Supplementary Materials and Methods and Supplementary Figures 1 through 5 from Multinuclear NMR and MRI Reveal an Early Metabolic Response to mTOR Inhibition in Sarcoma

Di Gialleonardo, Valentina; Aldeborgh, Hannah N.; Miloushev, Vesselin; Folkers, Kelly M.; Granlund, Kristin; Tap, William D.; Lewis, Jason S.; Weber, Wolfgang A.; Keshari, Kayvan R.

doi:10.1158/0008-5472.22412951.v1

Supplementary Materials and Methods and Supplementary Figures 1 through 5 from Multinuclear NMR and MRI Reveal an Early Metabolic Response to mTOR Inhibition in Sarcoma

journal contribution

posted on 2023-03-31, 00:24 authored by Valentina Di Gialleonardo, Hannah N. Aldeborgh, Vesselin Miloushev, Kelly M. Folkers, Kristin Granlund, William D. Tap, Jason S. Lewis, Wolfgang A. Weber, Kayvan R. Keshari

<p>Supplementary Materials and Methods. Figure S1. Major genetic alterations in of PI3K/AKT/mTOR pathway in 265 sarcoma tumors as summarized by TCGA using cBioPortal. Figure S2. Protein expression level detected with Western blot. Lactate Dehydrogenase (LDH), phosphorylated-LDH (p-LDH) and beta actin as a loading control. Figure S3. A) Metabolic profile of total and labeled intracellular B) and extracellular pool size of vehicle vs. rapamycin treated GIST-T1, DDLS, JJ012 and CS1. Last column on both panels shows the fractional enrichment of each metabolite analyzed. Figure S4. Bar plots represent the ratio of A) HP alanine/(HP total carbon) and B) HP bicarbonate/(HP total carbon) in CS1 tumor treated for 24 hours with 6% DMSO (black) and 15mg/kg of rapamycin (red). Figure S5. A) Intracellular concentration of ATP and B) NADPH of DMSO control (black) vs. rapamycin (red) treated GIST-T1, DDLS, JJ012 and CS1.</p>

Funding

NIH/NCI

NIH/NIBIB

Memorial Sloan Kettering's Center for Molecular Imaging and Nanotechnology

Cycle for Survival

American Italian Cancer Foundation

History

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DOI - Is supplement to Multinuclear NMR and MRI Reveal an Early Metabolic Response to mTOR Inhibition in Sarcoma

ARTICLE ABSTRACT

Biomarkers predicting rapalog responses in sarcomas where PI3K and mTOR are often hyperactivated could improve the suitable recruitment of responsive patients to clinical trials. PI3K/mTOR pathway activation drives energy production by regulating anaerobic glycolysis in cancer cells, suggesting a route toward a monitoring strategy. In this study, we took a multimodality approach to evaluate the phenotypic effects and metabolic changes that occur with inhibition of the PI3K/mTOR pathway. Its central role in regulating glycolysis in human sarcomas was evaluated by short- and long-term rapamycin treatment in sarcoma cell lines. We observed an overall decrease in lactate production in vitro, followed by cell growth inhibition. In vivo, we observed a similar quantitative reduction in lactate production as monitored by hyperpolarized MRI, also followed by tumor size changes. This noninvasive imaging method could distinguish reduced cell proliferation from induction of cell death. Our results illustrate the use of hyperpolarized MRI as a sensitive technique to monitor drug-induced perturbation of the PI3K/mTOR pathway in sarcomas. Cancer Res; 77(11); 3113–20. ©2017 AACR.