Identifying compounds that modulate autophagy using high-throughput screening and high-content imaging analysis

Authors

• L Mitchell
• M Ooka
• J Zhao
• L Zhang
• Z Wei
• R Huang
• M Xia

Abstract

Autophagy is a highly conserved eukaryotic cellular degradation process that maintains cellular homeostasis and prevents stress-induced damage through the removal of dysfunctional or aged proteins. Impaired autophagy has been implicated in the development of numerous neurodegenerative diseases characterized by the pathological aggregation of misfolded proteins. Thus, the pharmacological modulation of autophagy presents a promising therapeutic avenue for several diseases, including neurodegenerative diseases and cancer, due to its ability to sequester and degrade protein aggregates. In order to identify potentially therapeutic autophagy modulators, we evaluated 3,733 clinically approved or investigational drugs. High content imaging analysis using Mouse Embryonic Fibroblasts expressing GFP-labeled LC3, a biomarker for autophagy activation, identified 225 compounds as potential autophagy inducers in the primary screening, which were further narrowed down to 8 potential novel autophagy inducers through a confirmation study. Follow-up studies evaluated the autophagy modulation capabilities of the 8 potential novel autophagy inducers, focusing on targets both upstream and downstream of autophagy activation. Among these 8 compounds tested, 5 were found to activate downstream caspase pathways and subsequent apoptosis, while 4 compounds induced upstream p53 activation and 4 compounds activated endoplasmic reticulum stress. In this study, we identified several novel autophagy inducers that may be repurposed for the treatment of neurodegenerative diseases or cancer.

Scientific Focus Area: Molecular Biology and Biochemistry

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