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Robust detection of oncometabolic aberrations by 1H-13C heteronuclear single quantum correlation in intact biological specimens.

Communications biology (2020-06-27)
Yasaman Barekatain, Victoria C Yan, Kenisha Arthur, Jeffrey J Ackroyd, Sunada Khadka, John De Groot, Jason T Huse, Florian L Muller
ABSTRACT

Magnetic resonance (MR) spectroscopy has potential to non-invasively detect metabolites of diagnostic significance for precision oncology. Yet, many metabolites have similar chemical shifts, yielding highly convoluted 1H spectra of intact biological material and limiting diagnostic utility. Here, we show that hydrogen-carbon heteronuclear single quantum correlation (1H-13C HSQC) offers dramatic improvements in sensitivity compared to one-dimensional (1D) 13C NMR and significant signal deconvolution compared to 1D 1H spectra in intact biological settings. Using a standard NMR spectroscope with a cryoprobe but without specialized signal enhancing features such as magic angle spinning, metabolite extractions or 13C-isotopic enrichment, we obtain well-resolved 2D 1H-13C HSQC spectra in live cancer cells, in ex vivo freshly dissected xenografted tumors and resected primary tumors. This method can identify tumors with specific oncometabolite alterations such as IDH mutations by 2-hydroxyglutarate and PGD-deleted tumors by gluconate. Results suggest potential of 1H-13C HSQC as a non-invasive diagnostic in precision oncology.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
VitroCol®, protein from human, suitable for cell culture
Sigma-Aldrich
Deuterium oxide, 99.9 atom % D, contains 0.05 wt. % 3-(trimethylsilyl)propionic-2,2,3,3-d4 acid, sodium salt
Sigma-Aldrich
D-Gluconic acid sodium salt, ≥99%
Sigma-Aldrich
6-Phosphogluconic acid trisodium salt, ≥97% (enzymatic)
Sigma-Aldrich
L-α-Hydroxyglutaric acid disodium salt, ≥98.0% (GC)
Sigma-Aldrich
6-Phosphogluconic acid trisodium salt, ≥95%

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