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Related Experiment Video

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

Yuan Li1, Steven McGreal1, Jean Zhao2

  • 1Department of Pharmacology, Toxicology and Therapeutics, The University of Kansas Medical Center, Kansas City, KS 66160, USA.

Pharmacological Research
|May 12, 2016
PubMed
Summary
This summary is machine-generated.

This study used high-throughput screening to identify compounds that modulate autophagy, a key cellular process. Novel autophagy inducers and inhibitors were discovered, including psychiatric drugs affecting dopamine pathways.

Keywords:
AutophagyDopamine receptorGFP-LC3High-throughput screeningmTOR

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Pharmacology

Background:

  • Macroautophagy is a vital cellular degradation pathway crucial for maintaining homeostasis.
  • Dysregulation of autophagy is linked to neurodegenerative diseases, metabolic syndrome, and cancer.
  • Conversely, enhanced autophagy can protect against tissue injury and aging.

Purpose of the Study:

  • To identify novel modulators of autophagy using a quantitative high-throughput image screening (qHTS) approach.
  • To screen a library of pharmacologically active compounds for their effects on autophagic flux.
  • To investigate the mechanisms of action for identified autophagy modulators, particularly those related to dopamine pathways.

Main Methods:

  • Utilized mouse embryonic fibroblasts (MEFs) stably expressing GFP-LC3 for autophagy monitoring.
  • Employed a quantitative high-throughput image screening (qHTS) assay to measure autophagic flux.
  • Screened the Library of Pharmacologically Active Compounds (LOPAC) alone and with chloroquine (CQ).

Main Results:

  • Identified 173 compounds with >40% efficacy in modulating autophagy during primary screening.
  • Discovered novel autophagy inducers, inhibitors, and dual-effect modulators.
  • Found that psychiatric drugs targeting dopamine receptors (indatraline, chlorpromazine, fluphenazine) modulate autophagy, with distinct mTOR-dependent and independent mechanisms.

Conclusions:

  • Image-based autophagic flux qHTS is an efficient method for identifying autophagy modulators.
  • Psychiatric drugs offer potential avenues for therapeutic intervention in diseases involving autophagy dysregulation.
  • Further research into dopamine receptor-related compounds could yield new autophagy-targeting therapies.