Intact Cell Lipidomics Reveal Changes to the Ratio of Cardiolipins to Phosphatidylinositols in Response to Kanamycin in HeLa and Primary Cells

• Published in: Chemical research in toxicology Vol. 31; no. 8; pp. 688 - 696
• Main Authors: Rebollo-Ramirez, Sonia, Krokowski, Sina, Lobato-Márquez, Damian, Thomson, Michael, Pennisi, Ivana, Mostowy, Serge, Larrouy-Maumus, Gerald
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 20-08-2018
• Subjects: Adenosine Triphosphate - metabolism; Animals; Anti-Bacterial Agents - administration & dosage; Anti-Bacterial Agents - pharmacology; Cardiolipins - metabolism; Dose-Response Relationship, Drug; Ethambutol - pharmacology; HeLa Cells; Humans; Kanamycin - administration & dosage; Kanamycin - pharmacology; Levofloxacin - pharmacology; Lipid Metabolism; Membrane Potentials - drug effects; Mice; Mitochondria - drug effects; Phosphatidylinositols - metabolism; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
• ISSN: 0893-228X; 1520-5010
• DOI: 10.1021/acs.chemrestox.8b00038

Antimicrobial resistance is a major threat the world is currently facing. Development of new antibiotics and the assessment of their toxicity represent important challenges. Current methods for addressing antibiotic toxicity rely on measuring mitochondrial damage using ATP and/or membrane potential as a readout. In this study, we propose an alternative readout looking at changes in the lipidome on intact and unprocessed cells by matrix-assisted laser desorption ionization mass spectrometry. As a proof of principle, we evaluated the impact of known antibiotics (levofloxacin, ethambutol, and kanamycin) on the lipidome of HeLa cells and mouse bone marrow-derived macrophages. Our methodology revealed that clinically relevant concentrations of kanamycin alter the ratio of cardiolipins to phosphatidylinositols. Unexpectedly, only kanamycin had this effect even though all antibiotics used in this study led to a decrease in the maximal mitochondrial respiratory capacity. Altogether, we report that intact cell-targeted lipidomics can be used as a qualitative method to rapidly assess the toxicity of aminoglycosides in HeLa and primary cells. Moreover, these results demonstrate there is no direct correlation between the ratio of cardiolipins to phosphatidylinositols and the maximal mitochondrial respiratory capacity.