Abstract：
Exercise is a potent stimulus for combatting skeletal muscle ageing. To study the effects of exercise on muscle in a preclinical setting, we developed a combined endurance-resistance training stimulus for mice called progressive weighted wheel running (PoWeR). PoWeR improves molecular, biochemical, cellular and functional characteristics of skeletal muscle and promotes aspects of partial epigenetic reprogramming when performed late in life (22-24 months of age). In this investigation, we leveraged pan-mammalian DNA methylome arrays and tandem mass-spectrometry proteomics in skeletal muscle to provide detailed information on late-life PoWeR adaptations in female mice relative to age-matched sedentary controls (n = 7-10 per group). Differential CpG methylation at conserved promoter sites was related to transcriptional regulation genes as well as Nr4a3, Hes1 and Hox genes after PoWeR. Using a holistic method of -omics integration called binding and expression target analysis (BETA), methylome changes were associated with upregulated proteins related to global and mitochondrial translation after PoWeR (P = 0.03). Specifically, BETA implicated methylation control of ribosomal, mitoribosomal, and mitochondrial complex I protein abundance after training. DNA methylation may also influence LACTB, MIB1 and UBR4 protein induction with exercise - all are mechanistically linked to muscle health. Computational cistrome analysis predicted several transcription factors including MYC as regulators of the exercise trained methylome-proteome landscape, corroborating prior late-life PoWeR transcriptome data. Correlating the proteome to muscle mass and fatigue resistance revealed positive relationships with VPS13A and NPL levels, respectively. Our findings expose differential epigenetic and proteomic adaptations associated with translational regulation after PoWeR that could influence skeletal muscle mass and function in aged mice. KEY POINTS: Late-life combined endurance-resistance exercise training from 22-24 months of age in mice is shown to improve molecular, biochemical, cellular and in vivo functional characteristics of skeletal muscle and promote aspects of partial epigenetic reprogramming and epigenetic age mitigation. Integration of DNA CpG 36k methylation arrays using conserved sites (which also contain methylation ageing clock sites) with exploratory proteomics in skeletal muscle extends our prior work and reveals coordinated and widespread regulation of ribosomal, translation initiation, mitochondrial ribosomal (mitoribosomal) and complex I proteins after combined voluntary exercise training in a sizeable cohort of female mice (n = 7-10 per group and analysis). Multi-omics integration predicted epigenetic regulation of serine β-lactamase-like protein (LACTB - linked to tumour resistance in muscle), mind bomb 1 (MIB1 - linked to satellite cell and type 2 fibre maintenance) and ubiquitin protein ligase E3 component N-recognin 4 (UBR4 - linked to muscle protein quality control) after training. Computational cistrome analysis identified MYC as a regulator of the late-life training proteome, in agreement with prior transcriptional analyses. Vacuolar protein sorting 13 homolog A (VPS13A) was positively correlated to muscle mass, and the glycoprotein/glycolipid associated sialylation enzyme N-acetylneuraminate pyruvate lyase (NPL) was associated to in vivo muscle fatigue resistance.
摘要：
运动是对抗骨骼肌衰老的有效刺激。为了研究临床前运动对肌肉的影响，我们为小鼠开发了一种称为渐进式加权轮跑（PoWeR）的耐力-阻力组合训练刺激。PoWeR提高了分子，生物化学，骨骼肌的细胞和功能特征，并在生命后期（22-24个月大）进行时促进部分表观遗传重编程。在这次调查中,我们利用骨骼肌中的泛哺乳动物DNA甲基化组阵列和串联质谱蛋白质组学,提供了雌性小鼠相对于年龄匹配的久坐对照(每组n=7-10)的晚龄PoWeR适应的详细信息.PoWeR后保守启动子位点的差异CpG甲基化与转录调控基因以及Nr4a3，Hes1和Hox基因有关。使用称为结合和表达靶标分析（BETA）的整体组学整合方法，PoWeR后，甲基化组变化与整体和线粒体翻译相关的蛋白上调相关(P=0.03)。具体来说,BETA涉及核糖体的甲基化控制，有丝分裂体,训练后线粒体复合物I蛋白丰富。DNA甲基化也可能影响LACTB，MIB1和UBR4蛋白诱导与运动-都与肌肉健康有机械联系。计算cistrome分析预测了几种转录因子，包括MYC作为运动训练的甲基-蛋白质组景观的调节因子，证实先前的晚期PoWeR转录组数据。将蛋白质组与肌肉质量和抗疲劳性相关联，显示出与VPS13A和NPL水平呈正相关，分别。我们的发现揭示了PoWeR后与翻译调节相关的差异表观遗传和蛋白质组适应，这可能会影响老年小鼠的骨骼肌质量和功能。关键点：22-24月龄小鼠的晚年联合耐力-抵抗运动训练被证明可以改善分子水平，生物化学，骨骼肌的细胞和体内功能特征，并促进部分表观遗传重编程和表观遗传年龄缓解。使用骨骼肌中的保守位点（也包含甲基化老化时钟位点）与探索性蛋白质组学整合DNACpG36k甲基化阵列扩展了我们先前的工作，并揭示了核糖体的协调和广泛调节，翻译启动,在相当大的雌性小鼠队列中进行联合自愿运动训练后，线粒体核糖体（线粒体）和复合物I蛋白（每组n=7-10和分析）。多组学整合预测了丝氨酸β-内酰胺酶样蛋白（LACTB-与肌肉中的肿瘤抗性相关）的表观遗传调控，训练后的精神炸弹1（MIB1-与卫星细胞和2型纤维维持相关）和泛素蛋白连接酶E3成分N-识别蛋白4（UBR4-与肌肉蛋白质量控制相关）。计算cistrome分析确定MYC是晚期训练蛋白质组的调节剂，与先前的转录分析一致。液泡蛋白分选13同源物A(VPS13A)与肌肉质量呈正相关,糖蛋白/糖脂相关唾液酸酶N-乙酰神经氨酸丙酮酸裂解酶（NPL）与体内肌肉疲劳抗性有关。
