RT Journal Article
T1 Analysis of urinary exosomal metabolites identifies cardiovascular risk signatures with added value to urine analysis
A1 Agudiez, Marta
A1 Martínez, Paula J.
A1 Martin-Lorenzo, Marta
A1 Heredero, Angeles
A1 Santiago-Hernández, Aranzazu
A1 Molero Vilchez, María Dolores
A1 Garcia Segura, Juan Manuel
A1 Aldamiz-Echevarria, Gonzalo
A1 Álvarez Llamas, Gloria
AB Background: Subclinical atherosclerosis may result in fatal cardiovascular (CV) events, but the underlying mechanisms and molecular players leading to disease are not entirely understood. Thus, novel approaches capable of identifying the factors involved in pathological progression and providing a better understanding of the subjacent mechanisms are needed. Extracellular vesicles (EVs) have been shown to have numerous biological functions, and their metabolome has recently generated interest as a source of novel biomarkers. The metabolic content of the exosomes has been so far unexplored in cardiovascular disease (CVD), and here, we developed an analytical strategy aimed at probing urinary exosomal metabolite content and its association to CV risk. Results: Direct analysis of the exosomes without metabolite extraction was evaluated by high-resolution magic angle spinning (1 H HR-MAS). Other two methodologies for the analysis of exosomal metabolites by 1 H NMR were set up, based on methanol or organic solvents sequential extraction. The three methods were compared in terms of the number of detected signals and signal to noise ratio (S/N). The methanol method was applied to identify altered metabolites in the urinary exosomes of subjects with programmed coronary artery by-pass grafting (CABG) versus a control group. Target mass spectrometry (MS) was also performed for differential analysis. The clinical performance of exosomal metabolites of interest in CVD was investigated, and the added value of the exosomes compared to urine analysis was evaluated. Based on S/N ratio, simplicity, reproducibility, and quality of the spectrum, the methanol method was chosen for the study in CVD. A cardiometabolic signature composed by 4-aminohippuric acid, N-1-methylnicotinamide, and citric acid was identified in urinary exosomes. Directly in urine, 4-aminohippuric acid and citric acid do not show variation between groups and changes in N-1-methylnicotinamide are less pronounced, proving the added value of exosomes.Conclusions: We set up a novel methodology to analyze metabolic alterations in urinary exosomes and identified a cardiometabolic signature in these microvesicles. This study constitutes the first evidence of a role for the exosomal metabolism in CVD and demonstrates the possibility to evaluate the urinary exosomal metabolic content by NMR and MS.
PB BMC
SN Electronic: 1741-7007
YR 2020
FD 2020-12-14
LK https://hdl.handle.net/20.500.14352/8033
UL https://hdl.handle.net/20.500.14352/8033
LA eng
NO Instituto de Salud Carlos III/Fondo Europeo de Desarrollo Regional (FEDER)
NO Comunidad de Madrid
NO IDCSalud
NO Red de Investigación Renal (REDinREN)
NO Fundación SENEFRO/SEN
NO Fundación Conchita Rábago de Jiménez Díaz
DS Docta Complutense
RD 16 abr 2025