Screening with a Novel Cell-Based Assay for TAZ Activators Identifies a Compound That Enhances Myogenesis in C2C12 Cells and Facilitates Muscle Repair in a Muscle Injury Model
Screening with a Novel Cell-Based Assay for TAZ Activators Identifies a Compound That Enhances Myogenesis in C2C12 Cells and Facilitates Muscle Repair in a Muscle Injury Model
复制标题
DOI：
10.1128/mcb.01346-13
复制
发表时间：
2014-05-01
影响因子：
5.3
通讯作者：
Hata, Yutaka
中科院分区：
文献类型：
--
作者：
Yang, Zeyu;Nakagawa, Kentaro;Hata, Yutaka
具有PDZ结合基序（TAZ）的转录共激活因子与多种转录因子配合并发挥多种作用。当 TAZ 过度表达和激活时，永生化人类哺乳动物上皮 MCF10A 细胞形成球体。我们开发了一种基于细胞的检测方法，利用表达 TAZ 的 MCF10A 细胞形成的球体作为读数来筛选 18,458 种化合物中的 TAZ 激活剂。获得了 50 种化合物，其中 47 种被证实可激活 HEK293 细胞中 TAZ 依赖性 TEAD 响应报告活性。我们使用衍生的化合物子集作为 TAZ 激活剂候选迷你库，并寻找促进小鼠 C2C12 成肌细胞中肌生成的化合物。在这项研究中，我们重点关注一种化合物，IBS008738。 IBS008738 稳定 TAZ，增加非磷酸化 TAZ 水平，增强 MyoD 与肌生成素启动子的关联，上调 MyoD 依赖性基因转录，并与 C2C12 细胞中的肌生长抑制素竞争。 TAZ 敲低验证了 IBS008738 的作用取决于 C2C12 细胞中的内源性 TAZ。 IBS008738 促进心脏毒素引起的肌肉损伤中的肌肉修复，并防止地塞米松引起的肌肉萎缩。因此，这种基于细胞的测定可用于鉴定具有多种细胞输出的 TAZ 激活剂。我们的研究结果也支持 TAZ 是肌肉萎缩的潜在治疗靶点的观点。
The transcriptional coactivator with a PDZ-binding motif (TAZ) cooperates with various transcriptional factors and plays various roles. Immortalized human mammalian epithelial MCF10A cells form spheres when TAZ is overexpressed and activated. We developed a cell-based assay using sphere formation by TAZ-expressing MCF10A cells as a readout to screen 18,458 chemical compounds for TAZ activators. Fifty compounds were obtained, and 47 were confirmed to activate the TAZ-dependent TEAD-responsive reporter activity in HEK293 cells. We used the derived subset of compounds as a TAZ activator candidate minilibrary and searched for compounds that promote myogenesis in mouse C2C12 myoblast cells. In this study, we focused on one compound, IBS008738. IBS008738 stabilizes TAZ, increases the unphosphorylated TAZ level, enhances the association of MyoD with the myogenin promoter, upregulates MyoD-dependent gene transcription, and competes with myostatin in C2C12 cells. TAZ knockdown verifies that the effect of IBS008738 depends on endogenous TAZ in C2C12 cells. IBS008738 facilitates muscle repair in cardiotoxin- induced muscle injury and prevents dexamethasone-induced muscle atrophy. Thus, this cell-based assay is useful to identify TAZ activators with a variety of cellular outputs. Our findings also support the idea that TAZ is a potential therapeutic target for muscle atrophy.