Ferrostatin-1 specifically targets mitochondrial iron-sulfur clusters and aconitase to improve cardiac function in Sirtuin 3 cardiomyocyte knockout mice

Aubrey C Cantrell; Jessie Besanson; Quinesha Williams; Ngoc Hoang; Kristin Edwards; G Reid Bishop; Yingjie Chen; Heng Zeng; Jian-Xiong Chen

doi:10.1016/j.yjmcc.2024.05.003

Ferrostatin-1 specifically targets mitochondrial iron-sulfur clusters and aconitase to improve cardiac function in Sirtuin 3 cardiomyocyte knockout mice

J Mol Cell Cardiol. 2024 May 9:192:36-47. doi: 10.1016/j.yjmcc.2024.05.003. Online ahead of print.

Authors

Aubrey C Cantrell¹, Jessie Besanson¹, Quinesha Williams¹, Ngoc Hoang¹, Kristin Edwards¹, G Reid Bishop¹, Yingjie Chen², Heng Zeng³, Jian-Xiong Chen⁴

Affiliations

¹ Department of Pharmacology & Toxicology, University of Mississippi Medical Center, School of Medicine, Jackson, MS 39216, USA.
² Department of Physiology & Biophysics, University of Mississippi Medical Center, School of Medicine, Jackson, MS 39216, USA.
³ Department of Pharmacology & Toxicology, University of Mississippi Medical Center, School of Medicine, Jackson, MS 39216, USA. Electronic address: hzeng@umc.edu.
⁴ Department of Pharmacology & Toxicology, University of Mississippi Medical Center, School of Medicine, Jackson, MS 39216, USA. Electronic address: jchen3@umc.edu.

PMID: 38734062
DOI: 10.1016/j.yjmcc.2024.05.003

Abstract

Aims: Ferroptosis is a form of iron-regulated cell death implicated in ischemic heart disease. Our previous study revealed that Sirtuin 3 (SIRT3) is associated with ferroptosis and cardiac fibrosis. In this study, we tested whether the knockout of SIRT3 in cardiomyocytes (SIRT3cKO) promotes mitochondrial ferroptosis and whether the blockade of ferroptosis would ameliorate mitochondrial dysfunction.

Methods and results: Mitochondrial and cytosolic fractions were isolated from the ventricles of mice. Cytosolic and mitochondrial ferroptosis were analyzed by comparison to SIRT3loxp mice. An echocardiography study showed that SIRT3cKO mice developed heart failure as evidenced by a reduction of EF% and FS% compared to SIRT3loxp mice. Comparison of mitochondrial and cytosolic fractions of SIRT3cKO and SIRT3loxp mice revealed that, upon loss of SIRT3, mitochondrial, but not cytosolic, total lysine acetylation was significantly increased. Similarly, acetylated p53 was significantly upregulated only in the mitochondria. These data demonstrate that SIRT3 is the primary mitochondrial deacetylase. Most importantly, loss of SIRT3 resulted in significant reductions of frataxin, aconitase, and glutathione peroxidase 4 (GPX4) in the mitochondria. This was accompanied by a significant increase in levels of mitochondrial 4-hydroxynonenal. Treatment of SIRT3cKO mice with the ferroptosis inhibitor ferrostatin-1 (Fer-1) for 14 days significantly improved preexisting heart failure. Mechanistically, Fer-1 treatment significantly increased GPX4 and aconitase expression/activity, increased mitochondrial iron‑sulfur clusters, and improved mitochondrial membrane potential and Complex IV activity.

Conclusions: Inhibition of ferroptosis ameliorated cardiac dysfunction by specifically targeting mitochondrial aconitase and iron‑sulfur clusters. Blockade of mitochondrial ferroptosis may be a novel therapeutic target for mitochondrial cardiomyopathies.

Keywords: Cardiomyopathy; Ferroptosis; Friedreich's ataxia; Mitochondrial dysfunction; SIRT3.