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Quantitative proteomic analysis reveals AK2 as potential biomarker for late normal tissue radiotoxicity

BACKGROUND: Biomarkers for predicting late normal tissue toxicity to radiotherapy are necessary to personalize treatments and to optimize clinical benefit. Many radiogenomic studies have been published on this topic. Conversely, proteomics approaches are not much developed, despite their advantages....

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Autores principales: Lacombe, Jérôme, Brengues, Muriel, Mangé, Alain, Bourgier, Céline, Gourgou, Sophie, Pèlegrin, André, Ozsahin, Mahmut, Solassol, Jérôme, Azria, David
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6688300/
https://www.ncbi.nlm.nih.gov/pubmed/31399108
http://dx.doi.org/10.1186/s13014-019-1351-8
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author Lacombe, Jérôme
Brengues, Muriel
Mangé, Alain
Bourgier, Céline
Gourgou, Sophie
Pèlegrin, André
Ozsahin, Mahmut
Solassol, Jérôme
Azria, David
author_facet Lacombe, Jérôme
Brengues, Muriel
Mangé, Alain
Bourgier, Céline
Gourgou, Sophie
Pèlegrin, André
Ozsahin, Mahmut
Solassol, Jérôme
Azria, David
author_sort Lacombe, Jérôme
collection PubMed
description BACKGROUND: Biomarkers for predicting late normal tissue toxicity to radiotherapy are necessary to personalize treatments and to optimize clinical benefit. Many radiogenomic studies have been published on this topic. Conversely, proteomics approaches are not much developed, despite their advantages. METHODS: We used the isobaric tags for relative and absolute quantitation (iTRAQ) proteomic approach to analyze differences in protein expression levels in ex-vivo irradiated (8 Gy) T lymphocytes from patients with grade ≥ 2 radiation-induced breast fibrosis (grade ≥ 2 bf+) and patients with grade < 2 bf + after curative intent radiotherapy. Patients were selected from two prospective clinical trials (COHORT and PHRC 2005) and were used as discovery and confirmation cohorts. RESULTS: Among the 1979 quantified proteins, 23 fulfilled our stringent biological criteria. Immunoblotting analysis of four of these candidate proteins (adenylate kinase 2, AK2; annexin A1; heat shock cognate 71 kDa protein; and isocitrate dehydrogenase 2) confirmed AK2 overexpression in 8 Gy-irradiated T lymphocytes from patients with grade ≥ 2 bf + compared with patients with grade < 2 bf+. As these candidate proteins are involved in oxidative stress regulation, we also evaluated radiation-induced reactive oxygen species (ROS) production in peripheral blood mononuclear cells from patients with grade ≥ 2 bf + and grade < 2 bf+. Total ROS level, and especially superoxide anion level, increased upon ex-vivo 8 Gy-irradiation in all patients. Analysis of NADPH oxidases (NOXs), a major source of superoxide ion in the cell, showed a significant increase of NOX4 mRNA and protein levels after irradiation in both patient groups. Conversely, only NOX4 mRNA level was significantly different between groups (grade ≥ 2 bf + and grade < 2 bf+). CONCLUSION: These findings identify AK2 as a potential radiosensitivity candidate biomarker. Overall, our proteomic approach highlights the important role of oxidative stress in late radiation-induced toxicity, and paves the way for additional studies on NOXs and superoxide ion metabolism. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13014-019-1351-8) contains supplementary material, which is available to authorized users.
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spelling pubmed-66883002019-08-14 Quantitative proteomic analysis reveals AK2 as potential biomarker for late normal tissue radiotoxicity Lacombe, Jérôme Brengues, Muriel Mangé, Alain Bourgier, Céline Gourgou, Sophie Pèlegrin, André Ozsahin, Mahmut Solassol, Jérôme Azria, David Radiat Oncol Research BACKGROUND: Biomarkers for predicting late normal tissue toxicity to radiotherapy are necessary to personalize treatments and to optimize clinical benefit. Many radiogenomic studies have been published on this topic. Conversely, proteomics approaches are not much developed, despite their advantages. METHODS: We used the isobaric tags for relative and absolute quantitation (iTRAQ) proteomic approach to analyze differences in protein expression levels in ex-vivo irradiated (8 Gy) T lymphocytes from patients with grade ≥ 2 radiation-induced breast fibrosis (grade ≥ 2 bf+) and patients with grade < 2 bf + after curative intent radiotherapy. Patients were selected from two prospective clinical trials (COHORT and PHRC 2005) and were used as discovery and confirmation cohorts. RESULTS: Among the 1979 quantified proteins, 23 fulfilled our stringent biological criteria. Immunoblotting analysis of four of these candidate proteins (adenylate kinase 2, AK2; annexin A1; heat shock cognate 71 kDa protein; and isocitrate dehydrogenase 2) confirmed AK2 overexpression in 8 Gy-irradiated T lymphocytes from patients with grade ≥ 2 bf + compared with patients with grade < 2 bf+. As these candidate proteins are involved in oxidative stress regulation, we also evaluated radiation-induced reactive oxygen species (ROS) production in peripheral blood mononuclear cells from patients with grade ≥ 2 bf + and grade < 2 bf+. Total ROS level, and especially superoxide anion level, increased upon ex-vivo 8 Gy-irradiation in all patients. Analysis of NADPH oxidases (NOXs), a major source of superoxide ion in the cell, showed a significant increase of NOX4 mRNA and protein levels after irradiation in both patient groups. Conversely, only NOX4 mRNA level was significantly different between groups (grade ≥ 2 bf + and grade < 2 bf+). CONCLUSION: These findings identify AK2 as a potential radiosensitivity candidate biomarker. Overall, our proteomic approach highlights the important role of oxidative stress in late radiation-induced toxicity, and paves the way for additional studies on NOXs and superoxide ion metabolism. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13014-019-1351-8) contains supplementary material, which is available to authorized users. BioMed Central 2019-08-09 /pmc/articles/PMC6688300/ /pubmed/31399108 http://dx.doi.org/10.1186/s13014-019-1351-8 Text en © The Author(s). 2019 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Lacombe, Jérôme
Brengues, Muriel
Mangé, Alain
Bourgier, Céline
Gourgou, Sophie
Pèlegrin, André
Ozsahin, Mahmut
Solassol, Jérôme
Azria, David
Quantitative proteomic analysis reveals AK2 as potential biomarker for late normal tissue radiotoxicity
title Quantitative proteomic analysis reveals AK2 as potential biomarker for late normal tissue radiotoxicity
title_full Quantitative proteomic analysis reveals AK2 as potential biomarker for late normal tissue radiotoxicity
title_fullStr Quantitative proteomic analysis reveals AK2 as potential biomarker for late normal tissue radiotoxicity
title_full_unstemmed Quantitative proteomic analysis reveals AK2 as potential biomarker for late normal tissue radiotoxicity
title_short Quantitative proteomic analysis reveals AK2 as potential biomarker for late normal tissue radiotoxicity
title_sort quantitative proteomic analysis reveals ak2 as potential biomarker for late normal tissue radiotoxicity
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6688300/
https://www.ncbi.nlm.nih.gov/pubmed/31399108
http://dx.doi.org/10.1186/s13014-019-1351-8
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