Single-cell sequencing deconvolutes cellular responses to exercise in human skeletal muscle
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ABSTRACT: Regular physical activity is a key concept associated with a variety of health-related outcomes and successful ageing. While many of the beneficial effects of physical activity are undisputed, much of the adaptive mechanisms that lead to these benefits are not yet known. The skeletal muscle is a key contributor to physical performance and is also the target organ for many of the adaptive processes associated with exercise. Skeletal muscle is highly plastic, and changes in physical activity lead to a plethora of adaptive processes that, when repeated over time, result in structural and functional adaptations. Here we use single cell sequencing in humans to outline the effects of physical activity on cellular composition and cell type-specific processes in skeletal muscle. We show that myogenic cells in human skeletal muscle can be divided into three groups characterized by different degrees of cell maturation, and that exercise stimulates subpopulation of undifferentiated stem/progenitor myogenic cells to mature toward slow- or fast-twitch fibers. The cell type-specific adaptive mechanisms induced by exercise presented here contribute to the understanding of the skeletal muscle adaptations triggered by physical activity and may ultimately have implications for physiological and pathological processes affecting skeletal muscle, such as sarcopenia, cachexia, and glucose homeostasis.
Project description:Aging is a physiological process characterized by a progressive decline of biological functions and an increase in destructive processes in cells and organs. Physical activity and exercise positively affects the expression of skeletal muscle markers involved in longevity pathways. Recently, a new mechanism, autophagy, was introduced to the adaptations induced by acute and chronic exercise as responsible of positive metabolic modification and health-longevity promotion. However, the molecular mechanisms regulating autophagy in response to physical activity and exercise are sparsely described. We investigated the long-term adaptations resulting from lifelong recreational football training on the expression of skeletal muscle markers involved in autophagy signaling. We demonstrated that lifelong football training increased the expression of messengers: RAD23A, HSPB6, RAB1B, TRAP1, SIRT2 and HSBPB1, involved in the auto-lysosomal and proteasome-mediated protein degradation machinery; of RPL1, RPL4, RPL36, MRLP37, involved in cellular growth and differentiation processes; of the Bcl-2, HSP70, HSP90, PSMD13 and of the ATG5-ATG12 protein complex, involved in proteasome promotion and autophagy processes in muscle samples from lifelong trained subjects compared to age-matched untrained controls. In conclusion, our results indicated that lifelong football training positively influence exercise-induced autophagy processes and protein quality control in skeletal muscle, thus promoting healthy aging. The aim of the present research was to investigate, through a differential transcriptomic approach, the effects of lifelong recreational football training on the expression of key markers involved in the autophagy response for the maintenance of protein quality control, related to healthy longevity promotion, in skeletal muscle from VPG compared to elderly untrained control subjects.
Project description:Exercise stimulates systemic and tissue-specific adaptations that protect against lifestyle related diseases including obesity and type 2 diabetes. Exercise places high mechanical and energetic demands on contracting skeletal muscle, which require finely-tuned protein post-translational modifications involving signal transduction (e.g. phosphorylation) to elicit appropriate short- and long-term adaptive responses. To uncover the breadth of protein phosphorylation events underlying the adaptive responses to endurance exercise and skeletal muscle contraction, we performed global, unbiased mass spectrometry-based phosphoproteomic analyses of skeletal muscle from two rodent models, in situ muscle contraction in rats and treadmill-based endurance exercise in mice.
Project description:The PGC-1a gene is expressed as several transcriptional coactivator variants that regulate numerous adaptive processes. These include thermogenesis, hepatic metabolism, neuroprotection, and skeletal muscle adaptation to exercise training. To support such diverse functions, PGC-1 isoform expression and activation are under tight tissue- and context-specific control. Notably, PGC-1 isoforms generated by alternative gene promoter usage, and splicing are highly induced in skeletal muscle upon exercise. Here we show that PGC-12 and 3 are PGC-11 dimerization partners that limit its activity. Skeletal muscle PGC-12 and PGC-13 transgenics have reduced exercise performance and strength, which partially overlaps with PGC-1 loss-of-function. Mechanistically, PGC-11/3 dimerization precludes ERR recruitment and coactivation, so their co-expression in skeletal muscle impairs the innate bioenergetic adaptations characteristic of PGC-11 transgenics and reduces oxidative metabolism gene expression and exercise capacity. Removing this break to PGC-1a1 activity may have therapeutic applications in metabolic diseases and increase responsiveness to training.
Project description:Exercise induces skeletal muscle adaptation, and the p38 mitogen-activated protein kinase signaling pathway is thought to play an important role in the adaptive processes. We have obtained new evidence that the gamma isoform of p38 is required for exercise-induced metabolic adaptation in skeletal muscle; however, the neuromuscular activity-dependent target genes of p38gamma remain to be defined. We used microarrays to detail the global programme of gene expression underlying the skeletal muscle genetic reprogramming in response to increased contractile activity and identified distinct classes of up-regulated genes during this process that are dependent on the functional activity of the p38gamma isoform. Skeletal muscle-specific p38gamma knockout mice and the wild type littermates are subject to motor nerve stimulation of one of the tibialis anterior muscles followed by microarray analysis of both the stimulated and the contralateral control muscles.
Project description:We compared the short-term (4-day) adaptations in skeletal muscle mitochondrial function and gene expression between short-frequent bouts of physical activity and time matched single-continuous physical activity.
Project description:Exercise induces skeletal muscle adaptation, and the p38 mitogen-activated protein kinase signaling pathway is thought to play an important role in the adaptive processes. We have obtained new evidence that the gamma isoform of p38 is required for exercise-induced metabolic adaptation in skeletal muscle; however, the neuromuscular activity-dependent target genes of p38gamma remain to be defined. We used microarrays to detail the global programme of gene expression underlying the skeletal muscle genetic reprogramming in response to increased contractile activity and identified distinct classes of up-regulated genes during this process that are dependent on the functional activity of the p38gamma isoform.
Project description:Exercise late in life mitigates skeletal muscle epigenetic aging, providing evidence that physical activity is a "fountain of youth".
Project description:Exercise is an effective strategy in the prevention and treatment of metabolic diseases. Alterations in the skeletal muscle proteome, including post-translational modifications, regulate its metabolic adaptations to exercise. Here, we examined the effect of high-intensity interval training (HIIT) on the proteome and acetylome of human skeletal muscle, revealing the response of 3168 proteins and 1263 lysine acetyl-sites on 464 acetylated proteins. We identified novel protein adaptations to exercise training involved in metabolism and excitation-contraction coupling. Furthermore, HIIT increased the acetylation of mitochondrial proteins, particularly those of complex V, likely via non-enzymatic mechanisms. We also highlight the regulation of novel exercise-responsive histone acetyl-sites. These data demonstrate the plasticity of the skeletal muscle proteome and acetylome, providing insight into the regulation of contractile, metabolic and transcriptional processes within skeletal muscle. Herein, we provide a substantial hypothesis-generating resource to stimulate further mechanistic research investigating how exercise improves metabolic health.
Project description:How regular physical activity is able to improve health remains poorly understood, but release of substances from skeletal muscle following exercise is one proposed mechanism. Here we describe a novel mechanism through which physical activity initiates extracellular vesicle (EV)-mediated communication between skeletal muscle and adipose tissue causing increased adipocyte lipolysis as a result of enhanced catecholamine sensitivity. In response to a hypertrophic stimulus induced by mechanical overload (MOV), skeletal muscle released EVs containing muscle-specific miR-1 which were preferentially taken up by epididymal white adipose tissue (eWAT) and were associated with elevated adrenergic signaling and lipolysis. Inhibiting EV release by GW4869 treatment prevented the MOV-induced increase in eWAT miR-1 abundance and expression of lipolytic factors. miR-1 was shown to induce adrenergic receptor beta β3 (Adrβ3) expression in adipocytes by targeting transcription factor AP-2, alpha (Tfap2α, a known repressor of Adrβ3 expression. Ex vivo experiments showed enhanced catecholamine sensitivity and elevated fatty acid oxidation in eWAT following MOV. Following a bout of resistance exercise in humans, skeletal muscle miR-1 expression was decreased with a concomitant increase in EV miR-1 abundance, suggesting that a skeletal muscle-adipose tissue axis is operative in humans. Altogether, our highly novel findings demonstrate that skeletal muscle promotes metabolic adaptations in adipose tissue in response to MOV via EV-mediated delivery of miR-1.
Project description:Physical activity is associated with beneficial adaptations in human and rodent metabolism. We studied over 50 complex traits before and after exercise intervention in middle-aged men and a panel of 100 diverse strains of female mice. Candidate gene analyses in three brain regions, muscle, liver, heart, and adipose tissue of mice revealed genetic drivers of clinically relevant traits including volitional exercise volume, muscle metabolism, adiposity, and hepatic lipids. Although ~33% of genes differentially expressed in skeletal muscle following the exercise intervention were similar in mice and men independent of BMI, responsiveness of adipose tissue to exercise-stimulated weight loss appears impacted by species and underlying genotype. We leveraged genetic diversity to generate prediction models of metabolic trait responsiveness to volitional activity offering a framework for advancing personalized exercise prescription. Finally, we make the human and mouse data publicly available via user-friendly web-based application to enhance data mining and hypothesis development.