Multifaceted Analyses of Isolated Mitochondria Establish the Anticancer Drug 2-Hydroxyoleic Acid as an Inhibitor of Substrate Oxidation and an Activator of Complex IV-Dependent State 3 Respiration

Kumudesh Mishra, Mária Péter, Anna Maria Nardiello, Guy Keller, Victoria Llado, Paula Fernandez-Garcia, Ulf D. Kahlert, Dinorah Barasch, Ann Saada, Zsolt Török, Gábor Balogh, Pablo V. Escriba*, Stefano Piotto*, Or Kakhlon*

*Corresponding author for this work

    Research output: Contribution to journalArticlepeer-review

    2 Scopus citations

    Abstract

    The synthetic fatty acid 2-hydroxyoleic acid (2OHOA) has been extensively investigated as a cancer therapy mainly based on its regulation of membrane lipid composition and structure, activating various cell fate pathways. We discovered, additionally, that 2OHOA can uncouple oxi-dative phosphorylation, but this has never been demonstrated mechanistically. Here, we explored the effect of 2OHOA on mitochondria isolated by ultracentrifugation from U118MG glioblastoma cells. Mitochondria were analyzed by shotgun lipidomics, molecular dynamic simulations, spectrophotometric assays for determining respiratory complex activity, mass spectrometry for assessing beta oxidation and Seahorse technology for bioenergetic profiling. We showed that the main impact of 2OHOA on mitochondrial lipids is their hydroxylation, demonstrated by simulations to decrease co-enzyme Q diffusion in the liquid disordered membranes embedding respiratory complexes. This de-creased co-enzyme Q diffusion can explain the inhibition of disjointly measured complexes I-III activ-ity. However, it doesn’t explain how 2OHOA increases complex IV and state 3 respiration in intact mitochondria. This increased respiration probably allows mitochondrial oxidative phosphorylation to maintain ATP production against the 2OHOA-mediated inhibition of glycolytic ATP production. This work correlates 2OHOA function with its modulation of mitochondrial lipid composition, reflecting both 2OHOA anticancer activity and adaptation to it by enhancement of state 3 respiration.

    Original languageAmerican English
    Article number578
    JournalCells
    Volume11
    Issue number3
    DOIs
    StatePublished - 7 Feb 2022

    Bibliographical note

    Publisher Copyright:
    © 2022 by the authors. Li-censee MDPI, Basel, Switzerland.

    Keywords

    • 2-hydroxyoleic acid
    • Bioenergetics
    • Glycolysis
    • Membrane lipid therapy
    • Mitochondria
    • Molecular dynamics
    • Respiration
    • Shotgun lipidomics

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