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Powering single-cell genomics to unravel circulating tumour cell subpopulations in non-small cell lung cancer patients
BACKGROUND: Circulating tumour cells (CTCs) are attractive “liquid biopsy” candidates that could provide insights into the different phenotypes of tumours present within a patient. The epithelial-to-mesenchymal transition (EMT) of CTCs is considered a critical step in tumour metastasis; however, it...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer Berlin Heidelberg
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10097753/ https://www.ncbi.nlm.nih.gov/pubmed/35896898 http://dx.doi.org/10.1007/s00432-022-04202-y |
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author | Acheampong, Emmanuel Morici, Michael Abed, Afaf Bowyer, Samantha Asante, Du-Bois Lin, Weitao Millward, Michael Gray, Elin S. Beasley, Aaron B. |
author_facet | Acheampong, Emmanuel Morici, Michael Abed, Afaf Bowyer, Samantha Asante, Du-Bois Lin, Weitao Millward, Michael Gray, Elin S. Beasley, Aaron B. |
author_sort | Acheampong, Emmanuel |
collection | PubMed |
description | BACKGROUND: Circulating tumour cells (CTCs) are attractive “liquid biopsy” candidates that could provide insights into the different phenotypes of tumours present within a patient. The epithelial-to-mesenchymal transition (EMT) of CTCs is considered a critical step in tumour metastasis; however, it may confound traditional epithelial feature-based CTC isolation and detection. We applied single-cell copy number alteration (CNA) analysis for the identification of genomic alterations to confirm the neoplastic nature of circulating cells with only mesenchymal phenotypes. METHODS: We isolated CTCs from blood samples collected from 46 NSCLC patients using the Parsortix system. Enriched cells were subjected to immunofluorescent staining for CTC identification using a multi-marker panel comprising both epithelial and mesenchymal markers. A subset of isolated CTCs was subjected to whole genome amplification (WGA) and low-pass whole-genome sequencing (LP-WGS) for the analysis of copy number alterations (CNAs). RESULTS: CTCs were detected in 16/46 (34.8%) patients, inclusive of CK(+)/EpCAM(+) CTCs (3/46, 6.5%) and Vim(+) CTCs (13/46, 28.3%). Clusters of Vim(+) cells were detected in 8 samples, which constitutes 50% of the total number of NSCLC patients with CTCs. No patients had detectable hybrid CK(+)/EpCAM(+)/Vim(+) cells. All of the tested CK(+)/EpCAM(+) CTCs and 7/8 Vim(+) CTCs or CTC clusters carried CNAs confirming their neoplastic nature. Notably, the Vim(+) cluster with no CNAs was characterised by spindle morphology and, therefore, defined as normal mesenchymal circulating cells. CONCLUSION: Our results revealed that CK-negative, vimentin-expressing cells represent a large proportion of CTCs detected in NSCLC patients, which are likely missed by standard epithelial-marker-dependent CTC categorisation. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00432-022-04202-y. |
format | Online Article Text |
id | pubmed-10097753 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Springer Berlin Heidelberg |
record_format | MEDLINE/PubMed |
spelling | pubmed-100977532023-04-14 Powering single-cell genomics to unravel circulating tumour cell subpopulations in non-small cell lung cancer patients Acheampong, Emmanuel Morici, Michael Abed, Afaf Bowyer, Samantha Asante, Du-Bois Lin, Weitao Millward, Michael Gray, Elin S. Beasley, Aaron B. J Cancer Res Clin Oncol Original Article – Clinical Oncology BACKGROUND: Circulating tumour cells (CTCs) are attractive “liquid biopsy” candidates that could provide insights into the different phenotypes of tumours present within a patient. The epithelial-to-mesenchymal transition (EMT) of CTCs is considered a critical step in tumour metastasis; however, it may confound traditional epithelial feature-based CTC isolation and detection. We applied single-cell copy number alteration (CNA) analysis for the identification of genomic alterations to confirm the neoplastic nature of circulating cells with only mesenchymal phenotypes. METHODS: We isolated CTCs from blood samples collected from 46 NSCLC patients using the Parsortix system. Enriched cells were subjected to immunofluorescent staining for CTC identification using a multi-marker panel comprising both epithelial and mesenchymal markers. A subset of isolated CTCs was subjected to whole genome amplification (WGA) and low-pass whole-genome sequencing (LP-WGS) for the analysis of copy number alterations (CNAs). RESULTS: CTCs were detected in 16/46 (34.8%) patients, inclusive of CK(+)/EpCAM(+) CTCs (3/46, 6.5%) and Vim(+) CTCs (13/46, 28.3%). Clusters of Vim(+) cells were detected in 8 samples, which constitutes 50% of the total number of NSCLC patients with CTCs. No patients had detectable hybrid CK(+)/EpCAM(+)/Vim(+) cells. All of the tested CK(+)/EpCAM(+) CTCs and 7/8 Vim(+) CTCs or CTC clusters carried CNAs confirming their neoplastic nature. Notably, the Vim(+) cluster with no CNAs was characterised by spindle morphology and, therefore, defined as normal mesenchymal circulating cells. CONCLUSION: Our results revealed that CK-negative, vimentin-expressing cells represent a large proportion of CTCs detected in NSCLC patients, which are likely missed by standard epithelial-marker-dependent CTC categorisation. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00432-022-04202-y. Springer Berlin Heidelberg 2022-07-28 2023 /pmc/articles/PMC10097753/ /pubmed/35896898 http://dx.doi.org/10.1007/s00432-022-04202-y Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis 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 | Original Article – Clinical Oncology Acheampong, Emmanuel Morici, Michael Abed, Afaf Bowyer, Samantha Asante, Du-Bois Lin, Weitao Millward, Michael Gray, Elin S. Beasley, Aaron B. Powering single-cell genomics to unravel circulating tumour cell subpopulations in non-small cell lung cancer patients |
title | Powering single-cell genomics to unravel circulating tumour cell subpopulations in non-small cell lung cancer patients |
title_full | Powering single-cell genomics to unravel circulating tumour cell subpopulations in non-small cell lung cancer patients |
title_fullStr | Powering single-cell genomics to unravel circulating tumour cell subpopulations in non-small cell lung cancer patients |
title_full_unstemmed | Powering single-cell genomics to unravel circulating tumour cell subpopulations in non-small cell lung cancer patients |
title_short | Powering single-cell genomics to unravel circulating tumour cell subpopulations in non-small cell lung cancer patients |
title_sort | powering single-cell genomics to unravel circulating tumour cell subpopulations in non-small cell lung cancer patients |
topic | Original Article – Clinical Oncology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10097753/ https://www.ncbi.nlm.nih.gov/pubmed/35896898 http://dx.doi.org/10.1007/s00432-022-04202-y |
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