Bioenergetic Analyses of In Vitro and In Vivo Samples to Guide Toxicological Endpoints

Methods Mol Biol. 2020:2102:3-15. doi: 10.1007/978-1-0716-0223-2_1.

Abstract

Toxicology is a broad field that requires the translation of biochemical responses to xenobiotic exposures into useable information to ensure the safety of the public. Modern techniques are improving rapidly, both quantitatively and qualitatively, to provide the tools necessary to expand available toxicological datasets and refine our ability to translate that data into relevant information via bioinformatics. These new techniques can, and do, impact many of the current critical roles in toxicology, including the environmental, forensic, preclinical/clinical, and regulatory realms. One area of rapid expansion is our understanding of bioenergetics, or the study of the transformation of energy in living organisms, and new mathematical approaches are needed to interpret these large datasets. As bioenergetics are intimately involved in the regulation of how and when a cell responds to xenobiotics, monitoring these changes (i.e., metabolic fluctuations) in cells/tissues post-exposure provides an approach to define the temporal scale of pharmacodynamic responses, which can be used to guide additional toxicological techniques (e.g., "omics"). This chapter will summarize important in vitro assays and in vivo imaging techniques to take real-time measurements. Using this information, our laboratory has utilized bioenergetics to identify significant time points of pharmacodynamic relevance as well as forecast the cell's eventual fate.

Keywords: 18FDG; 2-NBDG; ATP; Bioenergetics; Forecasting; ICG; Live/Dead; MitoXpress; NADH.

MeSH terms

  • 4-Chloro-7-nitrobenzofurazan / analogs & derivatives
  • 4-Chloro-7-nitrobenzofurazan / metabolism
  • 4-Chloro-7-nitrobenzofurazan / pharmacology
  • Adenosine Triphosphate / metabolism
  • Animals
  • Biological Assay / methods*
  • Cell Survival / drug effects
  • Cell Survival / physiology
  • Deoxyglucose / analogs & derivatives
  • Deoxyglucose / metabolism
  • Deoxyglucose / pharmacology
  • Energy Metabolism / drug effects
  • Energy Metabolism / physiology*
  • Fluorodeoxyglucose F18 / metabolism
  • Humans
  • In Vitro Techniques
  • Indocyanine Green / pharmacology
  • Mitochondria / drug effects
  • Mitochondria / metabolism*
  • Mitochondria / physiology
  • NAD / metabolism
  • NADP / metabolism
  • Oxygen Consumption / drug effects
  • Oxygen Consumption / physiology
  • Positron Emission Tomography Computed Tomography
  • Toxicology / methods*
  • Workflow
  • Xenobiotics

Substances

  • Xenobiotics
  • NAD
  • Fluorodeoxyglucose F18
  • NADP
  • Adenosine Triphosphate
  • Deoxyglucose
  • 4-Chloro-7-nitrobenzofurazan
  • Indocyanine Green
  • 2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxyglucose