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Cellular organization and molecular differentiation model of breast cancer-associated fibroblasts
BACKGROUND: The role of cancer-associated fibroblasts (CAFs) during tumour progression is obscured by the inherently complex, heterotypic nature of fibroblast cells and behaviours in various subtypes of malignancies. Therefore, we sought to identify distinct fibroblast subpopulations at the single-c...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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BioMed Central
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5376683/ https://www.ncbi.nlm.nih.gov/pubmed/28372546 http://dx.doi.org/10.1186/s12943-017-0642-7 |
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author | Busch, Susann Andersson, Daniel Bom, Eva Walsh, Claire Ståhlberg, Anders Landberg, Göran |
author_facet | Busch, Susann Andersson, Daniel Bom, Eva Walsh, Claire Ståhlberg, Anders Landberg, Göran |
author_sort | Busch, Susann |
collection | PubMed |
description | BACKGROUND: The role of cancer-associated fibroblasts (CAFs) during tumour progression is obscured by the inherently complex, heterotypic nature of fibroblast cells and behaviours in various subtypes of malignancies. Therefore, we sought to identify distinct fibroblast subpopulations at the single-cell level. METHODS: Using single-cell quantitative PCR as a powerful tool to study heterogeneity and rare cell events, in a high-throughput manner a panel of gene targets are run simultaneously on transcripts isolated from single cells obtained by fluorescence-activated cell sort. Assessment of cells with stem-like characteristics was attained by anchorage-independent, anoikis-resistant culture. RESULTS: Single-cell analysis of fibroblasts and their tumour-activated counterparts demonstrated molecularly distinct cell types defined by differential expression of characteristic mesenchymal and fibroblast activation markers. Identified subpopulations presented overlapping gene expression patterns indicating transitional molecular states during fibroblast differentiation. Using single-cell resolution data we generated a molecular differentiation model which enabled the classification of patient-derived fibroblasts, validating our modelling approach. Remarkably, a subset of fibroblasts displayed expression of pluripotency markers, which was enriched for in non-adherent conditions. Yet the ability to form single-cell derived spheres was generally reduced in CAFs and upon fibroblast activation through TGFβ1 ligand and cancer cell-secreted factors. Hence, our data imply the existence of putative stem/progenitor cells as a physiological feature of undifferentiated fibroblasts. CONCLUSIONS: Within this comprehensive study we have identified distinct and intersecting molecular profiles defining fibroblast activation states and propose that underlying cellular heterogeneity, fibroblasts are hierarchically organized. Understanding the molecular make-up of cellular organization and differentiation routes will facilitate the discovery of more specific markers for stromal subtypes and targets for anti-stromal therapies. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12943-017-0642-7) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-5376683 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-53766832017-04-07 Cellular organization and molecular differentiation model of breast cancer-associated fibroblasts Busch, Susann Andersson, Daniel Bom, Eva Walsh, Claire Ståhlberg, Anders Landberg, Göran Mol Cancer Research BACKGROUND: The role of cancer-associated fibroblasts (CAFs) during tumour progression is obscured by the inherently complex, heterotypic nature of fibroblast cells and behaviours in various subtypes of malignancies. Therefore, we sought to identify distinct fibroblast subpopulations at the single-cell level. METHODS: Using single-cell quantitative PCR as a powerful tool to study heterogeneity and rare cell events, in a high-throughput manner a panel of gene targets are run simultaneously on transcripts isolated from single cells obtained by fluorescence-activated cell sort. Assessment of cells with stem-like characteristics was attained by anchorage-independent, anoikis-resistant culture. RESULTS: Single-cell analysis of fibroblasts and their tumour-activated counterparts demonstrated molecularly distinct cell types defined by differential expression of characteristic mesenchymal and fibroblast activation markers. Identified subpopulations presented overlapping gene expression patterns indicating transitional molecular states during fibroblast differentiation. Using single-cell resolution data we generated a molecular differentiation model which enabled the classification of patient-derived fibroblasts, validating our modelling approach. Remarkably, a subset of fibroblasts displayed expression of pluripotency markers, which was enriched for in non-adherent conditions. Yet the ability to form single-cell derived spheres was generally reduced in CAFs and upon fibroblast activation through TGFβ1 ligand and cancer cell-secreted factors. Hence, our data imply the existence of putative stem/progenitor cells as a physiological feature of undifferentiated fibroblasts. CONCLUSIONS: Within this comprehensive study we have identified distinct and intersecting molecular profiles defining fibroblast activation states and propose that underlying cellular heterogeneity, fibroblasts are hierarchically organized. Understanding the molecular make-up of cellular organization and differentiation routes will facilitate the discovery of more specific markers for stromal subtypes and targets for anti-stromal therapies. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12943-017-0642-7) contains supplementary material, which is available to authorized users. BioMed Central 2017-04-03 /pmc/articles/PMC5376683/ /pubmed/28372546 http://dx.doi.org/10.1186/s12943-017-0642-7 Text en © The Author(s). 2017 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 Busch, Susann Andersson, Daniel Bom, Eva Walsh, Claire Ståhlberg, Anders Landberg, Göran Cellular organization and molecular differentiation model of breast cancer-associated fibroblasts |
title | Cellular organization and molecular differentiation model of breast cancer-associated fibroblasts |
title_full | Cellular organization and molecular differentiation model of breast cancer-associated fibroblasts |
title_fullStr | Cellular organization and molecular differentiation model of breast cancer-associated fibroblasts |
title_full_unstemmed | Cellular organization and molecular differentiation model of breast cancer-associated fibroblasts |
title_short | Cellular organization and molecular differentiation model of breast cancer-associated fibroblasts |
title_sort | cellular organization and molecular differentiation model of breast cancer-associated fibroblasts |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5376683/ https://www.ncbi.nlm.nih.gov/pubmed/28372546 http://dx.doi.org/10.1186/s12943-017-0642-7 |
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