TY  - JOUR
AU  - Tretter, Jana Ylva
AU  - Schorpp, Kenji
AU  - Luxenburger, Elke
AU  - Trambauer, Johannes
AU  - Steiner, Harald
AU  - Hadian, Kamyar
AU  - Gires, Olivier
AU  - Niessing, Dierk
TI  - A high-content screen for small-molecule regulators of epithelial cell-adhesion molecule (EpCAM) cleavage yields a robust inhibitor.
JO  - The journal of biological chemistry
VL  - 293
IS  - 23
SN  - 0021-9258
CY  - Bethesda, Md.
PB  - Soc.60645
M1  - DZNE-2020-06323
SP  - 8994-9005
PY  - 2018
AB  - Epithelial cell-adhesion molecule (EpCAM) is a transmembrane protein that regulates cell cycle progression and differentiation and is overexpressed in many carcinomas. The EpCAM-induced mitogenic cascade is activated via regulated intramembrane proteolysis (RIP) of EpCAM by ADAM and γ-secretases, generating the signaling-active intracellular domain EpICD. Because of its expression pattern and molecular function, EpCAM is a valuable target in prognostic and therapeutic approaches for various carcinomas. So far, several immunotherapeutic strategies have targeted the extracellular domain of EpCAM. However, targeting the intracellular signaling cascade of EpCAM holds promise for specifically interfering with EpCAM's proliferation-stimulating signaling cascade. Here, using a yellow fluorescence protein-tagged version of the C-terminal fragment of EpCAM, we established a high-content screening (HCS) of a small-molecule compound library (n = 27,280) and characterized validated hits that target EpCAM signaling. In total, 128 potential inhibitors were initially identified, of which one compound with robust inhibitory effects on RIP of EpCAM was analyzed in greater detail. In summary, our study demonstrates that the development of an HCS for small-molecule inhibitors of the EpCAM signaling pathway is feasible. We propose that this approach may also be useful for identifying chemical compounds targeting other disorders involving membrane cleavage-dependent signaling pathways.
KW  - Cell Proliferation: drug effects
KW  - Drug Evaluation, Preclinical: methods
KW  - Epithelial Cell Adhesion Molecule: antagonists & inhibitors
KW  - Epithelial Cell Adhesion Molecule: metabolism
KW  - HEK293 Cells
KW  - High-Throughput Screening Assays: methods
KW  - Humans
KW  - Signal Transduction: drug effects
KW  - Small Molecule Libraries: chemistry
KW  - Small Molecule Libraries: pharmacology
KW  - Transcription, Genetic: drug effects
KW  - Epithelial Cell Adhesion Molecule (NLM Chemicals)
KW  - Small Molecule Libraries (NLM Chemicals)
LB  - PUB:(DE-HGF)16
C6  - pmid:29700109
C2  - pmc:PMC5995519
DO  - DOI:10.1074/jbc.RA118.002776
UR  - https://pub.dzne.de/record/140001
ER  -