Laboratory of Cell Biology, Genetics and Developmental Biology, Shaanxi Normal University College of Life Sciences, Xi'an 710062, China.
University Laboratory Animal Resources (ULAR), University of Pennsylvania School of Medicine, Philadelphia, PA 19144, USA.
Int J Mol Sci. 2023 Feb 4;24(4):3094. doi: 10.3390/ijms24043094.
Transgenic expression of Cre recombinase driven by a specific promoter is normally used to conditionally knockout a gene in a tissue- or cell-type-specific manner. In αMHC-Cre transgenic mouse model, expression of Cre recombinase is controlled by the myocardial-specific α-myosin heavy chain (αMHC) promoter, which is commonly used to edit myocardial-specific genes. Toxic effects of Cre expression have been reported, including intro-chromosome rearrangements, micronuclei formation and other forms of DNA damage, and cardiomyopathy was observed in cardiac-specific Cre transgenic mice. However, mechanisms associated with Cardiotoxicity of Cre remain poorly understood. In our study, our data unveiled that αMHC-Cre mice developed arrhythmias and died after six months progressively, and none of them survived more than one year. Histopathological examination showed that αMHC-Cre mice had aberrant proliferation of tumor-like tissue in the atrial chamber extended from and vacuolation of ventricular myocytes. Furthermore, the αMHC-Cre mice developed severe cardiac interstitial and perivascular fibrosis, accompanied by significant increase of expression levels of MMP-2 and MMP-9 in the cardiac atrium and ventricular. Moreover, cardiac-specific expression of Cre led to disintegration of the intercalated disc, along with altered proteins expression of the disc and calcium-handling abnormality. Comprehensively, we identified that the ferroptosis signaling pathway is involved in heart failure caused by cardiac-specific expression of Cre, on which oxidative stress results in cytoplasmic vacuole accumulation of lipid peroxidation on the myocardial cell membrane. Taken together, these results revealed that cardiac-specific expression of Cre recombinase can lead to atrial mesenchymal tumor-like growth in the mice, which causes cardiac dysfunction, including cardiac fibrosis, reduction of the intercalated disc and cardiomyocytes ferroptosis at the age older than six months in mice. Our study suggests that αMHC-Cre mouse models are effective in young mice, but not in old mice. Researchers need to be particularly careful when using αMHC-Cre mouse model to interpret those phenotypic impacts of gene responses. As the Cre-associated cardiac pathology matched mostly to that of the patients, the model could also be employed for investigating age-related cardiac dysfunction.
转基因表达的 Cre 重组酶由特定的启动子驱动,通常用于以组织或细胞类型特异性的方式条件性敲除基因。在αMHC-Cre 转基因小鼠模型中,Cre 重组酶的表达受心肌特异性α肌球蛋白重链(αMHC)启动子的控制,该启动子常用于编辑心肌特异性基因。已经报道了 Cre 表达的毒性作用,包括染色体内重排、微核形成和其他形式的 DNA 损伤,并且在心脏特异性 Cre 转基因小鼠中观察到心肌病。然而,与 Cre 相关的心脏毒性的机制仍知之甚少。在我们的研究中,我们的数据揭示了αMHC-Cre 小鼠在六个月后逐渐出现心律失常并死亡,没有一只存活超过一年。组织病理学检查显示,αMHC-Cre 小鼠的心房腔中出现了肿瘤样组织的异常增殖,并伴有心室肌细胞的空泡化。此外,αMHC-Cre 小鼠发生严重的心脏间质和血管周围纤维化,并伴有心房和心室中 MMP-2 和 MMP-9 表达水平的显著增加。此外,心脏特异性表达 Cre 导致闰盘解体,以及盘的改变蛋白表达和钙处理异常。综上所述,我们确定铁死亡信号通路参与了心脏特异性表达 Cre 引起的心力衰竭,其中氧化应激导致心肌细胞膜上脂质过氧化的细胞质空泡积累。总的来说,这些结果表明,心脏特异性表达 Cre 重组酶可导致小鼠心房间质肿瘤样生长,导致心脏功能障碍,包括心脏纤维化、闰盘减少和 6 个月以上小鼠的心肌细胞铁死亡。我们的研究表明,αMHC-Cre 小鼠模型在幼鼠中是有效的,但在老年鼠中则不然。研究人员在使用αMHC-Cre 小鼠模型来解释基因反应的那些表型影响时需要特别小心。由于 Cre 相关的心脏病理学与大多数患者的病理学最为匹配,因此该模型也可用于研究与年龄相关的心脏功能障碍。