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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Autophagy: Renovation of Cells and Tissues

Cell, Journal Year: 2011, Volume and Issue: 147(4), P. 728 - 741

Published: Nov. 1, 2011

Language: Английский

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Guidelines for the use and interpretation of assays for monitoring autophagy

Autophagy, Journal Year: 2012, Volume and Issue: 8(4), P. 445 - 544

Published: April 1, 2012

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. A key point needs emphasized a difference between measurements monitor numbers or volume autophagic elements (e.g., autophagosomes autolysosomes) at any stage process vs. those flux through pathway (i.e., complete process); thus, block macroautophagy results autophagosome accumulation differentiated from stimuli result increased activity, defined as 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 increase autolysosomes may reflect reduction degradative activity. 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 part question being asked system used. addition, emphasize no individual assay guaranteed one every situation, strongly recommend multiple 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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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 and Aging

Cell, Journal Year: 2011, Volume and Issue: 146(5), P. 682 - 695

Published: Sept. 1, 2011

Language: Английский

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Autophagy-Dependent Anticancer Immune Responses Induced by Chemotherapeutic Agents in Mice

Science, Journal Year: 2011, Volume and Issue: 334(6062), P. 1573 - 1577

Published: Dec. 15, 2011

Antineoplastic chemotherapies are particularly efficient when they elicit immunogenic cell death, thus provoking an anticancer immune response. Here we demonstrate that autophagy, which is often disabled in cancer, dispensable for chemotherapy-induced death but required its immunogenicity. In response to chemotherapy, autophagy-competent, not autophagy-deficient, cancers attracted dendritic cells and T lymphocytes into the tumor bed. Suppression of autophagy inhibited release adenosine triphosphate (ATP) from dying cells. Conversely, inhibition extracellular ATP-degrading enzymes increased pericellular ATP autophagy-deficient tumors, reestablished recruitment cells, restored chemotherapeutic responses only immunocompetent hosts. Thus, essential concentrations improve efficacy antineoplastic disabled.

Language: Английский

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Autophagy and Neurodegeneration: Pathogenic Mechanisms and Therapeutic Opportunities

Neuron, Journal Year: 2017, Volume and Issue: 93(5), P. 1015 - 1034

Published: March 1, 2017

Language: Английский

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mTORC1 Controls Mitochondrial Activity and Biogenesis through 4E-BP-Dependent Translational Regulation

Cell Metabolism, Journal Year: 2013, Volume and Issue: 18(5), P. 698 - 711

Published: Nov. 1, 2013

Language: Английский

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Lysosomes as a therapeutic target

Nature Reviews Drug Discovery, Journal Year: 2019, Volume and Issue: 18(12), P. 923 - 948

Published: Sept. 2, 2019

Language: Английский

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Metformin as adjunct antituberculosis therapy

Science Translational Medicine, Journal Year: 2014, Volume and Issue: 6(263)

Published: Nov. 19, 2014

The global burden of tuberculosis (TB) morbidity and mortality remains immense. A potential new approach to TB therapy is augment protective host immune responses. We report that the antidiabetic drug metformin (MET) reduces intracellular growth Mycobacterium (Mtb) in an AMPK (adenosine monophosphate-activated protein kinase)-dependent manner. MET controls drug-resistant Mtb strains, increases production mitochondrial reactive oxygen species, facilitates phagosome-lysosome fusion. In Mtb-infected mice, use ameliorated lung pathology, reduced chronic inflammation, enhanced specific response efficacy conventional drugs. Moreover, two separate human cohorts, treatment was associated with improved control infection decreased disease severity. Collectively, these data indicate a promising candidate host-adjunctive for improving effective TB.

Language: Английский

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