Effects of an acute exercise bout in hypoxia on extracellular vesicle release in healthy and prediabetic subjects

Warnier, Geoffrey; Groote, Estelle De; Britto, Florian; Delcorte, Ophélie; Nederveen, Joshua P.; Nilsson, Mats I.; Pierreux, Christophe E.; Tarnopolsky, Mark A.; Deldicque, Louise

doi:10.1152/ajpregu.00220.2021

Cited by 16 publications

(21 citation statements)

References 39 publications

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“…Interestingly, we found that CS-EVs had a higher expression of CD81 and HSP70, and trended towards an increase in Tsg101 compared to CON-EVs. Our observations are in line with previous studies that have shown that both acute/chronic exercise training increases EV protein levels of CD81 and HSP70 in mice and humans [24], [25], [30], [57], [58]. We did not find any difference in the expression of Alix and Flotillin-1 between CON-EV and CS-EV.…”

Section: Discussionsupporting

confidence: 93%

“…However, our result is similar to a recent study that showed no difference in the protein expression of TSG101 and CD81 in skeletal muscle from healthy humans with acute exercise, even though these proteins were increased in circulatory EVs with exercise. This same study also showed an increase in the mRNA levels of TSG101 in the skeletal muscle of prediabetic subjects with exercise [58]. It is possible that with longer exercise duration, intensity, or recovery, we may observe increase in TSG101 expression.…”

Section: Discussionmentioning

confidence: 62%

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Skeletal muscle-derived extracellular vesicles are altered with chronic contractile activity

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Seif

Bydak

et al. 2022

Preprint

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Extracellular vesicles (EVs) are small lipid membrane-bound structures that are secreted by all cells, and play a central role in cellular communication. EVs are released from skeletal muscle during exercise, but the effects of contractile activity on skeletal muscle-derived EVs (Skm-EVs) are poorly understood due to the challenges in distinguishing Skm-EVs derived from exercising muscle in vivo. To specifically characterize Skm-EVs, C2C12 myoblasts were differentiated into myotubes, and electrically paced (3h/day x 4days @14V, C-PACE EM, IonOptix) to mimic chronic exercise in vitro. EVs were isolated from conditioned media from control and stimulated myotubes using differential ultracentrifugation. Isolated EVs were characterized biophysically (size, zeta potential, yield, protein markers and by transmission electron microscopy; TEM). Chronic stimulation increased markers of mitochondrial biogenesis such as MitoTracker Red staining, cytochrome c oxidase activity and expression of cytochrome c (p<0.05, N=7-8) in stimulated vs. non-stimulated myotubes. The average size of EVs from chronically stimulated myotubes (CS-EVs, 132 nm) was 26% smaller than control (CON-EVs, 178 nm) (p<0.05, N=8). Size distribution analysis revealed that CS-EVs were enriched with 100-150 nm sized small EVs, while CON-EVs were largely composed of 200-250 nm sized vesicles (p<0.05, main interaction effect, N=8). TEM confirmed the presence of round-shaped vesicles of about 30-100 nm with an intact lipid bilayer in CON-EVs and CS-EVs. Zeta potential was 27% lower in CS-EVs vs. CON-EVs (p<0.05, N=8), and total EV protein yield remained unchanged between groups. Protein-based EV characterization showed that both CON-EVs/CS-EVs were CD81+ but CD63-, and expressed hallmark cytosolic proteins recovered in EVs: Tsg101, Flotillin-1, HSP70, and Alix. CD81 and HSP70 expression increased in CS-EVs vs. CON-EVs (p<0.05, N=9). We evaluated if chronic stimulation affected whole cell expression of transmembrane and cytosolic proteins used in EV origin/purity analysis. CD63 and ApoA1 were reduced with chronic stimulation in myotube lysates (p<0.05, N=7), whereas Tsg101, CD81, Flotillin-1 and HSP70 levels remained constant. Taken together, our study revealed that chronic stimulation triggers the release of more stable, smaller sized EVs, enriched with transmembrane and cytosolic protein markers of small EVs, and are CD81+/CD63- indicating the origin of these EVs might be ectosomal rather than endosomal. The upstream signaling cascades that regulate biogenesis of EVs with chronic stimulation, and whether skeletal muscle-EVs are released through endosomal or ectosomal pathways remains to be elucidated. Our findings support the role of chronic contractile activity in the modulation of EV biophysical characteristics. Whether this affects biological cargo recruitment into CS-EVs, and their subsequent biological activity remains to be established.

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Section: Discussionsupporting

confidence: 93%

Section: Discussionmentioning

confidence: 62%

Skeletal muscle-derived extracellular vesicles are altered with chronic contractile activity

Patience

Seif

Bydak

et al. 2022

Preprint

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“…We first investigated acute exercise‐induced alterations in circulating EVs. Several studies have reported that acute exercise promotes an immediate release of EVs into the circulation 4,26–31 . We observed an exercise‐induced increase in the EV tetraspanin CD63 in MEM‐purified samples and a tendency for increased CD9 and CD81.…”

Section: Discussionmentioning

confidence: 50%

“…Several studies have reported that acute exercise promotes an immediate release of EVs into the circulation. 4,[26][27][28][29][30][31] We observed an exerciseinduced increase in the EV tetraspanin CD63 in MEMpurified samples and a tendency for increased CD9 and CD81. However, we did not observe an increased particle concentration with the NTA method, which may be due to the presence of other particles such as protein aggregates and/or lipoprotein particles that mask the EV signal.…”

Section: Exercise-induced Evs In Tissue Crosstalkmentioning

confidence: 99%

Extracellular vesicles and microRNAs are altered in response to exercise, insulin sensitivity and overweight

et al. 2022

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Extracellular vesicles induced by exercise have emerged as potential mediators of tissue crosstalk. Extracellular vesicles and their cargo miRNAs have been linked to dysglycemia and obesity in animal models, but their role in humans is unclear. Aim:The aim of the study was to characterize the miRNA content in plasma extracellular vesicle isolates after acute and long-term exercise and to study associations between extracellular vesicle miRNAs, mRNA expression in skeletal muscle and adipose tissue, and cardiometabolic risk factors.Methods: Sedentary men with or without dysglycemia and overweight underwent an acute bicycle test and a 12-week exercise intervention with extensive metabolic phenotyping. Gene expression in m. vastus lateralis and subcutaneous adipose tissue was measured with RNA sequencing. Extracellular vesicles were purified from plasma with membrane affinity columns or size exclusion chromatography.Results: Extracellular vesicle miRNA profiling revealed a transient increase in the number of miRNAs after acute exercise. We identified miRNAs, such as miR-652-3p, that were associated to insulin sensitivity and adiposity. By performing explorative association analyses, we identified two miRNAs, miR-32-5p and miR-339-3p, that were strongly correlated to an adipose tissue macrophage signature. Conclusion:Numerous miRNAs in plasma extracellular vesicle isolates were increased by exercise, and several miRNAs correlated to insulin sensitivity and adiposity. Our findings warrant future studies to characterize exercise-induced extracellular vesicles and cargo miRNA to clarify where exercise-induced extracellular vesicles originate from, and to determine whether they influence metabolic health or exercise adaptation.

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“…Western blotting. An equal volume of vesicle isolates was loaded for all samples as usually performed with ELV protein homogenates (6,7,10,27). The proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to polyvinylidene difluoride membranes (Immobilon Transfer Membrane; Millipore).…”

Section: Methodsmentioning

confidence: 99%

Effects of a 6-wk Sprint Interval Training Protocol at Different Altitudes on Circulating Extracellular Vesicles

Warnier

Groote

Delcorte

et al. 2022

Medicine &Amp; Science in Sports &Amp; Exercise

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PurposeThis study aimed to investigate the modulation of circulating exosome-like extracellular vesicles (ELVs) after 6 wk of sprint interval training (SIT) at sea level and at 2000, 3000, and 4000 m.MethodsThirty trained endurance male athletes (18–35 yr) participated in a 6-wk SIT program (30-s all-out sprint, 4-min 30-s recovery; 4–9 repetitions, 2 sessions per week) at sea level (n = 8), 2000 m (fraction of inspired oxygen (Fio2) 0.167, n = 8), 3000 m (Fio2 0.145, n = 7), or 4000 m (Fio2 0.13, n = 7). Venous blood samples were taken before and after the training period. Plasma ELVs were isolated by size exclusion chromatography, counted by nanoparticle tracking analysis, and characterized according to international standards. Candidate ELV microRNAs (miRNAs) were quantified by real-time polymerase chain reaction.ResultsWhen the three hypoxic groups were analyzed separately, only very minor differences could be detected in the levels of circulating particles, ELV markers, or miRNA. However, the levels of circulating particles increased (+262%) after training when the three hypoxic groups were pooled, and tended to increase at sea level (+65%), with no difference between these two groups. A trend to an increase was observed for the two ELV markers, TSG101 (+65%) and HSP60 (+441%), at sea level, but not in hypoxia. Training also seemed to decrease the abundance of miR-23a-3p and to increase the abundance of miR-21-5p in hypoxia but not at sea level.ConclusionsA 6-wk SIT program tended to increase the basal levels of circulating ELVs when performed at sea level but not in hypoxia. In contrast, ELV miRNA cargo seemed to be modulated in hypoxic conditions only. Further research should explore the potential differences in the origin of ELVs between normoxic and local and systemic hypoxic conditions.

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Effects of an acute exercise bout in hypoxia on extracellular vesicle release in healthy and prediabetic subjects

Cited by 16 publications

References 39 publications

Skeletal muscle-derived extracellular vesicles are altered with chronic contractile activity

Skeletal muscle-derived extracellular vesicles are altered with chronic contractile activity

Extracellular vesicles and microRNAs are altered in response to exercise, insulin sensitivity and overweight

Effects of a 6-wk Sprint Interval Training Protocol at Different Altitudes on Circulating Extracellular Vesicles

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