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Assessment of the Mutational Status of NSCLC Using Hypermetabolic Circulating Tumor Cells

Molecular characterization is currently a key step in NSCLC therapy selection. Circulating tumor cells (CTC) are excellent candidates for downstream analysis, but technology is still lagging behind. In this work, we show that the mutational status of NSCLC can be assessed on hypermetabolic CTC, dete...

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Autores principales: Turetta, Matteo, Bulfoni, Michela, Brisotto, Giulia, Fasola, Gianpiero, Zanello, Andrea, Biscontin, Eva, Mariuzzi, Laura, Steffan, Agostino, Di Loreto, Carla, Cesselli, Daniela, Del Ben, Fabio
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6115779/
https://www.ncbi.nlm.nih.gov/pubmed/30110953
http://dx.doi.org/10.3390/cancers10080270
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author Turetta, Matteo
Bulfoni, Michela
Brisotto, Giulia
Fasola, Gianpiero
Zanello, Andrea
Biscontin, Eva
Mariuzzi, Laura
Steffan, Agostino
Di Loreto, Carla
Cesselli, Daniela
Del Ben, Fabio
author_facet Turetta, Matteo
Bulfoni, Michela
Brisotto, Giulia
Fasola, Gianpiero
Zanello, Andrea
Biscontin, Eva
Mariuzzi, Laura
Steffan, Agostino
Di Loreto, Carla
Cesselli, Daniela
Del Ben, Fabio
author_sort Turetta, Matteo
collection PubMed
description Molecular characterization is currently a key step in NSCLC therapy selection. Circulating tumor cells (CTC) are excellent candidates for downstream analysis, but technology is still lagging behind. In this work, we show that the mutational status of NSCLC can be assessed on hypermetabolic CTC, detected by their increased glucose uptake. We validated the method in 30 Stage IV NSCLC patients: peripheral blood samples were incubated with a fluorescent glucose analog (2-NBDG) and analyzed by flow cytometry. Cells with the highest glucose uptake were sorted out. EGFR and KRAS mutations were detected by ddPCR. In sorted cells, mutated DNA was found in 85% of patients, finding an exact match with primary tumor in 70% of cases. Interestingly, in two patients multiple KRAS mutations were detected. Two patients displayed different mutations with respect to the primary tumor, and in two out of the four patients with a wild type primary tumor, new mutations were highlighted: EGFR p.746_750del and KRAS p.G12V. Hypermetabolic CTC can be enriched without the need of dedicated equipment and their mutational status can successfully be assessed by ddPCR. Finally, the finding of new mutations supports the possibility of probing tumor heterogeneity.
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spelling pubmed-61157792018-08-31 Assessment of the Mutational Status of NSCLC Using Hypermetabolic Circulating Tumor Cells Turetta, Matteo Bulfoni, Michela Brisotto, Giulia Fasola, Gianpiero Zanello, Andrea Biscontin, Eva Mariuzzi, Laura Steffan, Agostino Di Loreto, Carla Cesselli, Daniela Del Ben, Fabio Cancers (Basel) Article Molecular characterization is currently a key step in NSCLC therapy selection. Circulating tumor cells (CTC) are excellent candidates for downstream analysis, but technology is still lagging behind. In this work, we show that the mutational status of NSCLC can be assessed on hypermetabolic CTC, detected by their increased glucose uptake. We validated the method in 30 Stage IV NSCLC patients: peripheral blood samples were incubated with a fluorescent glucose analog (2-NBDG) and analyzed by flow cytometry. Cells with the highest glucose uptake were sorted out. EGFR and KRAS mutations were detected by ddPCR. In sorted cells, mutated DNA was found in 85% of patients, finding an exact match with primary tumor in 70% of cases. Interestingly, in two patients multiple KRAS mutations were detected. Two patients displayed different mutations with respect to the primary tumor, and in two out of the four patients with a wild type primary tumor, new mutations were highlighted: EGFR p.746_750del and KRAS p.G12V. Hypermetabolic CTC can be enriched without the need of dedicated equipment and their mutational status can successfully be assessed by ddPCR. Finally, the finding of new mutations supports the possibility of probing tumor heterogeneity. MDPI 2018-08-14 /pmc/articles/PMC6115779/ /pubmed/30110953 http://dx.doi.org/10.3390/cancers10080270 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Turetta, Matteo
Bulfoni, Michela
Brisotto, Giulia
Fasola, Gianpiero
Zanello, Andrea
Biscontin, Eva
Mariuzzi, Laura
Steffan, Agostino
Di Loreto, Carla
Cesselli, Daniela
Del Ben, Fabio
Assessment of the Mutational Status of NSCLC Using Hypermetabolic Circulating Tumor Cells
title Assessment of the Mutational Status of NSCLC Using Hypermetabolic Circulating Tumor Cells
title_full Assessment of the Mutational Status of NSCLC Using Hypermetabolic Circulating Tumor Cells
title_fullStr Assessment of the Mutational Status of NSCLC Using Hypermetabolic Circulating Tumor Cells
title_full_unstemmed Assessment of the Mutational Status of NSCLC Using Hypermetabolic Circulating Tumor Cells
title_short Assessment of the Mutational Status of NSCLC Using Hypermetabolic Circulating Tumor Cells
title_sort assessment of the mutational status of nsclc using hypermetabolic circulating tumor cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6115779/
https://www.ncbi.nlm.nih.gov/pubmed/30110953
http://dx.doi.org/10.3390/cancers10080270
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