间充质干细胞作为携带 miR-124a 的外泌体的天然生物工厂在治疗神经胶质瘤中的应用。
Mesenchymal stem cells as natural biofactories for exosomes carrying miR-124a in the treatment of gliomas.
机构信息
Department of Neurosurgery and Brain Tumor Center, University of Texas MD Anderson Cancer Center, Houston, Texas.
出版信息
Neuro Oncol. 2018 Feb 19;20(3):380-390. doi: 10.1093/neuonc/nox152.
BACKGROUND
MicroRNAs (miRs) are promising new therapeutics for glioblastoma. However, which miRs are most effective against glioblastomas and how these miRs should be delivered are major unanswered problems.
METHODS
To identify potent antiglioma miRs, we selected 8 miRs based on a literature search and screened them against a panel of glioma stem cell (GSC) lines, representing all of the glioblastoma subtypes defined by The Cancer Genome Atlas. To address delivery, we tested the hypothesis that ex vivo cultured bone marrow-derived mesenchymal stem cells (MSCs) can package miRs into exosomes and that these engineered exosomes can systemically deliver antiglioma miRs to glioblastomas.
RESULTS
Of the screened miRs, we identified miR-124a as the most effective antiglioma agent against GSCs. We then transduced MSCs with lentivirus vectors containing miR-124a and isolated vesicles from the medium. Electron microscopy, western blotting, and Nanosight proved that the isolated vesicles were exosomes. Quantitative PCR documented that these exosomes contained high levels of miR-124a, which was not present in control exosomes. In vitro treatment of GSCs with exosomes containing miR-124a (Exo-miR124) resulted in a significant reduction in viability and clonogenicity of GSCs compared with controls. In vivo treatment of mice harboring intracranial GSC267 with systemically delivered Exo-miR124 resulted in 50% of animals living long term. No evidence of tumor was present on histological analysis of the survivors. Mechanistic studies showed that miR-124a acts by silencing Forkhead box (FOX)A2, resulting in aberrant intracellular lipid accumulation.
CONCLUSION
MSCs can be used as natural biofactories to produce Exo-miR124, which is an effective antiglioma agent worthy of further clinical evaluation.
背景
微小 RNA（miRs）是治疗脑胶质瘤的有前途的新疗法。然而，哪些 miR 对脑胶质瘤最有效，以及如何传递这些 miR 是尚未解决的主要问题。
方法
为了鉴定有效的抗脑胶质瘤 miR，我们根据文献检索选择了 8 个 miR，并对一组脑胶质瘤干细胞（GSC）系进行了筛选，这些细胞系代表了癌症基因组图谱定义的所有脑胶质瘤亚型。为了解决传递问题，我们测试了一个假设，即体外培养的骨髓间充质干细胞（MSCs）可以将 miR 包装到外泌体中，并且这些工程化的外泌体可以将抗脑胶质瘤 miR 系统地递送到脑胶质瘤中。
结果
在筛选的 miR 中，我们鉴定出 miR-124a 是针对 GSCs 最有效的抗脑胶质瘤药物。然后，我们用含有 miR-124a 的慢病毒载体转导 MSCs，并从培养基中分离出囊泡。电子显微镜、western blot 和 Nanosight 证明，分离出的囊泡是外泌体。定量 PCR 记录表明，这些外泌体含有高水平的 miR-124a，而对照外泌体中不存在。体外用含有 miR-124a 的外泌体（Exo-miR124）处理 GSCs，与对照组相比，GSCs 的活力和集落形成能力显著降低。用系统递送的 Exo-miR124 治疗携带颅内 GSC267 的小鼠，有 50%的动物长期存活。在幸存者的组织学分析中没有发现肿瘤的证据。机制研究表明，miR-124a 通过沉默叉头框（FOX）A2 起作用，导致细胞内脂质异常积累。
结论
MSCs 可以用作天然的生物工厂来产生 Exo-miR124，这是一种有效的抗脑胶质瘤药物，值得进一步的临床评估。
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