Asparagine (N)-linked glycosylation is a protein modification critical for glycoprotein folding, stability, and cellular localization. To identify small molecules that inhibit new targets in this biosynthetic pathway, we initiated a cell-based high-throughput screen and lead-compound-optimization campaign that delivered a cell-permeable inhibitor, NGI-1. NGI-1 targets oligosaccharyltransferase (OST), a hetero-oligomeric enzyme that exists in multiple isoforms and transfers oligosaccharides to recipient proteins. In non-small-cell lung cancer cells, NGI-1 blocks cell-surface localization and signaling of the epidermal growth factor receptor (EGFR) glycoprotein, but selectively arrests proliferation in only those cell lines that are dependent on EGFR (or fibroblast growth factor, FGFR) for survival. In these cell lines, OST inhibition causes cell-cycle arrest accompanied by induction of p21, autofluorescence, and cell morphology changes, all hallmarks of senescence. These results identify OST inhibition as a potential therapeutic approach for treating receptor-tyrosine-kinase-dependent tumors and provides a chemical probe for reversibly regulating N-linked glycosylation in mammalian cells.
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This work was funded by US National Institutes of Health (NIH) R03DA033178 and R01CA172391, and in part by a Research Scholar Grant from the American Cancer Society (J.N.C.) and by NIH RO1GM43768 (R.G.) and RO1GM038545 (M.A.L.). Additional funding was provided by the NIH-MLPCN program U54HG005031 (Kansas University) and U54HG005032 (Broad Institute). The mass spectrometry work was supported by NIH grant P41GM103490.
J.N.C. and J.E.G. are listed as inventors on a provisional patent application for the analogs reported in this manuscript.
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Lopez-Sambrooks, C., Shrimal, S., Khodier, C. et al. Oligosaccharyltransferase inhibition induces senescence in RTK-driven tumor cells. Nat Chem Biol 12, 1023–1030 (2016). https://doi.org/10.1038/nchembio.2194
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