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

项目总结/摘要 低运动能力是全因发病率和死亡率的重要危险因素。骨骼肌是一种 运动能力的关键组成部分，通过其通过线粒体底物氧化产生ATP的能力 (i.e.线粒体功能）。骨骼肌的特性也会影响全身的胰岛素敏感性， 影响代谢疾病的风险（例如2型糖尿病、肥胖症）。新生肌源性祖细胞是一种 骨骼肌可塑性的关键组成部分，对运动的反应，因为它们增殖，自我更新和通过 对子代群体进行表观遗传修饰微薄的证据表明，这些祖细胞是 通过生活方式干预“印记”，赋予随后的代谢适应。拟议的研究 将填补我们知识的空白，并直接评估运动训练对肌源性祖细胞的影响 细胞的方式导致提高运动能力和降低代谢疾病的风险。主 本提案的目标是利用交通运输和公共行政部的母项目，确定 短期（三个月）和终身（运动员）有氧训练（AT）和阻力训练（RT）的印记 通过表观遗传调节的关键肌细胞代谢特性（胰岛素敏感性、线粒体功能） (DNA甲基化，RNA表达）。我们 第二个目标是从MoTrPAC建立人原代骨骼肌细胞（HSkMC）培养库 其可用于未来的机械询问。我们假设运动训练 通过新运动的表观遗传学调节骨骼肌祖细胞的代谢特性 我们将建立HSkMC文化从肌肉活检， 325名个体（AT，RT，对照组）在三个月的干预前后和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的O2消耗），DNA甲基化和 RNA表达（RNAseq）与相关途径富集分析。目标1的目的是 确定短期和终身AT和RT标记关键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，研究了运动对胰岛素敏感性和线粒体功能的影响。 DNA甲基化和RNAseq数据的整合将识别新的运动反应基因，用于随后的研究。 体外遗传操作，以确定其对所述代谢终点的影响。的公开目的 目标2是为未来的调查创建一个HSkMC存储库。拟议研究的结果将 为运动训练赋予“代谢记忆”的能力提供了前所未有的机制性见解 通过特定运动反应基因的表观遗传修饰， 处方相对于优化从运动中获得的健康益处。

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.