Microvesicle-mediated RNA Molecule Delivery System Using Monocytes/Macrophages
Microvesicle-mediated RNA Molecule Delivery System Using Monocytes/Macrophages
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DOI：
10.1038/mt.2010.254
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发表时间：
2011-02-01
影响因子：
12.4
通讯作者：
Naoe, Tomoki
中科院分区：
文献类型：
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作者：
Akao, Yukihiro;Iio, Akio;Naoe, Tomoki
微泡（MV）和外泌体作为细胞间通讯工具从细胞脱落，是将RNA分子导航到身体组织的可能载体。认为静脉注射来自患者的此类MV或外泌体不会引起严重的非自身和毒性反应。以前，我们发现巨噬细胞摄取脂质体捕获的RNA分子，其中一些在细胞中保持未降解。在这里，我们证明了转染的RNA分子在人单核细胞白血病THP-1细胞脱落的THP-1巨噬细胞的内容物在无血清培养基中孵育过程中，脱落的生化分析，如定量逆转录（qRT）-PCR，TSG 101（膜相关的外泌体蛋白）的表达，和免疫电镜研究。与未修饰的miR-143转染后的量相比，miR-143 BP转染后THP-1巨噬细胞分泌更多的由MV（MV-miR-143 BPs）捕获的化学修饰的RNA分子（miR-143 BPs）。此外，我们表明，THP-1巨噬细胞，这是用miR-143 BP离体转染，分泌MV-miR-143 BP在异种移植裸鼠静脉注射后，因为miR-143水平显着增加，在血清中，肿瘤，和宿主动物的肾脏。这些数据表明，一些转染的miR-143 BP在体外和体内均作为MV-RNA从THP-1巨噬细胞分泌。
Microvesicles (MVs) and exosomes, which are shed from cells as a cell-to-cell communication tool, are possible vehicles for navigating RNA molecules to body tissues. It is considered that intravenous injection of such MVs or exosomes from patients would not cause severe not-self and toxic reactions. Previously, we found that macrophages take up liposome-entrapped RNA molecules, some of which remain undegraded in the cells. Here, we demonstrate that transfected RNA molecules in human monocytic leukemia THP-1 cells were shed from THP-1 macrophages as contents in MVs during incubation in serum-free medium, which shedding was shown by biochemical analyses such as quantitative reverse transcription (qRT)-PCR, expression of TSG101 (a membrane-associated exosomal protein), and immunoelectron microscopic study. More chemically modified RNA molecules (miR-143BPs) entrapped by MVs (MV-miR-143BPs) were secreted from THP-1 macrophages after miR-143BP transfection compared with the amount after transfection with nonmodified miR-143 transfection. Furthermore, we show that the THP-1 macrophages, which were transfected with the miR-143BP ex vivo, secreted MV-miR-143BPs in xenografted nude mice after intravenous injection, because miR-143 levels were significantly increased in the serum, tumor, and kidney of the host animals. These data suggest that some of the transfected miR-143BPs were secreted from THP-1 macrophages as MV-RNAs both in vitro and in vivo.