Abstract
Ferroptosis is an iron-dependent form of regulated necrosis. It is implicated in various human diseases, including ischemic organ damage and cancer. Here, we report the crucial role of autophagy, particularly autophagic degradation of cellular iron storage proteins (a process known as ferritinophagy), in ferroptosis. Using RNAi screening coupled with subsequent genetic analysis, we identified multiple autophagy-related genes as positive regulators of ferroptosis. Ferroptosis induction led to autophagy activation and consequent degradation of ferritin and ferritinophagy cargo receptor NCOA4. Consistently, inhibition of ferritinophagy by blockage of autophagy or knockdown of NCOA4 abrogated the accumulation of ferroptosis-associated cellular labile iron and reactive oxygen species, as well as eventual ferroptotic cell death. Therefore, ferroptosis is an autophagic cell death process, and NCOA4-mediated ferritinophagy supports ferroptosis by controlling cellular iron homeostasis.
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References
Danial NN, Korsmeyer SJ . Cell death: Critical control points. Cell 2004; 116:205–219.
Green DR, Kroemer G . The pathophysiology of mitochondrial cell death. Science 2004; 305:626–629.
Fuchs Y, Steller H . Programmed cell death in animal development and disease. Cell 2011; 147:742–758.
Bergsbaken T, Fink SL, Cookson BT . Pyroptosis: host cell death and inflammation. Nat Rev Microbiol 2009; 7:99–109.
Blum ES, Abraham MC, Yoshimura S, Lu Y, Shaham S . Control of nonapoptotic developmental cell death in Caenorhabditis elegans by a polyglutamine-repeat protein. Science 2012; 335:970–973.
Vanden Berghe T, Linkermann A, Jouan-Lanhouet S, Walczak H, Vandenabeele P . Regulated necrosis: the expanding network of non-apoptotic cell death pathways. Nat Rev Mol Cell Biol 2014; 15:135–147.
Yuan J, Kroemer G . Alternative cell death mechanisms in development and beyond. Genes Dev 2010; 24:2592–2602.
Yang WS, Stockwell BR . Ferroptosis: death by lipid peroxidation. Trends Cell Biol 2016; 26:165–176.
Moriwaki K, Chan FK . RIP3: a molecular switch for necrosis and inflammation. Genes Dev 2013; 27:1640–1649.
Vandenabeele P, Declercq W, Van Herreweghe F, Vanden Berghe T . The role of the kinases RIP1 and RIP3 in TNF-induced necrosis. Sci Signal 2010; 3:re4.
Dolma S, Lessnick SL, Hahn WC, Stockwell BR . Identification of genotype-selective antitumor agents using synthetic lethal chemical screening in engineered human tumor cells. Cancer Cell 2003; 3:285–296.
Dixon SJ, Lemberg KM, Lamprecht MR, et al. Ferroptosis: An iron-dependent form of nonapoptotic cell death. Cell 2012; 149:1060–1072.
Yang WS, SriRamaratnam R, Welsch ME, et al. Regulation of ferroptotic cancer cell death by GPX4. Cell 2014; 156:317–331.
Gao M, Monian P, Quadri N, Ramasamy R, Jiang X . Glutaminolysis and transferrin regulate ferroptosis. Mol Cell 2015; 59:298–308.
Gao M, Monian P, Jiang X . Metabolism and iron signaling in ferroptotic cell death. Oncotarget 2015; 6:35145–35146.
Friedmann Angeli JP, Schneider M, Proneth B, et al. Inactivation of the ferroptosis regulator Gpx4 triggers acute renal failure in mice. Nat Cell Biol 2014; 16:1180–1191.
Linkermann A, Skouta R, Himmerkus N, et al. Synchronized renal tubular cell death involves ferroptosis. Proc Natl Acad Sci USA 2014; 111:16836–16841.
Jiang L, Kon N, Li T, et al. Ferroptosis as a p53-mediated activity during tumour suppression. Nature 2015; 520:57–62.
Jiang X, Overholtzer M, Thompson CB . Autophagy in cellular metabolism and cancer. J Clin Invest 2015; 125:47–54.
Tsujimoto Y, Shimizu S . Another way to die: autophagic programmed cell death. Cell Death Differ 2005; 12:1528–1534.
Yu L, Wan F, Dutta S, et al. Autophagic programmed cell death by selective catalase degradation. Proc Natl Acad Sci USA 2006; 103:4952–4957.
Liu Y, Shoji-Kawata S, Sumpter RM Jr, et al. Autosis is a Na+,K+-ATPase-regulated form of cell death triggered by autophagy-inducing peptides, starvation, and hypoxia-ischemia. Proc Natl Acad Sci USA 2013; 110:20364–20371.
Liu Y, Levine B . Autosis and autophagic cell death: the dark side of autophagy. Cell Death Differ 2015; 22:367–376.
Dowdle WE, Nyfeler B, Nagel J, et al. Selective VPS34 inhibitor blocks autophagy and uncovers a role for NCOA4 in ferritin degradation and iron homeostasis in vivo. Nat Cell Biol 2014; 16:1069–1079.
Mancias JD, Wang X, Gygi SP, Harper JW, Kimmelman AC . Quantitative proteomics identifies NCOA4 as the cargo receptor mediating ferritinophagy. Nature 2014; 509:105–109.
Mancias JD, Pontano Vaites L, Nissim S, et al. Ferritinophagy via NCOA4 is required for erythropoiesis and is regulated by iron dependent HERC2-mediated proteolysis. Elife 2015; 4:e10308.
Bellelli R, Federico G, Matte A, et al. NCOA4 deficiency impairs systemic iron homeostasis. Cell Rep 2016; 14:411–421.
Mizushima N . Autophagy: process and function. Genes Dev 2007; 21:2861–2873.
Andrews NC, Schmidt PJ . Iron homeostasis. Annu Rev Physiol 2007; 69:69–85.
Hou W, Xie Y, Song X, et al. Autophagy promotes ferroptosis by degradation of ferritin. Autophagy 2016; 12:1425–1428.
Mathew R, Karantza-Wadsworth V, White E . Role of autophagy in cancer. Nat Rev Cancer 2007; 7:961–967.
Kroemer G, Marino G, Levine B . Autophagy and the integrated stress response. Mol Cell 2010; 40:280–293.
Dixon SJ, Patel DN, Welsch M, et al. Pharmacological inhibition of cystine-glutamate exchange induces endoplasmic reticulum stress and ferroptosis. Elife 2014; 3:e02523.
Rahman I, Kode A, Biswas SK . Assay for quantitative determination of glutathione and glutathione disulfide levels using enzymatic recycling method. Nat Protoc 2006; 1:3159–3165.
Prus E, Fibach E . Flow cytometry measurement of the labile iron pool in human hematopoietic cells. Cytometry A 2008; 73:22–27.
Padron D, Tall RD, Roth MG . Phospholipase D2 is required for efficient endocytic recycling of transferrin receptors. Mol Biol Cell 2006; 17:598–606.
Acknowledgements
We thank members of the Jiang lab for discussing and reading the manuscript. This work is partially supported by NIH grants (R01CA166413 and R01GM113013 to XJ), a Geoffrey Beene Cancer Research Foundation fund (to XJ), and NCI cancer center core grant P30 CA008748.
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( Supplementary information is linked to the online version of the paper on the Cell Research website.)
Supplementary information
Supplementary information, Figure S1
Autophagy inhibitor BafA1 inhibits cystine starvation-induced ferroptosis in both MEFs and HT1080 cells. (PDF 235 kb)
Supplementary information, Figure S2
The effect of autophagy inhibitors on ferroptosis is dose- and time-dependent. (PDF 850 kb)
Supplementary information, Figure S3
Autophagy is required for ferroptosis. (PDF 484 kb)
Supplementary information, Figure S4
Autophagy inhibitor BafA1 inhibits RSL3 starvation-induced ferroptosis in MEFs. (PDF 161 kb)
Supplementary information, Figure S5
Erastin does not alter transferrin internalization. (PDF 213 kb)
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Gao, M., Monian, P., Pan, Q. et al. Ferroptosis is an autophagic cell death process. Cell Res 26, 1021–1032 (2016). https://doi.org/10.1038/cr.2016.95
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DOI: https://doi.org/10.1038/cr.2016.95
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