Abstract
Neuronal ferroptosis plays a key role in neurologic deficits post intracerebral hemorrhage (ICH). However, the endogenous regulation of rescuing ferroptotic neurons is largely unexplored. Here, we analyzed the integrated alteration of metabolomic landscape after ICH using LC-MS and MALDI-TOF/TOF MS, and demonstrated that aconitate decarboxylase 1 (Irg1) and its product itaconate, a derivative of the tricarboxylic acid cycle, were protectively upregulated. Deficiency of Irg1 or depletion of neuronal Irg1 in striatal neurons was shown to exaggerate neuronal loss and behavioral dysfunction in an ICH mouse model using transgenic mice. Administration of 4-Octyl itaconate (4-OI), a cell-permeable itaconate derivative, and neuronal Irg1 overexpression protected neurons in vivo. In addition, itaconate inhibited ferroptosis in cortical neurons derived from mouse and human induced pluripotent stem cells in vitro. Mechanistically, we demonstrated that itaconate alkylated glutathione peroxidase 4 (GPx4) on its cysteine 66 and the modification allosterically enhanced GPx4’s enzymatic activity by using a bioorthogonal probe, itaconate-alkyne (ITalk), and a GPx4 activity assay using phosphatidylcholine hydroperoxide. Altogether, our research suggested that Irg1/itaconate-GPx4 axis may be a future therapeutic strategy for protecting neurons from ferroptosis post ICH.
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All the data used in this manuscript are available from the corresponding author upon reasonable request.
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Acknowledgements
The purified GPx4U46C mutant protein is generously gifted from Dr. Luhua Lai (Peking University). We thank for suggestions from Dr. Zufeng Guo (Chongqing Medical University) and Dr. Li Su (Peking University Health Science Center). We thank the Computing Platform of the Center for Life Science at Peking University for supporting the LC−MS/MS data analysis. We are grateful to Prof. Renxiao Wang’s group at the School of Pharmacy, Fudan University, for their technical aid in the virtual screening job described in this work. Graphical abstract has been created with BioRender (https://BioRender.com).
Funding
National Natural Science Foundation of China (32070735 and 82371321 to Q. Li, 81971037 and 82271240 to F.Yang, 21925701, 92153301 and 91953109 to C. Wang, 22207126 to Yanling Zhang). Beijing Natural Science Foundation Program and Scientific Research Key Program of Beijing Municipal Commission of Education (KZ202010025033 to Q. Li). The Special Research Fund for Central Universities, Peking Union Medical College (3332022042 to Yanling Zhang).
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Conceptualization: Wei C., Xiao Z., Zhang Yanling, Luo Z., Wang C., Yang F., and Li Q. In vitro experiments: Wei C., Lan T., Liu M., and Hu L. Established ICH animal models and performed behavior tests: Xiao Z., Wei C., Luo Z., Shen D., Liu J., and Wang X. Identified GPx4 itaconation: Zhang Yanling, Wei C., Liu D., Hu J., and Wang C. Collected data: Wei C., Xiao Z., Luo Z., Shen D., Liu J., Dai Q., Zhang Yurui, Sun Q., Shi L., and Wu W. Analyzed data: Wei C., Xiao Z., Luo Z., Shen D., Zhang C., Wang P., Wang C., Yang F., and Li Q. Writing–original draft: Wei C., Xiao Z., Zhang Yanling, Luo Z., Wang C., Yang F., and Li Q. Writing—review & editing: Wei C., Xiao Z., Zhang Yanling, Luo Z., Wang C., Yang F., and Li Q. All authors have agreed on the final version to be published.
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Wei, C., Xiao, Z., Zhang, Y. et al. Itaconate protects ferroptotic neurons by alkylating GPx4 post stroke. Cell Death Differ (2024). https://doi.org/10.1038/s41418-024-01303-8
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DOI: https://doi.org/10.1038/s41418-024-01303-8