Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, Dresden, 01307, Germany.
The Francis Crick Institute, 1 Midland Road, London, NW1 1AT, UK.
Nat Commun. 2017 Dec 1;8(1):1901. doi: 10.1038/s41467-017-01469-5.
Small molecule in vivo phenotypic screening is used to identify drugs or biological activities by directly assessing effects in intact organisms. However, current screening designs may not exploit the full potential of chemical libraries due to false negatives. Here, we demonstrate a modular small molecule screen in embryonic zebrafish that varies concentration, genotype and timing to target segmentation disorders, birth defects that affect the spinal column. By testing each small molecule in multiple interrelated ways, this screen recovers compounds that a standard screening design would have missed, increasing the hit frequency from the chemical library three-fold. We identify molecular pathways and segmentation phenotypes, which we share in an open-access annotated database. These hits provide insight into human vertebral segmentation disorders and myopathies. This modular screening strategy is applicable to other developmental questions and disease models, highlighting the power of relatively small chemical libraries to accelerate gene discovery and disease study.
小分子体内表型筛选用于通过直接评估完整生物体中的效应来鉴定药物或生物活性。然而，由于假阴性，当前的筛选设计可能无法充分利用化学文库的潜力。在这里，我们展示了一种在胚胎斑马鱼中模块化的小分子筛选方法，该方法通过改变浓度、基因型和时间来针对分节障碍进行筛选，分节障碍会影响脊柱的出生缺陷。通过以多种相互关联的方式测试每种小分子，该筛选方法恢复了标准筛选设计可能会遗漏的化合物，从而使化学文库中的命中频率增加了三倍。我们确定了分子途径和分节表型，并在一个开放获取的注释数据库中共享。这些命中结果为人类椎体分节障碍和肌肉疾病提供了新的见解。这种模块化的筛选策略适用于其他发育问题和疾病模型，突出了相对较小的化学文库在加速基因发现和疾病研究方面的强大功能。
