A High Throughput Image‐based Screening Reveals Novel Autophagy Inducers For Treating Drug‐Induced Liver Injury

• DOI: 10.1096/fasebj.29.1_supplement.148.6
• Publication Date: 2021-06-21T16:49:05Z
• Authors: Yuan Li; Steven McGreal; Jean Zhao; Ruili Huang; Menghang Xia; Wen‐Xing Ding
• Keywords: 0301 basic medicine; 0303 health sciences; 03 medical and health sciences; 3. Good health

ObjectiveTo identify novel autophagy modulators using a high throughput image‐based screening and to explore their mechanisms of regulation and the application for drug‐induced liver injury.MethodsThe library of pharmacologically active compounds (LOPAC) was screened in mouse embryonic fibroblasts (MEFs) stably expressing GFP‐LC3. Compounds were tested at different concentrations with or without lysosome inhibitor chloroquine (CQ), and GFP‐LC3 puncta were quantified to measure the autophagic flux. Mechanisms of autophagy induction of several selective positive compounds and the protection against acetaminophen (APAP)‐induced liver injury in mice were determined.ResultsThe primary screening revealed 173 compounds with sufficient hit efficacy. The confirmation study indicated several novel autophagy inducers, inhibitors, and compounds showing dual‐effect at different concentrations. Indatraline hydrochloride (IND), chlorpromazine hydrochloride (CPZ) and fluphenazine dihydrochloride (FPZ) were able to increase autophagy in a dose‐ and time‐dependent manner in MEF and other cancer cells. Mechanistically all three compounds inhibited mTOR, a key regulator of autophagy. Moreover, CPZ significantly suppressed APAP‐induced liver injury either co‐treated with or given 2 hours post APAP administration.ConclusionWe identified known and novel autophagy inducers through the high throughput image‐based screening. CPZ significantly suppressed APAP‐induced liver injury by inducing autophagy.[R01 AA020518, R01 DK 102142, P20 RR021940 (COBRE)]