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A rapid multiplex cell-free assay on biochip to evaluate functional aspects of double-strand break repair

The repair of DNA double-strand breaks (DSBs) involves interdependent molecular pathways, of which the choice is crucial for a cell’s fate when facing a damage. Growing evidence points toward the fact that DSB repair capacities correlate with disease aggressiveness, treatment response and treatment-...

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Autores principales: Tatin, Xavier, Muggiolu, Giovanna, Libert, Sarah, Béal, David, Maillet, Thierry, Breton, Jean, Sauvaigo, Sylvie
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9681732/
https://www.ncbi.nlm.nih.gov/pubmed/36414637
http://dx.doi.org/10.1038/s41598-022-23819-0
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author Tatin, Xavier
Muggiolu, Giovanna
Libert, Sarah
Béal, David
Maillet, Thierry
Breton, Jean
Sauvaigo, Sylvie
author_facet Tatin, Xavier
Muggiolu, Giovanna
Libert, Sarah
Béal, David
Maillet, Thierry
Breton, Jean
Sauvaigo, Sylvie
author_sort Tatin, Xavier
collection PubMed
description The repair of DNA double-strand breaks (DSBs) involves interdependent molecular pathways, of which the choice is crucial for a cell’s fate when facing a damage. Growing evidence points toward the fact that DSB repair capacities correlate with disease aggressiveness, treatment response and treatment-related toxicities in cancer. Scientific and medical communities need more easy-to-use and efficient tools to rapidly estimate DSB repair capacities from a tissue, enable routine-accessible treatment personalization, and hopefully, improve survival. Here, we propose a new functional biochip assay (NEXT-SPOT) that characterizes DSB repair-engaged cellular pathways and provides qualitative and quantitative information on the contribution of several pathways in less than 2 h, from 10 mg of cell lysates. We introduce the NEXT-SPOT technology, detail the molecular characterizations of different repair steps occurring on the biochip, and show examples of DSB repair profiling using three cancer cell lines treated or not with a DSB-inducer (doxorubicin) and/or a DNA repair inhibitor (RAD51 inhibitor; DNA-PK inhibitor; PARP inhibitor). Among others, we demonstrate that NEXT-SPOT can accurately detect decreased activities in strand invasion and end-joining mechanisms following DNA-PK or RAD51 inhibition in DNA-PK-proficient cell lines. This approach offers an all-in-one reliable strategy to consider DSB repair capacities as predictive biomarkers easily translatable to the clinic.
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spelling pubmed-96817322022-11-24 A rapid multiplex cell-free assay on biochip to evaluate functional aspects of double-strand break repair Tatin, Xavier Muggiolu, Giovanna Libert, Sarah Béal, David Maillet, Thierry Breton, Jean Sauvaigo, Sylvie Sci Rep Article The repair of DNA double-strand breaks (DSBs) involves interdependent molecular pathways, of which the choice is crucial for a cell’s fate when facing a damage. Growing evidence points toward the fact that DSB repair capacities correlate with disease aggressiveness, treatment response and treatment-related toxicities in cancer. Scientific and medical communities need more easy-to-use and efficient tools to rapidly estimate DSB repair capacities from a tissue, enable routine-accessible treatment personalization, and hopefully, improve survival. Here, we propose a new functional biochip assay (NEXT-SPOT) that characterizes DSB repair-engaged cellular pathways and provides qualitative and quantitative information on the contribution of several pathways in less than 2 h, from 10 mg of cell lysates. We introduce the NEXT-SPOT technology, detail the molecular characterizations of different repair steps occurring on the biochip, and show examples of DSB repair profiling using three cancer cell lines treated or not with a DSB-inducer (doxorubicin) and/or a DNA repair inhibitor (RAD51 inhibitor; DNA-PK inhibitor; PARP inhibitor). Among others, we demonstrate that NEXT-SPOT can accurately detect decreased activities in strand invasion and end-joining mechanisms following DNA-PK or RAD51 inhibition in DNA-PK-proficient cell lines. This approach offers an all-in-one reliable strategy to consider DSB repair capacities as predictive biomarkers easily translatable to the clinic. Nature Publishing Group UK 2022-11-21 /pmc/articles/PMC9681732/ /pubmed/36414637 http://dx.doi.org/10.1038/s41598-022-23819-0 Text en © The Author(s) 2022 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
Tatin, Xavier
Muggiolu, Giovanna
Libert, Sarah
Béal, David
Maillet, Thierry
Breton, Jean
Sauvaigo, Sylvie
A rapid multiplex cell-free assay on biochip to evaluate functional aspects of double-strand break repair
title A rapid multiplex cell-free assay on biochip to evaluate functional aspects of double-strand break repair
title_full A rapid multiplex cell-free assay on biochip to evaluate functional aspects of double-strand break repair
title_fullStr A rapid multiplex cell-free assay on biochip to evaluate functional aspects of double-strand break repair
title_full_unstemmed A rapid multiplex cell-free assay on biochip to evaluate functional aspects of double-strand break repair
title_short A rapid multiplex cell-free assay on biochip to evaluate functional aspects of double-strand break repair
title_sort rapid multiplex cell-free assay on biochip to evaluate functional aspects of double-strand break repair
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9681732/
https://www.ncbi.nlm.nih.gov/pubmed/36414637
http://dx.doi.org/10.1038/s41598-022-23819-0
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