Limits...
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

The immunophenotype of EF+ (A) and EFFib (C) cells as determined by flow cytometry. Cells were labeled with APC- and PE-labeled antibodies against CD34, CD73, CD99, CD105, c-Kit, CD271 and isotype-matched control antibodies. The isoptype control is shown in black and the specific antibody is shown in red. (B) Multiparameter flow cytometry with EFFib and EF+ cells for CD105 and CD271. (D) FACS analysis comparing expression of CD99 and c-Kit in EF+ cells (red line) and EF− cells (black line), four days after removal of doxycycline.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4829493&req=5

Figure 2: The immunophenotype of EF+ (A) and EFFib (C) cells as determined by flow cytometry. Cells were labeled with APC- and PE-labeled antibodies against CD34, CD73, CD99, CD105, c-Kit, CD271 and isotype-matched control antibodies. The isoptype control is shown in black and the specific antibody is shown in red. (B) Multiparameter flow cytometry with EFFib and EF+ cells for CD105 and CD271. (D) FACS analysis comparing expression of CD99 and c-Kit in EF+ cells (red line) and EF− cells (black line), four days after removal of doxycycline.

Mentions: Ewing sarcoma tumors can express both mesenchymal markers, including CD73 and CD105, and neural crest markers, including CD27126,27. We found that the EF+ cells, similar to Ewing sarcoma tumors, express the markers CD73, CD105 and CD271 (Figure 2A). Multiparameter flow cytometry was used to confirm that individual EF+ cells were positive for both CD105 and CD271 (Figure 2B). In contrast, the EFFib cells express the mesenchymal cell markers CD73 and CD105, consistent with the differentiation protocol used in this study and their fibroblast-like morphology (Figure 1D), but do not express the neural crest cell marker CD271. None of the cells express CD34, a hematopoietic cell marker.


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

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

The immunophenotype of EF+ (A) and EFFib (C) cells as determined by flow cytometry. Cells were labeled with APC- and PE-labeled antibodies against CD34, CD73, CD99, CD105, c-Kit, CD271 and isotype-matched control antibodies. The isoptype control is shown in black and the specific antibody is shown in red. (B) Multiparameter flow cytometry with EFFib and EF+ cells for CD105 and CD271. (D) FACS analysis comparing expression of CD99 and c-Kit in EF+ cells (red line) and EF− cells (black line), four days after removal of doxycycline.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4829493&req=5

Figure 2: The immunophenotype of EF+ (A) and EFFib (C) cells as determined by flow cytometry. Cells were labeled with APC- and PE-labeled antibodies against CD34, CD73, CD99, CD105, c-Kit, CD271 and isotype-matched control antibodies. The isoptype control is shown in black and the specific antibody is shown in red. (B) Multiparameter flow cytometry with EFFib and EF+ cells for CD105 and CD271. (D) FACS analysis comparing expression of CD99 and c-Kit in EF+ cells (red line) and EF− cells (black line), four days after removal of doxycycline.
Mentions: Ewing sarcoma tumors can express both mesenchymal markers, including CD73 and CD105, and neural crest markers, including CD27126,27. We found that the EF+ cells, similar to Ewing sarcoma tumors, express the markers CD73, CD105 and CD271 (Figure 2A). Multiparameter flow cytometry was used to confirm that individual EF+ cells were positive for both CD105 and CD271 (Figure 2B). In contrast, the EFFib cells express the mesenchymal cell markers CD73 and CD105, consistent with the differentiation protocol used in this study and their fibroblast-like morphology (Figure 1D), but do not express the neural crest cell marker CD271. None of the cells express CD34, a hematopoietic cell marker.

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