SoNar, a Highly Responsive NAD+/NADH Sensor, Allows High-Throughput Metabolic Screening of Anti-tumor Agents

Cell Metab. 2015 May 5;21(5):777-89. doi: 10.1016/j.cmet.2015.04.009.

Abstract

The altered metabolism of tumor cells confers a selective advantage for survival and proliferation, and studies have shown that targeting such metabolic shifts may be a useful therapeutic strategy. We developed an intensely fluorescent, rapidly responsive, pH-resistant, genetically encoded sensor of wide dynamic range, denoted SoNar, for tracking cytosolic NAD(+) and NADH redox states in living cells and in vivo. SoNar responds to subtle perturbations of various pathways of energy metabolism in real time, and allowed high-throughput screening for new agents targeting tumor metabolism. Among > 5,500 unique compounds, we identified KP372-1 as a potent NQO1-mediated redox cycling agent that produced extreme oxidative stress, selectively induced cancer cell apoptosis, and effectively decreased tumor growth in vivo. This study demonstrates that genetically encoded sensor-based metabolic screening could serve as a valuable approach for drug discovery.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antineoplastic Agents / pharmacology*
  • Antineoplastic Agents / therapeutic use
  • Apoptosis / drug effects
  • Cell Line, Tumor
  • Drug Screening Assays, Antitumor / methods*
  • Heterocyclic Compounds, 4 or More Rings / pharmacology*
  • Heterocyclic Compounds, 4 or More Rings / therapeutic use
  • Humans
  • Mice, Nude
  • NAD / metabolism*
  • NAD(P)H Dehydrogenase (Quinone) / metabolism
  • Neoplasms / drug therapy*
  • Neoplasms / metabolism
  • Neoplasms / pathology
  • Oxidation-Reduction / drug effects
  • Oxidative Stress / drug effects
  • Pentose Phosphate Pathway / drug effects
  • Tetrazoles / pharmacology*
  • Tetrazoles / therapeutic use

Substances

  • Antineoplastic Agents
  • Heterocyclic Compounds, 4 or More Rings
  • KP372-1
  • Tetrazoles
  • NAD
  • NAD(P)H Dehydrogenase (Quinone)
  • NQO1 protein, human