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Image-Based Morphological Profiling Identifies a Lysosomotropic, Iron-Sequestering Autophagy Inhibitor.

Luca Laraia^1,2, Guillaume Garivet^1,3, Daniel J Foley^1,4

¹Department of Chemical Biology, Max-Planck-Institute of Molecular Physiology, Otto-Hahn-Strasse 11, 44227, Dortmund, Germany.

Angewandte Chemie (International Ed. in English)

|November 27, 2019

Summary

Researchers discovered the autophagy inhibitor autoquin and its mechanism using cell painting. Autoquin accumulates in lysosomes, inhibits fusion, and sequesters iron, leading to cell death, showcasing cell painting

Keywords:

autophagy cell painting lysosome proteomics target identification

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The Lactate Dehydrogenase Sequestration Assay — A Simple and Reliable Method to Determine Bulk Autophagic Sequestration Activity in Mammalian Cells

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Area of Science:

Chemical biology
Cellular imaging
Drug discovery

Background:

Chemical proteomics is a standard method for small-molecule target identification.
Current methods often fail to identify non-protein targets, creating a need for alternative approaches.
Understanding small molecule mechanisms is crucial for drug development.

Purpose of the Study:

To discover novel small molecules targeting autophagy.
To identify the molecular mechanism of action for newly discovered compounds.
To evaluate the utility of image-based morphological profiling in chemical biology.

Main Methods:

Discovery of the autophagy inhibitor autoquin.
Utilized cell painting assay for image-based morphological profiling.
Analyzed compound-induced cellular fingerprints across 579 parameters.

Main Results:

Autoquin was identified as an autophagy inhibitor.
The compound accumulates in lysosomes, inhibiting lysosome-autophagosome fusion.
Autoquin sequesters Fe2+ in lysosomes, increasing reactive oxygen species and causing cell death.

Conclusions:

The cell painting assay effectively deconvolutes the mode of action for small molecules.
Autoquin's mechanism involves lysosomal iron sequestration and subsequent oxidative stress.
This study highlights the potential of morphological profiling for broader chemical biology applications.