由于无效的药物和不良的给药方法，胶质母细胞瘤（GBM）的预后较差。 MicroRNA (miR) 已被探索作为胶质母细胞瘤的新型疗法，但最佳的 miR 和理想的递送策略仍未解决。在这项研究中，我们试图找出最有效的泛亚型抗 GBM miR，并为这些 miR 开发改进的递送系统。我们对代表所有 GBM 的 7 个胶质瘤干细胞 (GSC) 系的 600 多个 miR 进行了无偏见筛选-亚型，以确定一组泛亚型特异性抗 GBM miR，然后使用可用的 TCGA GBM 患者结果和 miR 表达数据来磨练最有可能在临床上有效的 miR。为了增强 miR 的递送和表达，我们生成了编码所有三种 miR 的多顺反子质粒 (pPolymiR)，并使用 HEK293T 细胞作为生物工厂，将 pPolymiR 包装到工程外泌体 (eExos) 中，该外泌体在其结构中掺入了病毒蛋白 (Gag/VSVg) (eExos) pPolymiR) 来增强功能。我们的逐步筛选确定 miR-124-2、miR-135a-2 和 let-7i 是所有 GBM 亚型中具有临床相关性的最有效的 miR。 eExos pPolymiR 的递送导致 GSC 中所有三种 miR 的高表达，并显着降低体外 GSC 增殖。与单独携带每种 miR 或作为混合物的 eExos 相比，eExos pPolymiR 延长了携带 GSC 的小鼠的体内存活率。 eExos pPolymiR，其中包括在多顺反子质粒中编码的泛亚型抗神经胶质瘤特异性 miR 组合和一种新型外泌体递送平台代表了一种新的、潜在的强大抗 GBM 疗法。© 作者 2023。由牛津大学出版社代表神经肿瘤学会出版。版权所有。如需权限，请发送电子邮件至：journals.permissions@oup.com。

Glioblastoma (GBM) has poor prognosis due to ineffective agents and poor delivery methods. MicroRNAs (miRs) have been explored as novel therapeutics for glioblastoma, but the optimal miRs and the ideal delivery strategy remain unresolved. In this study, we sought to identify the most effective pan-subtype anti-GBM miRs and to develop an improved delivery system for these miRs.We conducted an unbiased screen of over 600 miRs against 7 Glioma Stem Cell (GSCs) lines representing all GBM-subtypes to identify a set of pan-subtype-specific anti-GBM miRs and then used available TCGA GBM patient outcomes and miR expression data to hone in on miRs that were most likely to be clinically effective. To enhance delivery and expression of the miRs, we generated a polycistronic plasmid encoding all three miRs (pPolymiR) and used HEK293T cells as biofactories to package pPolymiR into engineered exosomes (eExos) that incorporate viral proteins (Gag/ VSVg) in their structure (eExos+pPolymiR) to enhance function.Our stepwise screen identified miR-124-2, miR-135a-2, and let-7i as the most effective miRs across all GBM subtypes with clinical relevance. Delivery of eExos+pPolymiR resulted in high expression of all three miRs in GSCs, and significantly decreased GSC proliferation in vitro. eExos+pPolymiR prolonged survival of GSC-bearing mice in vivo when compared with eExos carrying each of the miRs individually or as a cocktail.eExos+pPolymiR, which includes a pan-subtype anti-glioma-specific miR combination encoded in a polycistronic plasmid and a novel exosome delivery platform represents a new and potentially powerful anti-GBM therapeutic.© The Author(s) 2023. Published by Oxford University Press on behalf of the Society for Neuro-Oncology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.