The Comparative Methylome and Transcriptome After Change of Direction Compared to Straight Line Running Exercise in Human Skeletal Muscle

Abstract

The methylome and transcriptome signatures following exercise that are physiologically and metabolically relevant to sporting contexts such as team sports or health prescription scenarios (e.g., high intensity interval training/HIIT) has not been investigated. To explore this, we performed two different sport/exercise relevant high-intensity running protocols in five male sport team members using a repeated measures design of: (1) change of direction (COD) versus; (2) straight line (ST) running exercise with a wash-out period of at least 2 weeks between trials. Skeletal muscle biopsies collected from the vastus lateralis 30 min and 24 h post exercise, were assayed using 850K methylation arrays and a comparative analysis with recent (subject-unmatched) sprint and acute aerobic exercise meta-analysis transcriptomes was performed. Despite COD and ST exercise being matched for classically defined intensity measures (speed × distance and number of accelerations/decelerations), COD exercise elicited greater movement (GPS-Playerload), physiological (HR), metabolic (lactate) as well as central and peripheral (differential RPE) exertion measures compared with ST exercise, suggesting COD exercise evoked a higher exercise intensity. The exercise response alone across both conditions evoked extensive alterations in the methylome 30 min and 24 h post exercise, particularly in MAPK, AMPK and axon guidance pathways. COD evoked a considerably greater hypomethylated signature across the genome compared with ST exercise, particularly at 30 min post exercise, enriched in: Protein binding, MAPK, AMPK, insulin, and axon guidance pathways. Comparative methylome analysis with sprint running transcriptomes identified considerable overlap, with 49% of genes that were altered at the expression level also differentially methylated after COD exercise. After differential methylated region analysis, we observed that VEGFA and its downstream nuclear transcription factor, NR4A1 had enriched hypomethylation within their promoter regions. VEGFA and NR4A1 were also significantly upregulated in the sprint transcriptome and meta-analysis of exercise transcriptomes. We also confirmed increased gene expression of VEGFA, and considerably larger increases in the expression of canonical metabolic genes PPARGC1A (that encodes PGC1-α) and NR4A3 in COD vs. ST exercise. Overall, we demonstrate that increased physiological/metabolic load via COD exercise in human skeletal muscle evokes considerable epigenetic modifications that are associated with changes in expression of genes responsible for adaptation to exercise.

Skeletal Muscle Possesses an Epigenetic Memory of Exercise: Role of Nucleus Type-Specific DNA Methylation

“… 6 Despite this general similarity, the hypomethylated signatures seems to be enriched in divergent gene pathways between exercise types, for example, in growth, actin and ECM related pathways following resistance exercise, 4 compared with MAPK, AMPK, and Insulin signalling after running exercise. 5 At the pathway level in myonuclei in the present study there was enriched hypomethylation of promoter CpGs particularly in Wnt signalling and growth-related pathways, with hypomethylation and upregulation of gene expression in growth-related pathways also observed after resistance exercise in humans. 4 Interestingly, Wen et al, also identified differential regulation of the Wnt pathway in interstitial nuclei, where they demonstrated opposite profiles to myonuclei, that is, hypermethylation of Wnt pathway genes in interstitial compared with hypomethylation in the myonuclei.…”

“…Indeed, Wen et al identified that promoter hypomethylation slightly outweighed hypermethylation signatures in myonuclei after training, and promoter hypomethylation was also predominant in interstitial nuclei, with these data like those signatures observed in human muscle tissue after the first training period. 3 Indeed, exercise as a “hypomethylating stimuli” has also been supported by studies using different types of exercise (not exclusively resistance exercise), such as sprint interval running exercise 5 and at the candidate gene level after aerobic exercise. 6 Despite this general similarity, the hypomethylated signatures seems to be enriched in divergent gene pathways between exercise types, for example, in growth, actin and ECM related pathways following resistance exercise, 4 compared with MAPK, AMPK, and Insulin signalling after running exercise.…”

Recurrent training rejuvenates and enhances transcriptome and methylome responses in young and older human muscle

“…[10][11][12][13] More recently, methylome modifications comparing change of direction to straight line running also identified a more explicit hypomethylated signature for change of direction running in genes within protein binding, MAPK, AMPK, insulin, and axon guidance pathways. 14 These beneficial methylome adaptations could even be passed on to later generations, as has been observed in mice. 15 In older muscle, lifelong physical activity induced promoter hypomethylation of genes involved in energy metabolism, myogenesis, contractile properties, and oxidative stress.…”

Impact of Physical Activity and Exercise on the Epigenome in Skeletal Muscle and Effects on Systemic Metabolism

“…PGC1-α is a key regulatory gene for mitochondrial biogenesis, fatty acid oxidation, and skeletal muscle sensitivity to insulin. The PGC-1α gene is hypomethylated after an intense exercise session [ 32 , 67 , 68 ]. Barrès et al performed biopsies of the vastus lateralis , in which they found a different methylation state of the promoter of this gene, specifically, 10% less methylated compared to the resting state.…”