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Pharmacol Res. 2016 Aug;110:35-49. doi: 10.1016/j.phrs.2016.05.004. Epub 2016 May 7.

A cell-based quantitative high-throughput image screening identified novel autophagy modulators.

Author information

1
Department of Pharmacology, Toxicology and Therapeutics, The University of Kansas Medical Center, Kansas City, KS 66160, USA.
2
National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, MD 20892, USA.
3
Department of Pulmonary, Shanghai Chest Hospital, Shanghai Jiaotong University, Shanghai, 200030 China.
4
Department of Pharmacology, Toxicology and Therapeutics, The University of Kansas Medical Center, Kansas City, KS 66160, USA. Electronic address: wxding@kumc.edu.

Abstract

Macroautophagy is a major cellular degradation pathway for long-lived proteins and cellular organelles to maintain cellular homeostasis. Reduced autophagy has been implicated in neurodegenerative diseases, metabolic syndrome, and tumorigenesis. In contrast, increased autophagy has been shown to protect against tissue injury and aging. Here we employed a cell-based quantitative high-throughput image screening (qHTS) for autophagy modulators using mouse embryonic fibroblasts (MEFs) that are stably expressing GFP-LC3. The library of pharmacologically active compounds (LOPAC) was used to screen for the autophagy modulators in compounds alone or in combination with the lysosome inhibitor chloroquine (CQ). The GFP-LC3 puncta were then quantified to measure autophagic flux. The primary screening revealed 173 compounds with efficacy more than 40%. These compounds were cherry-picked and re-tested at multiple different concentrations using the same assay. A number of novel autophagy inducers, inhibitors, and modulators with dual-effects on autophagy were identified from the cherry-pick screening. Interestingly, we found a group of compounds that induce autophagy are related to dopamine receptors and are commonly used as clinical psychiatric drugs. Among them, indatraline hydrochloride (IND), a dopamine inhibitor, and chlorpromazine hydrochloride (CPZ) and fluphenazine dihydrochloride (FPZ), two dopamine receptor antagonists, were further evaluated. We found that FPZ-induced autophagy through mTOR inhibition but IND and CPZ induced autophagy in an mTOR-independent manner. Our data suggest that image-based autophagic flux qHTS can efficiently identify autophagy inducers and inhibitors.

KEYWORDS:

Autophagy; Dopamine receptor; GFP-LC3; High-throughput screening; mTOR

PMID:
27168224
PMCID:
PMC4995889
DOI:
10.1016/j.phrs.2016.05.004
[Indexed for MEDLINE]
Free PMC Article

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