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Modeling the initiation of Ewing sarcoma tumorigenesis in differentiating human embryonic stem cells.

Gordon DJ, Motwani M, Pellman D - Oncogene (2015)

Bottom Line: Here, we report a novel approach to model the initiation of Ewing sarcoma tumorigenesis that exploits the developmental and pluripotent potential of human embryonic stem cells.The inducible expression of EWS-FLI1 in embryoid bodies, or collections of differentiating stem cells, generates cells with properties of Ewing sarcoma tumors, including characteristics of transformation.Furthermore, these cells also demonstrate a requirement for the persistent expression of EWS-FLI1 for cell survival and growth, which is a hallmark of Ewing sarcoma tumors.

View Article: PubMed Central - PubMed

Affiliation: Department of Pediatric Oncology, Dana-Farber Cancer Institute, Boston, MA, USA.

ABSTRACT
Oncogenic transformation in Ewing sarcoma tumors is driven by the fusion oncogene EWS-FLI1. However, despite the well-established role of EWS-FLI1 in tumor initiation, the development of models of Ewing sarcoma in human cells with defined genetic elements has been challenging. Here, we report a novel approach to model the initiation of Ewing sarcoma tumorigenesis that exploits the developmental and pluripotent potential of human embryonic stem cells. The inducible expression of EWS-FLI1 in embryoid bodies, or collections of differentiating stem cells, generates cells with properties of Ewing sarcoma tumors, including characteristics of transformation. These cell lines exhibit anchorage-independent growth, a lack of contact inhibition and a strong Ewing sarcoma gene expression signature. Furthermore, these cells also demonstrate a requirement for the persistent expression of EWS-FLI1 for cell survival and growth, which is a hallmark of Ewing sarcoma tumors.

No MeSH data available.


Related in: MedlinePlus

EF+ cells exhibit an Ewing sarcoma gene expression signature. (A) Unsupervised hierarchical clustering of 88 cancer cell lines, across 12 different tumor types, and the EF+ cells. The Ewing sarcoma cell line cluster is outlined in blue and the EF+ cell line cluster is outlined in red. (B) RT-qPCR of select upregulated and downregulated genes. The error bars indicate the standard error of the mean from three experiments. The upper panel shows genes that are upregulated in the EF+ cells and the lower panel shows genes upregulated in the EF− cells. (C and D) Gene set enrichment analysis (GSEA) shows enrichment of the Hancock et al. upregulated and downregulated gene sets in the gene expression data from the EF+ versus EF− (C) and EF+ versus EFFib (D) comparisons. GSEA plots are also shown for the curated gene sets RIGGI_EWING and ZHANG_TARGETS_OF_EWSR1_FLI1 from the MSigDB collection. The normalized enrichment scores (NES) and FDR q-values are shown.
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Figure 5: EF+ cells exhibit an Ewing sarcoma gene expression signature. (A) Unsupervised hierarchical clustering of 88 cancer cell lines, across 12 different tumor types, and the EF+ cells. The Ewing sarcoma cell line cluster is outlined in blue and the EF+ cell line cluster is outlined in red. (B) RT-qPCR of select upregulated and downregulated genes. The error bars indicate the standard error of the mean from three experiments. The upper panel shows genes that are upregulated in the EF+ cells and the lower panel shows genes upregulated in the EF− cells. (C and D) Gene set enrichment analysis (GSEA) shows enrichment of the Hancock et al. upregulated and downregulated gene sets in the gene expression data from the EF+ versus EF− (C) and EF+ versus EFFib (D) comparisons. GSEA plots are also shown for the curated gene sets RIGGI_EWING and ZHANG_TARGETS_OF_EWSR1_FLI1 from the MSigDB collection. The normalized enrichment scores (NES) and FDR q-values are shown.

Mentions: Next we addressed whether the EF+ cells exhibited an Ewing sarcoma gene expression signature and, if so, whether our doxycycline-inducible, isogenic system would be advantageous in identifying gene expression changes related to EWS-FLI1. Unsupervised hierarchical clustering was performed with the EF+ cells and 88 cancer cell lines across 12 different tumor types (Figure 5A). Notably, the EF+ cells clustered most closely with the Ewing sarcoma cell lines relative to the other sarcomas and tumor types.


Modeling the initiation of Ewing sarcoma tumorigenesis in differentiating human embryonic stem cells.

Gordon DJ, Motwani M, Pellman D - Oncogene (2015)

EF+ cells exhibit an Ewing sarcoma gene expression signature. (A) Unsupervised hierarchical clustering of 88 cancer cell lines, across 12 different tumor types, and the EF+ cells. The Ewing sarcoma cell line cluster is outlined in blue and the EF+ cell line cluster is outlined in red. (B) RT-qPCR of select upregulated and downregulated genes. The error bars indicate the standard error of the mean from three experiments. The upper panel shows genes that are upregulated in the EF+ cells and the lower panel shows genes upregulated in the EF− cells. (C and D) Gene set enrichment analysis (GSEA) shows enrichment of the Hancock et al. upregulated and downregulated gene sets in the gene expression data from the EF+ versus EF− (C) and EF+ versus EFFib (D) comparisons. GSEA plots are also shown for the curated gene sets RIGGI_EWING and ZHANG_TARGETS_OF_EWSR1_FLI1 from the MSigDB collection. The normalized enrichment scores (NES) and FDR q-values are shown.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4829493&req=5

Figure 5: EF+ cells exhibit an Ewing sarcoma gene expression signature. (A) Unsupervised hierarchical clustering of 88 cancer cell lines, across 12 different tumor types, and the EF+ cells. The Ewing sarcoma cell line cluster is outlined in blue and the EF+ cell line cluster is outlined in red. (B) RT-qPCR of select upregulated and downregulated genes. The error bars indicate the standard error of the mean from three experiments. The upper panel shows genes that are upregulated in the EF+ cells and the lower panel shows genes upregulated in the EF− cells. (C and D) Gene set enrichment analysis (GSEA) shows enrichment of the Hancock et al. upregulated and downregulated gene sets in the gene expression data from the EF+ versus EF− (C) and EF+ versus EFFib (D) comparisons. GSEA plots are also shown for the curated gene sets RIGGI_EWING and ZHANG_TARGETS_OF_EWSR1_FLI1 from the MSigDB collection. The normalized enrichment scores (NES) and FDR q-values are shown.
Mentions: Next we addressed whether the EF+ cells exhibited an Ewing sarcoma gene expression signature and, if so, whether our doxycycline-inducible, isogenic system would be advantageous in identifying gene expression changes related to EWS-FLI1. Unsupervised hierarchical clustering was performed with the EF+ cells and 88 cancer cell lines across 12 different tumor types (Figure 5A). Notably, the EF+ cells clustered most closely with the Ewing sarcoma cell lines relative to the other sarcomas and tumor types.

Bottom Line: Here, we report a novel approach to model the initiation of Ewing sarcoma tumorigenesis that exploits the developmental and pluripotent potential of human embryonic stem cells.The inducible expression of EWS-FLI1 in embryoid bodies, or collections of differentiating stem cells, generates cells with properties of Ewing sarcoma tumors, including characteristics of transformation.Furthermore, these cells also demonstrate a requirement for the persistent expression of EWS-FLI1 for cell survival and growth, which is a hallmark of Ewing sarcoma tumors.

View Article: PubMed Central - PubMed

Affiliation: Department of Pediatric Oncology, Dana-Farber Cancer Institute, Boston, MA, USA.

ABSTRACT
Oncogenic transformation in Ewing sarcoma tumors is driven by the fusion oncogene EWS-FLI1. However, despite the well-established role of EWS-FLI1 in tumor initiation, the development of models of Ewing sarcoma in human cells with defined genetic elements has been challenging. Here, we report a novel approach to model the initiation of Ewing sarcoma tumorigenesis that exploits the developmental and pluripotent potential of human embryonic stem cells. The inducible expression of EWS-FLI1 in embryoid bodies, or collections of differentiating stem cells, generates cells with properties of Ewing sarcoma tumors, including characteristics of transformation. These cell lines exhibit anchorage-independent growth, a lack of contact inhibition and a strong Ewing sarcoma gene expression signature. Furthermore, these cells also demonstrate a requirement for the persistent expression of EWS-FLI1 for cell survival and growth, which is a hallmark of Ewing sarcoma tumors.

No MeSH data available.


Related in: MedlinePlus