Identification of a novel FUS/ETV4 fusion and comparative analysis with other Ewing sarcoma fusion proteins

Ewing sarcoma is an aggressive pediatric bone cancer defined by a chromosomal translocation fusing one of the FET family members to a member of the ETS transcription factor family. To date, there have been seven reported translocations, with the most recent translocation reported over a decade ago. We now report the first identification of a novel translocation occurring between the FUS gene and ETS family member ETV4 detected in a neonatal patient with Ewing sarcoma. Given its apparent rarity, we conducted an initial characterization of FUS/ETV4 function by performing genomic localization and transcriptional regulatory studies. We knocked down endogenous EWS/FLI in the A673 cell line, and expressed FUS/ETV4 in its stead, and performed CUT&Tag and RNA-sequencing analyses. We compared these data to similar knock-down/rescue analyses of other rare (non-EWS/FLI) Ewing sarcoma-associated translocation products. Through this comparative analysis in the same genetic background, we demonstrate significant similarities across these fusions, and in doing so, validate this novel FUS/ETV4 translocation as a bona fide Ewing sarcoma translocation. This study presents the first genomic comparisons of the rare Ewing sarcoma-associated translocation products, and reveals that the FET/ETS fusions share highly similar, but not identical, genomic localization and transcriptional regulation patterns. These data provide insights into the roles of both the FET and ETS sides of these fusions, and provide a generic strategy to provide further strength to the notion that FET/ETS fusions are key drivers of, and thus pathognomonic for, Ewing sarcoma.

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EWS-FLI-1 creates a cell surface microenvironment conducive to IGF signaling by inducing pappalysin-1

10.1101/195693 ◽

2017 ◽

Author(s):

Panneerselvam Jayabal ◽

Peter J. Houghton ◽

Yuzuru Shiio

Keyword(s):

Cell Surface ◽

Ewing Sarcoma ◽

Chromosomal Translocation ◽

Gene Promoter ◽

Igf Binding Proteins ◽

A Cell ◽

Ets Family ◽

Novel Target ◽

Sarcoma Cells ◽

Igf Signaling

AbstractEwing sarcoma is an aggressive cancer of bone and soft tissue in children with poor prognosis. It is characterized by the chromosomal translocation between EWS and an Ets family transcription factor, most commonly FLI-1. EWS-FLI-1 fusion accounts for 85% of Ewing sarcoma cases. EWS-FLI-1 regulates the expression of a number of genes important for sarcomagenesis, can transform NIH3T3 and C3H10T1/2 cells, and is necessary for proliferation and tumorigenicity of Ewing sarcoma cells, suggesting that EWS-FLI-1 is the causative oncoprotein.Here we report that EWS-FLI-1 induces the expression of pappalysin-1 (PAPPA), a cell surface protease that degrades IGF binding proteins (IGFBPs) and increases the bioavailability of IGF. EWS-FLI-1 binds to the pappalysin-1 gene promoter and stimulates the expression of pappalysin-1, leading to degradation of IGFBPs and enhanced IGF signaling. Silencing of pappalysin-1 strongly inhibited anchorage-dependent and anchorage-independent growth as well as xenograft tumorigenicity of Ewing sarcoma cells. These results suggest that EWS-FLI-1 creates a cell surface microenvironment conducive to IGF signaling by inducing pappalysin-1, which emerged as a novel target to inhibit IGF signaling in Ewing sarcoma.

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Genomic alterations and associated pathway abnormalities in Ewing sarcoma.

Journal of Clinical Oncology ◽

10.1200/jco.2021.39.15_suppl.11532 ◽

2021 ◽

Vol 39 (15_suppl) ◽

pp. 11532-11532

Author(s):

Adam Rock ◽

Warren Allen Chow ◽

An Uche ◽

Sherri Z. Millis

Keyword(s):

Ewing Sarcoma ◽

Clinical Care ◽

Clinical Status ◽

Growth Factor Receptor ◽

Chromosomal Translocation ◽

Bone Cancer ◽

Genomic Alterations ◽

Patient Pathways ◽

Metastatic Setting ◽

Risk Of Cancer

11532 Background: Ewing Sarcoma (ES) is an aggressive, translocation-associated, bone cancer associated with a poor prognosis in the recurrent or metastatic setting. ES is identified by the canonical balanced reciprocal chromosomal translocation involving EWSR1 and ETS transcription factors. Secondary somatic alterations in ES are rarely described and genomic alterations (GA) affecting various molecular pathways may work synergistically with ETS-FL1 translocations to promote oncogenesis. Alterations in fibroblast growth factor receptor 4 (FGFR4), a receptor tyrosine kinase protein that functions in cellular processes, have been observed to affect carcinogenesis. Moreover, the FGFR4-Gly388Arg (G388R) single nucleotide polymorphism (SNP) is found to increase the risk of cancer with mouse embryonic fibroblasts derived from knock-in strain of homologous Fgfr4 G385R mice exhibiting a transformed phenotype. We sought to further evaluate the frequency of FGFR4 G388R SNP in relation to other identifiable pathway alterations observed with Comprehensive Genomic Profiling (CGP). Methods: Next generation sequencing (NGS) analysis was obtained in the context of clinical care with clinical status, outcomes, and source acquisition (primary tumor, metastasis, or recurrence) unknown to Foundation Medicine. CGP, FoundationOne Heme, evaluated GAs including base substitutions, indels, amplifications, copy number alterations, gene fusions and rearrangements. 189 samples were assayed by hybrid-capture based CGP, including 406 DNA-sequenced genes in addition to 265 RNA-sequenced genes commonly reported to be rearranged in cancer, as previously described. Tumor mutational burden was assessed from a minimum 1.4 Mb sequenced DNA. Microsatellite instability (MSI) status was determined by a novel algorithm analyzing 114 specific loci. Results: The median age of evaluated patients was 20 (range 0-70) with the number of alterations averaging 7 per patient. Pathways noted to be altered in the presence of FGFR388R SNP occurred frequently with the following pathways most observed: MAPK (33%), WNT (32%), NOTCH1 (20%), HRR (19%), Histone/chromatin remodeling (18%). FGFR388R SNP was observed in more than half (51%) of evaluated samples. Most affected pathways irrespective of FGFR388R SNP status included: MAPK (n = 89), HRR (n = 75), and PIK3 (n = 64). All evaluated samples were TMB low ( < 10 mut/mb) and Microsatellite Stable. Conclusions: Secondary GAs affecting major pathways were observed in high frequency, often co-occurring with the FGFR4 G388R SNP. Secondary alteration of known oncogenic pathways may contribute to sarcoma formation in ES potentially informing further therapeutic strategies in the future.

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Molecular dissection of the mechanism by which EWS/FLI expression compromises actin cytoskeletal integrity and cell adhesion in Ewing sarcoma

Molecular Biology of the Cell ◽

10.1091/mbc.e14-01-0007 ◽

2014 ◽

Vol 25 (18) ◽

pp. 2695-2709 ◽

Cited By ~ 26

Author(s):

Aashi Chaturvedi ◽

Laura M. Hoffman ◽

Christopher C. Jensen ◽

Yi-Chun Lin ◽

Allie H. Grossmann ◽

...

Keyword(s):

Cell Adhesion ◽

Ewing Sarcoma ◽

Transcriptional Repression ◽

Morphological Changes ◽

Xenograft Model ◽

Chromosomal Translocation ◽

Bone Cancer ◽

Selective Advantage ◽

Altered Expression ◽

Genes Encoding

Ewing sarcoma is the second-most-common bone cancer in children. Driven by an oncogenic chromosomal translocation that results in the expression of an aberrant transcription factor, EWS/FLI, the disease is typically aggressive and micrometastatic upon presentation. Silencing of EWS/FLI in patient-derived tumor cells results in the altered expression of hundreds to thousands of genes and is accompanied by dramatic morphological changes in cytoarchitecture and adhesion. Genes encoding focal adhesion, extracellular matrix, and actin regulatory proteins are dominant targets of EWS/FLI-mediated transcriptional repression. Reexpression of genes encoding just two of these proteins, zyxin and α5 integrin, is sufficient to restore cell adhesion and actin cytoskeletal integrity comparable to what is observed when the EWS/FLI oncogene expression is compromised. Using an orthotopic xenograft model, we show that EWS/FLI-induced repression of α5 integrin and zyxin expression promotes tumor progression by supporting anchorage-independent cell growth. This selective advantage is paired with a tradeoff in which metastatic lung colonization is compromised.

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Analysis of the principles of reference inventory today

Geodesy and Cartography ◽

10.22389/0016-7126-2017-924-6-43-48 ◽

2017 ◽

Vol 924 (6) ◽

pp. 43-48 ◽

Cited By ~ 1

Author(s):

V.N. Klyushnichenko

Keyword(s):

Comparative Analysis ◽

Real Estate ◽

Developed Countries ◽

Real Property ◽

Property Owner ◽

State Register ◽

Information Object ◽

Bona Fide ◽

Source Of Information ◽

Lost Object

A comparative analysis of the principles of the cadastre in the most developed countries and in Russia. It is shown that some of the principles of cadastre, it is advisable to introduce into the Russian legislation. Such principles include the principle of Renzenberger, as well as the principles of Ruoff and Kuranda. The Russian inventory has more than twenty years, however, it cannot be considered complete, as registered in cadastre only 60 % of real estate. Full filling of the cadastre information on real estate is possible, if we abandon the application of the principle of reference. Unlike foreign domestic inventory the inventory contains errors that complicate the procedure of registration of immovable property. In addition, the domestic inventory is not the only source of information about the property that causes the ambiguity of the information about the same object. Important is also that the damage caused inaccurate inventory information bona fide buyer or seller of real property under current law, does not exceed one million rubles, regardless of the value of the lost object. Foreign inventory recognizes the property owner the main participant of the changes, however, the Russian legislation allows for the adjustment of the information object without the application of the property owner. See principles of the foreign inventory is useful for the maintenance of the national cadastre. This will simplify the process of state cadastral accounting of real estate, reduce the time of its formation and to increase the reliability of materials of the Unified state register of real estate.

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Cloning and characterization of the rat apobec-1 gene: a comparative analysis of gene structure and promoter usage in rat and mouse

The Journal of Lipid Research ◽

10.1016/s0022-2275(20)37193-5 ◽

1997 ◽

Vol 38 (6) ◽

pp. 1103-1119

Author(s):

K Hirano ◽

J Min ◽

T Funahashi ◽

N O Davidson

Keyword(s):

Comparative Analysis ◽

Gene Structure ◽

Promoter Usage

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Metastasis of Ewing Sarcoma to the Pancreas: Case Report and Literature Review

Case Reports in Oncological Medicine ◽

10.1155/2020/7075048 ◽

2020 ◽

Vol 2020 ◽

pp. 1-9

Author(s):

Hyma Polimera ◽

Prashanth Moku ◽

Shady Piedra Abusharar ◽

Monali Vasekar ◽

Jayakrishna Chintanaboina

Keyword(s):

Ewing Sarcoma ◽

Rare Case ◽

Bone Cancer ◽

Positive Outcome ◽

Tumor Burden ◽

Response To Treatment ◽

Effective Management ◽

Review Of The Literature ◽

Appropriate Treatment ◽

Prompt Diagnosis

Ewing sarcoma (ES) is a highly aggressive malignant bone cancer. ES is part of the Ewing sarcoma family of tumors (ESFT), which express characteristic t(11;22) translocation as well as higher levels of CD99. Given that metastasis and tumor burden are significant prognostic factors in patient’s response to treatment, prompt diagnosis is needed to effectively treat ESFT patients. However, the challenges in classifying and characterizing ESFT complicate effective management and treatment of ES. In this report, we present a rare case of ES metastasis to the pancreas. Upon review of the literature, we found 39 cases of ESFT involving the pancreas, but only 3 were metastatic to the pancreas while the remaining cases of ESFT primarily originated from the pancreas. Given the rarity of such metastasis, the positive outcome in our patient’s case may explain the importance of prompt diagnosis in order to initiate appropriate treatment.

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A comparative analysis of neural and fuzzy cluster techniques applied to the characterization of electric load in substations

2004 IEEE/PES Transmision and Distribution Conference and Exposition: Latin America (IEEE Cat. No. 04EX956) ◽

10.1109/tdc.2004.1432503 ◽

2005 ◽

Cited By ~ 4

Author(s):

D.Z. Marques ◽

K.A. de Almeida ◽

A.M. de Deus ◽

A.R.G. da Silva Paulo ◽

W. da Silva Lima

Keyword(s):

Comparative Analysis ◽

Fuzzy Cluster ◽

Electric Load

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A novel long intergenic non-coding RNA, Nostrill, regulates iNOS gene transcription and neurotoxicity in microglia

Journal of Neuroinflammation ◽

10.1186/s12974-020-02051-5 ◽

2021 ◽

Vol 18 (1) ◽

Author(s):

Nicholas W. Mathy ◽

Olivia Burleigh ◽

Andrew Kochvar ◽

Erin R. Whiteford ◽

Matthew Behrens ◽

...

Keyword(s):

Nitric Oxide ◽

No Production ◽

Cortical Tissue ◽

Non Coding Rna ◽

Transcriptional Regulatory ◽

Lncrna Locus ◽

Rna Immunoprecipitation ◽

Long Non Coding Rna

Abstract Background Microglia are resident immunocompetent and phagocytic cells in the CNS. Pro-inflammatory microglia, stimulated by microbial signals such as bacterial lipopolysaccharide (LPS), viral RNAs, or inflammatory cytokines, are neurotoxic and associated with pathogenesis of several neurodegenerative diseases. Long non-coding RNAs (lncRNA) are emerging as important tissue-specific regulatory molecules directing cell differentiation and functional states and may help direct proinflammatory responses of microglia. Characterization of lncRNAs upregulated in proinflammatory microglia, such as NR_126553 or 2500002B13Rik, now termed Nostrill (iNOS Transcriptional Regulatory Intergenic LncRNA Locus) increases our understanding of molecular mechanisms in CNS innate immunity. Methods Microglial gene expression array analyses and qRT-PCR were used to identify a novel long intergenic non-coding RNA, Nostrill, upregulated in LPS-stimulated microglial cell lines, LPS-stimulated primary microglia, and LPS-injected mouse cortical tissue. Silencing and overexpression studies, RNA immunoprecipitation, chromatin immunoprecipitation, chromatin isolation by RNA purification assays, and qRT-PCR were used to study the function of this long non-coding RNA in microglia. In vitro assays were used to examine the effects of silencing the novel long non-coding RNA in LPS-stimulated microglia on neurotoxicity. Results We report here characterization of intergenic lncRNA, NR_126553, or 2500002B13Rik now termed Nostrill (iNOS Transcriptional Regulatory Intergenic LncRNA Locus). Nostrill is induced by LPS stimulation in BV2 cells, primary murine microglia, and in cortical tissue of LPS-injected mice. Induction of Nostrill is NF-κB dependent and silencing of Nostrill decreased inducible nitric oxide synthase (iNOS) expression and nitric oxide (NO) production in BV2 and primary microglial cells. Overexpression of Nostrill increased iNOS expression and NO production. RNA immunoprecipitation assays demonstrated that Nostrill is physically associated with NF-κB subunit p65 following LPS stimulation. Silencing of Nostrill significantly reduced NF-κB p65 and RNA polymerase II recruitment to the iNOS promoter and decreased H3K4me3 activating histone modifications at iNOS gene loci. In vitro studies demonstrated that silencing of Nostrill in microglia reduced LPS-stimulated microglial neurotoxicity. Conclusions Our data indicate a new regulatory role of the NF-κB-induced Nostrill and suggest that Nostrill acts as a co-activator of transcription of iNOS resulting in the production of nitric oxide by microglia through modulation of epigenetic chromatin remodeling. Nostrill may be a target for reducing the neurotoxicity associated with iNOS-mediated inflammatory processes in microglia during neurodegeneration.

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