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The copper transporter CTR1 and cisplatin accumulation at the single-cell level by LA-ICP-TOFMS

More than a decade ago, studies on cellular cisplatin accumulation via active membrane transport established the role of the high affinity copper uptake protein 1 (CTR1) as a main uptake route besides passive diffusion. In this work, CTR1 expression, cisplatin accumulation and intracellular copper c...

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Autores principales: Schoeberl, Anna, Gutmann, Michael, Theiner, Sarah, Corte-Rodríguez, Mario, Braun, Gabriel, Vician, Petra, Berger, Walter, Koellensperger, Gunda
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9742377/
https://www.ncbi.nlm.nih.gov/pubmed/36518851
http://dx.doi.org/10.3389/fmolb.2022.1055356
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author Schoeberl, Anna
Gutmann, Michael
Theiner, Sarah
Corte-Rodríguez, Mario
Braun, Gabriel
Vician, Petra
Berger, Walter
Koellensperger, Gunda
author_facet Schoeberl, Anna
Gutmann, Michael
Theiner, Sarah
Corte-Rodríguez, Mario
Braun, Gabriel
Vician, Petra
Berger, Walter
Koellensperger, Gunda
author_sort Schoeberl, Anna
collection PubMed
description More than a decade ago, studies on cellular cisplatin accumulation via active membrane transport established the role of the high affinity copper uptake protein 1 (CTR1) as a main uptake route besides passive diffusion. In this work, CTR1 expression, cisplatin accumulation and intracellular copper concentration was assessed for single cells revisiting the case of CTR1 in the context of acquired cisplatin resistance. The single-cell workflow designed for in vitro experiments enabled quantitative imaging at resolutions down to 1 µm by laser ablation-inductively coupled plasma-time-of-flight mass spectrometry (LA-ICP-TOFMS). Cisplatin-sensitive ovarian carcinoma cells A2780 as compared to the cisplatin-resistant subline A2780cis were investigated. Intracellular cisplatin and copper levels were absolutely quantified for thousands of individual cells, while for CTR1, relative differences of total CTR1 versus plasma membrane-bound CTR1 were determined. A markedly decreased intracellular cisplatin concentration accompanied by reduced copper concentrations was observed for single A2780cis cells, along with a distinctly reduced (total) CTR1 level as compared to the parental cell model. Interestingly, a significantly different proportion of plasma membrane-bound versus total CTR1 in untreated A2780 as compared to A2780cis cells was observed. This proportion changed in both models upon cisplatin exposure. Statistical analysis revealed a significant correlation between total and plasma membrane-bound CTR1 expression and cisplatin accumulation at the single-cell level in both A2780 and A2780cis cells. Thus, our study recapitulates the crosstalk of copper homeostasis and cisplatin uptake, and also indicates a complex interplay between subcellular CTR1 localization and cellular cisplatin accumulation as a driver for acquired resistance development.
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spelling pubmed-97423772022-12-13 The copper transporter CTR1 and cisplatin accumulation at the single-cell level by LA-ICP-TOFMS Schoeberl, Anna Gutmann, Michael Theiner, Sarah Corte-Rodríguez, Mario Braun, Gabriel Vician, Petra Berger, Walter Koellensperger, Gunda Front Mol Biosci Molecular Biosciences More than a decade ago, studies on cellular cisplatin accumulation via active membrane transport established the role of the high affinity copper uptake protein 1 (CTR1) as a main uptake route besides passive diffusion. In this work, CTR1 expression, cisplatin accumulation and intracellular copper concentration was assessed for single cells revisiting the case of CTR1 in the context of acquired cisplatin resistance. The single-cell workflow designed for in vitro experiments enabled quantitative imaging at resolutions down to 1 µm by laser ablation-inductively coupled plasma-time-of-flight mass spectrometry (LA-ICP-TOFMS). Cisplatin-sensitive ovarian carcinoma cells A2780 as compared to the cisplatin-resistant subline A2780cis were investigated. Intracellular cisplatin and copper levels were absolutely quantified for thousands of individual cells, while for CTR1, relative differences of total CTR1 versus plasma membrane-bound CTR1 were determined. A markedly decreased intracellular cisplatin concentration accompanied by reduced copper concentrations was observed for single A2780cis cells, along with a distinctly reduced (total) CTR1 level as compared to the parental cell model. Interestingly, a significantly different proportion of plasma membrane-bound versus total CTR1 in untreated A2780 as compared to A2780cis cells was observed. This proportion changed in both models upon cisplatin exposure. Statistical analysis revealed a significant correlation between total and plasma membrane-bound CTR1 expression and cisplatin accumulation at the single-cell level in both A2780 and A2780cis cells. Thus, our study recapitulates the crosstalk of copper homeostasis and cisplatin uptake, and also indicates a complex interplay between subcellular CTR1 localization and cellular cisplatin accumulation as a driver for acquired resistance development. Frontiers Media S.A. 2022-11-28 /pmc/articles/PMC9742377/ /pubmed/36518851 http://dx.doi.org/10.3389/fmolb.2022.1055356 Text en Copyright © 2022 Schoeberl, Gutmann, Theiner, Corte-Rodríguez, Braun, Vician, Berger and Koellensperger. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Molecular Biosciences
Schoeberl, Anna
Gutmann, Michael
Theiner, Sarah
Corte-Rodríguez, Mario
Braun, Gabriel
Vician, Petra
Berger, Walter
Koellensperger, Gunda
The copper transporter CTR1 and cisplatin accumulation at the single-cell level by LA-ICP-TOFMS
title The copper transporter CTR1 and cisplatin accumulation at the single-cell level by LA-ICP-TOFMS
title_full The copper transporter CTR1 and cisplatin accumulation at the single-cell level by LA-ICP-TOFMS
title_fullStr The copper transporter CTR1 and cisplatin accumulation at the single-cell level by LA-ICP-TOFMS
title_full_unstemmed The copper transporter CTR1 and cisplatin accumulation at the single-cell level by LA-ICP-TOFMS
title_short The copper transporter CTR1 and cisplatin accumulation at the single-cell level by LA-ICP-TOFMS
title_sort copper transporter ctr1 and cisplatin accumulation at the single-cell level by la-icp-tofms
topic Molecular Biosciences
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9742377/
https://www.ncbi.nlm.nih.gov/pubmed/36518851
http://dx.doi.org/10.3389/fmolb.2022.1055356
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