Metabolomics analysis of metabolic effects of nicotinamide phosphoribosyltransferase (NAMPT) inhibition on human cancer cells

PLoS One. 2014 Dec 8;9(12):e114019. doi: 10.1371/journal.pone.0114019. eCollection 2014.

Abstract

Nicotinamide phosphoribosyltransferase (NAMPT) plays an important role in cellular bioenergetics. It is responsible for converting nicotinamide to nicotinamide adenine dinucleotide, an essential molecule in cellular metabolism. NAMPT has been extensively studied over the past decade due to its role as a key regulator of nicotinamide adenine dinucleotide-consuming enzymes. NAMPT is also known as a potential target for therapeutic intervention due to its involvement in disease. In the current study, we used a global mass spectrometry-based metabolomic approach to investigate the effects of FK866, a small molecule inhibitor of NAMPT currently in clinical trials, on metabolic perturbations in human cancer cells. We treated A2780 (ovarian cancer) and HCT-116 (colorectal cancer) cell lines with FK866 in the presence and absence of nicotinic acid. Significant changes were observed in the amino acids metabolism and the purine and pyrimidine metabolism. We also observed metabolic alterations in glycolysis, the citric acid cycle (TCA), and the pentose phosphate pathway. To expand the range of the detected polar metabolites and improve data confidence, we applied a global metabolomics profiling platform by using both non-targeted and targeted hydrophilic (HILIC)-LC-MS and GC-MS analysis. We used Ingenuity Knowledge Base to facilitate the projection of metabolomics data onto metabolic pathways. Several metabolic pathways showed differential responses to FK866 based on several matches to the list of annotated metabolites. This study suggests that global metabolomics can be a useful tool in pharmacological studies of the mechanism of action of drugs at a cellular level.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acids / metabolism
  • Cell Line, Tumor
  • Citric Acid Cycle
  • Cluster Analysis
  • Creatine / metabolism
  • Glycolysis
  • Humans
  • Lipid Metabolism
  • Mass Spectrometry / methods
  • Metabolic Networks and Pathways
  • Metabolome*
  • Metabolomics* / methods
  • Neoplasms / metabolism*
  • Nicotinamide Phosphoribosyltransferase / antagonists & inhibitors*
  • Nicotinamide Phosphoribosyltransferase / metabolism
  • Pentose Phosphate Pathway
  • Purines / metabolism
  • Pyrimidines / metabolism

Substances

  • Amino Acids
  • Purines
  • Pyrimidines
  • Nicotinamide Phosphoribosyltransferase
  • pyrimidine
  • Creatine
  • purine

Grant support

This work was supported by Eli Lilly and Company. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.