Molecular imaging of N-linked glycosylation suggests glycan biosynthesis is a novel target for cancer therapy.
OBJECTIVE
Redundant receptor tyrosine kinase (RTK) signaling is a mechanism for therapeutic resistance to epidermal growth factor receptor (EGFR) inhibition. A strategy to reduce parallel signaling by coexpressed RTKs is inhibition of N-linked glycosylation (NLG), an endoplasmic reticulum (ER) cotranslational protein modification required for receptor maturation and cell surface expression. We therefore investigated the feasibility of blocking NLG in vivo to reduce overexpression of RTKs.


METHODS
We developed a model system to dynamically monitor NLG in vitro and in vivo using bioluminescent imaging techniques. Functional imaging of NLG is accomplished with a luciferase reporter (ER-LucT) modified for endoplasmic reticulum translation and glycosylation. After in vitro validation, this reporter was integrated with D54 glioma xenografts to do noninvasive imaging of tumors, and inhibition of NLG was correlated with RTK protein levels and tumor growth.


RESULTS
The ER-LucT reporter shows the ability to sensitively and specifically detect NLG inhibition. Using this molecular imaging approach we carried out serial imaging studies to determine safe and efficacious in vivo dosing of the GlcNAc-1-phosphotransferase inhibitor tunicamycin, which blocks N-glycan precursor biosynthesis. Molecular analyses of tunicamycin-treated tumors showed reduced levels of EGFR and Met, two RTKs overexpressed in gliomas. Furthermore, D54 and U87MG glioma xenograft tumor experiments showed significant reductions in tumor growth following NLG inhibition and radiation therapy, consistent with an enhancement in tumor radiosensitivity.


CONCLUSIONS
This study suggests that NLG inhibition is a novel therapeutic strategy for targeting EGFR and RTK signaling in both gliomas and other malignant tumors.
Contessa
Joseph N
JN
Department of Radiation Oncology, The University of Michigan, Ann Arbor, Michigan, USA. joseph.contessa@yale.edu
Bhojani
Mahaveer S
MS
Freeze
Hudson H
HH
Ross
Brian D
BD
Rehemtulla
Alnawaz
A
Lawrence
Theodore S
TS
D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D045744:Cell Line, Tumor; D004721:Endoplasmic Reticulum / Q000378:metabolism; D066246:ErbB Receptors / Q000037:antagonists & inhibitors* / Q000378:metabolism; D005910:Glioma / Q000188:drug therapy / Q000378:metabolism*; D006031:Glycosylation / Q000187:drug effects*; D006801:Humans; D051379:Mice; D057054:Molecular Imaging / Q000379:methods*; D011134:Polysaccharides / Q000096:biosynthesis*; D019859:Proto-Oncogene Proteins c-met / Q000378:metabolism; D011897:Random Allocation; D020794:Receptor Protein-Tyrosine Kinases / Q000378:metabolism; D017978:Receptors, Growth Factor / Q000378:metabolism; D012680:Sensitivity and Specificity; D015398:Signal Transduction / Q000187:drug effects*; D014415:Tunicamycin / Q000494:pharmacology; D023041:Xenograft Model Antitumor Assays
D000970:Antineoplastic Agents; D011134:Polysaccharides; D017978:Receptors, Growth Factor; D014415:Tunicamycin; D066246:ErbB Receptors; C491743:MET protein, human; D019859:Proto-Oncogene Proteins c-met; D020794:Receptor Protein-Tyrosine Kinases