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

DOI

10.1208/s12249-020-01892-w

Authors

Kandarp M. Dave, Graduate School of Pharmaceutical Sciences, Duquesne University, 453 Mellon Hall, 600 Forbes Avenue, Pittsburgh, Pennsylvania, 15282, USA.
Wanzhu Zhao, Graduate School of Pharmaceutical Sciences, Duquesne University, 453 Mellon Hall, 600 Forbes Avenue, Pittsburgh, Pennsylvania, 15282, USA.
Catherine Hoover, Department of Chemistry and Physics, Mansfield University, Mansfield, Pennsylvania, USA.
Anisha D'Souza, Graduate School of Pharmaceutical Sciences, Duquesne University, 453 Mellon Hall, 600 Forbes Avenue, Pittsburgh, Pennsylvania, 15282, USA.
Devika S Manickam, Graduate School of Pharmaceutical Sciences, Duquesne University, 453 Mellon Hall, 600 Forbes Avenue, Pittsburgh, Pennsylvania, 15282, USA. soundaramanickd@duq.edu.

Document Type

Journal Article

Publication Date

1-3-2021

Publication Title

AAPS PharmSciTech

Volume

22

Issue

1

First Page

18

Keywords

ATP increase, brain endothelial cells, exosomes and microvesicles, ischemic stroke, mitochondria and mtDNA

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.

Open Access

Green Accepted

Repository Citation

Dave, K. M., Zhao, W., Hoover, C., D'Souza, A., & S Manickam, D. (2021). Extracellular Vesicles Derived from a Human Brain Endothelial Cell Line Increase Cellular ATP Levels. AAPS PharmSciTech, 22 (1), 18. https://doi.org/10.1208/s12249-020-01892-w