Direct isolation of circulating extracellular vesicles from blood for vascular risk profiling in type 2 diabetes mellitus. Issue 13 (27th May 2021)

• Record Type: Journal Article
• Authors: Tay, Hui Min; Leong, Sheng Yuan; Xu, Xiaohan; Kong, Fang; Upadya, Megha; Dalan, Rinkoo; Tay, Chor Yong; Dao, Ming; Suresh, Subra; Hou, Han Wei
• Is Part Of: Lab on a chip. Volume 21:Issue 13(2021)
• Page Start: 2511
• Page End: 2523
• Publication Date: 2021-05-27
• Subjects: Miniature electronic equipment -- Periodicals; Combinatorial chemistry -- Periodicals; Biotechnology -- Periodicals; 543.0813
• DOI: 10.1039/d1lc00333j
• Languages: English
• ISSNs: 1473-0197

Abstract

Abstract : A simple, economical and scalable microfluidic separation technology (ExoDFF) for label-free isolation of circulating extracellular vesicles (EVs) from whole blood. Abstract : Extracellular vesicles (EVs) are key mediators of communication among cells, and clinical utilities of EVs-based biomarkers remain limited due to difficulties in isolating EVs from whole blood reliably. We report a novel inertial-based microfluidic platform for direct isolation of nanoscale EVs (exosomes, 50 to 200 nm) and medium-sized EVs (microvesicles, 200 nm to 1 μm) from blood with high efficiency (three-fold increase in EV yield compared to ultracentrifugation). In a pilot clinical study of healthy ( n = 5) and type 2 diabetes mellitus (T2DM, n = 9) subjects, we detected higher EV levels in T2DM patients ( P < 0.05), and identified a subset of "high-risk" T2DM subjects with abnormally high (∼10-fold to 50-fold) amounts of platelet (CD41a+) or leukocyte-derived (CD45+) EVs. Our in vitro endothelial cell assay further revealed that EVs from "high-risk" T2DM subjects induced significantly higher vascular inflammation (ICAM-1 expression) ( P < 0.05) as compared to healthy and non-"high-risk" T2DM subjects, reflecting a pro-inflammatory phenotype. Overall, the EV isolation tool is scalable, and requires less manual labour, cost and processing time. This enables further development of EV-based diagnostics, whereby a combined immunological and functional phenotyping strategy can potentially be used for rapid vascular risk stratification in T2DM.