-siRNA). Moreover, to overcome the immunosuppressive microenvironment within the glioblastoma (GBM) tissues, a checkpoint inhibitor named as anti-NKG2A monoclonal antibody (aNKG2A), which was able of promoting anti-tumour immunity by unleashing both T and NK Cells, was further conjugated on the surface of siLSINCT5-loaded nanoparticles via the pH-sensitive linkage. Therefore, the developed dual functional and siLSINCT5-loaded dendrimer nanoparticles (tLyp/aNKNP-siRNA) was supposed to have the ability to efficiently cross the BBB and inhibit GBM by simultaneously inhibit the LSINCT5-activated signalling pathways and activate the anti-tumour immunity. The hypothesis was thoroughly confirmed by in vitro cellular and in vivo animal experiments, and provided a novel strategy for combating glioblastoma.

摘要

血脑屏障 (BBB) 和复杂的肿瘤免疫抑制微环境对对抗胶质母细胞瘤等脑肿瘤提出了严峻的挑战。在这项研究中，我们开发了一种由 PAMAM 开发的新型双功能树枝状大分子药物递送系统 (DDS)，并装载有 siLSINCT5 (NP-siRNA)，可有效穿过 BBB 以抑制胶质母细胞瘤。为了实现血脑屏障穿越的目标，在NP-siRNA的表面装饰有细胞穿透肽tLyp-1（tLypNP-siRNA）。此外，为了克服胶质母细胞瘤 (GBM) 组织内的免疫抑制微环境，一种名为抗 NKG2A 单克隆抗体 (aNKG2A) 的检查点抑制剂能够通过释放 T 细胞和 NK 细胞来促进抗肿瘤免疫，进一步结合在载有 siLSINCT5 的纳米粒子的表面通过pH 敏感的连接。因此，开发的双功能和载有 siLSINCT5 的树枝状大分子纳米粒子 (tLyp/aNKNP-siRNA) 被认为具有有效穿过 BBB 并通过同时抑制 LSINCT5 激活的信号通路和激活抗肿瘤免疫来抑制 GBM 的能力。这一假设得到了体外细胞和体内动物实验的彻底证实，并为对抗胶质母细胞瘤提供了一种新的策略。