Discovery and characterisation of novel myokines using innovative proteomic analysis

肥胖与代谢紊乱的发展有关，包括胰岛素抵抗和II型糖尿病。尽管运动在预防和治疗这些疾病方面的效果已知，但令人惊讶的是，这种治疗方法和对相关机制的研究被忽视了。近年来，在东道主实验室和其他实验室的工作中，产生了肌动因子的概念，这是一种通过收缩骨骼肌产生和释放的蛋白质，目的是协调运动对代谢紊乱的有益影响。自从鉴定出典型的肌细胞因子IL-6以来，人们已经描述了一些肌细胞因子，但往往是偶然发现的。因此，本研究旨在确定骨骼肌对运动的反应所分泌的新的肌动蛋白，并重要的是，确定它们的生物学相关性。为此，在第一步中，将运动小鼠的蛋白质组与休息小鼠的蛋白质组进行比较。为了便于在体内检测释放的肌细胞因子，将采用创新的蛋白质组稳定同位素标记与细胞培养中的氨基酸(SILAC)掺入方法，使用在宿主实验室进行的体外和体内SILAC标记实验中制备的肌肉裂解物。表现出运动反应的蛋白质随后将通过生物信息学分析被询问分泌序列。然后，将通过代谢筛选实验在体外和体内评估推定的肌动蛋白对代谢功能的意义。这项研究项目确定的可能的新型肌动蛋白将提供治疗靶点，并有助于了解运动对胰岛素抵抗和II型糖尿病等疾病的有益影响。

Obesity is associated with the development of metabolic disorders, including insulin resistance and type II diabetes. Despite the known efficacy of exercise in preventing and treating these conditions, this therapeutic approach and research into the mechanisms involved, is surprisingly overlooked. In recent years, work in the host laboratory and others, has given rise to the concept of the myokine, a protein produced and released from contracting skeletal muscle in order to orchestrate the beneficial effects of exercise on metabolic disorders. Since the identification of the proto-typical myokine IL-6, a select number of myokines have been described, but often discovered serendipitously. Hence, this study aims to identify novel myokines which are secreted by the skeletal muscle in response to exercise and importantly, determine their biological relevance. To this end, in a first step the proteome of an exercising mouse will be compared with the proteome of a rested mouse. To facilitate the in vivo detection of released myokines, the innovative proteomic stable isotope labelling with amino acids in cell culture (SILAC) spike-in approach will be adopted, using prepared muscle lysates from in vitro and in vivo SILAC labelling experiments carried out at the host laboratory. Proteins exhibiting an exercise response will then be interrogated for secretory sequences via bioinformatical analyses. The significance of putative myokines on metabolic function will then be assessed in vitro and in vivo by metabolic screening experiments. The putative novel myokines identified by this research project will provide therapeutic targets and help to understand the beneficial effects of exercise on disease conditions, such as insulin resistance and type II diabetes.