The Immunosuppressant Mycophenolic Acid Alters Nucleotide and Lipid Metabolism in an Intestinal Cell Model.
The study objective was to elucidate the molecular mechanisms underlying the negative effects of mycophenolic acid (MPA) on human intestinal cells. Effects of MPA exposure and guanosine supplementation on nucleotide concentrations in LS180 cells were assessed using liquid chromatography-mass spectrometry. Proteomics analysis was carried out using stable isotope labeling by amino acids in cell culture combined with gel-based liquid chromatography-mass spectrometry and lipidome analysis using1H nuclear magnetic resonance spectroscopy. Despite supplementation, depletion of guanosine nucleotides (p < 0.001 at 24 and 72 h; 5, 100, and 250 μM MPA) and upregulation of uridine and cytidine nucleotides (p < 0.001 at 24 h; 5 μM MPA) occurred after exposure to MPA. MPA significantly altered 35 proteins mainly related to nucleotide-dependent processes and lipid metabolism. Cross-reference with previous studies of MPA-associated protein changes widely corroborated these results, but showed differences that may be model- and/or method-dependent. MPA exposure increased intracellular concentrations of fatty acids, cholesterol, and phosphatidylcholine (p < 0.01 at 72 h; 100 μM MPA) which corresponded to the changes in lipid-metabolizing proteins. MPA affected intracellular nucleotide levels, nucleotide-dependent processes, expression of structural proteins, fatty acid and lipid metabolism in LS180 cells. These changes may compromise intestinal membrane integrity and contribute to gastrointestinal toxicity.
Department of Anesthesiology, School of Medicine, University of Colorado Denver, Anschutz Medical Campus, Bioscience 2, 12705 East Montview Boulevard, Suite 200, Aurora, CO 80045, USA.|Department of Organic Chemistry and Instrumental Analytics, University of Bremen, Leobener Strasse NW2 C0041, 28359 Bremen, Germany.
Svenja
S
Heischmann
Department of Biochemistry and Molecular Genetics, School of Medicine, University of Colorado Denver, Anschutz Medical Campus, Biological Mass Spectrometry Facility, 12801 East 17th Avenue, Aurora, CO 80045, USA.
Monika
M
Dzieciatkowska
Department of Biochemistry and Molecular Genetics, School of Medicine, University of Colorado Denver, Anschutz Medical Campus, Biological Mass Spectrometry Facility, 12801 East 17th Avenue, Aurora, CO 80045, USA.
Kirk
K
Hansen
Department of Biochemistry and Molecular Genetics, School of Medicine, University of Colorado Denver, Anschutz Medical Campus, Biological Mass Spectrometry Facility, 12801 East 17th Avenue, Aurora, CO 80045, USA.
Dieter
D
Leibfritz
Department of Anesthesiology, School of Medicine, University of Colorado Denver, Anschutz Medical Campus, Bioscience 2, 12705 East Montview Boulevard, Suite 200, Aurora, CO 80045, USA.
Uwe
U
Christians
D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D002853:Chromatography, Liquid; D006801:Humans; D007166:Immunosuppressive Agents / Q000494:pharmacology*; D007413:Intestinal Mucosa / Q000187:drug effects* / Q000378:metabolism*; D050356:Lipid Metabolism / Q000187:drug effects*; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D009173:Mycophenolic Acid / Q000494:pharmacology*; D009711:Nucleotides / Q000378:metabolism*; D020543:Proteome; D040901:Proteomics / Q000379:methods; D051381:Rats; D015398:Signal Transduction
D007166:Immunosuppressive Agents; D009711:Nucleotides; D020543:Proteome; D009173:Mycophenolic Acid