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

By D. Klionsky, A. K. Abdel-Aziz, Sara Abdelfatah, M. Abdellatif, A. Abdoli, S. Abel, H. Abeliovich, Marie H Abildgaard, Y. P. Abudu, A. Acevedo‐Arozena and 2,910 more. Autophagy, 2021

3,841 citations recorded by the source corpus.

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

Subfields: Epidemiology, Molecular Biology

View Paper: https://www.semanticscholar.org/paper/6737c2627bcad064410a48f8077e3ca455fa0639

How researchers see this paper

The researchers whose trusted networks rank it highest, strongest advocate first. Every researcher gets a different order.

In Epidemiology

1. Guido Kroemer
2. D. Tang
3. R. Kang
4. D. Klionsky
5. Jiao Liu
6. Srinivisa Reddy Bonam
7. S. Muller
8. Fengjuan Wang

In Molecular Biology

1. Guido Kroemer
2. D. Tang
3. R. Kang
4. D. Klionsky
5. Jiao Liu
6. Beth Levine
7. Xin Chen
8. L. Galluzzi

Citations in the corpus

• Lysosomes as a therapeutic target (2019, background): “In this regard, the tremendous work in recent years to establish international guidelines for standardizing research in autophagy -and, in particular, to propose relevant methodologies for monitoring autophagy that are accepted by the whole community -is unique 231, 232 .”
• Recent advances in Alzheimer’s disease: mechanisms, clinical trials and new drug development strategies (2024, background): “220,223,224 226 Macroautophagy, the main subtype, is primarily regulated by mTORC1 for activating the unc-51-like autophagy activating kinase 1 (ULK1) complex and dephosphor-ylating transcription factor EB (TFEB) to induce autophagy.”
• Autophagy-Dependent Ferroptosis: Machinery and Regulation (2020, methodology): “, bafilomycin A1 and chloroquine) stage of autophagy (Table S1), as denoted in specific guidelines (Klionsky et al., 2016).”