Endurance-dependent urinary extracellular vesicle signature: shape, metabolic miRNAs, and purine content distinguish triathletes from inactive people

Abstract

Extracellular vesicles (EVs) enriched with bioactive molecules have gained considerable attention in nanotechnology because they are critical to intercellular communication while maintaining low immunological impact. Among biological matrices, urine has emerged as a noninvasive source of extracellular-contained liquid biopsy, currently of interest as a readout for physiological adaptations. Therefore, we aimed to evaluate chronic adaptations of endurance sport practice in terms of urinary EV parameters and evaluated by food consumption assessment. Two balanced groups of 13 inactive controls vs. triathlon athletes were enrolled; their urinary EVs were obtained by differential ultracentrifugation and analyzed by dynamic light scattering and transmission electron and atomic force microscopy. The cargo was analyzed by means of purine and miRNA content through HPLC-UV and qRT-PCR. Specific urinary EV signatures differentiated inactive versus endurance-trained in terms of peculiar shape. Particularly, a spheroid shape, smaller size, and lower roughness characterize EVs from triathletes. Metabolic and regulatory miRNAs often associated with skeletal muscle (i.e., miR378a-5p, miR27a-3p, miR133a, and miR206) also accounted for a differential signature. These miRNAs and guanosine in urinary EVs can be used as a readout for metabolic status along with the shape and roughness of EVs, novel informative parameters that are rarely considered. The network models allow scholars to entangle nutritional and exercise factors related to EVs’ miRNA and purine content to depict metabolic signatures. All in all, multiplex biophysical and molecular analyses of urinary EVs may serve as promising prospects for research in exercise physiology.

Extracellular Vesicles in Environmental Toxicological Studies: Association between Urinary Concentrations of Phthalate Metabolites and Exosomal miRNA Expression Profiles

“…The extraction of urinary ELV by differential ultracentrifugation has been deeply described in Pietrangelo et al [16]. Briefly, urines were centrifuged at 300× g (4 • C) for 10 min and then at 2000× g (4 • C) for 20 min.…”

“…Exosomal miRs were isolated from urinary exosomes using the miRNeasy mini kit (Qiagen, Germantown, MD, USA), as described by Pietrangelo et al [16]. MiRs were polyadenylated using a Poly(A) polymerase Tailing Kit (Diatech LabLine, Jesi, Italy).…”

“…Most recently, we demonstrated that the differential expression of metabolic and regulatory miRs, found in EVs, can be used for differentiating endurance athletes from physically inactive controls [16]. Similarly, several environmental studies have linked the alteration in miR expression profiles to the exposure to various environmental stressors.…”

Exploring the role of urinary extracellular vesicles in kidney physiology, aging, and disease progression

“…2 ) ( 48 ). Another very recent paper highlights differences in several uEV parameters isolated from inactive subjects and triathlon athletes, studying chronic adaptations of endurance practice ( 49 ). uEVs isolated from triathletes resulted in smaller and lower roughness; moreover, miRNAs associated with skeletal muscle activity and metabolic pathways as well as guanosine were differentially expressed by uEVs from the two cohorts of enrolled subjects ( 49 ).…”

“…Another very recent paper highlights differences in several uEV parameters isolated from inactive subjects and triathlon athletes, studying chronic adaptations of endurance practice ( 49 ). uEVs isolated from triathletes resulted in smaller and lower roughness; moreover, miRNAs associated with skeletal muscle activity and metabolic pathways as well as guanosine were differentially expressed by uEVs from the two cohorts of enrolled subjects ( 49 ). The study of uEV modulation by sports exercise seems to be of interest by the scientific community with further implications in training, diet of professional athletes, and also in relationship with drugs and doping.…”