College of Pharmacy, Jinan University, Guangzhou 510632, China.
Post-Doctoral Innovation Base, Jinan University Affiliation, Yuanzhi Health Technology Co., Ltd., Hengqin New District, Zhuhai, Guangdong 51900, China.
Int J Mol Sci. 2021 Oct 6;22(19):10806. doi: 10.3390/ijms221910806.
Heart failure is the end-stage of all cardiovascular diseases with a ~25% 5-year survival rate, and insufficient mitochondrial energy production to meet myocardial demand is the hallmark of heart failure. Mitochondrial components involved in the regulation of ATP production remain to be fully elucidated. Recently, roles of 2',3'-cyclic nucleotide-3'-phosphodiesterase (CNPase) in the pathophysiological processes of heart diseases have emerged, implicated by evidence that mitochondrial CNPase proteins are associated with mitochondrial integrity under metabolic stress. In this study, a zebrafish heart failure model was established, by employing antisense morpholino oligonucleotides and the CRISPR-Cas9 gene-editing system, which recapitulates heart failure phenotypes including heart dysfunction, pericardial edema, ventricular enlargement, bradycardia, and premature death. The translational implications of CNPase in the pathophysiological process of heart failure were tested in a pressure overload-induced heart hypertrophy model, which was carried out in rats through transverse abdominal aorta constriction (TAAC). AAV9-mediated myocardial delivery of CNPase mitigated the hypertrophic response through the specific hydrolysis of 2'-3'-cyclic nucleotides, supported by the decrease of cardiac hypertrophy and fibrosis, the integrity of mitochondrial ultrastructure, and indicators of heart contractility in the AAV9-TAAC group. Finally, the biometrics of a mitochondrial respiration assay carried out on a Seahorse cellular energy analyzer demonstrated that CNPase protects mitochondrial respiration and ATP production from AngII-induced metabolic stress. In summary, this study provides mechanistic insights into CNPase-2',3'-cyclic nucleotide metabolism that protects the heart from energy starvation and suggests novel therapeutic approaches to treat heart failure by targeting CNPase activity.
心力衰竭是所有心血管疾病的终末期阶段，5 年生存率约为 25%，心肌对能量的需求得不到满足是心力衰竭的标志。目前仍未完全阐明参与调节 ATP 产生的线粒体成分。最近，2'，3'-环核苷酸 3'-磷酸二酯酶（CNPase）在心脏疾病的病理生理过程中的作用已经显现出来，有证据表明，在代谢应激下，线粒体 CNPase 蛋白与线粒体的完整性有关。在这项研究中，通过反义寡核苷酸和 CRISPR-Cas9 基因编辑系统建立了一种斑马鱼心力衰竭模型，该模型再现了心力衰竭表型，包括心脏功能障碍、心包水肿、心室扩大、心动过缓和过早死亡。通过在大鼠中进行横腹主动脉缩窄（TAAC），建立压力超负荷诱导的心脏肥厚模型，测试了 CNPase 在心力衰竭病理生理过程中的转化意义。AAV9 介导的心肌 CNPase 传递通过特异性水解 2'，3'-环核苷酸，减轻了肥厚反应，AAV9-TAAC 组的心脏肥大和纤维化减少，线粒体超微结构的完整性以及心脏收缩性指标得到改善。最后，在 Seahorse 细胞能量分析仪上进行的线粒体呼吸测定的生物统计学表明，CNPase 可保护线粒体呼吸和 ATP 产生免受 AngII 引起的代谢应激。总之，这项研究提供了 CNPase-2'，3'-环核苷酸代谢保护心脏免受能量饥饿的机制见解，并提出了通过靶向 CNPase 活性治疗心力衰竭的新治疗方法。
