Modeling Initiation of Ewing Sarcoma in Human Neural Crest Cells

Levetzow, Cornelia von; Jiang, Xiaohua; Gwye, Ynnez; Levetzow, Gregor von; Hung, Long; Cooper, Aaron; Hsu, Jessie Hao-Ru; Lawlor, Elizabeth R.

doi:10.1371/journal.pone.0019305

Cited by 153 publications

(168 citation statements)

References 44 publications

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“…We previously demonstrated DKK2 to be an agonist of the canonical WNT/β-catenin pathway and critical mediator of osteolytic tumor growth in ES [5]. Here, analysis of mesenchymal and neuroectodermal stem cells demonstrated an up-regulation of DKK2 in EWS-FLI1 expressing neuroectodermal stem cells [27]. Long-term exposure in differentiation media resulted in a significant further up-regulation of DKK2 and HOXD13 in EWS- Previous results already demonstrated agonists of WNT/β-catenin signaling to support skeletogenesis [40] or to play a role in terminal osteoblast differentiation into mineralized bone matrices [41].…”

Section: Discussionmentioning

confidence: 65%

“…Consistent with these results, suppression of DKK2 by specific shRNA [5] in different ES lines resulted in a significant down-regulation of HOXD10, HOXD11 and HOXD13 ( Figure 1F). Further, down-regulation of TCF4, a transcription factor in the canonical WNT/β-catenin pathway, resulted in a partial inhibition of posterior www.impactjournals.com/oncotarget Posterior HOXD genes contribute to chondrogenic as well as bone associated gene expression in ES ES are bone or soft tissue neoplasms with a prominent immature stemness phenotype maintained by epigenetic repressors BMI1 and EZH2 [27,28]. Further, posterior HOXD genes regulated by EZH2 during development are known to influence the ossification pattern of bones [22], so it seemed relevant to investigate whether the expression of posterior HOXD genes in ES may influence their differentiation capacity.…”

Section: Hoxd Genes Are Stimulated Via Dkk2mentioning

confidence: 99%

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The posterior HOXD locus: Its contribution to phenotype and malignancy of Ewing sarcoma

Heyking¹,

Roth²,

Ertl³

et al. 2016

European Journal of Cancer

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Section: Discussionmentioning

confidence: 65%

Section: Hoxd Genes Are Stimulated Via Dkk2mentioning

confidence: 99%

The posterior HOXD locus: Its contribution to phenotype and malignancy of Ewing sarcoma

Heyking¹,

Roth²,

Ertl³

et al. 2016

European Journal of Cancer

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“…Individual probeset expression from ESTS was normalized by overall gene expression and then compared between neural crest (NC), hMSC, and TC32 with/without EWS-FLI1. Both NC and hMSC have been putatively evaluated as the ES cell of origin (28,37,38). Using the paired cell line models, we were able to calculate ANOVA P values for each probeset to measure the significance of a consistent effect of EWS-FLI1 across models.…”

Section: Tumors From Es Patients Demonstrate Similar Ews-fli1-influencedmentioning

confidence: 99%

Oncogenic fusion protein EWS-FLI1 is a network hub that regulates alternative splicing

Selvanathan

Graham

Erkizan

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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The synthesis and processing of mRNA, from transcription to translation initiation, often requires splicing of intragenic material. The final mRNA composition varies based on proteins that modulate splice site selection. EWS-FLI1 is an Ewing sarcoma (ES) oncoprotein with an interactome that we demonstrate to have multiple partners in spliceosomal complexes. We evaluate the effect of EWS-FLI1 on posttranscriptional gene regulation using both exon array and RNAseq. Genes that potentially regulate oncogenesis, including CLK1, CASP3, PPFIBP1, and TERT, validate as alternatively spliced by EWS-FLI1. In a CLIP-seq experiment, we find that EWS-FLI1 RNA-binding motifs most frequently occur adjacent to intron-exon boundaries. EWS-FLI1 also alters splicing by directly binding to known splicing factors including DDX5, hnRNP K, and PRPF6. Reduction of EWS-FLI1 produces an isoform of γ-TERT that has increased telomerase activity compared with wild-type (WT) TERT. The small molecule YK-4-279 is an inhibitor of EWS-FLI1 oncogenic function that disrupts specific protein interactions, including helicases DDX5 and RNA helicase A (RHA) that alters RNA-splicing ratios. As such, YK-4-279 validates the splicing mechanism of EWS-FLI1, showing alternatively spliced gene patterns that significantly overlap with EWS-FLI1 reduction and WT human mesenchymal stem cells (hMSC). Exon array analysis of 75 ES patient samples shows similar isoform expression patterns to cell line models expressing EWS-FLI1, supporting the clinical relevance of our findings. These experiments establish systemic alternative splicing as an oncogenic process modulated by EWS-FLI1. EWS-FLI1 modulation of mRNA splicing may provide insight into the contribution of splicing toward oncogenesis, and, reciprocally, EWS-FLI1 interactions with splicing proteins may inform the splicing code.T he alternative splicing of mRNA expands the diversity of the human proteome through evolution and ontogeny (1, 2). Spliceosomal network interactions, including proteins that recognize splice enhancer and silencer regions, are critical for the regulation of alternative splicing leading to protein isoforms with disparate functionality (3). Alternative splicing provides both a method by which to categorize subsets of cancers and an avenue for more effective targeted treatments (4). However, the spliceosomal protein interaction networks that are specific to cancer have not been systematically defined; alternative splicing can also change protein-protein interactions within networks (5). A systems biology approach can be used to study the relationship between splicing and oncogenesis by using tumor models with chromosomal translocations whose expressed fusion proteins have putative roles in splicing (6, 7).Fusion proteins produced by chromosomal translocations in sarcomas often contain the amino-terminal portion of EWS and are classified as transcription factors due to the presence of a canonical carboxyl-terminal DNA-binding domain (6). EWS-FLI1 is a well-established ES oncoprotein and is re...

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“…Ewing sarcoma is a highly malignant bone and soft tissue neoplasia of still enigmatic histogenesis with a prominent stemness phenotype (1,2). Histogenesis may be endothelial, neuroectodermal (3)(4)(5), or osteochondrogenic (6,7).…”

Section: Introductionmentioning

confidence: 99%

Deep Sequencing in Conjunction with Expression and Functional Analyses Reveals Activation of FGFR1 in Ewing Sarcoma

Agelopoulos

Richter

Schmidt

et al. 2015

Clinical Cancer Research

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PURPOSE: A low mutation rate seems to be a general feature of pediatric cancers, in particular in oncofusion gene-driven tumors. Genetically, Ewing sarcoma is defined by balanced chromosomal EWS/ETS translocations, which give rise to oncogenic chimeric proteins (EWS-ETS). Other contributing somatic mutations involved in disease development have only been observed at low frequency. EXPER-IMENTAL DESIGN: Tumor samples of 116 Ewing sarcoma patients were analyzed here. Whole-genome sequencing was performed on two patients with normal, primary, and relapsed tissue. Whole-exome sequencing was performed on 50 Ewing sarcoma and 22 matched normal tissues. A discovery dataset of 14 of these tumor/normal pairs identified 232 somatic mutations. Recurrent nonsynonymous mutations were validated in the 36 remaining exomes. Transcriptome analysis was performed in a subset of 14 of 50 Ewing sarcomas and DNA copy number gain and expression of FGFR1 in 63 of 116 Ewing sarcomas. RESULTS: Relapsed tumors consistently showed a 2-to 3-fold increased number of mutations. We identified several recurrently mutated genes at low frequency (ANKRD30A, CCDC19, KIAA0319, KIAA1522, LAMB4, SLFN11, STAG2, TP53, UNC80, ZNF98). An oncogenic fibroblast growth factor receptor 1 (FGFR1) mutation (N546K) was detected, and the FGFR1 locus frequently showed copy number gain (31.7%) in primary tumors. Furthermore, high-level FGFR1 expression was noted as a characteristic feature of Ewing sarcoma. RNA interference of FGFR1 expression in Ewing sarcoma lines blocked proliferation and completely suppressed xenograft tumor growth. FGFR1 tyrosine kinase inhibitor (TKI) therapy in a patient with Ewing sarcoma relapse significantly reduced 18-FDG-PET activity. CONCLUSIONS: FGFR1 may constitute a promising target for novel therapeutic approaches in Ewing sarcoma.

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Modeling Initiation of Ewing Sarcoma in Human Neural Crest Cells

Cited by 153 publications

References 44 publications

The posterior HOXD locus: Its contribution to phenotype and malignancy of Ewing sarcoma

The posterior HOXD locus: Its contribution to phenotype and malignancy of Ewing sarcoma

Oncogenic fusion protein EWS-FLI1 is a network hub that regulates alternative splicing

Deep Sequencing in Conjunction with Expression and Functional Analyses Reveals Activation of FGFR1 in Ewing Sarcoma

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