Skip to main content
main-content

01.12.2017 | Research | Ausgabe 1/2017 Open Access

Molecular Cancer 1/2017

Cellular organization and molecular differentiation model of breast cancer-associated fibroblasts

Zeitschrift:
Molecular Cancer > Ausgabe 1/2017
Autoren:
Susann Busch, Daniel Andersson, Eva Bom, Claire Walsh, Anders Ståhlberg, Göran Landberg
Wichtige Hinweise

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.

Abstract

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.
Zusatzmaterial
Additional file 1: Characterization of patient-derived fibroblasts and clinical information of used tumours. Fibroblast morphology and SMAα expression levels of isolated normal and cancer-associated fibroblasts. Clinical information of all analysed tumours. (PDF 1033 kb)
12943_2017_642_MOESM1_ESM.pdf
Additional file 2: Gene correlation analysis. Heatmaps depicting gene correlation analyses according to fibroblast subgroups (normal versus cancer-activated) for CAF cell line model and primary fibroblasts. (PDF 2716 kb)
12943_2017_642_MOESM2_ESM.pdf
Literatur
Über diesen Artikel

Weitere Artikel der Ausgabe 1/2017

Molecular Cancer 1/2017 Zur Ausgabe

Neu im Fachgebiet Onkologie

Mail Icon II Newsletter

Bestellen Sie unseren kostenlosen Newsletter Update Onkologie und bleiben Sie gut informiert – ganz bequem per eMail.

Bildnachweise