Differential fates of biomolecules delivered to target cells via extracellular vesicles
Differential fates of biomolecules delivered to target cells via extracellular vesicles
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DOI：
10.1073/pnas.1418401112
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发表时间：
2015-03-24
影响因子：
11.1
通讯作者：
Contag, Christopher H.
中科院分区：
文献类型：
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作者：
Kanada, Masamitsu;Bachmann, Michael H.;Contag, Christopher H.
细胞外囊泡（EV），特别是外泌体和微囊泡（MV），被认为是通过细胞之间的生物分子转移在细胞-细胞通信中发挥关键作用。这两种类型的EV的生物起源不同，因为它们起源于内体（外泌体）或血浆（MV）膜。为了阐明EV介导细胞间通讯的主要手段，我们表征了它们从瞬时转染细胞中封装和递送不同类型大分子的能力。两种EV类型均包封报告蛋白和mRNA，但只有MV将报告功能转移到受体细胞。受体细胞中的从头报告蛋白表达仅由经由MV递送后的质粒DNA（pDNA）引起。两种EV均将报告基因mRNA递送至受体细胞，但在未翻译的情况下迅速降解。MV还介导编码Cre重组酶的功能性pDNA在体内递送至转基因Crelox报告小鼠中的组织。在本研究的参数内，MV递送功能性pDNA，但不递送RNA，而来自相同来源的外来体不递送功能性核酸。这些结果对于理解电动汽车在蜂窝通信中的作用以及电动汽车作为交付工具的发展具有重要意义。此外，使用来自瞬时转染细胞的EV的研究可能被pDNA转移的优势所混淆。
Extracellular vesicles (EVs), specifically exosomes and microvesicles (MVs), are presumed to play key roles in cell-cell communication via transfer of biomolecules between cells. The biogenesis of these two types of EVs differs as they originate from either the endosomal (exosomes) or plasma (MVs) membranes. To elucidate the primary means through which EVs mediate intercellular communication, we characterized their ability to encapsulate and deliver different types of macromolecules from transiently transfected cells. Both EV types encapsulated reporter proteins and mRNA but only MVs transferred the reporter function to recipient cells. De novo reporter protein expression in recipient cells resulted only from plasmid DNA (pDNA) after delivery via MVs. Reporter mRNA was delivered to recipient cells by both EV types, but was rapidly degraded without being translated. MVs also mediated delivery of functional pDNA encoding Cre recombinase in vivo to tissues in transgenic Crelox reporter mice. Within the parameters of this study, MVs delivered functional pDNA, but not RNA, whereas exosomes from the same source did not deliver functional nucleic acids. These results have significant implications for understanding the role of EVs in cellular communication and for development of EVs as delivery tools. Moreover, studies using EVs from transiently transfected cells may be confounded by a predominance of pDNA transfer.