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Metabolic Imaging of 3-Phosphoglycerate as a Biomarker in Enolase1 Deleted Cells

Y Barekatain1*, F Muller2 , (1) MD Anderson Cancer Center UTHealth, Houston, TX, (2) ,Houston,


(Sunday, 7/14/2019) 4:30 PM - 5:00 PM

Room: Exhibit Hall | Forum 8

Purpose: Targeting therapies against tumor metabolomic vulnerabilities have gained increasing attention in the precision oncology. This presents the opportunity for the magnetic resonance spectroscopy to identify the subsets of patients with tumors that exhibit such metabolomic vulnerabilities. We have previously demonstrated that the deletion of enolase1 gene presents this metabolomic vulnerabilities. Our mass spectroscopy data shows that enolase1 deleted cells and tumors accumulate 3-phosphoglycerate and glycerate. We propose heteronuclear single quantum correlation (HSQC) technique to detect 3-phosphoglycerate. Here, we successfully demonstrate the measurement of 3-phosphoglycerate with 2D-HSQC using 500 MHz NMR machine in live glioma cells carrying the deletion of enolase1 gene.

Methods: We placed suspension cells of enolase1 deleted and wildtype cells in 5mm NMR tubes in D2O phosphate buffer. (1H-13C) heteronuclear single quantum correlation (HSQC) were acquired with HSQCEDETGPSISP pulse program. We used consecutive scans, each 8 minutes long. We acquired one scan before adding 1-13C labeled glucose. We then added 1-13C labeled glucose and followed the fate of the labeled carbon atom.

Results: Signals corresponding to known metabolites were detectable with natural abundance 13C testifying the powerful sensitivity of this techniques as well as resolving power to identify individual metabolites as compared to 1D proton spectrum. Upon adding 1-13C glucose, the 3-phosphoglycerate peak starts to appear. Later during the scan, the 3-phosphoglycerate peak disappeared while the glycerate peak appeared. Under the same experimental condition, detection of 3-phosphoglycerate and glycerate were not observed in the wildtype enolase1 cell lines, as we expected based on our mass spectroscopy data.

Conclusion: Our results suggests that even in highly heterogenous system, 2D heteronuclear spectroscopy is readily applicable and supports the use of this technique for the purposes of identification of metabolomic vulnerabilities in patients in vivo.


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