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Measuring NQO1 Bioactivation Using [2H7]Glucose.

Rohit Mahar1, Mario C Chang1, Matthew E Merritt1

  • 1Department of Biochemistry and Molecular Biology, College of Medicine, University of Florida, Gainesville, FL 32610, USA.

Cancers
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Beta-lapachone treatment generates reactive oxygen species (ROS) in NQO1-expressing cancer cells, leading to cell death. Measuring changes in glycolytic rates via 2H-NMR can assess treatment efficacy and reduce toxicity.

Area of Science:

  • Biochemistry
  • Oncology
  • Metabolic Engineering

Background:

  • Beta-lapachone is a cancer therapeutic that relies on NAD(P)H: quinone oxidoreductase 1 (NQO1) for bioactivation.
  • NQO1 bioactivation generates reactive oxygen species (ROS), leading to DNA damage, ATP depletion, and cell death via NAD+-keresis.
  • Assessing cancer cell glycolysis changes can indicate treatment efficacy and potentially reduce chemotherapy dosage and toxicity.

Purpose of the Study:

  • To evaluate the utility of measuring glycolytic flux changes for assessing beta-lapachone efficacy in NQO1-expressing cancers.
  • To establish a sensitive method for detecting NQO1-mediated metabolic alterations in cancer cells.

Main Methods:

  • Utilized 2H-NMR spectroscopy with [2H7]glucose administration to quantify deuterated metabolic products (2H-lactate and HDO) in A549, MiaPaCa2, and HCT-116 cancer cell lines.
Keywords:
2H-NMRHDO[2H7]glucosecancercell cultureβ-lapachone

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  • Analyzed the relationship between glucose consumption and the production of 2H-lactate and HDO.
  • Confirmed NMR findings using gas chromatography-mass spectrometry (GC-MS) to assess energy metabolism post-beta-lapachone treatment.

    • Main Results:

    • NQO1 bioactivation by beta-lapachone was successfully detected through changes in glycolytic flux in lung, pancreatic, and colon cancer cell lines.
    • Linear correlations were observed between glucose consumption and the production of 2H-lactate and HDO.
    • HDO was found to be a more sensitive marker for glycolytic flux than 2H-lactate due to its higher concentration.
    • GC-MS analysis corroborated NMR results, indicating downregulated energy metabolism in NQO1-positive cells after treatment.

    Conclusions:

    • Measuring glycolytic rates, particularly HDO production, is a sensitive method for assessing NQO1-mediated bioactivation of beta-lapachone.
    • This approach can be used to monitor the effects of chemotherapeutics targeting glycolysis.
    • The demonstrated method holds potential for in vivo translation for cancer treatment monitoring.