Development of a human skeletal muscle cell-based high throughput screen for musc

描述（由申请人提供）：骨骼肌萎缩是一种严重的疾病，普遍存在于老龄化人口（肌肉减少症）和各种特定疾病中。这种情况通过肌肉再生受损导致肌肉功能丧失，导致跌倒和受伤增加，丧失独立性，生活质量下降。这些变化产生了巨大的保健负担，据估计，2000年老龄人口的保健负担将达到185亿美元。由于发病率和死亡率与老年受试者的活动能力直接相关，促进肌肉再生的新药应通过减少住院和养老院护理来降低医疗保健费用。肌肉再生是由卫星细胞控制的，这是一种成体肌肉干细胞。当肌肉受到损伤时，卫星细胞被激活增殖，随后分化为最终的成肌细胞。本申请的总体目标是建立一个基于人类卫星细胞的系统，该系统可以支持：1)识别控制卫星细胞增殖的信号通路和因子，2)研究肌肉萎缩背后的变化，以及3)肌肉再生的药物发现项目。在Aim1中，我们将使用人体肌肉活检材料，建立384孔格式的卫星细胞测定，以确定细胞增殖能力，维持其卫星细胞表型，并允许测量不同药物对这些特性的影响。该平台将建立一个强大的筛选系统，可用于II期工作，以确定改变细胞复制和分化能力的药物。Aim 2的工作将通过检查一组已知对骨骼肌有影响的药物来验证该分析。该分析的实用性将通过筛选高度注释的化合物的小文库来证明。重要的是，这项工作将是确定促进肌肉再生的新机制和分子的第一步。该筛选试验将在II期工作中完善后成为Zen Bio合同分析服务的一部分，使制药，生物技术和学术机构能够分析化合物和生物制剂对人类卫星细胞的作用。第二阶段的努力还将帮助Zen Bio开发一个知识产权组合，涵盖肌肉再生的化学和目标驱动信息。

DESCRIPTION (provided by applicant): Skeletal muscle wasting is a serious condition prevalent in the aging population (sarcopenia) and in a variety of specific diseases. This condition results in loss of muscle function through impaired muscle regeneration, resulting in an increase in falls and injuries, a loss of independence, and a reduced quality of life. These changes produce a large health care burden estimated in the year 2000 to be $18.5 billion in the aged population. Since the morbidity and mortality rates are directly correlated to the mobility of elderly subjects, new medicines that promote muscle regeneration should lower health care costs through less hospitalization and nursing home care. Muscle regeneration is controlled by the satellite cell, an adult muscle stem cell. When muscle is injured, the satellite cell becomes activated to proliferate and subsequently differentiate to terminally committed myoblasts. The overall objective of this application is to establish a human satellite cell-based system that can support: 1) identification of the signaling pathways and agents that control the proliferation of satellite cells, 2) studies of the alterations that underlie muscle wasting, and 3) drug discovery programs on muscle regeneration. In Aim1 using human muscle biopsy material, we will establish a satellite cell assay in a 384-well format that can determine the capacity of the cells to proliferate, maintain their satellite cell phenotype and allow measurement of the effects of different agents on these properties. This platform will establish a robust screening system that can be used in Phase II work to identify agents that modify the cells' capacity to replicate and differentiate. Efforts in Aim 2 will validate the assay by examining a set of agents with known effects on skeletal muscle. The utility of the assay will be demonstrated by screening small libraries of highly annotated compounds. Importantly, this work will be the first step toward identifying novel mechanisms and molecules that promote muscle regeneration. The screening assay will become part of Zen Bio's contract assay service when it is refined by work in Phase II, allowing pharmaceutical, biotech and academic institutions to profile the actions of compounds and biological agents on human satellite cells. Phase II efforts will also help Zen Bio develop an intellectual property portfolio that covers chemical and target-driven information on muscle regeneration. PUBLIC HEALTH RELEVANCE: Muscle wasting is a major problem in the elderly and in a variety of specific diseases (e.g. chronic obstruction pulmonary disease, cancer related cachexia, end stage renal disease). This application describes the generation of a cell-based assay system using the human satellite cell, an adult stem cell that controls skeletal muscle regeneration. This assay will provide a method for determining the controls that regulate human satellite cell proliferation, examine the cause of the poor muscle regeneration seen in various states such as elderly subjects and the disease states mentioned above, and facilitate the identification of new drugs that can reverse the age and disease-related decline in muscle mass and function.