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群(骨质疏松症)和各种特定的疾病中。这种情况会导致肌肉再生受损，从而导致肌肉功能丧失，导致跌倒和受伤的增加，丧失独立性，降低生活质量。这些变化造成了巨大的保健负担，据估计，2000年老年人口的保健负担为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该会通过减少住院和养老院护理来降低医疗成本。肌肉再生是由卫星细胞控制的，卫星细胞是一种成年肌肉干细胞。当肌肉受损时，卫星细胞被激活增殖，随后分化为终末成肌细胞。这项应用的总体目标是建立一个以人类卫星细胞为基础的系统，该系统可以支持：1)识别控制卫星细胞增殖的信号通路和试剂，2)研究肌肉损耗背后的变化，以及3)肌肉再生的药物发现计划。在使用人类肌肉活检材料的AIM1中，我们将建立一种384孔格式的卫星细胞分析，该方法可以确定细胞的增殖能力，保持其卫星细胞表型，并允许测量不同因素对这些特性的影响。该平台将建立一个强大的筛选系统，可用于第二阶段的工作，以确定改变细胞复制和分化能力的试剂。AIM 2的努力将通过检查一组对骨骼肌有已知影响的药物来验证检测结果。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。筛选化验将成为Zen Bio合同化验服务的一部分，当它在第二阶段进行完善时，允许制药、生物技术和学术机构描述化合物和生物制剂对人类卫星细胞的作用。第二阶段的工作还将帮助Zen Bio开发一个知识产权组合，涵盖肌肉再生方面的化学和目标驱动信息。 公共卫生相关性：肌肉萎缩是老年人和各种特定疾病(如慢性阻塞性肺病、癌症相关恶病质、终末期肾病)的主要问题。这项申请描述了使用人类卫星细胞生成基于细胞的分析系统，人类卫星细胞是一种控制骨骼肌再生的成年干细胞。这一检测将提供一种方法来确定调节人类卫星细胞增殖的控制因素，检查在不同状态下出现的肌肉再生不良的原因，如老年受试者和上述疾病状态，并有助于发现可以逆转年龄和疾病相关的肌肉质量和功能下降的新药。

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.