Morphological profiling is a method to classify target pathways of antibacterials based on how bacteria respond to treatment through changes to cellular shape and spatial organization. Here we utilized the cell-to-cell variation in morphological features of Mycobacterium tuberculosis bacilli to develop a rapid profiling platform called Morphological Evaluation and Understanding of Stress (MorphEUS). MorphEUS classified 94% of tested drugs correctly into broad categories according to modes of action previously identified in the literature. In the other 6%, MorphEUS pointed to key off-target activities. We observed cell wall damage induced by bedaquiline and moxifloxacin through secondary effects downstream from their main target pathways. We implemented MorphEUS to correctly classify three compounds in a blinded study and identified an off-target effect for one compound that was not readily apparent in previous studies. We anticipate that the ability of MorphEUS to rapidly identify pathways of drug action and the proximal cause of cellular damage in tubercle bacilli will make it applicable to other pathogens and cell types where morphological responses are subtle and heterogeneous.

形态特征分析是一种根据细菌如何通过改变细胞形状和空间组织来响应治疗的方法，对抗菌药物的靶途径进行分类的方法。在这里，我们利用了结核分枝杆菌形态特征中的细胞间差异杆菌可以开发一个称为“形态评估和压力理解”（MorphEUS）的快速分析平台。根据先前文献中确定的作用方式，MorphEUS将94％的测试药物正确分类为大类。在另外的6％中，MorphEUS指出了关键的脱靶活动。我们观察到苯达喹啉和莫西沙星通过其主要靶途径下游的次级作用诱导细胞壁损伤。我们实施了MorphEUS，以在盲法研究中正确地对三种化合物进行分类，并确定了一种化合物的脱靶效应，这在以前的研究中尚不明显。