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

Cells

Kumudesh Mishra¹  Or Kakhlon¹  Anna Maria Nardiello²  Stefano Piotto²  Guy Keller³  Ann Saada³  Dinorah Barasch³  Victoria Llado⁴  Pablo V. Escriba⁴  Paula Fernandez-Garcia⁴  Mária Péter⁵  Zsolt Török⁵  Gábor Balogh⁵  Ulf D. Kahlert⁶ 

[1] Department of Neurology, Hadassah-Hebrew University Medical Center, Ein Kerem, Jerusalem 9112102, Israel;Department of Pharmacy, University of Salerno, Via Giovanni Paolo II, 132, 84084 Fisciano, SA, Italy;Faculty of Medicine, Hebrew University of Jerusalem, Ein Kerem, Jerusalem 9112102, Israel;Laminar Pharmaceuticals, Ctra. de Valldemossa Km. 7, 4 Parc BIT Ed. Naorte Bolque A-1°-3, 07121 Palma de Mallorca, Spain;Lipodom Ltd., Dorottya Utca 35-37, 6726 Szeged, Hungary;Molecular and Experimental Surgery, Clinic for General, Visceral, Vascular, and Transplant Surgery, Medical Faculty, University Hospital Magdeburg, 39120 Magdeburg, Germany;

关键词: 2-hydroxyoleic acid;    mitochondria;    molecular dynamics;    respiration;    glycolysis;    shotgun lipidomics;   

DOI  :  10.3390/cells11030578

来源: DOAJ

【 摘 要 】

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 oxidative 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 decreased co-enzyme Q diffusion can explain the inhibition of disjointly measured complexes I–III activity. 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.

【 授权许可】

Unknown