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Extracellular Vesicles Derived from a Human Brain Endothelial Cell Line Increase Cellular ATP Levels

  • Research Article
  • Theme: NIPTE Research and Perspective: Advances in Nanotechnology-Based Drug Delivery
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A Correction to this article was published on 09 February 2023

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Abstract

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 (1) differences in exogenous pDNA loading into EVs as a function of cell type using brain endothelial and macrophage cell lines and (2) EV-mediated increases in the intracellular ATP levels in the recipient hCMEC/D3 monolayers.

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Acknowledgments

The authors express their deep appreciation to Dr. Lauren O’Donnell (DU), Mss. Manisha Chandwani, and Yashika Kamte for flow cytometry support. We thank Dr. Jelena Janjic for allowing use of the Malvern Zetasizer Nano.

Funding

The study was supported using start-up funds for the Manickam laboratory from the School of Pharmacy at Duquesne University (DU). We acknowledge funding for CH through the Neurodegeneration Undergraduate Research Program NIH R25NS100118 (Drs. Benedict Kolber, Kevin Tidgewell, Michael Cascio, and Rita Mihailescu, DU).

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  1. Kandarp M. Dave and Wanzhu Zhao contributed equally to this work.

Authors and Affiliations

  1. Graduate School of Pharmaceutical Sciences, Duquesne University, 453 Mellon Hall, 600 Forbes Avenue, Pittsburgh, Pennsylvania, 15282, USA

    Kandarp M. Dave, Wanzhu Zhao, Anisha D’Souza & Devika S Manickam

  2. Department of Chemistry and Physics, Mansfield University, Mansfield, Pennsylvania, USA

    Catherine Hoover

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  1. Kandarp M. Dave
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Correspondence to Devika S Manickam.

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Dave, K.M., Zhao, W., Hoover, C. et al. Extracellular Vesicles Derived from a Human Brain Endothelial Cell Line Increase Cellular ATP Levels. AAPS PharmSciTech 22, 18 (2021). https://doi.org/10.1208/s12249-020-01892-w

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