Oligosaccharyltransferase inhibition induces senescence in RTK-driven tumor cells.
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.
Department of Therapeutic Radiology, Yale School of Medicine, New Haven, Connecticut, USA.
Cecilia
C
Lopez-Sambrooks
Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
Shiteshu
S
Shrimal
Center for the Science of Therapeutics, Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
Carol
C
Khodier
University of Kansas Specialized Chemistry Center, University of Kansas, Lawrence, Kansas, USA.
Daniel P
http://orcid.org/0000-0002-8305-0606
DP
Flaherty
Department of Therapeutic Radiology, Yale School of Medicine, New Haven, Connecticut, USA.
Natalie
N
Rinis
Department of Therapeutic Radiology, Yale School of Medicine, New Haven, Connecticut, USA.
Jonathan C
JC
Charest
Department of Pharmacology, UT Southwestern Medical Center at Dallas, Dallas, Texas, USA.
Ningguo
N
Gao
Complex Carbohydrate Research Center, Department of Biochemistry and Molecular Biology, University of Georgia, Athens, Georgia, USA.
Peng
P
Zhao
Complex Carbohydrate Research Center, Department of Biochemistry and Molecular Biology, University of Georgia, Athens, Georgia, USA.
Lance
L
Wells
Center for the Science of Therapeutics, Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
Timothy A
TA
Lewis
Department of Pharmacology, UT Southwestern Medical Center at Dallas, Dallas, Texas, USA.
Mark A
MA
Lehrman
Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
Reid
R
Gilmore
University of Kansas Specialized Chemistry Center, University of Kansas, Lawrence, Kansas, USA.
Jennifer E
JE
Golden
Department of Therapeutic Radiology, Yale School of Medicine, New Haven, Connecticut, USA.|Department of Pharmacology, Yale School of Medicine, New Haven, Connecticut, USA.
Joseph N
JN
Contessa
D001549:Benzamides / Q000737:chemistry / Q000494:pharmacology*; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D016922:Cellular Senescence / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D006602:Hexosyltransferases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D057166:High-Throughput Screening Assays; D006801:Humans; D008565:Membrane Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D015394:Molecular Structure; D020794:Receptor Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D013329:Structure-Activity Relationship; D013449:Sulfonamides / Q000737:chemistry / Q000494:pharmacology*
D001549:Benzamides; C000618319:CID2519269; D004791:Enzyme Inhibitors; D008565:Membrane Proteins; D013449:Sulfonamides; D006602:Hexosyltransferases; C025620:dolichyl-diphosphooligosaccharide - protein glycotransferase; D020794:Receptor Protein-Tyrosine Kinases
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