Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition)

Autophagy, Journal Year: 2016, Volume and Issue: 12(1), P. 1 - 222

Published: Jan. 2, 2016

In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, on this topic has continued to accelerate, and many new scientists have entered field. Our knowledge base relevant technologies also been expanding. Accordingly, it is important update these monitoring autophagy different organisms. Various reviews described range assays that used purpose. Nevertheless, there continues be confusion regarding acceptable methods measure autophagy, especially multicellular eukaryotes. For example, a key point needs emphasized difference between measurements monitor numbers or volume autophagic elements (e.g., autophagosomes autolysosomes) at any stage process versus those flux through pathway (i.e., complete including amount rate cargo sequestered degraded). particular, block macroautophagy results autophagosome accumulation must differentiated from stimuli increase activity, defined as increased induction coupled with delivery to, degradation within, lysosomes (in most higher eukaryotes some protists such Dictyostelium) vacuole plants fungi). other words, investigators field understand appearance more does not necessarily equate fact, cases, accumulate because trafficking without concomitant change biogenesis, whereas an autolysosomes may reflect reduction degradative activity. It worth emphasizing here lysosomal digestion evaluating its competence crucial part evaluation flux, autophagy. Here, present selection interpretation use by who aim examine related processes, well reviewers need provide realistic reasonable critiques papers are focused processes. These meant formulaic rules, appropriate depend question being asked system used. addition, emphasize no individual assay guaranteed one every situation, strongly recommend multiple Along lines, potential pleiotropic effects due blocking genetic manipulation, imperative target gene knockout RNA interference than autophagy-related protein. Atg proteins, groups involved cellular pathways implying all proteins can specific marker process. guidelines, consider various assessing what information can, cannot, obtained them. Finally, discussing merits limits particular assays, hope encourage technical innovation

Language: Английский

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Ferroptosis: A Regulated Cell Death Nexus Linking Metabolism, Redox Biology, and Disease

Cell, Journal Year: 2017, Volume and Issue: 171(2), P. 273 - 285

Published: Oct. 1, 2017

Language: Английский

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Targeting autophagy in cancer

Nature reviews. Cancer, Journal Year: 2017, Volume and Issue: 17(9), P. 528 - 542

Published: July 28, 2017

Language: Английский

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Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)¹

Autophagy, Journal Year: 2021, Volume and Issue: 17(1), P. 1 - 382

Published: Jan. 2, 2021

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered field. Our knowledge base relevant new technologies also been expanding. Thus, it is important to formulate on a regular basis updated monitoring autophagy different organisms. Despite numerous reviews, there continues be confusion regarding acceptable methods evaluate autophagy, especially multicellular eukaryotes. Here, present investigators select interpret examine related processes, reviewers provide realistic reasonable critiques reports that are focused these processes. These not meant dogmatic rules, because appropriateness any assay largely depends question being asked system used. Moreover, no individual perfect every situation, calling use multiple techniques properly monitor each experimental setting. Finally, several core components machinery implicated distinct autophagic processes (canonical noncanonical autophagy), implying genetic approaches block should rely targeting two or more autophagy-related genes ideally participate steps pathway. Along similar lines, proteins involved regulate other cellular pathways including apoptosis, all them can used as specific marker bona fide responses. critically discuss current assessing information they can, cannot, provide. ultimate goal encourage intellectual technical innovation

Language: Английский

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Autophagy promotes ferroptosis by degradation of ferritin

Autophagy, Journal Year: 2016, Volume and Issue: 12(8), P. 1425 - 1428

Published: June 1, 2016

Macroautophagy/autophagy is an evolutionarily conserved degradation pathway that maintains homeostasis. Ferroptosis, a novel form of regulated cell death, characterized by production reactive oxygen species from accumulated iron and lipid peroxidation. However, the relationship between autophagy ferroptosis at genetic level remains unclear. Here, we demonstrated contributes to ferritin in fibroblasts cancer cells. Knockout or knockdown Atg5 (autophagy-related 5) Atg7 limited erastin-induced with decreased intracellular ferrous levels, Remarkably, NCOA4 (nuclear receptor coactivator 4) was selective cargo for autophagic turnover (namely ferritinophagy) ferroptosis. Consistently, inhibition inhibited suppressed In contrast, overexpression increased promoted These findings provide insight into interplay death.

Language: Английский

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Reactive Oxygen Species-Induced Lipid Peroxidation in Apoptosis, Autophagy, and Ferroptosis

Oxidative Medicine and Cellular Longevity, Journal Year: 2019, Volume and Issue: 2019, P. 1 - 13

Published: Oct. 13, 2019

Reactive oxygen species- (ROS-) induced lipid peroxidation plays a critical role in cell death including apoptosis, autophagy, and ferroptosis. This fundamental conserved mechanism is based on an excess of ROS which attacks biomembranes, propagates chain reactions, subsequently induces different types death. A highly evolved sophisticated antioxidant system exists that acts to protect the cells from oxidative damage. In this review, we discussed how propagate reactions products initiate apoptosis autophagy current models. We also during ferroptosis, summarized pathological conditions illness. aim bring more global integrative sight know ROS-induced occurs among

Language: Английский

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NRF2 plays a critical role in mitigating lipid peroxidation and ferroptosis

Redox Biology, Journal Year: 2019, Volume and Issue: 23, P. 101107 - 101107

Published: Jan. 12, 2019

The transcription factor nuclear erythroid 2-related 2 (NRF2) is a key regulator of the cellular antioxidant response, controlling expression genes that counteract oxidative and electrophilic stresses. Many pathological conditions are linked to imbalances in redox homeostasis, illustrating important role defense systems preventing pathogenic effects associated with accumulation reactive species. In particular, it becoming increasingly apparent lipid peroxides has an driving pathogenesis multiple disease states. A example this recent discovery novel form cell death termed ferroptosis. Ferroptosis iron-dependent, peroxidation-driven cascade become target development anti-cancer therapies, as well prevention neurodegenerative cardiovascular diseases. review, we will provide brief overview peroxidation, components involved ferroptotic cascade. We also highlight NRF2 signaling pathway mediating peroxidation ferroptosis, focusing on established mitigate these pathways, relevance NRF2-lipid peroxidation-ferroptosis axis disease.

Language: Английский

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Ferroptosis is an autophagic cell death process

Cell Research, Journal Year: 2016, Volume and Issue: 26(9), P. 1021 - 1032

Published: Aug. 12, 2016

Language: Английский

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Molecular definitions of autophagy and related processes

The EMBO Journal, Journal Year: 2017, Volume and Issue: 36(13), P. 1811 - 1836

Published: June 8, 2017

Language: Английский

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Ferroptosis: machinery and regulation

Autophagy, Journal Year: 2020, Volume and Issue: 17(9), P. 2054 - 2081

Published: Aug. 19, 2020

Ferroptosis is an iron-dependent, non-apoptotic form of regulated cell death caused by lipid peroxidation, which controlled integrated oxidation and antioxidant systems. The iron-containing enzyme lipoxygenase the main promoter ferroptosis producing hydroperoxides, its function relies on activation ACSL4-dependent biosynthesis. In contrast, selenium-containing GPX4 currently recognized as a central repressor ferroptosis, activity depends glutathione produced from cystine-glutamate antiporter SLC7A11. Many metabolic (especially involving iron, lipids, amino acids) degradation pathways (macroautophagy/autophagy ubiquitin-proteasome system) orchestrate complex ferroptotic response through direct or indirect regulation iron accumulation peroxidation. Although detailed mechanism membrane injury during remains mystery, ESCRT III-mediated plasma repair can make cells resistant to ferroptosis. Here, we review recent rapid progress in understanding molecular mechanisms focus epigenetic, transcriptional, posttranslational this process.Abbreviations: 2ME: beta-mercaptoethanol; α-KG: α-ketoglutarate; ccRCC: clear renal carcinoma; EMT: epithelial-mesenchymal transition; FAO: fatty acid beta-oxidation; GSH: glutathione; MEFs: mouse embryonic fibroblasts; MUFAs: monounsaturated acids; NO: nitric oxide; NOX: NADPH oxidase; PPP: pentose phosphate pathway; PUFA: polyunsaturated acid; RCD: death; RNS: reactive nitrogen species; ROS: oxygen RTAs: radical-trapping antioxidants; UPS: system; UTR: untranslated region.

Language: Английский

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