Urinary extracellular vesicles as a source of protein-based biomarkers in feline chronic kidney disease and hypertension
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Authors
Lawson, Jack
Syme, Harriet
Karttunen, Jenni
Stewart, Sarah
Antrobus, Robin
Karet, Fiona
Journal Title
Journal of Small Animal Practice
ISSN
0022-4510
Publisher
Wiley
Type
Article
This Version
AM
Metadata
Show full item recordCitation
Lawson, J., Syme, H., Karttunen, J., Stewart, S., Antrobus, R., Karet, F., & Williams, T. Urinary extracellular vesicles as a source of protein-based biomarkers in feline chronic kidney disease and hypertension. Journal of Small Animal Practice https://doi.org/10.17863/CAM.85550
Abstract
Objectives
To validate a methodology for isolating feline urinary extracellular vesicles (uEVs) and characterise the uEV population and proteome in cats with normal renal function and cats with normotensive or hypertensive chronic kidney disease
Methods
Feline uEVs were isolated using three different methods (precipitation alone, precipitation followed by size exclusion chromatography, and ultrafiltration followed by size exclusion chromatography, which were compared via transmission electron microscopy and nanoparticle tracking analysis . Cats with normal renal function (n = 9), normotensive CKD (n = 10) and hypertensive CKD (n = 9) were identified and uEVs isolated from patient urine samples via ultrafiltration followed by size exclusion chromatography. EV size and concentration were determined using nanoparticle tracking analysis, and subsequently underwent proteomic analysis using LC-MS/MS to identify differences in protein expression between categories.
Results
uEV preparations contained particles of the expected size and morphology, and those obtained by UF+SEC had a significantly higher purity (highest particle: protein ratio). The uEV proteomes contained EV markers and proteins originating from all nephron segments. UEV concentration and size were unaffected by renal disease or hypertension. There were no differentially expressed proteins detected when comparing uEVs derived from cats in the healthy category with the combined CKD category, but five differentially expressed proteins were identified between the normotensive CKD and hypertensive CKD categories.
Clinical Significance
Feline uEVs can be successfully isolated from stored urine samples. Differentially expressed uEV proteins were discovered in cats with hypertensive CKD, and warrant further investigation into their utility as biomarkers or therapeutic targets.
Embargo Lift Date
2025-06-15
Identifiers
This record's DOI: https://doi.org/10.17863/CAM.85550
This record's URL: https://www.repository.cam.ac.uk/handle/1810/338141
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