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A genetically encoded tool to increase cellular NADH/NAD+ ratio in living cells

Xingxiu Pan1, Mina L Heacock1,2, Evana N Abdulaziz1,3

  • 1Laboratory of Redox Biology and Metabolism, Scintillon Institute, San Diego, CA, USA.

Nature Chemical Biology
|October 26, 2023
PubMed

Insights

Researchers developed a new genetic tool to induce reductive stress, an overabundance of NADH, in cells. This tool helps study diseases linked to redox imbalance and reveals that reductive stress effects vary by cell type.

Area of Science:

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Background:

  • Impaired redox metabolism contributes to diseases like cancer, neurodegeneration, and aging.
  • Studying redox imbalance is difficult due to limited perturbation strategies.
  • Reductive stress (NADH overabundance) is less understood than oxidative stress.

Purpose of the Study:

  • Introduce a novel genetically encoded tool to induce reductive stress.
  • Investigate the cellular and transcriptomic consequences of reductive stress.
  • Provide a new method for studying redox metabolism in various cellular contexts.

Main Methods:

  • Genetically encoded expression of soluble transhydrogenase from Escherichia coli (EcSTH) in mammalian cells.
  • Utilized a mitochondrially targeted version (mitoEcSTH) for compartment-specific effects.
  • Analyzed metabolic and transcriptomic changes in response to induced reductive stress.

Main Results:

  • EcSTH and mitoEcSTH successfully elevated the NADH/NAD+ ratio in a compartment-specific manner.
  • Demonstrated the feasibility of using a genetic tool to induce and study reductive stress.
  • Observed that metabolic and transcriptomic signatures of reductive stress are cell-type dependent.

Conclusions:

  • A novel genetically encoded tool (EcSTH) enables robust induction of reductive stress in living cells.
  • This tool offers an orthogonal strategy to study the mechanisms of redox imbalance.
  • Findings highlight the cellular context-specific nature of reductive stress responses.

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