Affinity enrichment of extracellular vesicles from plasma reveals mRNA changes associated with acute ischemic stroke

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Issue Date
2020-10-26Author
Wijerathne, Harshani
Witek, Malgorzata A.
Jackson, Joshua M.
Brown, Virginia
Hupert, Mateusz L.
Herrera, Kristina
Kramer, Cameron
Davidow, Abigail E.
Li, Yan
Baird, Alison E.
Murphy, Michael C.
Soper, Steven A.
Publisher
Nature Research
Type
Article
Article Version
Scholarly/refereed, publisher version
Rights
© The Author(s) 2020. This article is licensed under a Creative Commons Attribution 4.0 International License.
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Show full item recordAbstract
Currently there is no in vitro diagnostic test for acute ischemic stroke (AIS), yet rapid diagnosis is crucial for effective thrombolytic treatment. We previously demonstrated the utility of CD8(+) T-cells’ mRNA expression for AIS detection; however extracellular vesicles (EVs) were not evaluated as a source of mRNA for AIS testing. We now report a microfluidic device for the rapid and efficient affinity-enrichment of CD8(+) EVs and subsequent EV’s mRNA analysis using droplet digital PCR (ddPCR). The microfluidic device contains a dense array of micropillars modified with anti-CD8α monoclonal antibodies that enriched 158 ± 10 nm sized EVs at 4.3 ± 2.1 × 109 particles/100 µL of plasma. Analysis of mRNA from CD8(+) EVs and their parental T-cells revealed correlation in the expression for AIS-specific genes in both cell lines and healthy donors. In a blinded study, 80% test positivity for AIS patients and controls was revealed with a total analysis time of 3.7 h.
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Citation
Wijerathne, H., Witek, M.A., Jackson, J.M. et al. Affinity enrichment of extracellular vesicles from plasma reveals mRNA changes associated with acute ischemic stroke. Commun Biol 3, 613 (2020). https://doi.org/10.1038/s42003-020-01336-y
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