Discovery of autophagy modulators through the construction of a high-content screening platform via monitoring of lipid droplets

Given its importance in various cellular processes, autophagy has attracted growing attention as a promising therapeutic target in many human diseases. Herein, we report the construction of a high-content screening platform for the discovery of autophagy modulators via monitoring of cellular lipid droplets as a late-stage marker of autophagy. Notably, our platform discriminates the effects of chemical modulators on autophagic flux without additional transfection, washing, or fixation. This approach allows the simultaneous identification of completely different types of autophagy regulators in a single operation. Mode-of-action studies of 2 representative hit compounds confirmed 1 compound as an mTOR-independent autophagy activator and the other as an autophagy inhibitor that blocks autophagic flux.