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Published March 9, 2016 | Accepted Version + Supplemental Material
Journal Article Open

Supramolecular Probes for Assessing Glutamine Uptake Enable Semi-Quantitative Metabolic Models in Single Cells


We describe a supramolecular surface competition assay for quantifying glutamine uptake from single cells. Cy3-labeled cyclodextrins were immobilized on a glass surface as a supramolecular host/FRET donor, and adamantane-BHQ2 conjugates were employed as the guest/quencher. An adamantane-labeled glutamine analog was selected through screening a library of compounds and validated by cell uptake experiments. When integrated onto a single cell barcode chip with a multiplex panel of 15 other metabolites, associated metabolic enzymes, and phosphoproteins, the resultant data provided input for a steady-state model that describes energy potential in single cells and correlates that potential with receptor tyrosine kinase signaling. We utilize this integrated assay to interrogate a dose-dependent response of model brain cancer cells to EGFR inhibition. We find that low-dose (1 μM erlotinib) drugging actually increases cellular energy potential even as glucose uptake and phosphoprotein signaling is repressed. We also identify new interactions between phosphoprotein signaling and cellular energy processes that may help explain the facile resistance exhibited by certain cancer patients to EGFR inhibitors.

Additional Information

© 2016 American Chemical Society. Received: November 20, 2015. Publication Date (Web): February 26, 2016. We thank Dr. David Nathanson for the suggestions on glioblastoma cell line culturing and drug treatments. We acknowledge the following agencies and foundations for support: The National Cancer Institute (1U54 CA199090-01 J.R.H. PI, W.W. and R01-CA170689 J.R.H. PI), the Ben and Catherine Ivy Foundation (J.R.H.), the Jean Perkins Foundation (J.R.H. PI), and the Phelps Family Foundation (J.R.H. and W.W.). The authors declare the following competing financial interest(s): J.R.H. is a board member of IsoPlexis, which is a company seeking to commercialize the SCBC technology.

Attached Files

Accepted Version - nihms-787683.pdf

Supplemental Material - ja5b12187_si_001.pdf


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August 20, 2023
October 17, 2023