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Open Access 01-12-2023 | Stroke | Research

Plasma derived extracellular vesicle biomarkers of microglia activation in an experimental stroke model

Authors: A. D. Roseborough, S. J. Myers, R. Khazaee, Y. Zhu, L. Zhao, E. Iorio, F. M. Elahi, S. H. Pasternak, S. N. Whitehead

Published in: Journal of Neuroinflammation | Issue 1/2023

Abstract

Chronic microglia activation post-stroke is associated with worse neurological and cognitive outcomes. However, measurement of microglia activation in vivo is currently limited. Plasma derived extracellular vesicles (EVs) are cell-specific indicators that may allow for non-invasive measurement of microglia phenotype. The aim of this study was to identify activation-state specific microglia EVs (MEVs) in vitro followed by validation in an experimental stroke model. Following pro-inflammatory activation, MEVs contain the microglia protein TMEM119 alongside increased expression of the Toll-like receptor 4 co-receptor CD14. Immunoprecipitation followed by fluorescent nanoparticle tracking analysis (ONI Nanoimager) was used to confirm the isolation of TMEM119⁺/CD14⁺ EVs from rat plasma. Electron microscopy confirmed that TMEM119 and CD14 localize to the MEV membrane. To model ischemia, plasma was collected from 3-month wildtype Fischer344 rats prior to, 7 and 28 days after endothelin-1 or saline injection into the dorsal right striatum. Fluorescently labelled MEVs were directly measured in the plasma using nanoflow cytometry (Apogee A60 Microplus). We report a significant increase in circulating TMEM119⁺/CD14⁺ EVs 28-days post-stroke in comparison to baseline levels and saline-injected rats, which correlated weakly with stroke volume. TMEM119⁺/MHC-II⁺ EVs were also increased post-stroke in comparison to baseline and saline-injected animals. This study is the first to describe an EV biomarker of activated microglia detected directly in plasma following stroke and represents a future tool for the measurement of microglia activity in vivo.

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Title: Plasma derived extracellular vesicle biomarkers of microglia activation in an experimental stroke model
Authors: A. D. Roseborough
S. J. Myers
R. Khazaee
Y. Zhu
L. Zhao
E. Iorio
F. M. Elahi
S. H. Pasternak
S. N. Whitehead
Publication date: 01-12-2023
Publisher: BioMed Central
Keywords: Stroke
Biomarkers
Published in: Journal of Neuroinflammation / Issue 1/2023
Electronic ISSN: 1742-2094
DOI: https://doi.org/10.1186/s12974-023-02708-x

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Plasma derived extracellular vesicle biomarkers of microglia activation in an experimental stroke model

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