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A high-content screen for small-molecule regulators of epithelial cell-adhesion molecule (EpCAM) cleavage yields a

Jana Ylva Tretter1, Kenji Schorpp2, Elke Luxenburger3

  • 1From the Institute of Structural Biology and.

The Journal of Biological Chemistry
|April 28, 2018
PubMed

Insights

Researchers developed a high-content screening method to find small molecules that inhibit the Epithelial cell-adhesion molecule (EpCAM) signaling pathway. This approach successfully identified inhibitors targeting EpCAM

Area of Science:

  • Oncology
  • Molecular Biology
  • Biochemistry

Background:

  • Epithelial cell-adhesion molecule (EpCAM) is a transmembrane protein crucial for cell cycle progression and differentiation.
  • Overexpressed in many carcinomas, EpCAM drives proliferation via a mitogenic cascade involving regulated intramembrane proteolysis (RIP).
  • Targeting EpCAM's intracellular signaling offers a promising therapeutic strategy beyond current extracellular domain-focused immunotherapies.

Purpose of the Study:

  • To establish a high-content screening (HCS) assay for identifying small-molecule inhibitors of the EpCAM signaling pathway.
  • To characterize compounds that interfere with EpCAM's regulated intramembrane proteolysis (RIP).
  • To validate the feasibility of HCS for discovering inhibitors of membrane cleavage-dependent signaling.

Main Methods:

  • Development of an HCS assay using a yellow fluorescence protein-tagged EpCAM C-terminal fragment.
  • Screening of a small-molecule compound library (n = 27,280).
  • Characterization of validated hits, with detailed analysis of one potent inhibitor of EpCAM RIP.

Main Results:

  • Identification of 128 potential EpCAM signaling inhibitors from the compound library.
  • Discovery of one compound exhibiting robust inhibitory effects on EpCAM regulated intramembrane proteolysis (RIP).
  • Demonstration of the feasibility of using HCS to find inhibitors for the EpCAM signaling pathway.

Conclusions:

  • The developed HCS assay is effective for identifying small-molecule inhibitors of the EpCAM signaling pathway.
  • Targeting EpCAM's intracellular signaling cascade via RIP inhibition is a viable therapeutic strategy.
  • This HCS approach can be adapted to discover inhibitors for other signaling pathways involving membrane cleavage.
Keywords:
anticancer drugcell signalingepithelial cell adhesion molecule (EpCAM)high-throughput screening (HTS)intramembrane proteolysis

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