Abstract 219: Acyloxy Nitroso Compounds Inhibit LIF Signaling in Endothelial Cells and Cardiac Myocytes: Evidence That STAT3 Signaling is Redox-Sensitive
Nitroxyl donors have positive inotropic and lusitropic effects that may have therapeutic potential for treating heart failure, but their effects on other signaling pathways are not well studied. Previously we showed that leukemia inhibitory factor (LIF)-induced signaling is inhibited by oxidative stress, which targets JAK1 activation. In addition, we recently reported evidence for a cysteine-based redox switch in the JH1 catalytic domain of JAK 1 and 2. Since nitroxyl is thiophylic, we postulated that nitroxyl donors would inhibit LIF-induced JAK-STAT3 activation. Pretreatment of human microvascular endothelial cells (HMEC-1) or neonatal rat ventricular myocytes with the nitroxyl donors Angeli’s salt and the recently-described nitrosocyclohexyl acetate (NCA) inhibited LIF-induced STAT3 activation as indexed by Y705 phosphorylation. Pretreatment of HMEC-1 with NCA also blocked LIF-induced expression of the inflammation-related genes, ICAM-1 and CEBPD. The related acyloxy nitroso compound 1-nitrosocyclohexyl pivalate (NCP), which is not a nitroxyl donor, was equally effective in inhibiting STAT3 activation, suggesting that these compounds were acting as electrophiles on a thiolate residue. NCA had no effect on the catalytic activity of JAK1 and modestly affected JAK1-induced tyrosine phosphorylation of the LIF receptor. Thus, the JH1 redox switch is not a direct target of the acyloxy nitroso compound. However, pretreatment of recombinant human STAT3 with NCA or NCP reduced the labeling of free sulfhydryl residues with fluorescein-5-maleimide. In addition, we document by dimedone labeling that oxidation of STAT3 is associated with formation of sulfenic acid residues, a hallmark of redox-sensitive proteins. Altogether our evidence indicates that STAT3 has redox-sensitive cysteines that regulate its activation and are targeted by nitroxyl donors and related acyloxy nitroso compounds. These findings raise the possibility of new therapeutic strategies to target STAT3 signaling.
- © 2012 by American Heart Association, Inc.