Cirbp suppression compromises DHODH-mediated ferroptosis defense and attenuates hypothermic cardioprotection in an aged donor transplantation model
Yifan Zhu, … , Hao Zhang, Yiwei Liu
Yifan Zhu, … , Hao Zhang, Yiwei Liu
Published May 1, 2024
Citation Information: J Clin Invest. 2024;134(9):e175645. https://doi.org/10.1172/JCI175645.
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Research Article Transplantation

Cirbp suppression compromises DHODH-mediated ferroptosis defense and attenuates hypothermic cardioprotection in an aged donor transplantation model

Abstract

Hypothermia is commonly used to protect donor hearts during transplantation. However, patients transplanted with aged donor hearts still have severe myocardial injury and decreased survival rates, but the underlying mechanism remains unknown. Because aged hearts are not considered suitable for donation, the number of patients awaiting heart transplants is increasing. In this study, we examined whether hypothermic cardioprotection was attenuated in aged donor hearts during transplantation and evaluated potential therapeutic targets. Using a rat heart transplantation model, we found that hypothermic cardioprotection was impaired in aged donor hearts but preserved in young donor hearts. RNA-Seq showed that cold-inducible RNA-binding protein (Cirbp) expression was decreased in aged donor hearts, and these hearts showed severe ferroptosis after transplantation. The young donor hearts from Cirbp-KO rats exhibited attenuated hypothermic cardioprotection, but Cirbp overexpression in aged donor hearts ameliorated hypothermic cardioprotection. Cardiac proteomes revealed that dihydroorotate dehydrogenase (DHODH) expression was significantly decreased in Cirbp-KO donor hearts during transplantation. Consequently, DHODH-mediated ubiquinone reduction was compromised, thereby exacerbating cardiac lipid peroxidation and triggering ferroptosis after transplantation. A cardioplegic solution supplemented with CIRBP agonists improved hypothermic cardioprotection in aged donor hearts, indicating that this method has the potential to broaden the indications for using aged donor hearts in transplantation.

Authors

Yifan Zhu, Chenyu Jiang, Jian He, Chen He, Xingliang Zhou, Xu Huang, Yi Shen, Liwei Wu, Yongnan Li, Bei Feng, Yi Yan, Jun Li, Hao Zhang, Yiwei Liu

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Figure 4

Decreased expression of SP1 suppresses CIRBP expression in aged donor heart.

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Decreased expression of SP1 suppresses CIRBP expression in aged donor he...
(A) Volcano plots showing the differentially expressed genes between young and aged donor hearts after transplantation. Green dots represent genes whose expression was significantly decreased (DOWN) in aged donor hearts. Red dots represent genes whose expression was significantly increased (UP) in aged donor hearts. Gray dots represent genes whose expression did not reach statistical significance (NO). (B) Western blotting and quantification of CIRBP in young and aged donor hearts after transplantation. (C) Real-time qPCR analysis of Cirbp mRNA expression in young and aged donor hearts before transplantation. (D) Western blotting and quantification of nuclear CIRBP in young and aged donor hearts before transplantation. (E) Immunofluorescence staining for CIRBP in young and aged donor hearts before transplantation and fluorescence intensity of nuclear CIRBP. Scale bars: 100 μm. (F) Western blotting and quantitation of nuclear CIRBP in young and aged donor hearts after transplantation. (G) Western blotting and quantitation of cytoplasmic CIRBP in young and aged donor hearts after transplantation. (H) Immunofluorescence staining for CIRBP in young and aged donor hearts after transplantation and fluorescence intensity of CIRBP. Scale bars: 50 μm. (I) Western blotting and quantitation of nuclear SP1 in young and aged donor hearts before transplantation. (J) Immunofluorescence staining for SP1 in young and aged donor hearts before transplantation and fluorescence intensity of SP1. Scale bars: 100 μm. (K) Integrative Genomics Viewer (IGV) screenshot of SP1 ChIP-Seq data for the Cirbp gene. (L) The degree to which SP1 binds the Cirbp promoter region was determined by ChIP-qPCR (n = 3 in each group). Quantitative data are shown as the mean ± standard deviation, with individual values presented as a dot plot. *P < 0.05, **P < 0.01, and ***P < 0.001, by 2-sided Student’s t test.