Review

. 2024 Jun;20(6):1213-1246.

doi: 10.1080/15548627.2024.2319901. Epub 2024 Mar 24.

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

Xin Chen¹, Andrey S Tsvetkov², Han-Ming Shen³, Ciro Isidoro⁴, Nicholas T Ktistakis⁵, Andreas Linkermann^{6

7}, Werner J H Koopman^{8

9}, Hans-Uwe Simon^{10

11}, Lorenzo Galluzzi^{12

13

14}, Shouqing Luo¹⁵, Daqian Xu¹⁶, Wei Gu¹⁷, Olivier Peulen¹⁸, Qian Cai¹⁹, David C Rubinsztein^{20

21}, Jen-Tsan Chi²², Donna D Zhang²³, Changfeng Li²⁴, Shinya Toyokuni^{25

26}, Jinbao Liu²⁷, Jong-Lyel Roh²⁸, Enyong Dai²⁹, Gabor Juhasz^{30

31}, Wei Liu³², Jianhua Zhang³³, Minghua Yang^{34

35}, Jiao Liu³⁶, Ling-Qiang Zhu³⁷, Weiping Zou³⁸, Mauro Piacentini^{39

40}, Wen-Xing Ding⁴¹, Zhenyu Yue⁴², Yangchun Xie⁴³, Morten Petersen⁴⁴, David A Gewirtz⁴⁵, Michael A Mandell⁴⁶, Charleen T Chu⁴⁷, Debasish Sinha⁴⁸, Eftekhar Eftekharpour^{49

50}, Boris Zhivotovsky^{51

52

53}, Sébastien Besteiro^{54

55}, Dmitry I Gabrilovich⁵⁶, Do-Hyung Kim⁵⁷, Valerian E Kagan⁵⁸, Hülya Bayir⁵⁹, Guang-Chao Chen⁶⁰, Scott Ayton⁶¹, Jan D Lünemann⁶², Masaaki Komatsu⁶³, Stefan Krautwald⁶⁴, Ben Loos⁶⁵, Eric H Baehrecke⁶⁶, Jiayi Wang^{67

68

69}, Jon D Lane⁷⁰, Junichi Sadoshima⁷¹, Wan Seok Yang⁷², Minghui Gao⁷³, Christian Münz⁷⁴, Michael Thumm⁷⁵, Martin Kampmann^{76

77}, Di Yu^{78

79}, Marta M Lipinski⁸⁰, Jace W Jones⁸¹, Xuejun Jiang⁸², Herbert J Zeh⁸³, Rui Kang⁸³, Daniel J Klionsky⁸⁴, Guido Kroemer^{85

50

55}, Daolin Tang⁸³

Affiliations

¹ Guangzhou Municipal and Guangdong Provincial Key Laboratory of Protein Modification and Degradation, School of Basic Medical Sciences, Guangzhou Medical University, Guangzhou, China.
² Department of Neurology, The University of Texas McGovern Medical School at Houston, Houston, TX, USA.
³ Department of Biomedical Sciences, Faculty of Health Sciences, University of Macau, Macau, China.
⁴ Department of Health Sciences, University of Piemonte Orientale, Novara, Italy.
⁵ Babraham Institute, Cambridge, UK.
⁶ Division of Nephrology, Department of Internal Medicine 3, University Hospital Carl Gustav Carus at the Technische Universität Dresden, Germany.
⁷ Division of Nephrology, Department of Medicine, Albert Einstein College of Medicine, Bronx, New York, USA.
⁸ Department of Pediatrics, Radboud Center for Mitochondrial Medicine, Radboud University Medical Center, Nijmegen, The Netherlands.
⁹ Human and Animal Physiology, Wageningen University, Wageningen, The Netherlands.
¹⁰ Institute of Pharmacology, University of Bern, Bern, Switzerland.
¹¹ Institute of Biochemistry, Brandenburg Medical School, Neuruppin, Germany.
¹² Department of Radiation Oncology, Weill Cornell Medical College, New York, NY, USA.
¹³ Sandra and Edward Meyer Cancer Center, New York, NY, USA.
¹⁴ Caryl and Israel Englander Institute for Precision Medicine, New York, NY, USA.
¹⁵ Peninsula Medical School, University of Plymouth, Plymouth, UK.
¹⁶ Zhejiang Provincial Key Laboratory of Pancreatic Disease, The First Affiliated Hospital, Institute of Translational Medicine, Zhejiang University School of Medicine, Zhejiang University, Hangzhou, China.
¹⁷ Institute for Cancer Genetics, and Department of Pathology and Cell Biology, Vagelos College of Physicians & Surgeons, Columbia University, New York, NY, USA.
¹⁸ Metastasis Research Laboratory, GIGA Cancer-University of Liège, Liège, Belgium.
¹⁹ Department of Cell Biology and Neuroscience, Rutgers, The State University of New Jersey, Piscataway, NJ, USA.
²⁰ Department of Medical Genetics, Cambridge Institute for Medical Research, University of Cambridge, Cambridge, UK.
²¹ UK Dementia Research Institute, University of Cambridge, Cambridge, UK.
²² Department of Molecular Genetics and Microbiology, Duke University, Durham, NC, USA.
²³ Pharmacology and Toxicology, R. Ken Coit College of Pharmacy, University of Arizona, Tucson, AZ, USA.
²⁴ Department of Endoscopy Center, China-Japan Union Hospital of Jilin University, Changchun, China.
²⁵ Department of Pathology and Biological Response, Nagoya University Graduate School of Medicine, Nagoya, Japan.
²⁶ Center for Low-temperature Plasma Sciences, Nagoya University, Nagoya, Japan.
²⁷ Affiliated Cancer Hospital & Institute of Guangzhou Medical University, Guangzhou Municipal and Guangdong Provincial Key Laboratory of Protein Modification and Degradation, State Key Laboratory of Respiratory Disease, School of Basic Medical Sciences, Guangzhou Medical University, Guangzhou, China.
²⁸ Department of Otorhinolaryngology-Head and Neck Surgery, CHA Bundang Medical Center, CHA University, Seongnam, Republic of Korea.
²⁹ The Second Department of Hematology and Oncology, China-Japan Union Hospital of Jilin University, Changchun, Jilin, China.
³⁰ Biological Research Center, Institute of Genetics, Szeged, Hungary.
³¹ Department of Anatomy, Cell and Developmental Biology, Eotvos Lorand University, Budapest, Hungary.
³² Department of Orthopedics, Changzheng Hospital, Second Affiliated Hospital of Naval Medical University, Shanghai, China.
³³ Department of Pathology, University of Alabama at Birmingham, Birmingham, AL, USA.
³⁴ Department of Pediatrics, The Third Xiangya Hospital, Central South University, Changsha, Hunan, China.
³⁵ Hunan Clinical Research Center of Pediatric Cancer, Changsha, China.
³⁶ DAMP Laboratory, Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, China.
³⁷ Department of Pathophysiology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
³⁸ Departments of Surgery and Pathology, University of Michigan Medical School, Ann Arbor, USA.
³⁹ Department of Biology, University of Rome "Tor Vergata", Rome, Italy.
⁴⁰ National Institute for Infectious Diseases IRCCS "Lazzaro Spallanzani", Rome, Italy.
⁴¹ Department of Pharmacology, Toxicology and Therapeutics, The University of Kansas Medical Center, Kansas City, KS, USA.
⁴² Department of Neurology, Friedman Brain Institute, Icahn School of Medicine at Mount Sinai, New York, NY, USA.
⁴³ Department of Oncology, Central South University, Changsha, Hunan, China.
⁴⁴ Functional genomics, Department of Biology, Copenhagen University, Denmark.
⁴⁵ Department of Pharmacology and Toxicology, Virginia Commonwealth University, Massey Cancer Center, Richmond, VA, USA.
⁴⁶ Department of Molecular Genetics and Microbiology, University of New Mexico, Albuquerque, USA.
⁴⁷ Pathology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA.
⁴⁸ Department of Ophthalmology, University of Pittsburgh, Pittsburgh, PA, USA; Wilmer Eye lnstitute, The Johns Hopkins University School of Medicine, Baltimore, MD, USA.
⁴⁹ Department of Physiology and Pathophysiology, University of Manitoba, Winnipeg, Canada.
⁵⁰ Metabolomics and Cell Biology Platforms, Gustave Roussy Cancer, Villejuif, France; Gustave Roussy Cancer, Villejuif, France.
⁵¹ Institute of Environmental Medicine, Karolinska Institutet, Stockholm, Sweden, Europe.
⁵² Faculty of Medicine, Lomonosov Moscow State University, Moscow, Russia.
⁵³ Engelhardt Institute of Molecular Biology, Moscow, Russia.
⁵⁴ LPHI, University Montpellier, CNRS, Montpellier, France.
⁵⁵ Institut du Cancer Paris CARPEM, Department of Biology, Hôpital Européen Georges Pompidou, AP-HP, Paris, France.
⁵⁶ AstraZeneca, Early Oncology R&D, Gaithersburg, MD, USA.
⁵⁷ Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Minneapolis, MN, USA.
⁵⁸ Department of Environmental Health, University of Pittsburgh, Pittsburgh, PA, USA.
⁵⁹ Department of Pediatrics, Columbia University, New York, USA.
⁶⁰ Institute of Biological Chemistry, Academia Sinica, Taipei, Taiwan.
⁶¹ Florey Institute, University of Melbourne, Parkville, Australia.
⁶² Department of Neurology with Institute of Translational Neurology, University of Münster, Münster, Germany.
⁶³ Department of Physiology, Juntendo University School of Medicine, Bunkyo-ku Tokyo, Japan.
⁶⁴ Department of Nephrology and Hypertension, University Hospital Schleswig-Holstein, Kiel, Germany.
⁶⁵ Department of Physiological Sciences, Stellenbosch University, Stellenbosch, South Africa.
⁶⁶ Department of Molecular, Cell and Cancer Biology, University of Massachusetts Chan Medical School, Worcester, MA, USA.
⁶⁷ Department of Clinical Laboratory, Shanghai Chest Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
⁶⁸ Shanghai Institute of Thoracic Oncology Shanghai Chest Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
⁶⁹ College of Medical Technology, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
⁷⁰ School of Biochemistry, University of Bristol, Bristol, UK.
⁷¹ Rutgers New Jersey Medical School, Department of Cell Biology and Molecular Medicine, Newark, USA.
⁷² Department of Biological Sciences, St. John's University, New York City, NY, USA.
⁷³ The HIT Center for Life Sciences, School of Life Science and Technology, Harbin Institute of Technology, Harbin, China.
⁷⁴ Institute of Experimental Immunology, University of Zürich, Zürich, Switzerland.
⁷⁵ Department of Cellular Biochemistry, University Medical Center Goettingen, Goettingen, Germany.
⁷⁶ Department of Biochemistry & Biophysics, University of California, San Francisco, USA.
⁷⁷ Institute for Neurodegenerative Diseases, University of California, San Francisco, USA.
⁷⁸ Faculty of Medicine, Frazer Institute, University of Queensland, Brisbane, Australia.
⁷⁹ Faculty of Medicine, Ian Frazer Centre for Children's Immunotherapy Research, Child Health Research Centre, University of Queensland, Brisbane, Australia.
⁸⁰ Department of Anesthesiology & Department of Neurobiology, University of Maryland School of Medicine, Baltimore, MD, USA.
⁸¹ Department of Pharmaceutical Sciences, School of Pharmacy, University of Maryland, Baltimore, MD, USA.
⁸² Cell Biology Program, Memorial Sloan Kettering Cancer Center, New York, NY, USA.
⁸³ Department of Surgery, UT Southwestern Medical Center, Dallas, TX, USA.
⁸⁴ Life Sciences Institute and Department of Molecular, Cellular and Developmental Biology, University of Michigan, Ann Arbor, MI, USA.
⁸⁵ Centre de Recherche des Cordeliers, Université de Paris, Sorbonne Université, INSERM U1138, Institut Universitaire de France, Paris, France.

PMID: 38442890
PMCID: PMC11210914
DOI: 10.1080/15548627.2024.2319901

Review

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

Xin Chen et al. Autophagy. 2024 Jun.

. 2024 Jun;20(6):1213-1246.

doi: 10.1080/15548627.2024.2319901. Epub 2024 Mar 24.

Authors

Affiliations

¹ Guangzhou Municipal and Guangdong Provincial Key Laboratory of Protein Modification and Degradation, School of Basic Medical Sciences, Guangzhou Medical University, Guangzhou, China.
² Department of Neurology, The University of Texas McGovern Medical School at Houston, Houston, TX, USA.
³ Department of Biomedical Sciences, Faculty of Health Sciences, University of Macau, Macau, China.
⁴ Department of Health Sciences, University of Piemonte Orientale, Novara, Italy.
⁵ Babraham Institute, Cambridge, UK.
⁶ Division of Nephrology, Department of Internal Medicine 3, University Hospital Carl Gustav Carus at the Technische Universität Dresden, Germany.
⁷ Division of Nephrology, Department of Medicine, Albert Einstein College of Medicine, Bronx, New York, USA.
⁸ Department of Pediatrics, Radboud Center for Mitochondrial Medicine, Radboud University Medical Center, Nijmegen, The Netherlands.
⁹ Human and Animal Physiology, Wageningen University, Wageningen, The Netherlands.
¹⁰ Institute of Pharmacology, University of Bern, Bern, Switzerland.
¹¹ Institute of Biochemistry, Brandenburg Medical School, Neuruppin, Germany.
¹² Department of Radiation Oncology, Weill Cornell Medical College, New York, NY, USA.
¹³ Sandra and Edward Meyer Cancer Center, New York, NY, USA.
¹⁴ Caryl and Israel Englander Institute for Precision Medicine, New York, NY, USA.
¹⁵ Peninsula Medical School, University of Plymouth, Plymouth, UK.
¹⁶ Zhejiang Provincial Key Laboratory of Pancreatic Disease, The First Affiliated Hospital, Institute of Translational Medicine, Zhejiang University School of Medicine, Zhejiang University, Hangzhou, China.
¹⁷ Institute for Cancer Genetics, and Department of Pathology and Cell Biology, Vagelos College of Physicians & Surgeons, Columbia University, New York, NY, USA.
¹⁸ Metastasis Research Laboratory, GIGA Cancer-University of Liège, Liège, Belgium.
¹⁹ Department of Cell Biology and Neuroscience, Rutgers, The State University of New Jersey, Piscataway, NJ, USA.
²⁰ Department of Medical Genetics, Cambridge Institute for Medical Research, University of Cambridge, Cambridge, UK.
²¹ UK Dementia Research Institute, University of Cambridge, Cambridge, UK.
²² Department of Molecular Genetics and Microbiology, Duke University, Durham, NC, USA.
²³ Pharmacology and Toxicology, R. Ken Coit College of Pharmacy, University of Arizona, Tucson, AZ, USA.
²⁴ Department of Endoscopy Center, China-Japan Union Hospital of Jilin University, Changchun, China.
²⁵ Department of Pathology and Biological Response, Nagoya University Graduate School of Medicine, Nagoya, Japan.
²⁶ Center for Low-temperature Plasma Sciences, Nagoya University, Nagoya, Japan.
²⁷ Affiliated Cancer Hospital & Institute of Guangzhou Medical University, Guangzhou Municipal and Guangdong Provincial Key Laboratory of Protein Modification and Degradation, State Key Laboratory of Respiratory Disease, School of Basic Medical Sciences, Guangzhou Medical University, Guangzhou, China.
²⁸ Department of Otorhinolaryngology-Head and Neck Surgery, CHA Bundang Medical Center, CHA University, Seongnam, Republic of Korea.
²⁹ The Second Department of Hematology and Oncology, China-Japan Union Hospital of Jilin University, Changchun, Jilin, China.
³⁰ Biological Research Center, Institute of Genetics, Szeged, Hungary.
³¹ Department of Anatomy, Cell and Developmental Biology, Eotvos Lorand University, Budapest, Hungary.
³² Department of Orthopedics, Changzheng Hospital, Second Affiliated Hospital of Naval Medical University, Shanghai, China.
³³ Department of Pathology, University of Alabama at Birmingham, Birmingham, AL, USA.
³⁴ Department of Pediatrics, The Third Xiangya Hospital, Central South University, Changsha, Hunan, China.
³⁵ Hunan Clinical Research Center of Pediatric Cancer, Changsha, China.
³⁶ DAMP Laboratory, Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, China.
³⁷ Department of Pathophysiology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
³⁸ Departments of Surgery and Pathology, University of Michigan Medical School, Ann Arbor, USA.
³⁹ Department of Biology, University of Rome "Tor Vergata", Rome, Italy.
⁴⁰ National Institute for Infectious Diseases IRCCS "Lazzaro Spallanzani", Rome, Italy.
⁴¹ Department of Pharmacology, Toxicology and Therapeutics, The University of Kansas Medical Center, Kansas City, KS, USA.
⁴² Department of Neurology, Friedman Brain Institute, Icahn School of Medicine at Mount Sinai, New York, NY, USA.
⁴³ Department of Oncology, Central South University, Changsha, Hunan, China.
⁴⁴ Functional genomics, Department of Biology, Copenhagen University, Denmark.
⁴⁵ Department of Pharmacology and Toxicology, Virginia Commonwealth University, Massey Cancer Center, Richmond, VA, USA.
⁴⁶ Department of Molecular Genetics and Microbiology, University of New Mexico, Albuquerque, USA.
⁴⁷ Pathology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA.
⁴⁸ Department of Ophthalmology, University of Pittsburgh, Pittsburgh, PA, USA; Wilmer Eye lnstitute, The Johns Hopkins University School of Medicine, Baltimore, MD, USA.
⁴⁹ Department of Physiology and Pathophysiology, University of Manitoba, Winnipeg, Canada.
⁵⁰ Metabolomics and Cell Biology Platforms, Gustave Roussy Cancer, Villejuif, France; Gustave Roussy Cancer, Villejuif, France.
⁵¹ Institute of Environmental Medicine, Karolinska Institutet, Stockholm, Sweden, Europe.
⁵² Faculty of Medicine, Lomonosov Moscow State University, Moscow, Russia.
⁵³ Engelhardt Institute of Molecular Biology, Moscow, Russia.
⁵⁴ LPHI, University Montpellier, CNRS, Montpellier, France.
⁵⁵ Institut du Cancer Paris CARPEM, Department of Biology, Hôpital Européen Georges Pompidou, AP-HP, Paris, France.
⁵⁶ AstraZeneca, Early Oncology R&D, Gaithersburg, MD, USA.
⁵⁷ Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Minneapolis, MN, USA.
⁵⁸ Department of Environmental Health, University of Pittsburgh, Pittsburgh, PA, USA.
⁵⁹ Department of Pediatrics, Columbia University, New York, USA.
⁶⁰ Institute of Biological Chemistry, Academia Sinica, Taipei, Taiwan.
⁶¹ Florey Institute, University of Melbourne, Parkville, Australia.
⁶² Department of Neurology with Institute of Translational Neurology, University of Münster, Münster, Germany.
⁶³ Department of Physiology, Juntendo University School of Medicine, Bunkyo-ku Tokyo, Japan.
⁶⁴ Department of Nephrology and Hypertension, University Hospital Schleswig-Holstein, Kiel, Germany.
⁶⁵ Department of Physiological Sciences, Stellenbosch University, Stellenbosch, South Africa.
⁶⁶ Department of Molecular, Cell and Cancer Biology, University of Massachusetts Chan Medical School, Worcester, MA, USA.
⁶⁷ Department of Clinical Laboratory, Shanghai Chest Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
⁶⁸ Shanghai Institute of Thoracic Oncology Shanghai Chest Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
⁶⁹ College of Medical Technology, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
⁷⁰ School of Biochemistry, University of Bristol, Bristol, UK.
⁷¹ Rutgers New Jersey Medical School, Department of Cell Biology and Molecular Medicine, Newark, USA.
⁷² Department of Biological Sciences, St. John's University, New York City, NY, USA.
⁷³ The HIT Center for Life Sciences, School of Life Science and Technology, Harbin Institute of Technology, Harbin, China.
⁷⁴ Institute of Experimental Immunology, University of Zürich, Zürich, Switzerland.
⁷⁵ Department of Cellular Biochemistry, University Medical Center Goettingen, Goettingen, Germany.
⁷⁶ Department of Biochemistry & Biophysics, University of California, San Francisco, USA.
⁷⁷ Institute for Neurodegenerative Diseases, University of California, San Francisco, USA.
⁷⁸ Faculty of Medicine, Frazer Institute, University of Queensland, Brisbane, Australia.
⁷⁹ Faculty of Medicine, Ian Frazer Centre for Children's Immunotherapy Research, Child Health Research Centre, University of Queensland, Brisbane, Australia.
⁸⁰ Department of Anesthesiology & Department of Neurobiology, University of Maryland School of Medicine, Baltimore, MD, USA.
⁸¹ Department of Pharmaceutical Sciences, School of Pharmacy, University of Maryland, Baltimore, MD, USA.
⁸² Cell Biology Program, Memorial Sloan Kettering Cancer Center, New York, NY, USA.
⁸³ Department of Surgery, UT Southwestern Medical Center, Dallas, TX, USA.
⁸⁴ Life Sciences Institute and Department of Molecular, Cellular and Developmental Biology, University of Michigan, Ann Arbor, MI, USA.
⁸⁵ Centre de Recherche des Cordeliers, Université de Paris, Sorbonne Université, INSERM U1138, Institut Universitaire de France, Paris, France.

PMID: 38442890
PMCID: PMC11210914
DOI: 10.1080/15548627.2024.2319901

Abstract

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.

Keywords: Cell death; ferritinophagy; iron; lipid peroxidation; lipophagy; lysosome.

PubMed Disclaimer

Conflict of interest statement

L.G. is/has been holding research contracts with Lytix Biopharma, Promontory and Onxeo, has received consulting/advisory honoraria from Boehringer Ingelheim, AstraZeneca, OmniSEQ, Onxeo, The Longevity Labs, Inzen, Imvax, Sotio, Promontory, Noxopharm, EduCom, and the Luke Heller TECPR2 Foundation, and holds Promontory stock options. D.C.R. is a consultant for Aladdin Healthcare Technologies Ltd., Mindrank AI, Nido Biosciences, Drishti Discoveries, Retro Biosciences and PAQ Therapeutics. D.I. G. is an employee and shareholder of AstraZeneca. M. K. serves on the Scientific Advisory Boards of Engine Biosciences, Casma Therapeutics, Cajal Neuroscience, Alector, and Montara Therapeutics, and is an advisor to Modulo Bio and Recursion Therapeutics. X.J. holds inventorship of patents related to autophagy and cell death, and holds equity as well as consults for Exarta Therapeutics and Lime Therapeutics. G.K. has been holding research contracts with Daiichi Sankyo, Eleor, Kaleido, Lytix Pharma, PharmaMar, Osasuna Therapeutics, Samsara Therapeutics, Sanofi, Tollys, and Vascage. G.K. is on the Board of Directors of the Bristol Myers Squibb Foundation France. G.K. is a scientific co-founder of everImmune, Osasuna Therapeutics, Samsara Therapeutics and Therafast Bio. G.K. is in the scientific advisory boards of Hevolution, Institut Servier and Longevity Vision Funds. G.K. is the inventor of patents covering therapeutic targeting of aging, cancer, cystic fibrosis and metabolic disorders. G.K.’wife, Laurence Zitvogel, has held research contracts with Glaxo Smyth Kline, Incyte, Lytix, Kaleido, Innovate Pharma, Daiichi Sankyo, Pilege, Merus, Transgene, 9 m, Tusk and Roche, was on the on the Board of Directors of Transgene, is a cofounder of everImmune, and holds patents covering the treatment of cancer and the therapeutic manipulation of the microbiota. G.K.’s brother, Romano Kroemer, was an employee of Sanofi and now consults for Boehringer-Ingelheim. The other authors declare no conflicts of interest or financial interests. The funders had no role in the writing of the manuscript.

Figures

**Figure 1.**
Overview of ferroptosis. The SLC7A11-GSH-GPX4 pathway and GPX4-independent pathways (such as AIFM2, GCH, DHODH, and MGST1) inhibit lipid peroxidation, whereas ACSL4, ALOX, and POR promote peroxidation of PUFA-containing phospholipids (PUFA-PL). Cystine is transported into cells by SLC7A11 and rapidly reduced to cysteine, which is utilized for GSH synthesis. GPX4 inhibits lipid peroxidation, the primary driver of ferroptosis, by converting GSH to GSSG. The transcription factor NFE2L2 serves as a key antioxidant system by upregulating genes involved in both GPX4-dependent and GPX4-independent pathways. Lipid peroxidation generates 4HNE, which can induce membrane damage and release DAMPs. Conversely, the endosomal sorting complex required for transport (ESCRT-III) machinery acts as a protective mechanism that delays membrane damage. TFRC increases intracellular iron levels, thereby promoting lipid peroxidation and ferroptosis.

**Figure 2.**
Classification of ferroptosis based on autophagic response. Autophagy-dependent ferroptosis: this process relies on the autophagic machinery to induce ferroptosis. Autophagy-independent ferroptosis: In this case, autophagy occurs alongside ferroptosis but does not directly induce ferroptosis or may even have a protective role in the context of ferroptosis.

**Figure 3.**
Ferritinophagy in ferroptosis. Ferritin is a cytosolic protein involved in iron storage, and NCOA4 serves as a receptor for the autophagic degradation of ferritin, a process referred to as ferritinophagy. Conditions and factors such as hypoxia, TRIM7, and HERC2 decrease NCOA4 expression, thereby inhibiting ferritinophagy-dependent ferroptosis. Conversely, PTBP1 and ATM promote NCOA4-mediated ferritinophagy. Additionally, ALDH1A3 can bind to MAP1LC3 to enhance ferritinophagy-dependent ferroptosis.

**Figure 4.**
Lipophagy in ferroptosis. Lipophagy refers to the autophagic clearance of lipid droplets, which function as storage organelles for neutral lipids. RAB7A, associated with lipid droplets, drives lipophagy and promotes subsequent ferroptosis. The formation of lipid droplets, mediated by TPD52 and PLTP, antagonizes lipophagy-dependent ferroptosis. In contrast, PGRMC1 triggers lipophagy-dependent ferroptosis. PUFA, polyunsaturated fatty acid.

**Figure 5.**
Mitophagy in ferroptosis. Mitophagy is a selective process that involves the autophagic clearance of damaged or dysfunctional mitochondria. The PINK1-PRKN pathway is a well-studied regulatory pathway for mitophagy. In this pathway, PINK1 recruits PRKN, which facilitates the degradation of mitochondria through mitophagy receptors such as FUNDC1. Mitochondrial fission induced by DNM1L/Drp1 can promote mitophagy-dependent ferroptosis. Additionally, the mitochondrial protein FTMT inhibits ROS production and ferroptosis.

**Figure 6.**
Clockophagy in ferroptosis. BMAL1/ARNTL, a transcription factor involved in the circadian clock, undergoes selective degradation through autophagy, a process known as clockophagy. The autophagy receptor SQSTM1/p62 plays a role in recognizing and degrading BMAL1 during clockophagy. Degradation of BMAL1 results in the upregulation of *EGLN2*, a target gene of BMAL1. EGLN2, in turn, inhibits the function of HIF1A, which acts as a suppressor of ferroptosis by inducing the expression of FABP3 and FABP7. These proteins regulate fatty acid uptake and lipid storage, thereby limiting the availability of polyunsaturated fatty acid (PUFA) and influencing the susceptibility to ferroptosis.

**Figure 7.**
CMA- or TAX1BP1-mediated GPX4 degradation in ferroptosis. CMA serves as a pathway for GPX4 protein degradation during ferroptosis. LAMP2 plays a key role in transporting GPX4 across the lysosomal membrane, and HSPA8/HSC70 interacts with GPX4 and LAMP2, facilitating CMA. CMA-mediated degradation of GPX4 can be inhibited by CKB-mediated GPX4 phosphorylation. LGMN assists in CMA-mediated GPX4 degradation. Furthermore, macroautophagy/autophagy also contributes to GPX4 degradation during ferroptosis. SMPD1/ASM promotes the autophagic degradation of GPX4, whereas MTOR inhibits GPX4 protein degradation and subsequent ferroptosis. Copper directly binds to GPX4, leading to its autophagic degradation, and, in this process, TAX1BP1 acts as an autophagic receptor.

**Figure 8.**
Autophagy-dependent ferroptosis in diseases. Autophagy-dependent ferroptosis plays a role in various diseases, encompassing both cancer and non-cancerous conditions.

**Figure 9.**
Reaction between MDA and TBA to form the MDA-TBA adduct.

**Figure 10.**
Fluorescent probe-based assays for monitoring lipid peroxidation. (A) BODIPY − 581/591-C11 assay; (B) click-iT LAA assay; (C) LiperFluo assay.

**Figure 11.**
Assessing Fe²⁺ with the FerroOrange assay.

**Figure 12.**
Assessing Fe²⁺ with the phen green SK assay.

See this image and copyright information in PMC

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

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

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