Non-invasive in-cell determination of free cytosolic [NAD+]/[NADH] ratios using hyperpolarized glucose show large variations in metabolic phenotypes

Accumulating evidence suggest that the pyridine nucleotide NAD has far wider biological functions than its classical role in energy metabolism. NAD is used by hundreds of enzymes that catalyze substrate oxidation and, as such, it plays a key role in various biological processes such as aging, cell d...

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Published in:The Journal of biological chemistry 2014-01, Vol.289 (4), p.2344-2352
Main Authors: Christensen, Caspar Elo, Karlsson, Magnus, Winther, Jakob R, Jensen, Pernille Rose, Lerche, Mathilde H
Format: Article
Language:English
Subjects:
Breast Neoplasms - metabolism
Breast Neoplasms - pathology
Cell Line, Tumor
Female
Glucose - metabolism
Glycolysis
Humans
Lactic Acid - metabolism
Male
Metabolism
NAD - metabolism
Prostatic Neoplasms - metabolism
Prostatic Neoplasms - pathology
Pyruvic Acid - metabolism
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Summary:Accumulating evidence suggest that the pyridine nucleotide NAD has far wider biological functions than its classical role in energy metabolism. NAD is used by hundreds of enzymes that catalyze substrate oxidation and, as such, it plays a key role in various biological processes such as aging, cell death, and oxidative stress. It has been suggested that changes in the ratio of free cytosolic [NAD(+)]/[NADH] reflects metabolic alterations leading to, or correlating with, pathological states. We have designed an isotopically labeled metabolic bioprobe of free cytosolic [NAD(+)]/[NADH] by combining a magnetic enhancement technique (hyperpolarization) with cellular glycolytic activity. The bioprobe reports free cytosolic [NAD(+)]/[NADH] ratios based on dynamically measured in-cell [pyruvate]/[lactate] ratios. We demonstrate its utility in breast and prostate cancer cells. The free cytosolic [NAD(+)]/[NADH] ratio determined in prostate cancer cells was 4 times higher than in breast cancer cells. This higher ratio reflects a distinct metabolic phenotype of prostate cancer cells consistent with previously reported alterations in the energy metabolism of these cells. As a reporter on free cytosolic [NAD(+)]/[NADH] ratio, the bioprobe will enable better understanding of the origin of diverse pathological states of the cell as well as monitor cellular consequences of diseases and/or treatments.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.M113.498626