A novel function of ATF3 in suppression of ferroptosis in mouse heart suffered ischemia/reperfusion
A novel function of ATF3 in suppression of ferroptosis in mouse heart suffered ischemia/reperfusion
复制标题
ATF3抑制缺血/再灌注小鼠心脏铁死亡的新功能
DOI：
10.1016/j.freeradbiomed.2022.07.006
复制
发表时间：
2022-08-03
影响因子：
7.4
通讯作者：
Ling, Yuanna
中科院分区：
文献类型：
--
作者：
Liu, Haiqiong;Mo, Huaqiang;Ling, Yuanna
前言：铁下垂是一种新发现的程序性细胞死亡类型，已被证明与心肌缺血/再灌注(I/R)损伤的进展有关。目的：我们确认激活转录因子3(ATF3)是I/R引起的铁下垂的重要调节因子，并探讨ATF3在心肌铁下垂中的作用及其可能机制。方法：本研究通过对不同I/R方案的小鼠心脏进行动态RNA测序(RNA-seq)分析，确定ATF3在心肌I/R损伤中是一个重要的调节分子。采用基因敲除、挽救和过表达等方法，观察ATF3对心肌I/R损伤的影响。采用体内和体外功能丧失/获得技术，探讨ATF3对I/R损伤后铁性下垂的影响。进一步应用染色质免疫沉淀序列(ChIP-seq)分析ATF3对AC16人心肌细胞的调控作用。结果：ATF3在再灌注早期表达水平最高，敲除ATF3可明显加重心肌I/R损伤，ATF3再表达可明显减轻心肌I/R损伤。ATF3基因的敲除和重新表达改变了多种铁下垂基因的转录水平。此外，结果表明，ATF3的过表达抑制了erastin和RSL3引发的心肌细胞铁下垂。CHIP-SEQ和双荧光素酶活性分析表明，ATF3能与FANCD2基因的转录起始点结合，提高FANCD2启动子的活性。结论：ATF3可抑制AC16细胞缺氧再灌注损伤后的心肌细胞铁脂性死亡，其机制可能与调节FANCD2有关。我们的研究为心肌I/R损伤治疗的分子靶点提供了新的视角。
Introduction: Ferroptosis, a newly identified type of programmed cell death type, has been proven to contribute to the progression of myocardial ischemia/reperfusion (I/R) injury. However, little is known about ferroptosis regulation in I/R injury.Objectives: We identified activating transcription factor 3 (ATF3) as a vital regulator of I/R induced ferroptosis and investigated the effects and potential mechanism of ATF3 in cardiac ferroptosis.Methods: In this study, the dynamic RNA-sequencing (RNA-seq) analysis were performed on mouse hearts exposed to different I/R schedules to identify that ATF3 represents an important modulatory molecule in myocardial I/R injury. Then knockout, rescue and overexpression methods were used in mice and neonatal mouse cells (NMCs) to illustrate the effect of ATF3 on myocardial I/R injury. Loss/gain of function techniques were used both in vivo and in vitro to explore the effects of ATF3 on ferroptosis in I/R injury. Furthermore, chromatin immunoprecipitation sequence (ChIP-seq) analysis was performed in the AC16 human cardiomyocyte cell line to investigate potential genes regulated by ATF3.Results: ATF3 expression reached highest level at early stage of reperfusion, knockout of ATF3 significantly aggravated I/R injury, which could be rescued by ATF3 re-expression. Knockout and the re-expression of ATF3 changed the transcription levels of multiple ferroptosis genes. In addition, results showed that overexpression of ATF3 inhibits cardiomyocyte ferroptosis triggered by erastin and RSL3. Lastly, ChIP-seq and dual luciferase activity analysis revealed ATF3 could bind to the transcription start site of Fanconi anaemia complementation group D2 (FANCD2) and increased the FANCD2 promoter activity. Furthermore, we first demonstrated that overexpression of FANCD2 exerts significant anti-ferroptosis and cardioprotective effect on AC16 cell H/R injury.Conclusion: ATF3 inhibits cardiomyocyte ferroptotic death in I/R injury, which might be related with regulating FANCD2. Our study provides new insight into the molecular target for the therapy of myocardial I/R injury.