Stanford Cardiovascular Institute and Division of Cardiovascular Medicine, Department of Medicine, Stanford University School of Medicine, Stanford, California, USA. Electronic address: https://twitter.com/gentaro_ikeda.
Stanford Cardiovascular Institute and Division of Cardiovascular Medicine, Department of Medicine, Stanford University School of Medicine, Stanford, California, USA. Electronic address: https://twitter.com/meeshsantoso.
J Am Coll Cardiol. 2021 Mar 2;77(8):1073-1088. doi: 10.1016/j.jacc.2020.12.060.
Mitochondrial dysfunction results in an imbalance between energy supply and demand in a failing heart. An innovative therapy that targets the intracellular bioenergetics directly through mitochondria transfer may be necessary.
The purpose of this study was to establish a preclinical proof-of-concept that extracellular vesicle (EV)-mediated transfer of autologous mitochondria and their related energy source enhance cardiac function through restoration of myocardial bioenergetics.
Human-induced pluripotent stem cell-derived cardiomyocytes (iCMs) were employed. iCM-conditioned medium was ultracentrifuged to collect mitochondria-rich EVs (M-EVs). Therapeutic effects of M-EVs were investigated using in vivo murine myocardial infarction (MI) model.
Electron microscopy revealed healthy-shaped mitochondria inside M-EVs. Confocal microscopy showed that M-EV-derived mitochondria were transferred into the recipient iCMs and fused with their endogenous mitochondrial networks. Treatment with 1.0 × 10/ml M-EVs significantly restored the intracellular adenosine triphosphate production and improved contractile profiles of hypoxia-injured iCMs as early as 3 h after treatment. In contrast, isolated mitochondria that contained 300× more mitochondrial proteins than 1.0 × 10/ml M-EVs showed no effect after 24 h. M-EVs contained mitochondrial biogenesis-related messenger ribonucleic acids, including proliferator-activated receptor γ coactivator-1α, which on transfer activated mitochondrial biogenesis in the recipient iCMs at 24 h after treatment. Finally, intramyocardial injection of 1.0 × 10 M-EVs demonstrated significantly improved post-MI cardiac function through restoration of bioenergetics and mitochondrial biogenesis.
M-EVs facilitated immediate transfer of their mitochondrial and nonmitochondrial cargos, contributing to improved intracellular energetics in vitro. Intramyocardial injection of M-EVs enhanced post-MI cardiac function in vivo. This therapy can be developed as a novel, precision therapeutic for mitochondria-related diseases including heart failure.
线粒体功能障碍导致衰竭心脏的能量供应和需求失衡。一种通过线粒体转移直接靶向细胞内生物能量学的创新疗法可能是必要的。
本研究旨在建立一个临床前概念验证，即通过恢复心肌生物能量学，细胞外囊泡（EV）介导的自体线粒体及其相关能量源转移可增强心脏功能。
使用人诱导多能干细胞衍生的心肌细胞（iCMs）。iCM 条件培养基经超速离心收集富含线粒体的 EV（M-EVs）。使用体内小鼠心肌梗死（MI）模型研究 M-EV 的治疗效果。
电子显微镜显示 M-EVs 内有形状健康的线粒体。共焦显微镜显示 M-EV 衍生的线粒体被转移到受体 iCMs 中，并与它们的内源性线粒体网络融合。用 1.0×10/ml M-EV 处理可显著恢复缺氧损伤 iCMs 的细胞内三磷酸腺苷产生，并在治疗后 3 小时即可改善收缩轮廓。相比之下，尽管含有比 1.0×10/ml M-EV 多 300 倍的线粒体蛋白，但分离的线粒体在 24 小时后没有效果。M-EVs 包含与线粒体生物发生相关的信使 RNA，包括激活蛋白-1α辅激活因子-1，在治疗后 24 小时转移到受体 iCMs 中可激活线粒体生物发生。最后，心肌内注射 1.0×10 M-EVs 通过恢复生物能量和线粒体生物发生，显著改善 MI 后的心脏功能。
M-EVs 促进其线粒体和非线粒体货物的即时转移，有助于改善体外细胞内能量学。心肌内注射 M-EVs 增强了体内 MI 后的心脏功能。这种治疗方法可以作为一种新的、精准的治疗方法，用于治疗包括心力衰竭在内的与线粒体相关的疾病。
