Lee Youngki, Kim Minkyung, Ha Junkyu, Lee Minhyung
Department of Bioengineering College of Engineering, Hanyang University Seoul Korea.
Bioeng Transl Med. 2022 Oct 18;8(2):e10426. doi: 10.1002/btm2.10426. eCollection 2023 Mar.
The brain-targeted delivery of therapeutic oligonucleotides has been investigated as a new treatment modality for various brain diseases, such as brain tumors. However, delivery efficiency into the brain has been limited due to the blood-brain barrier. In this research, brain-targeted exosome-mimetic cell membrane nanovesicles (CMNVs) were designed to enhance the delivery of therapeutic oligonucleotides into the brain. First, CMNVs were produced by extrusion with isolated C6 cell membrane fragments. Then, CMNVs were decorated with cholesterol-linked T7 peptides as a targeting ligand by hydrophobic interaction, producing T7-CMNV. T7-CMNV was in aqueous solution maintained its nanoparticle size for over 21 days. The targeting and delivery effects of T7-CMNVs were evaluated in an orthotopic glioblastoma animal model. 2'--metyl and cholesterol-TEG modified anti-microRNA-21 oligonucleotides (AMO21c) were loaded into T7-CMNVs, and biodistribution experiments indicated that T7-CMNVs delivered AMO21c more efficiently into the brain than CMNVs, scrambled T7-CMNVs, lipofectamine, and naked AMO21c after systemic administration. In addition, AMO21c down-regulated miRNA-21 (miR-21) levels in glioblastoma tissue most efficiently in the T7-CMNVs group. This enhanced suppression of miR-21 resulted in the up-regulation of PDCD4 and PTEN. Eventually, brain tumor size was reduced in the T7-CMNVs group more efficiently than in the other control groups. With stability, low toxicity, and targeting efficiency, T7-CMNVs may be useful to the development of oligonucleotide therapy for brain tumors.
治疗性寡核苷酸的脑靶向递送已被研究作为治疗各种脑部疾病（如脑肿瘤）的一种新的治疗方式。然而，由于血脑屏障的存在，进入大脑的递送效率一直受到限制。在本研究中，设计了脑靶向的外泌体模拟细胞膜纳米囊泡（CMNVs）以增强治疗性寡核苷酸向大脑的递送。首先，通过用分离的C6细胞膜片段挤压制备CMNVs。然后，通过疏水相互作用用胆固醇连接的T7肽作为靶向配体修饰CMNVs，产生T7-CMNV。T7-CMNV在水溶液中在21天以上保持其纳米颗粒大小。在原位胶质母细胞瘤动物模型中评估了T7-CMNVs的靶向和递送效果。将2'-甲基和胆固醇-TEG修饰的抗微小RNA-21寡核苷酸（AMO21c）装载到T7-CMNVs中，生物分布实验表明，全身给药后，T7-CMNVs比CMNVs、 scrambled T7-CMNVs、脂质体和游离AMO21c更有效地将AMO21c递送至大脑。此外，在T7-CMNVs组中，AMO21c最有效地降低了胶质母细胞瘤组织中的微小RNA-21（miR-21）水平。这种对miR-调节21的增强抑制导致了PDCD4和PTEN的上调。最终，T7-CMNVs组中的脑肿瘤大小比其他对照组更有效地减小。由于具有稳定性、低毒性和靶向效率，T7-CMNVs可能有助于脑肿瘤寡核苷酸治疗的发展。
