Extracellular superoxide dismutase (EC-SOD) R213G variant reduces mitochondrial ROS and preserves mitochondrial function in bleomycin-induced lung injury

• Published in: Advances in redox research : an official journal of the Society for Redox Biology and Medicine and the Society for Free Radical Research-Europe Vol. 5; p. 100035
• Main Authors: Elajaili, Hanan, Hernandez-Lagunas, Laura, Harris, Peter, Sparagna, Genevieve C., Jonscher, Raleigh, Ohlstrom, Denis, Sucharov, Carmen C., Bowler, Russell P., Suliman, Hagir, Fritz, Kristofer S., Roede, James R., Nozik, Eva S.
• Format: Journal Article
• Language: English
• Published: Elsevier B.V 01.07.2022; Elsevier
• Subjects: Cardiolipin; EC-SOD; Mitochondria; R213G; Redox potential; PBS; DTPA; GSSG; bleomycin; KHB; PTFE; Mito-TEMPO-H; Mitochondria; SNP; CPH; EhGSSG; Redox potential; Cardiolipin; EC-SOD; GSH; WT; R213G; HPLC; CMH
• ISSN: 2667-1379; 2667-1379
• DOI: 10.1016/j.arres.2022.100035

•The R213G variant of extracellular superoxide dismutase, which lowers matrix binding affinity and releases EC-SOD into extracellular fluids, lowers local superoxide levels in plasma and bronchoalveolar lavage fluid following bleomycin•The R213G EC-SOD variant protects the lung intracellular redox state, exemplified by less oxidized Eh GSSG, lower mitochondrial ROS level, and less mitochondrial cardiolipin oxidation•The R213G EC-SOD variant protects mitochondrial function in the setting of bleomycin-induced inflammation Extracellular superoxide dismutase (EC-SOD) is highly expressed in the lung and vasculature. A common human single nucleotide polymorphism (SNP) in the matrix binding region of EC-SOD leads to a single amino acid substitution, R213G, and alters EC-SOD tissue binding affinity. The change in tissue binding affinity redistributes EC-SOD from tissue to extracellular fluids. Mice (R213G mice) expressing a knock-in of this EC-SOD SNP exhibit elevated plasma and reduced lung EC-SOD content and activity and are protected against bleomycin-induced lung injury and inflammation. It is unknown how the redistribution of EC-SOD alters site-specific redox-regulated molecules relevant for protection. In this study, we tested the hypothesis that the change in the local EC-SOD content would influence not only the extracellular redox microenvironment where EC-SOD is localized but also protect the intracellular redox status of the lung. Mice were treated with bleomycin and harvested 7 days post-treatment. Superoxide levels, measured by electron paramagnetic resonance (EPR), were lower in plasma and Bronchoalveolar lavage fluid (BALF) cells in R213G mice compared to wild-type (WT) mice, while lung cellular superoxide levels in R213G mice were not elevated post-bleomycin compared to WT mice despite low lung EC-SOD levels. Lung glutathione redox potential (EhGSSG), determined by HPLC and fluorescence, was more oxidized in WT compared to R213G mice. In R213G mice, lung mitochondrial oxidative stress was reduced shown by mitochondrial superoxide level measured by EPR in lung and the resistance to bleomycin-induced cardiolipin oxidation. Bleomycin treatment suppressed mitochondrial respiration in WT mice. Mitochondrial function was impaired at baseline in R213G mice but did not exhibit further suppression in respiration post-bleomycin.  Collectively, the results indicate that R213G variant preserves intracellular redox state and protects mitochondrial function in the setting of bleomycin-induced inflammation. [Display omitted]