Proteomic Profile of Urinary Extracellular Vesicles Identifies AGP1 as a Potential Biomarker of Primary Aldosteronism
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Date
2021
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Abstract
Context: Primary aldosteronism (PA) represents 6% to 10% of all essential hypertension patients and is diagnosed using the aldosterone-to-renin ratio (ARR) and confirmatory studies. The complexity of PA diagnosis encourages the identification of novel PA biomarkers. Urinary extracellular vesicles (uEVs) are a potential source of biomarkers, considering that their cargo reflects the content of the parent cell.
Objective: We aimed to evaluate the proteome of uEVs from PA patients and identify potential biomarker candidates for PA.
Methods: Second morning spot urine was collected from healthy controls (n = 8) and PA patients (n = 7). The uEVs were isolated by ultracentrifugation and characterized. Proteomic analysis on uEVs was performed using LC-MS Orbitrap.
Results: Isolated uEVs carried extracellular vesicle markers, showed a round shape and sizes between 50 and 150 nm. The concentration of uEVs showed a direct correlation with urinary creatinine (r = 0.6357; P= 0.0128).The uEV size mean (167 +/- 6 vs 183 +/- 4nm) and mode (137 +/- 7 vs 171 +/- 11nm) was significantly smaller in PA patients than in control subjects, but similar in concentration. Proteomic analysis of uEVs from PA patients identified an upregulation of alpha-1-acid glycoprotein 1 (AGP1) in PA uEVs, which was confirmed using immunoblot. A receiver operating characteristic curve analysis showed an area under the curve of 0.92 (0.82 to 1; P. 0.0055).
Conclusion: Proteomic and further immunoblot analyses of uEVs highlights AGP1 as potential biomarker for PA.
Objective: We aimed to evaluate the proteome of uEVs from PA patients and identify potential biomarker candidates for PA.
Methods: Second morning spot urine was collected from healthy controls (n = 8) and PA patients (n = 7). The uEVs were isolated by ultracentrifugation and characterized. Proteomic analysis on uEVs was performed using LC-MS Orbitrap.
Results: Isolated uEVs carried extracellular vesicle markers, showed a round shape and sizes between 50 and 150 nm. The concentration of uEVs showed a direct correlation with urinary creatinine (r = 0.6357; P= 0.0128).The uEV size mean (167 +/- 6 vs 183 +/- 4nm) and mode (137 +/- 7 vs 171 +/- 11nm) was significantly smaller in PA patients than in control subjects, but similar in concentration. Proteomic analysis of uEVs from PA patients identified an upregulation of alpha-1-acid glycoprotein 1 (AGP1) in PA uEVs, which was confirmed using immunoblot. A receiver operating characteristic curve analysis showed an area under the curve of 0.92 (0.82 to 1; P. 0.0055).
Conclusion: Proteomic and further immunoblot analyses of uEVs highlights AGP1 as potential biomarker for PA.
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Keywords
primary aldosteronism urine, extracellular vesicles, proteomics, hypertension, aldosterone-to-renin ratio