De Rycker Manu, Rigoreau Laurent, Dowding Sarah, Parker Peter J
Protein Phosphorylation Laboratory, London Research Institute, Cancer Research UK, London, UK.
Chem Biol Drug Des. 2009 Jun;73(6):599-610. doi: 10.1111/j.1747-0285.2009.00817.x.
High-content cell-based screens provide a powerful tool to identify new chemicals that interfere with complex biological processes. Here, we describe the identification of a new inhibitor of microtubule dynamics (micropolyin) using a high-content screen. Integrated high-resolution imaging allowed for fast selection of hits and progression to target identification. Treatment of cells with micropolyin efficiently causes a pro-metaphase arrest, with abnormal spindle morphology and with the spindle assembly checkpoint activated. The arrest appears to result from interference of micropolyin with microtubule dynamics. We show in vitro that tubulin is indeed the target of micropolyin and that micropolyin inhibits microtubule polymerization. Our results demonstrate the power of high-content image- and cell-based screening approaches to identify potential new drug candidates. As our approach is unbiased, it should allow for discovery of new targets that may otherwise be overlooked.
基于细胞的高内涵筛选提供了一个强大的工具，用于识别干扰复杂生物过程的新化学物质。在此，我们描述了使用高内涵筛选鉴定一种新的微管动力学抑制剂（微聚菌素）的过程。集成的高分辨率成像能够快速筛选出命中物并推进到靶点鉴定。用微聚菌素处理细胞可有效导致前中期停滞，纺锤体形态异常且纺锤体组装检查点被激活。这种停滞似乎是由于微聚菌素干扰了微管动力学所致。我们在体外表明，微管蛋白确实是微聚菌素的靶点，并且微聚菌素抑制微管聚合。我们的结果证明了基于高内涵图像和细胞的筛选方法在识别潜在新药候选物方面的强大作用。由于我们的方法是无偏向性的，它应该能够发现那些可能被忽视的新靶点。
