Asensio-Lopez Maria Del Carmen, Ruiz-Ballester Miriam, Pascual-Oliver Silvia, Bastida-Nicolas Francisco Jose, Sassi Yassine, Fuster Jose Javier, Pascual-Figal Domingo, Soler Fernando, Lax Antonio
Centro Nacional de Investigaciones Cardiovasculares (CNIC), Madrid, Spain.
R&D Department, Biocardio S.L, El Palmar, Murcia, Spain.
Mol Med. 2025 May 15;31(1):189. doi: 10.1186/s10020-025-01242-1.
The causal relationship between the activation of nuclear factor erythroid 2-related factor 2 (NRF2) and the preservation of SERCA2a function in mitigating myocardial ischemia-reperfusion (mI/R) injury, along with the associated regulatory mechanisms, remains incompletely understood. This study aims to unravel how NRF2 directly or indirectly influences SERCA2a function and its regulators, phospholamban (PLN) and Dwarf Open Reading Frame (DWORF), by testing the pharmacological repositioning of AEOL-10150 (AEOL) in the context of mI/R injury.
C57BL6/J, Nrf2 knockout (Nrf2), and wild-type (Nrf2) mice, as well as human induced pluripotent stem cell-derived cardiomyocytes (hiPSCMs) were subjected to I/R injury. Gain/loss of function techniques, RT-qPCR, western blotting, LC/MS/MS, and fluorescence spectroscopy were utilized. Cardiac dimensions and function were assessed by echocardiography.
In the early stages of mI/R injury, AEOL administration reduced mitochondrial ROS production, decreased myocardial infarct size, and improved cardiac function. These effects were due to NRF2 activation, leading to the overexpression of the micro-peptide DWORF, consequently enhancing SERCA2a activity. The cardioprotective effect induced by AEOL was diminished in Nrf2 mice and in Nrf2/Dworf knockdown models in hiPSCMs subjected to simulated I/R injury. Our data show that AEOL-induced NRF2-mediated upregulation of DWORF disrupts the phospholamban-SERCA2a interaction, leading to enhanced SERCA2a activation and improved cardiac function.
Taken together, our study reveals that AEOL-induced NRF2-mediated overexpression of DWORF enhances myocardial function through the activation of the SERCA2a offering promising therapeutic avenues for mI/R injury.
核因子红细胞2相关因子2（NRF2）的激活与肌浆网钙ATP酶2a（SERCA2a）功能在减轻心肌缺血再灌注（mI/R）损伤中的保护作用之间的因果关系，以及相关的调节机制，仍未完全明确。本研究旨在通过在mI/R损伤背景下测试AEOL-10150（AEOL）的药物重新定位，来阐明NRF2如何直接或间接影响SERCA2a功能及其调节因子——受磷蛋白（PLN）和矮小开放阅读框（DWORF）。
对C57BL6/J、Nrf2基因敲除（Nrf2）和野生型（Nrf2）小鼠，以及人诱导多能干细胞衍生的心肌细胞（hiPSCMs）进行缺血再灌注损伤。采用功能获得/丧失技术、实时定量聚合酶链反应（RT-qPCR）、蛋白质免疫印迹法、液相色谱-质谱联用（LC/MS/MS）和荧光光谱法。通过超声心动图评估心脏大小和功能。
在mI/R损伤的早期阶段，给予AEOL可减少线粒体活性氧生成，减小心肌梗死面积，并改善心脏功能。这些作用归因于NRF2激活，导致微小肽DWORF过表达，从而增强SERCA2a活性。在遭受模拟缺血再灌注损伤的hiPSCMs的Nrf2小鼠和Nrf2/Dworf基因敲低模型中，AEOL诱导的心脏保护作用减弱。我们的数据表明，AEOL诱导的NRF2介导的DWORF上调破坏了受磷蛋白-SERCA2a相互作用，导致SERCA2a激活增强和心脏功能改善。
综上所述，我们的研究表明，AEOL诱导的NRF2介导的DWORF过表达通过激活SERCA2a增强心肌功能，为mI/R损伤提供了有前景的治疗途径。
