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Proteome characterisation of extracellular vesicles isolated from heart

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Version 1 2021-08-04, 05:34
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posted on 2023-11-08, 00:41 authored by Bethany ClaridgeBethany Claridge, Alin RaiAlin Rai, H Fang, A Matsumoto, J Luo, Julie McMullenJulie McMullen, David GreeningDavid Greening
Cardiac intercellular communication is critical for heart function and often dysregulated in cardiovascular diseases. While cardiac extracellular vesicles (cEVs) are emerging mediators of signalling, their isolation remains a technical challenge hindering our understanding of cEV protein composition. Here, we utilised Langendorff-collagenase-based enzymatic perfusion and differential centrifugation to isolate cEVs from mouse heart (yield 3–6 μg/heart). cEVs are ∼200 nm, express classical EV markers (Cd63/81/9+, Tsg101+, Pdcd6ip/Alix+), and are depleted of blood (Alb/Fga/Hba) and cardiac damage markers (Mb, Tnnt2, Ldhb). Comparison with mechanically-derived EVs revealed greater detection of EV markers and decreased cardiac damage contaminants. Mass spectrometry-based proteomic profiling revealed 1721 proteins in cEVs, implicated in proteasomal and autophagic proteostasis, glycolysis, and fatty acid metabolism; essential functions often disrupted in cardiac pathologies. There was striking enrichment of 942 proteins in cEVs compared to mouse heart tissue - implicated in EV biogenesis, antioxidant activity, and lipid transport, suggesting active cargo selection and specialised function. Interestingly, cEVs contain marker proteins for cardiomyocytes, cardiac progenitors, B-cells, T-cells, macrophages, smooth muscle cells, endothelial cells, and cardiac fibroblasts, suggesting diverse cellular origin. We present a method of cEV isolation and provide insight into potential functions, enabling future studies into EV roles in cardiac physiology and disease.

Funding

National Health and Medical Research Council, Grant/Award Numbers: 1139489, 1201805

History

Publication Date

2021-07-01

Journal

Proteomics

Volume

21

Issue

13-14

Article Number

e2100026

Pagination

11p.

Publisher

Wiley

ISSN

1615-9853

Rights Statement

This is the peer reviewed version of the following article: Claridge, B, Rai, A, Fang, H. et al. Proteome characterisation of extracellular vesicles isolated from heart. Proteomics. 2021; 21:e2100026, which has been published in final form at https://doi.org/10.1002/pmic.202100026. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions. This article may not be enhanced, enriched or otherwise transformed into a derivative work, without express permission from Wiley or by statutory rights under applicable legislation. Copyright notices must not be removed, obscured or modified. The article must be linked to Wiley’s version of record on Wiley Online Library and any embedding, framing or otherwise making available the article or pages thereof by third parties from platforms, services and websites other than Wiley Online Library must be prohibited.

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