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Related Concept Videos

Autophagy01:27

Autophagy

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Autophagy is a self-digesting process by which a cell protects itself from threats both within and outside the cell, ranging from abnormal proteins to invading bacteria. In this process, obsolete components of the cell and invading microbes are degraded by hydrolytic enzymes active in an acidic environment of the lysosomal lumen.
An autophagic pathway consists of a series of signaling events activated in response to diverse stress and physiological conditions such as food deprivation,...
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Related Experiment Video

Updated: May 2, 2026

Author Spotlight: A Selective Luciferase-Based Assay for Monitoring ATG4B 27 Activity in Cells
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Identifying Small Molecules which Inhibit Autophagy: a Phenotypic Screen Using Image-Based High-Content Cell

J V Peppard1, C Rugg1, M Smicker1

  • 1Lead Generation and Candidate Realization, R&D, Bridgewater, NJ 07059, U.S.A.

Current Chemical Genomics and Translational Medicine
|March 6, 2014
PubMed
Summary
This summary is machine-generated.

Researchers screened 250,000 compounds to find small-molecule autophagy inhibitors for cancer therapy. They identified promising compounds targeting ULK and Vsp34, validating their high-content screening approach.

Keywords:
HTSLC3autophagyhigh-content screening.image-based screeninginhibitorsphenotypic screening

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In Vitro and In Vivo Detection of Mitophagy in Human Cells, C. Elegans, and Mice
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In Vitro and In Vivo Detection of Mitophagy in Human Cells, C. Elegans, and Mice

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Area of Science:

  • Oncology
  • Cell Biology
  • Drug Discovery

Background:

  • Autophagy is a crucial cellular process implicated in cancer development and progression.
  • It can act as both a tumor suppressor and a pro-survival mechanism, aiding cancer cells in resisting metabolic stress and chemotherapy.
  • Understanding and modulating autophagy is a key strategy for developing novel cancer therapies.

Purpose of the Study:

  • To identify novel small-molecule autophagy inhibitors using a high-throughput screening (HTS) and high-content analysis (HCA) approach.
  • To validate the efficacy of phenotypic, cell image-based assays for simultaneous screening of multiple biological targets in a physiologically relevant context.
  • To discover compounds that can overcome chemoresistance by targeting the pro-survival functions of autophagy.

Main Methods:

  • Utilized HeLa cells stably expressing EGFP-LC3 for a primary phenotypic screen, monitoring autophagosome formation.
  • Conducted a large-scale screen of approximately 250,000 compounds using HTS/HCA.
  • Employed a multi-step triage process involving secondary cellular assays, cheminformatics, and bioinformatics to filter and select promising compounds.

Main Results:

  • The primary screen identified approximately 10,500 initial positive compounds.
  • A rigorous triage process successfully narrowed down the candidates to about 400 fully qualified compounds.
  • The screening identified chemical series targeting autophagy-related kinases ULK and Vsp34, validating the phenotypic screening strategy.

Conclusions:

  • The developed HTS/HCA phenotypic screening approach is effective for identifying small-molecule autophagy inhibitors.
  • The study successfully identified specific compounds with activity against key autophagy regulators, providing potential leads for cancer drug development.
  • This work validates the utility of image-based screening for discovering modulators of complex cellular pathways like autophagy in cancer research.