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

Year: 2020
Type: article
Abstract: 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.
DOI: 10.1038/s42003-020-01336-y
Authors: Harshani Wijerathne, Malgorzata A. Witek, Joshua M. Jackson, Virginia Brown, Mateusz L. Hupert, Kristina Herrera, Cameron Kramer, Abigail E. Davidow, Yan Li, Alison E. Baird, Michael C. Murphy, Steven A. Soper
Institutions: University of Kansas, University of North Carolina at Chapel Hill, The University of Kansas Cancer Center, University of Kansas Medical Center, BioFluidica (United States), State University of New York, SUNY Downstate Health Sciences University, Louisiana State University
Language: English
Open Access status: gold
Estimated APC paid: $3,290
Funders: U.S. Department of Health and Human Services, National Institutes of Health, Cancer Center, University of Kansas, National Cancer Institute, National Institute of General Medical Sciences, National Institute of Biomedical Imaging and Bioengineering, Office of Extramural Research, National Institutes of Health
Awards: G1411983911, G1725766855, G1779537631, G204892779, G2234905489, spFRET for Expression Profiling mRNA: Discovering Markers for Stroke Diagnosis, Parallel and high throughput isolation of cancer cell derived extracellular vesicles, G2607785744, Biotechnology Resource Center of Biomodular Multi scale Systems CBM2 for Precision Molecular Diagnostics, G2952127986, G303123301, G3120329307, G3345231741, The University of Kansas Cancer Center, G4827751652, G5277976840, G6679460133, Molecular Analysis of Disease Pathways, G6976316766, G700021213, G8014481505, G8205639842, G9000053679, G9000081616, G9000129762, G9000180596, G9000248580, The Kansas Institute for Precision Medicine
OpenAlex ID: W3093701411
FWCI: 1.658
Cites: 40
Cited by: 37
Related to: 10