Dataset Information

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

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 TGF?1 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.

SUBMITTER: Busch S

PROVIDER: S-EPMC5376683 | biostudies-literature | 2017 Apr

REPOSITORIES: biostudies-literature

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Publications

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

Busch Susann S Andersson Daniel D Bom Eva E Walsh Claire C Ståhlberg Anders A Landberg Göran G

Molecular cancer 20170403 1

<h4>Background</h4>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.<h4>Methods</h4>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 simultan ...[more]

PMID: 28372546

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Project description:Simple Summary Primary, organ-confined prostate cancer is treatable with surgery, chemotherapy, radiation, and active surveillance; however, a subset of patients will develop metastatic disease. The main issue in prostate cancer is the need of prognostic tools for identifying patients at risk for lethal metastatic tumor, as well as a lack of curative therapies for such patients. The tissues surrounding the prostate cancer cells consist of a mixture of stromal cells that are structurally and functionally different from stromal cells in normal prostate. The stromal component, as a source of prognostic information for metastatic disease progression, has received little attention. In the present study, we characterized stromal cells from cancer tissues as compared to stromal cells from normal adjacent tumor tissue or human benign prostate. The stroma’s specific profile will allow to predict the presence of aggressive tumors, therapy resistance, and metastasis development. Abstract Background: Stromal components surrounding epithelial cancer cells seem to play a pivotal role during epithelial-to-mesenchymal transition (EMT), tumor invasion, and metastases. To identify the molecular mechanisms underlying tumor–stroma interactions may yield novel therapeutic targets for prostate cancer. Methods: Gene expression profile of prostate-cancer associated fibroblast (PCAF) and prostate non-cancer associated fibroblast (PNAF) cells isolated from radical prostatectomy was performed by Illumina, analyzed, and further processed by Ingenuity®: IPA® software. qRT-PCR was performed on an independent set of 17 PCAF, 12 PNAF, and 12 fibroblast cell lines derived from patients with benign prostatic hyperplasia (BPHF). Results: Using microarray analysis, we found six upregulated genes and two downregulated genes in PCAFs compared to PNAFs. To validate microarray results, we performed qRT-PCR for the most significantly regulated genes involved in the modulation of proliferation and androgen resistance on an independent set of PNAF, PCAF, and BHPF samples. We confirmed the increased expression of SCARB1, MAPK3K1, and TGF-β as well as the decreased expression of S100A10 in PCAFs compared to PNAFs and BPHFs. Conclusions: These results provide strong evidence that the observed changes in the gene expression profile of PCAFs can contribute to functional alteration of adjacent prostate cancer cells.

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Dataset Information

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

Background

Methods

Results

Conclusions

Publications

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

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