Extracellular Vesicles Derived from a Human Brain Endothelial Cell Line Increase Cellular ATP Levels.
Extracellular Vesicles Derived from a Human Brain Endothelial Cell Line Increase Cellular ATP Levels.
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DOI：
10.1208/s12249-020-01892-w
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发表时间：
2021-01-03
影响因子：
3.3
通讯作者：
S Manickam D
中科院分区：
文献类型：
--
作者：
Dave KM;Zhao W;Hoover C;D'Souza A;S Manickam D
工程化的细胞衍生的细胞外囊泡（EV）如外来体和微囊泡由于其较低的免疫原性和固有的靶细胞归巢能力而具有作为安全有效的药物载体的巨大潜力。除了固有的囊泡货物如脂质、蛋白质和核酸之外，还已知EV含有功能性线粒体/线粒体DNA，其可以转移到受体细胞以增加细胞生物能量。在这项概念验证研究中，我们分离了幼稚EV和装载有编码来自人脑内皮细胞系hCMEC/D3和RAW 264.7巨噬细胞的脑源性神经营养因子（BDNF-EV）的外源性质粒DNA的工程EV。我们测试了天然或工程EV中的线粒体组分是否可以增加受体脑内皮细胞中的ATP水平。EV（例如，外来体和微泡;使用差速离心法从未处理的（未处理的）或pDNA转染的（Luc-DNA或BDNF-DNA）细胞的条件培养基中分离EXO和MV。与hCMEC/D3细胞系衍生的EV相比，RAW 264.7细胞系衍生的EV在72 h时在受体hCMEC/D3细胞中显示出显著更高的DNA加载和增加的荧光素酶表达。来自hCMEC/D3细胞的幼稚EV和来自RAW 264.7细胞的BDNF-EV在暴露后24和48小时显示受体hCMEC/D3细胞的ATP水平的小但显著更大的增加。总之，我们已经使用脑内皮细胞和巨噬细胞系证明了作为细胞类型的函数的外源性pDNA加载到EV中的差异，以及受体hCMEC/D3单层中EV介导的细胞内ATP水平的增加。
Engineered cell-derived extracellular vesicles (EVs) such as exosomes and microvesicles hold immense potential as safe and efficient drug carriers due to their lower immunogenicity and inherent homing capabilities to target cells. In addition to innate vesicular cargo such as lipids, proteins, and nucleic acids, EVs are also known to contain functional mitochondria/mitochondrial DNA that can be transferred to recipient cells to increase cellular bioenergetics. In this proof-of-concept study, we isolated naïve EVs and engineered EVs loaded with an exogenous plasmid DNA encoding for brain-derived neurotrophic factor (BDNF-EVs) from hCMEC/D3, a human brain endothelial cell line, and RAW 264.7 macrophages. We tested whether mitochondrial components in naïve or engineered EVs can increase ATP levels in the recipient brain endothelial cells. EVs (e.g., exosomes and microvesicles; EXOs and MVs) were isolated from the conditioned medium of either untreated (naïve) or pDNA-transfected (Luc-DNA or BDNF-DNA) cells using a differential centrifugation method. RAW 264.7 cell line–derived EVs showed a significantly higher DNA loading and increased luciferase expression in the recipient hCMEC/D3 cells at 72 h compared with hCMEC/D3 cell line–derived EVs. Naïve EVs from hCMEC/D3 cells and BDNF-EVs from RAW 264.7 cells showed a small, but a significantly greater increase in the ATP levels of recipient hCMEC/D3 cells at 24 and 48 h post-exposure. In summary, we have demonstrated differences in exogenous pDNA loading into EVs as a function of cell type using brain endothelial and macrophage cell lines and EV-mediated increases in the intracellular ATP levels in the recipient hCMEC/D3 monolayers.
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影响因子：
46.9
作者：
Alvarez-Erviti, Lydia;Seow, Yiqi;Wood, Matthew J. A.
通讯作者：
Wood, Matthew J. A.
影响因子：
7.3
作者：
Andreu Z;Yáñez-Mó M
通讯作者：
Yáñez-Mó M
影响因子：
3.1
作者：
Cipolla, MJ;Crete, R;Rix, RD
通讯作者：
Rix, RD
影响因子：
64.5
作者：
KING, MP;ATTARDI, G
通讯作者：
ATTARDI, G
影响因子：
11.4
作者：
Hough KP;Trevor JL;Strenkowski JG;Wang Y;Chacko BK;Tousif S;Chanda D;Steele C;Antony VB;Dokland T;Ouyang X;Zhang J;Duncan SR;Thannickal VJ;Darley-Usmar VM;Deshane JS
通讯作者：
Deshane JS