Artesunate-induced mitophagy alters cellular redox status
Jianbin Zhang,
Xin Sun,
Liming Wang,
Yin Kwan Wong,
Yew Mun Lee,
Chao Zhou,
Guoqing Wu,
Tongwei Zhao,
Liu Yang,
Liqin Lu,
Jianing Zhong,
Dongsheng Huang,
Jigang Wang
Affiliations
Jianbin Zhang
Department of Oncology, Zhejiang Provincial People's Hospital, People's Hospital of Hangzhou Medical College, Hangzhou 310014, China; Clinical Research Institute, Key Laboratory of Tumor Molecular Diagnosis and Individual Medicine of Zhejiang Province, Zhejiang Provincial People's Hospital, People's Hospital of Hangzhou Medical College, Hangzhou 310014, China; Key Laboratory of Cardio-cerebrovascular disease prevention & therapy, Gannan Medical University, Ganzhou 341000, China; Correspondence to: Department of Oncology, Clinical Research Institute, Key Laboratory of Tumor Molecular Diagnosis and Individualized Medicine of Zhejiang Province, Zhejiang Provincial People's Hospital, People's Hospital of Hangzhou Medical College, Hangzhou 310014, China.
Xin Sun
Department of Oncology, Zhejiang Provincial People's Hospital, People's Hospital of Hangzhou Medical College, Hangzhou 310014, China
Liming Wang
Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, 117593, Singapore
Yin Kwan Wong
Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, 117593, Singapore; Artemisinin Research Center, China Academy of Chinese Medical Sciences, Beijing 100700, China
Yew Mun Lee
Department of Pharmacology, National University of Singapore, 117600, Singapore
Chao Zhou
Department of Oncology, Zhejiang Provincial People's Hospital, People's Hospital of Hangzhou Medical College, Hangzhou 310014, China
Guoqing Wu
Department of Oncology, Zhejiang Provincial People's Hospital, People's Hospital of Hangzhou Medical College, Hangzhou 310014, China
Tongwei Zhao
Department of Oncology, Zhejiang Provincial People's Hospital, People's Hospital of Hangzhou Medical College, Hangzhou 310014, China
Liu Yang
Clinical Research Institute, Key Laboratory of Tumor Molecular Diagnosis and Individual Medicine of Zhejiang Province, Zhejiang Provincial People's Hospital, People's Hospital of Hangzhou Medical College, Hangzhou 310014, China
Liqin Lu
Department of Oncology, Zhejiang Provincial People's Hospital, People's Hospital of Hangzhou Medical College, Hangzhou 310014, China
Jianing Zhong
Key Laboratory of Cardio-cerebrovascular disease prevention & therapy, Gannan Medical University, Ganzhou 341000, China
Dongsheng Huang
Clinical Research Institute, Key Laboratory of Tumor Molecular Diagnosis and Individual Medicine of Zhejiang Province, Zhejiang Provincial People's Hospital, People's Hospital of Hangzhou Medical College, Hangzhou 310014, China; Correspondence to: Clinical Research Institute, Key Laboratory of Tumor Molecular Diagnosis and Individualized Medicine of Zhejiang Province, Zhejiang Provincial People's Hospital, Hangzhou 310014, China.
Jigang Wang
Artemisinin Research Center, China Academy of Chinese Medical Sciences, Beijing 100700, China; Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China; Department of Pharmacology, National University of Singapore, 117600, Singapore; Key Laboratory of Cardio-cerebrovascular disease prevention & therapy, Gannan Medical University, Ganzhou 341000, China; Corresponding author at: Artemisinin Research Center, Institute of Chinese Materia Medica, and China Academy of Chinese Medical Sciences, Beijing 100700, China.
Artesunate (ART) is a prominent anti-malarial with significant anti-cancer properties. Our previous studies showed that ART enhances lysosomal function and ferritin degradation, which was necessary for its anti-cancer properties. ART targeting to mitochondria also significantly improved its efficacy, but the effect of ART on mitophagy, an important cellular pathway that facilitates the removal of damaged mitochondria, remains unknown. Here, we first observed that ART mainly localizes in the mitochondria and its probe labeling revealed that it binds to a large number of mitochondrial proteins and causes mitochondrial fission. Second, we found that ART treatment leads to autophagy induction and the decrease of mitochondrial proteins. When autophagy is inhibited, the decrease of mitochondrial proteins could be reversed, indicating that the degradation of mitochondrial proteins is through mitophagy. Third, our results showed that ART treatment stabilizes the full-length form of PTEN induced putative kinase 1 (PINK1) on the mitochondria and activates the PINK1-dependent pathway. This in turn leads to the recruitment of Parkin, sequestosome 1 (SQSTM1), ubiquitin and microtubule-associated proteins 1A/1B light chain 3 (LC3) to the mitochondria and culminates in mitophagy. When PINK1 is knocked down, ART-induced mitophagy is markedly suppressed. Finally, we investigated the effect of mitophagy by ART on mitochondrial functions and found that knockdown of PINK1 alters the cellular redox status in ART-treated cells, which is accompanied with a significant decrease in glutathione (GSH) and increase in mitochondrial reactive oxidative species (mROS) and cellular lactate levels. Additionally, knockdown of PINK1 leads to a significant increase of mitochondrial depolarization and more cell apoptosis by ART, suggesting that mitophagy protects from ART-induced cell death. Taken together, our findings reveal the molecular mechanism that ART induces cytoprotective mitophagy through the PINK1-dependent pathway, suggesting that mitophagy inhibition could enhance the anti-cancer activity of ART. Keywords: Artesunate, Artemisinin, Mitophagy, PINK1, Parkin, ROS
