A Novel Strategy to Identify Substances that Improve Mitochnodrial Fitness

项目摘要/摘要 线粒体功能障碍会导致线粒体疾病，并与衰老和神经退行性变密切相关 帕金森氏症和阿尔茨海默氏症等疾病。最近的发现支持线粒体功能障碍可以 被克服来治疗与年龄相关的衰退。赛特金的假说是运动诱导血液分泌-- 携带的蛋白质系统地作用于刺激移除受损的线粒体和丰富健康的 线粒体(线粒体适合度)。该公司的目标是识别这些蛋白质，以开发成生物制品 这将成为治疗与线粒体功能障碍相关的无数疾病的平台。这个 Discovery平台建立在运动逆转小鼠模型线粒体功能障碍的现有工作基础上 治疗线粒体疾病。来自突变小鼠的细胞为识别因子提供了一个有吸引力的工具 改善线粒体的适合性。在此应用中，测试了基于细胞的检测平台的可行性。 该平台测量集落形成、mtDNA拷贝数、线粒体呼吸和 线粒体质量控制蛋白，并将在目标1中使用已知的线粒体激动剂进行评估 生物发生[苯扎贝特和格列酮(PPAR激动剂)；二甲双胍和AICAR(AMPK)；以及白藜芦醇 (SIRT1)]。这些结果将被综合起来，为每种激动剂制定一个“健康评分”。无论化验平台 能够根据激动剂的适应度分数快速区分激动剂对线粒体生物发生的影响 下定决心。第二部分将评估该分析平台在发现分泌因子方面的效用。 瞄准。Cytegen发现，至少有一种因素在锻炼而不是久坐时显著上调， 突变小鼠将被重组生产并在测试平台上进行测试。体能得分将为 与对照组相比。这项工作的完成将确定分析平台可以在多大程度上使用 出于发现的目的。如果成功，这个平台将有助于识别增强线粒体的物质 可在最终转化为临床试验的第二阶段申请中进一步追求的适合性。

Project Summary/Abstract Mitochondrial dysfunction causes mitochondrial diseases and is tightly linked to aging and neurodegenerative disorders such as Parkinson's and Alzheimer's. Recent discoveries support that mitochondrial dysfunction can be overcome to treat age-related decline. Cytegen's hypothesis is that exercise induces the secretion of blood- borne proteins that act systemically to stimulate removal of damaged mitochondria and enrichment of healthy mitochondria (mitochondrial fitness). The company's goal is to identify these proteins to develop into biologics that would serve as a platform to treat the myriad of diseases associated with mitochondrial dysfunction. The discovery platform builds on existing work that exercise reversed mitochondrial dysfunction in a mouse model for mitochondrial disease. Cells derived from the mutant mouse offer an attractive tool for identifying factors that improve mitochondrial fitness. In this application, the feasibility of a cell-based assay platform is tested. This platform measures colony formation, mtDNA copy number, mitochondrial respiration, and expression of mitochondrial quality control proteins, and will be evaluated in Aim 1 using known agonists of mitochondrial biogenesis [bezafibrate and the glitazones (PPAR agonists); metformin and AICAR (AMPK); and resveratrol (Sirt1)]. The results will be integrated to develop a "fitness score" for each agonist. Whether the assay platform is capable of rapidly differentiating agonists' impact on mitochondrial biogenesis by their fitness score will be determined. The utility of the assay platform for discovery of secreted factors will be evaluated in the second aim. At least one factor identified by Cytegen to be significantly upregulated in exercised, not sedentary, mutant mice will be recombinantly produced and tested in the assay platform. The fitness score will be compared to controls. The completion of this work will identify to what extent the assay platform can be used for discovery purposes. If successful, this platform will help identify substances that enhance mitochondrial fitness that can be further pursued in a Phase II application for ultimate translation into clinical trials.