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

Xue, Min and Wei, Wei and Su, Yapeng and Johnson, Dazy and Heath, James R. (2016) Supramolecular Probes for Assessing Glutamine Uptake Enable Semi-Quantitative Metabolic Models in Single Cells. Journal of the American Chemical Society, 138 (9). pp. 3085-3093. ISSN 0002-7863. PMCID PMC4887079. http://resolver.caltech.edu/CaltechAUTHORS:20160307-080722516

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Abstract

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.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1021/jacs.5b12187DOIArticle
http://pubs.acs.org/doi/abs/10.1021/jacs.5b12187PublisherArticle
http://pubs.acs.org/doi/suppl/10.1021/jacs.5b12187PublisherSupporting Information
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4887079/PubMed CentralArticle
ORCID:
AuthorORCID
Heath, James R.0000-0001-5356-4385
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.
Funders:
Funding AgencyGrant Number
National Cancer Institute1U54 CA199090-01
National Cancer InstituteR01-CA170689
Ben and Catherine Ivy FoundationUNSPECIFIED
Jean Perkins FoundationUNSPECIFIED
Phelps Family FoundationUNSPECIFIED
PubMed Central ID:PMC4887079
Record Number:CaltechAUTHORS:20160307-080722516
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20160307-080722516
Official Citation:Supramolecular Probes for Assessing Glutamine Uptake Enable Semi-Quantitative Metabolic Models in Single Cells Min Xue, Wei Wei, Yapeng Su, Dazy Johnson, and James R. Heath Journal of the American Chemical Society 2016 138 (9), 3085-3093 DOI: 10.1021/jacs.5b12187
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:65093
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:08 Mar 2016 18:03
Last Modified:25 Apr 2017 03:47

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