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Morpho‐metabotyping the oxidative stress response

Oxidative stress and reactive oxygen species (ROS) are central to many physiological and pathophysiological processes. However, due to multiple technical challenges, it is hard to capture a comprehensive readout of the cell, involving both biochemical and functional status. We addressed this problem...

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Autores principales: Rusz, Mate, Del Favero, Giorgia, El Abiead, Yasin, Gerner, Christopher, Keppler, Bernhard K., Jakupec, Michael A., Koellensperger, Gunda
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8322264/
https://www.ncbi.nlm.nih.gov/pubmed/34326354
http://dx.doi.org/10.1038/s41598-021-94585-8
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author Rusz, Mate
Del Favero, Giorgia
El Abiead, Yasin
Gerner, Christopher
Keppler, Bernhard K.
Jakupec, Michael A.
Koellensperger, Gunda
author_facet Rusz, Mate
Del Favero, Giorgia
El Abiead, Yasin
Gerner, Christopher
Keppler, Bernhard K.
Jakupec, Michael A.
Koellensperger, Gunda
author_sort Rusz, Mate
collection PubMed
description Oxidative stress and reactive oxygen species (ROS) are central to many physiological and pathophysiological processes. However, due to multiple technical challenges, it is hard to capture a comprehensive readout of the cell, involving both biochemical and functional status. We addressed this problem by developing a fully parallelized workflow for metabolomics (providing absolute quantities for > 100 metabolites including TCA cycle, pentose phosphate pathway, purine metabolism, glutathione metabolism, cysteine and methionine metabolism, glycolysis and gluconeogenesis) and live cell imaging microscopy. The correlative imaging strategy was applied to study morphological and metabolic adaptation of cancer cells upon short-term hydrogen peroxide (H(2)O(2)) exposure in vitro. The combination provided rich metabolic information at the endpoint of exposure together with imaging of mitochondrial effects. As a response, superoxide concentrations were elevated with a strong mitochondrial localization, and multi-parametric image analysis revealed a shift towards fragmentation. In line with this, metabolism reflected both the impaired mitochondrial function and shifts to support the first-line cellular defense and compensate for energy loss. The presented workflow combining high-end technologies demonstrates the applicability for the study of short-term oxidative stress, but it can be suitable for the in-depth study of various short-term oxidative and other cellular stress-related phenomena.
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spelling pubmed-83222642021-07-30 Morpho‐metabotyping the oxidative stress response Rusz, Mate Del Favero, Giorgia El Abiead, Yasin Gerner, Christopher Keppler, Bernhard K. Jakupec, Michael A. Koellensperger, Gunda Sci Rep Article Oxidative stress and reactive oxygen species (ROS) are central to many physiological and pathophysiological processes. However, due to multiple technical challenges, it is hard to capture a comprehensive readout of the cell, involving both biochemical and functional status. We addressed this problem by developing a fully parallelized workflow for metabolomics (providing absolute quantities for > 100 metabolites including TCA cycle, pentose phosphate pathway, purine metabolism, glutathione metabolism, cysteine and methionine metabolism, glycolysis and gluconeogenesis) and live cell imaging microscopy. The correlative imaging strategy was applied to study morphological and metabolic adaptation of cancer cells upon short-term hydrogen peroxide (H(2)O(2)) exposure in vitro. The combination provided rich metabolic information at the endpoint of exposure together with imaging of mitochondrial effects. As a response, superoxide concentrations were elevated with a strong mitochondrial localization, and multi-parametric image analysis revealed a shift towards fragmentation. In line with this, metabolism reflected both the impaired mitochondrial function and shifts to support the first-line cellular defense and compensate for energy loss. The presented workflow combining high-end technologies demonstrates the applicability for the study of short-term oxidative stress, but it can be suitable for the in-depth study of various short-term oxidative and other cellular stress-related phenomena. Nature Publishing Group UK 2021-07-29 /pmc/articles/PMC8322264/ /pubmed/34326354 http://dx.doi.org/10.1038/s41598-021-94585-8 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Rusz, Mate
Del Favero, Giorgia
El Abiead, Yasin
Gerner, Christopher
Keppler, Bernhard K.
Jakupec, Michael A.
Koellensperger, Gunda
Morpho‐metabotyping the oxidative stress response
title Morpho‐metabotyping the oxidative stress response
title_full Morpho‐metabotyping the oxidative stress response
title_fullStr Morpho‐metabotyping the oxidative stress response
title_full_unstemmed Morpho‐metabotyping the oxidative stress response
title_short Morpho‐metabotyping the oxidative stress response
title_sort morpho‐metabotyping the oxidative stress response
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8322264/
https://www.ncbi.nlm.nih.gov/pubmed/34326354
http://dx.doi.org/10.1038/s41598-021-94585-8
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