Frontiers in Cell and Developmental Biology ( IF 5.5 ) Pub Date : 2020-12-16 , DOI: 10.3389/fcell.2020.618552 Jenny Ostrop , Rosalie T. Zwiggelaar , Marianne Terndrup Pedersen , François Gerbe , Korbinian Bösl , Håvard T. Lindholm , Alberto Díez-Sánchez , Naveen Parmar , Silke Radetzki , Jens Peter von Kries , Philippe Jay , Kim B. Jensen , Cheryl Arrowsmith , Menno J. Oudhoff
Intestinal organoids are an excellent model to study epithelial biology. Yet, the selection of analytical tools to accurately quantify heterogeneous organoid cultures remains limited. Here, we developed a semi-automated organoid screening method, which we applied to a library of highly specific chemical probes to identify epigenetic regulators of intestinal epithelial biology. The role of epigenetic modifiers in adult stem cell systems, such as the intestinal epithelium, is still undefined. Based on this resource dataset, we identified several targets that affected epithelial cell differentiation, including HDACs, EP300/CREBBP, LSD1, and type I PRMTs, which were verified by complementary methods. For example, we show that inhibiting type I PRMTs, which leads enhanced epithelial differentiation, blocks the growth of adenoma but not normal organoid cultures. Thus, epigenetic probes are powerful tools to study intestinal epithelial biology and may have therapeutic potential.
中文翻译:
半自动类器官筛选方法证明了肠上皮分化的表观遗传控制。
肠类器官是研究上皮生物学的绝佳模型。然而,用于精确定量异质类器官培养物的分析工具的选择仍然受到限制。在这里,我们开发了一种半自动化类器官筛查方法,将其应用于高度特异性的化学探针库中,以鉴定肠上皮生物学的表观遗传调控因子。表观遗传修饰剂在成体干细胞系统(例如肠上皮)中的作用仍然不确定。基于此资源数据集,我们确定了影响上皮细胞分化的几个靶标,包括HDAC,EP300 / CREBBP,LSD1和I型PRMT,这些靶标已通过补充方法进行了验证。例如,我们表明抑制I型PRMTs会导致上皮分化增强,阻止腺瘤的生长,但不阻止正常的类器官培养。因此,表观遗传学探针是研究肠道上皮生物学的强大工具,可能具有治疗潜力。