RVG制导外泌体递送si-linc-POU3F3靶向清除残余胶质瘤

Glioma is the most common type of intercranial malignant tumor,surgery combined with radiation and chemotherapy has been the major therapeutic option.However,this kind of tumor cells are so aggressive and with special anatomical location that it can not be removed entirely,becoming a recurrent source.Exploring a drug delivery system that based on novel mechanism and target is crucial to eradicate residual glioma selectively.Our previous study confirmed that the downregulation of long non-coding RNA（linc-RNA POU3F3）can induce tumor cell apoptosis effectively,and its small interfering RNA(si-linc-POU3F3) has a good potential for targeted drugs,in the meantime, we found that exosomes can be used as its carrier.In this study, we are going to use the exosomes that the surface of their membrane is carrying rabies virus glycoprotein RVG elements as carrier,since the RVG has a speciality of addicting to neurons，they can enter the brain tissue with si-linc-POU3F3 and work on glioma cells, eradicating residual glioma entirely.RVG guided-exosomes are made restruction of lentivirus and electroporation,observing cell proliferation,migration and apoptosis in vitro,and test the ability of combining and to observe tumor cells through confocal microscopy and pathology in vivo.Finally,it can provide a novel strategy of eradicating residual glioma effectively.

胶质瘤是颅内最常见的恶性肿瘤，手术合并放化疗为其主要治疗手段。但瘤细胞的侵袭生长特性和特殊解剖位置，致其难以被彻底清除，成为复发根源。探索基于新机制和新靶点的给药体系以靶向清除残余胶质瘤细胞具有重要意义。我们前期研究证实下调长链非编码RNA linc-POU3F3能有效诱导瘤细胞凋亡，其干扰RNA（si-linc-POU3F3）具备良好的靶向药物潜能，同时我们发现外泌体可作为其优良载体。本研究拟以膜表面携带狂犬病毒糖蛋白RVG片段的外泌体为载体，利用RVG嗜神经元特性，导向性携带si-linc-POU3F3进入脑组织靶向作用于胶质瘤细胞，实现残余瘤细胞的彻底清除。本项目以重组慢病毒、电转染等技术制备RVG制导外泌体，体外共培养观察瘤细胞增殖、迁移、凋亡；体内实验通过共聚焦显微镜、病理检验等技术观察其对瘤细胞的靶向结合及清除能力。最终为探索高效清除术后残余胶质瘤提供新思路。

MicroRNA（miRNA）是小的非编码RNA，可在转录和翻译过程中调节基因表达，其失调可导致肿瘤发生。外泌体通过其分子货物在细胞通讯中起重要作用。在这项研究中，我们证明了骨髓间充质干细胞（BMSC）衍生的外泌体可以通过传递miR-512-5p改变肿瘤微环境的通讯中的肿瘤表型。我们的研究结果发现，胶质母细胞（GBM）组织和细胞中miR-512-5p被下调，而Jagged 1（JAG1）是miR-512-5p的靶基因。同时，我们阐明了在GBM中miR-512-5p和JAG1的表达模式以及它们之间的相互作用。此外，我们观察到，BMSC衍生的外来体可能包含miR-512-5p并在体外将其转运至GBM细胞。结果表明，BMSC衍生的外泌体miR-512-5p可以抑制GBM细胞增殖，并通过抑制JAG1表达诱导细胞周期停滞。此外，进行了体内测定以验证体外发现。数据表明，源自BMSC的外泌体可将miR-512-5p转运至GBM，并通过靶向JAG1抑制GBM进程。简而言之，这项研究揭示了一种治疗GBM的新分子机制，并且进一步验证了miRNA可以包装到外泌体中，参与GBM中的细胞通讯。