HDO production from [²H₇]glucose Quantitatively Identifies Warburg Metabolism

Area of Science:

• Biochemistry
• Medical Imaging
• Metabolic Engineering

Background:

• Cancer cells exhibit increased glucose uptake and aerobic glycolysis (Warburg effect).
• Current diagnostic methods for cancer metabolism are often costly, impractical, or have adverse effects.
• Sensitive detection of metabolic shifts is crucial for early cancer diagnosis and monitoring.

Purpose of the Study:

• To develop and validate a sensitive 1H/2H NMR method for measuring lactate isotopomer and HDO production kinetics.
• To utilize a deuterated glucose tracer for quantitative analysis of metabolic pathways in cancer cells.
• To explore the potential of HDO production as a diagnostic marker for Warburg metabolism.

Main Methods:

• Incubation of HUH-7 (hepatocellular carcinoma) and AML12 (normal hepatocytes) cells with [2H7]glucose.
• Acquisition of 1H/2H NMR data from cell media over time.
• Calculation of lactate isotopomer efflux rates and analysis of HDO production.

Main Results:

• Differential production of lactate isotopomers (lactate-CH3, lactate-CH2D, lactate-CHD2) was observed due to deuterium loss during glycolysis.
• A strong correlation between glucose uptake and HDO production was found in HUH-7 cells.
• Quantitative deuterium mass balance confirmed the link between [2H7]glucose uptake and 2H-lactate/HDO production.

Conclusions:

• Monitoring HDO production via 1H/2H NMR is a sensitive and potentially non-invasive diagnostic feature for Warburg metabolism in cancers.
• The kinetics of lactate isotopomer and HDO production can be accurately measured using NMR.
• Increased T1 relaxation times of 2H-lactate isotopomers suggest potential for metabolism-based contrast generation in MRI.