International consensus guidelines for the definition, detection, and interpretation of autophagy-dependent ferroptosis

Chen, Xin; Tsvetkov, Andrey S.; Shen, Han Ming; Isidoro, Ciro; Ktistakis, Nicholas T.; Linkermann, Andreas; Koopman, Werner J.H.; Simon, Hans Uwe; Galluzzi, Lorenzo; Luo, Shouqing; Xu, Daqian; Gu, Wei; Peulen, Olivier; Cai, Qian; Rubinsztein, David C.; Chi, Jen Tsan; Zhang, Donna D.; Li, Changfeng; Toyokuni, Shinya; Liu, Jinbao; Roh, Jong Lyel; Dai, Enyong; Juhasz, Gabor; Liu, Wei; Zhang, Jianhua; Yang, Minghua; Liu, Jiao; Zhu, Ling Qiang; Zou, Weiping; Piacentini, Mauro; Ding, Wen Xing; Yue, Zhenyu; Xie, Yangchun; Petersen, Morten; Gewirtz, David A.; Mandell, Michael A.; Chu, Charleen T.; Sinha, Debasish; Eftekharpour, Eftekhar; Zhivotovsky, Boris; Besteiro, Sébastien; Gabrilovich, Dmitry I.; Kim, Do Hyung; Kagan, Valerian E.; Bayir, Hülya; Chen, Guang Chao; Ayton, Scott; Lünemann, Jan D.; Komatsu, Masaaki; Krautwald, Stefan; Loos, Ben; Baehrecke, Eric H.; Wang, Jiayi; Lane, Jon D.; Sadoshima, Junichi; Yang, Wan Seok; Gao, Minghui; Münz, Christian; Thumm, Michael; Kampmann, Martin; Yu, Di; Lipinski, Marta M.; Jones, Jace W.; Jiang, Xuejun; Zeh, Herbert J.; Kang, Rui; Klionsky, Daniel J.; Kroemer, Guido; Tang, Daolin


Macroautophagy/autophagy is a complex degradation process with a dual role in cell death that is influenced by the cell types that are involved and the stressors they are exposed to. Ferroptosis is an iron-dependent oxidative form of cell death characterized by unrestricted lipid peroxidation in the context of heterogeneous and plastic mechanisms. Recent studies have shed light on the involvement of specific types of autophagy (e.g. ferritinophagy, lipophagy, and clockophagy) in initiating or executing ferroptotic cell death through the selective degradation of anti-injury proteins or organelles. Conversely, other forms of selective autophagy (e.g. reticulophagy and lysophagy) enhance the cellular defense against ferroptotic damage. Dysregulated autophagy-dependent ferroptosis has implications for a diverse range of pathological conditions. This review aims to present an updated definition of autophagy-dependent ferroptosis, discuss influential substrates and receptors, outline experimental methods, and propose guidelines for interpreting the results. Abbreviation: 3-MA:3-methyladenine; 4HNE: 4-hydroxynonenal; ACD: accidentalcell death; ADF: autophagy-dependentferroptosis; ARE: antioxidant response element; BH2:dihydrobiopterin; BH4: tetrahydrobiopterin; BMDMs: bonemarrow-derived macrophages; CMA: chaperone-mediated autophagy; CQ:chloroquine; DAMPs: danger/damage-associated molecular patterns; EMT,epithelial-mesenchymal transition; EPR: electronparamagnetic resonance; ER, endoplasmic reticulum; FRET: Försterresonance energy transfer; GFP: green fluorescent protein;GSH: glutathione;IF: immunofluorescence; IHC: immunohistochemistry; IOP, intraocularpressure; IRI: ischemia-reperfusion injury; LAA: linoleamide alkyne;MDA: malondialdehyde; PGSK: Phen Green™ SK;RCD: regulatedcell death; PUFAs: polyunsaturated fatty acids; RFP: red fluorescentprotein;ROS: reactive oxygen species; TBA: thiobarbituricacid; TBARS: thiobarbituric acid reactive substances; TEM:transmission electron microscopy.