Investigating the effects of aerobic and resistance training in vivo on skeletal muscle metabolism in vitro in primary human muscle cells (MoTrMyo)

项目摘要/摘要 低运动量是导致全因发病率和死亡率的重要危险因素。骨骼肌是一种 通过线粒体底物氧化产生三磷酸腺苷的运动能力的关键组成部分 (即线粒体功能)。骨骼肌的特性也会影响全身的胰岛素敏感性 影响代谢性疾病(如2型糖尿病、肥胖症)的风险。新生的肌源性祖细胞是一种 骨骼肌在增殖、自我更新和传递过程中对运动反应可塑性的关键组成部分 关于对女儿群体的表观遗传修饰。极少的证据表明这些祖细胞 “印记”的生活方式干预，使随后的新陈代谢适应。建议进行的研究 将填补我们知识的空白，直接评估运动训练的能力，印记成肌祖细胞 以提高运动能力和降低代谢性疾病风险的方式。初级阶段 本提案的目标是利用MoTrPAC父项目，并确定 短期(三个月)和终身(运动员)有氧训练(AT)和耐力训练(RT)的印记 表观遗传调控的关键心肌细胞代谢特性(胰岛素敏感性、线粒体功能) (DNA甲基化，RNA表达)骨骼肌祖细胞中新的运动反应基因。我们的 第二个目标是从MoTrPAC建立人原代骨骼肌细胞(HSkMC)培养库 可以用来进行未来的机械审讯。我们假设运动训练会给人留下深刻印象 新型运动的表观遗传调控对骨骼肌祖细胞代谢特性的影响 反应基因在AT和RT之间会有所不同。我们将通过肌肉活组织检查建立HSkMC培养。 干预前后325人(AT、RT、对照组)和75名运动员(AT 和RT)，并评估胰岛素作用(胰岛素刺激的糖原合成、胰岛素信号转导)， 线粒体功能(使用碳水化合物和脂肪酸底物消耗氧气)，DNA甲基化和 RNA表达(RNAseq)和相关的途径浓缩分析。目标1声明的目的是 确定短期和终身AT和RT印记关键心肌细胞代谢的不同能力 通过运动反应基因的表观遗传调节的特性(胰岛素敏感性、线粒体功能)。 DNA甲基化和RNAseq数据的整合将为后续的研究确定新的运动反应基因 体外基因操作，以确定其对所述代谢终点的影响。声明的目的是 目标2是创建一个HSkMC储存库，用于未来的调查。拟议研究的结果将 对运动训练赋予“新陈代谢记忆”的能力提供了前所未有的机械洞察力 通过对特定运动反应基因的表观遗传修饰并通知后续运动 与优化运动所获得的健康益处相关的处方。

Project Summary/Abstract Low exercise capacity is an important risk factor for all-cause morbidity and mortality. Skeletal muscle is a critical component of exercise capacity through its ability to generate ATP via mitochondrial substrate oxidation (i.e. mitochondrial function). Skeletal muscle properties also influence whole-body insulin sensitivity which affects risk for metabolic diseases (e.g. type 2 diabetes, obesity). Nascent myogenic progenitor cells are a critical component of skeletal muscle plasticity in response to exercise as they proliferate, self‐renew and pass on epigenetic modifications to daughter populations. Meager evidence suggests that these progenitor cells are “imprinted” by lifestyle interventions which confers subsequent metabolic adaptations. The proposed studies will fill a gap in our knowledge and directly assess the ability of exercise training to imprint myogenic progenitor cells in a manner leading to improved exercise capacity and reduced risk for metabolic disease. The primary goal of the present proposal is to leverage the MoTrPAC parent project and determine the distinct abilities of short-term (three months) and life-long (athletes) aerobic training (AT) and resistance training (RT) to imprint key myocellular metabolic properties (insulin sensitivity, mitochondrial function) through epigenetic regulation (DNA methylation, RNA expression) of novel exercise response genes in skeletal muscle progenitor cells. Our secondary goal is to establish a human primary skeletal muscle cell (HSkMC) culture repository from MoTrPAC which can be utilized for future mechanistic interrogations. We hypothesize that exercise training will imprint metabolic properties in skeletal muscle progenitor cells through epigenetic regulation of novel exercise response genes that will differ between AT and RT. We will establish HSkMC cultures from muscle biopsies of 325 individuals (AT, RT, control groups) before and following three months of intervention and 75 athletes (AT and RT) and assess insulin action (insulin-stimulated glycogen synthesis, insulin signal transduction), mitochondrial function (O2 consumption using carbohydrate and fatty acid substrates), DNA methylation and RNA expressions (RNAseq) with associated pathway enrichment analyses. The stated purpose of Aim 1 is to determine the distinct abilities of short-term and life-long AT and RT to imprint key myocellular metabolic properties (insulin sensitivity, mitochondrial function) through epigenetic regulation of exercise response genes. Integration of DNA methylation and RNAseq data will identify novel exercise response genes for subsequent genetic manipulation in vitro to determine its effects on the stated metabolic endpoints. The stated purpose of Aim 2 is to create an HSkMC repository for future investigations. Findings from the proposed studies will provide unprecedented mechanistic insight into the ability of exercise training to confer a “metabolic memory” through epigenetic modification of specific exercise response genes and inform subsequent exercise prescriptions relative to optimizing health benefits gained from exercise.