Mitochondria-containing extracellular vesicles (EV) reduce mouse brain infarct sizes and EV/HSP27 protect ischemic brain endothelial cultures

DOI

10.1016/j.jconrel.2023.01.025

Kandarp M. Dave, Graduate School of Pharmaceutical Sciences, Duquesne University, Pittsburgh, PA, USA.
Donna B. Stolz, Center for Biologic Imaging, University of Pittsburgh Medical School, Pittsburgh, PA, USA.
Venugopal R. Venna, Department of Neurology, McGovern Medical School, University of Texas Health Science Center, Houston, TX, USA.
Victoria A. Quaicoe, Department of Neurology, McGovern Medical School, University of Texas Health Science Center, Houston, TX, USA.
Michael E. Maniskas, Department of Neurology, McGovern Medical School, University of Texas Health Science Center, Houston, TX, USA.
Michael John Reynolds, Pittsburgh Heart Lung Blood Vascular Institute, University of Pittsburgh Medical School, Pittsburgh, PA, USA.
Riyan Babidhan, Graduate School of Pharmaceutical Sciences, Duquesne University, Pittsburgh, PA, USA.
Duncan X. Dobbins, Graduate School of Pharmaceutical Sciences, Duquesne University, Pittsburgh, PA, USA.
Maura N. Farinelli, Graduate School of Pharmaceutical Sciences, Duquesne University, Pittsburgh, PA, USA; Department of Biochemistry and Molecular Biology, Gettysburg College, Gettysburg, PA, USA.
Abigail Sullivan, Graduate School of Pharmaceutical Sciences, Duquesne University, Pittsburgh, PA, USA; Psychological and Brain Sciences, Villanova University, Villanova, PA, USA.
Tarun N. Bhatia, Graduate School of Pharmaceutical Sciences, Duquesne University, Pittsburgh, PA, USA.
Hannah Yankello, Departments of Chemical and Biomedical Engineering, Carnegie Mellon University, Pittsburgh, PA, USA.
Rohan Reddy, Graduate School of Pharmaceutical Sciences, Duquesne University, Pittsburgh, PA, USA.
Younsoo Bae, Department of Pharmaceutical Sciences, College of Pharmacy, The University of Kentucky, Lexington, KY, USA.
Rehana K. Leak, Graduate School of Pharmaceutical Sciences, Duquesne University, Pittsburgh, PA, USA.
Sruti S. Shiva, Pittsburgh Heart Lung Blood Vascular Institute, University of Pittsburgh Medical School, Pittsburgh, PA, USA; Department of Pharmacology & Chemical Biology, University of Pittsburgh Medical School, Pittsburgh, PA, USA.
Louise D. McCullough, Department of Neurology, McGovern Medical School, University of Texas Health Science Center, Houston, TX, USA.
Devika S. Manickam, Graduate School of Pharmaceutical Sciences, Duquesne University, Pittsburgh, PA, USA. Electronic address: soundaramanickd@duq.edu.

Document Type

Journal Article

Publication Date

2-1-2023

Publication Title

Journal of controlled release : official journal of the Controlled Release Society

Volume

354

First Page

368

Last Page

393

Keywords

BBB protection, Extracellular vesicles, Heat shock protein, Ischemic stroke, Mitochondria, Paracellular permeability

Abstract

Ischemic stroke causes brain endothelial cell (BEC) death and damages tight junction integrity of the blood-brain barrier (BBB). We harnessed the innate mitochondrial load of BEC-derived extracellular vesicles (EVs) and utilized mixtures of EV/exogenous 27 kDa heat shock protein (HSP27) as a one-two punch strategy to increase BEC survival (via EV mitochondria) and preserve their tight junction integrity (via HSP27 effects). We demonstrated that the medium-to-large (m/lEV) but not small EVs (sEV) transferred their mitochondrial load, that subsequently colocalized with the mitochondrial network of the recipient primary human BECs. Recipient BECs treated with m/lEVs showed increased relative ATP levels and mitochondrial function. To determine if the m/lEV-meditated increase in recipient BEC ATP levels was associated with m/lEV mitochondria, we isolated m/lEVs from donor BECs pre-treated with oligomycin A (OGM, mitochondria electron transport complex V inhibitor), referred to as OGM-m/lEVs. BECs treated with naïve m/lEVs showed a significant increase in ATP levels compared to untreated OGD cells, OGM-m/lEVs treated BECs showed a loss of ATP levels suggesting that the m/lEV-mediated increase in ATP levels is likely a function of their innate mitochondrial load. In contrast, sEV-mediated ATP increases were not affected by inhibition of mitochondrial function in the donor BECs. Intravenously administered m/lEVs showed a reduction in brain infarct sizes compared to vehicle-injected mice in a mouse middle cerebral artery occlusion model of ischemic stroke. We formulated binary mixtures of human recombinant HSP27 protein with EVs: EV/HSP27 and ternary mixtures of HSP27 and EVs with a cationic polymer, poly (ethylene glycol)-b-poly (diethyltriamine): (PEG-DET/HSP27)/EV. (PEG-DET/HSP27)/EV and EV/HSP27 mixtures decreased the paracellular permeability of small and large molecular mass fluorescent tracers in oxygen glucose-deprived primary human BECs. This one-two punch approach to increase BEC metabolic function and tight junction integrity may be a promising strategy for BBB protection and prevention of long-term neurological dysfunction post-ischemic stroke.

Open Access

36642252 (pubmed); NIHMS1866826 (mid); PMC9974867 (pmc); 10.1016/j.jconrel.2023.01.025 (doi); S0168-3659(23)00026-3 (pii)

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