Direct isolation of circulating extracellular vesicles from blood for vascular risk profiling in type 2 diabetes mellitus
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 (exo...
| Main Authors: | , , , , , , , , , |
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| Format: | Journal Article |
| Language: | English |
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2022
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| Online Access: | https://hdl.handle.net/10356/150958 |
| _version_ | 1826120494353481728 |
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| author | Tay, Hui Min Leong, Sheng Yuan Xu, Xiaohan Kong, Fang Upadya, Megha Dalan, Rinkoo Tay, Chor Yong Dao, Ming Suresh, Subra Hou, Han Wei |
| author2 | School of Biological Sciences |
| author_facet | School of Biological Sciences Tay, Hui Min Leong, Sheng Yuan Xu, Xiaohan Kong, Fang Upadya, Megha Dalan, Rinkoo Tay, Chor Yong Dao, Ming Suresh, Subra Hou, Han Wei |
| author_sort | Tay, Hui Min |
| collection | NTU |
| description | 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. |
| first_indexed | 2024-10-01T05:17:42Z |
| format | Journal Article |
| id | ntu-10356/150958 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T05:17:42Z |
| publishDate | 2022 |
| record_format | dspace |
| spelling | ntu-10356/1509582024-04-26T16:02:56Z Direct isolation of circulating extracellular vesicles from blood for vascular risk profiling in type 2 diabetes mellitus Tay, Hui Min Leong, Sheng Yuan Xu, Xiaohan Kong, Fang Upadya, Megha Dalan, Rinkoo Tay, Chor Yong Dao, Ming Suresh, Subra Hou, Han Wei School of Biological Sciences School of Mechanical and Aerospace Engineering School of Materials Science and Engineering Lee Kong Chian School of Medicine (LKCMedicine) Tan Tock Seng Hospital Medicine, Health and Life Sciences Biomechanics Nanotechnology 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. Ministry of Education (MOE) Nanyang Technological University Singapore-MIT Alliance for Research and Technology (SMART) Submitted/Accepted version H. W. H. would like to acknowledge the kind financial support from SMART Innovation Centre (ING-000539 BIO IGN and ING-001058 BIO IGN), Singapore Ministry of Education (MOE) Academic Research Fund Tier 1 (RG53/18), as well as A. Menarini Biomarkers Singapore Pte Ltd. S. Y. L would like to acknowledge support from the NTU Research Scholarship. 2022-02-22T07:53:08Z 2022-02-22T07:53:08Z 2021 Journal Article Tay, H. M., Leong, S. Y., Xu, X., Kong, F., Upadya, M., Dalan, R., Tay, C. Y., Dao, M., Suresh, S. & Hou, H. W. (2021). Direct isolation of circulating extracellular vesicles from blood for vascular risk profiling in type 2 diabetes mellitus. Lab On a Chip, 21(13), 2511-2523. https://dx.doi.org/10.1039/d1lc00333j 1473-0197 https://hdl.handle.net/10356/150958 10.1039/d1lc00333j 13 21 2511 2523 en ING-000539 BIO IGN ING-001058 BIO IGN RG53/18 Lab on a Chip © 2021 The Author(s). Published by The Royal Society of Chemistry. All rights reserved. This article may be downloaded for personal use only. Any other use requires prior permission of the copyright holder. The Version of Record is available online at http://doi.org/10.1039/D1LC00333J. application/pdf |
| spellingShingle | Medicine, Health and Life Sciences Biomechanics Nanotechnology Tay, Hui Min Leong, Sheng Yuan Xu, Xiaohan Kong, Fang Upadya, Megha Dalan, Rinkoo Tay, Chor Yong Dao, Ming Suresh, Subra Hou, Han Wei Direct isolation of circulating extracellular vesicles from blood for vascular risk profiling in type 2 diabetes mellitus |
| title | Direct isolation of circulating extracellular vesicles from blood for vascular risk profiling in type 2 diabetes mellitus |
| title_full | Direct isolation of circulating extracellular vesicles from blood for vascular risk profiling in type 2 diabetes mellitus |
| title_fullStr | Direct isolation of circulating extracellular vesicles from blood for vascular risk profiling in type 2 diabetes mellitus |
| title_full_unstemmed | Direct isolation of circulating extracellular vesicles from blood for vascular risk profiling in type 2 diabetes mellitus |
| title_short | Direct isolation of circulating extracellular vesicles from blood for vascular risk profiling in type 2 diabetes mellitus |
| title_sort | direct isolation of circulating extracellular vesicles from blood for vascular risk profiling in type 2 diabetes mellitus |
| topic | Medicine, Health and Life Sciences Biomechanics Nanotechnology |
| url | https://hdl.handle.net/10356/150958 |
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