High-content screening of human primary muscle satellite cells for new therapies for muscular atrophy/dystrophy.
High-content screening of human primary muscle satellite cells for new therapies for muscular atrophy/dystrophy.
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DOI：
10.2174/2213988501307010021
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发表时间：
2013
期刊：
影响因子：
--
通讯作者：
Sexton JZ
中科院分区：
文献类型：
--
作者：
Nierobisz LS;Cheatham B;Buehrer BM;Sexton JZ
成肌细胞的增殖和分化是骨骼肌正常生长和修复所必需的。肌肉的恢复依赖于静止的肌肉干细胞--卫星细胞。在肌肉损伤期间，卫星细胞变得有丝分裂活跃，并通过相互融合和/或与肌纤维融合来开始修复过程。衰老、长期不活动、肥胖、恶病质和其他肌肉衰减性疾病与静止和增殖的卫星细胞数量减少有关，这阻碍了修复过程。开发了一个高含量/高通量的平台，用于对体外培养的人原代卫星细胞进行稳健的表型评估，以发现可能改善肌肉恢复的化学探针。使用两个高度注释的小分子文库开发了1600化合物中试屏幕。这一筛选产生了15种剂量响应性化合物，这些化合物增加了来自单一肥胖人类捐赠者的卫星细胞的增殖率。其中两种化合物在3供体肥胖大批量反筛选时仍有剂量反应。以ALK-5抑制剂LY364947为阳性对照，评价卫星细胞增殖/分化延迟。使用多变量方法进行探索性数据分析，以发现细胞表型的增殖与分化相关的变化。初步筛选工作成功地确定了一些表型结果，这些结果与刺激增殖和延迟分化的预期效果有关。
Myoblast proliferation and differentiation are essential for normal skeletal muscle growth and repair. Muscle recovery is dependent on the quiescent population of muscle stem cells - satellite cells. During muscle injury, satellite cells become mitotically active and begin the repair process by fusing with each other and/or with myofibers. Aging, prolonged inactivity, obesity, cachexia and other muscle wasting diseases are associated with a decreased number of quiescent and proliferating satellite cells, which impedes the repair process. A high-content/high-throughput platform was developed and utilized for robust phenotypic evaluation of human primary satellite cells in vitro for the discovery of chemical probes that may improve muscle recovery. A 1600 compound pilot screen was developed using two highly annotated small molecule libraries. This screen yielded 15 dose responsive compounds that increased proliferation rate in satellite cells derived from a single obese human donor. Two of these compounds remained dose responsive when counter-screened in 3-donor obese superlot. The Alk-5 inhibitor LY364947, was used as a positive control for assessing satellite cell proliferation/delayed differentiation. A multivariate approach was utilized for exploratory data analysis to discover proliferation vs. differentiation-dependent changes in cellular phenotype. Initial screening efforts successfully identified a number of phenotypic outcomes that are associated with desired effect of stimulation of proliferation and delayed differentiation.