Peng Fei, Chen Xiaoniao, Wu Lingling, He Jiayi, Li Zongjin, Hong Quan, Zhao Qiang, Qian Meng, Wang Xu, Shen Wanjun, Qi Tingting, Huang Yiyu, Cai Guangyan, Zhang Chuyue, Chen Xiangmei
School of Medicine, Nankai University, Tianjin 300071, China.
Department of Nephrology, First Medical Center of Chinese PLA General Hospital, National Key Laboratory of Kidney Diseases, National Clinical Research Center for Kidney Diseases, Beijing Key Laboratory of Kidney Diseases Research, Beijing 100853, China.
Theranostics. 2025 Apr 13;15(11):5499-5517. doi: 10.7150/thno.113741. eCollection 2025.
The disruption of mitochondrial homeostasis in acute kidney injury (AKI) is an important factor that drives persistent renal dysfunction. Mesenchymal stem cell-derived extracellular vesicles (MSC-EVs) have shown great therapeutic potential in AKI, but insufficient specificity of targeting the impaired mitochondrial function. Herein, we developed an engineered nitric oxide (NO)-primed MSC-EVs (pEVs) to restore mitochondrial homeostasis for AKI therapy. A cisplatin-induced AKI model was established to investigate the therapeutic effects of MSC-EVs. Proteomic and Western blot analyses compared mitochondrial cargos and functional assays included mitochondrial complex I activity and Adenosine triphosphate (ATP) quantification. Mitochondrial transfer was tracked using flow cytometry and confocal imaging. Mitochondrial dynamics, oxidative stress, and apoptosis were evaluated through ATP measurement, western blotting and rotenone-mediated respiratory chain inhibition. Our data indicated that pEVs outperformed cEVs in restoring renal function and histopathology. Additionally, a reduction in mitochondria-associated oxidative stress and cell death was observed. Proteomic profiling revealed that NO priming enriched pEVs with mitochondrial complex I components, which directly enhanced their bioenergetic capacity, as evidenced by higher mitochondrial complex I activity and elevated ATP production compared to cEVs. In vivo tracking confirmed targeted delivery of pEV-derived mitochondrial contents to renal tubular cells, reducing mitochondrial reactive oxygen species (ROS) and restoring mitochondrial mass. Crucially, mitochondria-depleted pEVs abolished these therapeutic effects, establishing mitochondrial cargos as the primary therapeutic driver. Furthermore, pEVs activated a pro-survival cascade in recipient cells, showing superior efficacy in promoting mitochondrial biogenesis, dynamics, and mitophagy, thereby restoring renal mitochondrial homeostasis. Our study elucidated a mitochondria-targeted therapeutic strategy enabled by engineered EVs that deliver functional cargo to restore mitochondrial homeostasis. These advances provide transformative potential for AKI and other mitochondrial disorders.
急性肾损伤（AKI）中线粒体稳态的破坏是导致持续性肾功能障碍的重要因素。间充质干细胞衍生的细胞外囊泡（MSC-EVs）在AKI治疗中显示出巨大的治疗潜力，但靶向受损线粒体功能的特异性不足。在此，我们开发了一种工程化一氧化氮（NO）预处理的MSC-EVs（pEVs），以恢复线粒体稳态用于AKI治疗。建立了顺铂诱导的AKI模型来研究MSC-EVs的治疗效果。蛋白质组学和蛋白质印迹分析比较了线粒体货物，功能测定包括线粒体复合物I活性和三磷酸腺苷（ATP）定量。使用流式细胞术和共聚焦成像追踪线粒体转移。通过ATP测量、蛋白质印迹和鱼藤酮介导的呼吸链抑制评估线粒体动力学、氧化应激和细胞凋亡。我们的数据表明，pEVs在恢复肾功能和组织病理学方面优于cEVs。此外，观察到线粒体相关氧化应激和细胞死亡减少。蛋白质组学分析表明，NO预处理使pEVs富含线粒体复合物I成分，这直接增强了它们的生物能量能力，与cEVs相比，线粒体复合物I活性更高和ATP产量升高证明了这一点。体内追踪证实pEV衍生的线粒体内容物靶向递送至肾小管细胞，减少线粒体活性氧（ROS）并恢复线粒体质量。至关重要的是，线粒体耗尽的pEVs消除了这些治疗效果，确定线粒体货物是主要的治疗驱动因素。此外，pEVs在受体细胞中激活了促生存级联反应，在促进线粒体生物发生、动力学和线粒体自噬方面显示出卓越的功效，从而恢复肾线粒体稳态。我们的研究阐明了一种由工程化细胞外囊泡实现的线粒体靶向治疗策略，该策略递送功能性货物以恢复线粒体稳态。这些进展为AKI和其他线粒体疾病提供了变革潜力。
