DOE OSTI · 3024702
Robust Rapid Cellular Metabolite Sensing Using Benchtop NMR and SABRE-Hyperpolarized [1- 13 C]Pyruvate
Abstract
Hyperpolarized NMR has emerged as a powerful analytical technique to significantly enhance targeted NMR signals, improving the sensitivity for investigations of unique chemical and biological dynamics. Here, we demonstrate the use of a hyperpolarization strategy based on Signal Amplification By Reversible Exchange (SABRE) to generate highly reproducible doses of a hyperpolarized [1- 13 C]pyruvate probe for benchtop characterization of yeast metabolism. This method allows rapid, scalable, and benchtop preparation of biocompatible hyperpolarized solutions suitable for live-cell experiments. We show that this production can be dove-tailed into a modular, compact workflow to characterize real-time metabolism in cell cultures, using Saccharomyces cerevisiae (Baker’s yeast) as a model organism. With high temporal resolution, we show that this method can resolve the conversion of hyperpolarized [1- 13 C]pyruvate into oxidative decarboxylation products CO 2 and bicarbonate. This conversion exhibits sustained and detectable metabolic activity for over 300 s after introduction of the agent to the cells. We model the metabolite kinetics to show decarboxylation activity and derive estimates of the pH over time from the CO 2 and bicarbonate (carbonic acid buffer system) equilibrium to probe changes in the cellular environment during active metabolism. These results highlight the utility of benchtop SABRE-hyperpolarized [1- 13 C]pyruvate as a scalable, specific probe for metabolic phenotyping of living cells using compact, low-cost instrumentation well-suited for future high-throughput applications across microbial engineering, drug response profiling, and dynamic metabolic screening.
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Gyesi, Joseph [Wayne State Univ., Detroit, MI (United States). Karmanos Cancer Institute (KCI)], TomHon, Patrick [Wayne State Univ., Detroit, MI (United States). Karmanos Cancer Institute (KCI)] (ORCID:0000000332029812), Abdurraheem, Abubakar [Wayne State Univ., Detroit, MI (United States). Karmanos Cancer Institute (KCI)], Samoilenko, Anna [Wayne State Univ., Detroit, MI (United States). Karmanos Cancer Institute (KCI)], Scofield, Sydney [Wayne State Univ., Detroit, MI (United States)], Oladun, Clementinah [Wayne State Univ., Detroit, MI (United States). Karmanos Cancer Institute (KCI)], McBride, Stephen [North Carolina State University, Raleigh, NC (United States)], Curran, Erica [North Carolina State University, Raleigh, NC (United States)] (ORCID:0009000513765541), Pike, Megan [North Carolina State University, Raleigh, NC (United States)], Kadari, Kamal [Wayne State Univ., Detroit, MI (United States). Karmanos Cancer Institute (KCI)], Welch, Sydney D. [Wayne State Univ., Detroit, MI (United States). Karmanos Cancer Institute (KCI)], Lipka, Sam [Wayne State Univ., Detroit, MI (United States). Karmanos Cancer Institute (KCI)], Balboa, Steven [Wayne State Univ., Detroit, MI (United States). Karmanos Cancer Institute (KCI)], Fehl, Charlie [Wayne State Univ., Detroit, MI (United States). Karmanos Cancer Institute (KCI)] (ORCID:0000000161820879), Sadagurski, Marianna [Wayne State Univ., Detroit, MI (United States)], Hövener, Jan-Bernd [Kiel University (Germany). University Medical Center] (ORCID:0000000172557252), Theis, Thomas [North Carolina State University, Raleigh, NC (United States)] (ORCID:0000000167799978), Goodson, Boyd M. [Southern Illinois Univ., Carbondale, IL (United States)] (ORCID:0000000160795077), Chekmenev, Eduard Y. [Wayne State Univ., Detroit, MI (United States). Karmanos Cancer Institute (KCI)] (ORCID:0000000287458801). 2026-03-10. Robust Rapid Cellular Metabolite Sensing Using Benchtop NMR and SABRE-Hyperpolarized [1- 13 C]Pyruvate. https://doi.org/10.1021/acs.analchem.5c05076
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