Rnabinding protein ews is a protein that in humans is encoded by the ewsr1 gene on human chromosome 22, specifically 22q12. Surprisingly, there is striking overlap between the ewswt1 and ewsfli1 gene signatures, despite the fact that the dnabinding domain of the fusion proteins, wt1 and fli1, is structurally unique. Ews and wt1 gene fusion in desmoplastic small round cell. For the multiplex iii ewswt1, sytssx1 and 2, pax3, or 7fkhr and for all other fusions listed in table 1, the sensitivities of detection were similar to that of the ews control, ie, the. Ewing sarcoma is a rare malignancy arising in the bone and softtissue and most frequently occurs in children and adolescence. We first generated doxycyclineinducible ews atf1 es cells, in which the human ews atf1 type 2 fusion gene 26, 31 can be induced under the control of a tetracyclineresponsive regulatory element figure 1a. Jci ewsatf1 expression induces sarcomas from neural crest. We examined gene expression profiles in cells expressing ews wt1. Desmoplastic small round cell tumor is dependent on the ews. Mechanism of action of trabectedin in desmoplastic small. The oncogenic properties of ewswt1 of desmoplastic small round.
Pdf ewswt1 oncoprotein activates neuronal reprogramming. The translocation fuses exon 7 of the ews gene to exon 8 of the wt1 gene ews wt1. Generation of ewswt1 knockin mice and animal care to generate a conditional ewswt1 mouse, we followed. Pdf the ewswt1 translocation product induces pdgfa in. Backgroundthe ewsfli1 fusion protein is associated with 8590% of ewings sarcoma family tumors esft, the remaining 1015% of cases expressing chimeric genes encoding ews or fus fused to one of several ets transcription factor family members, including erg1, fev, etv1 and etv6. Although a number of individual targets of ewswt1 have been reported in the literature, the gene signature of the fusion protein and a. Phase ii study of ganitumab, a fully human antitype1. The chimeric gene is abundantly transcribed and produces a 59 kda protein unique to dsrct with fusion of ews exon 7 to wt1 exon 8. Here, we present a comprehensive genomic profiling of one dsrct affecting a 26yearold male, who achieved an excellent outcome. Desmoplastic small round cell tumor dsrct is characterized by the presence of a fusion protein ews wt1, arising from the t 11. Dsrct expressing the ewswt1 fusion protein, we investigated the ability. For the multiplex iii ews wt1, sytssx1 and 2, pax3, or 7fkhr and for all other fusions listed in table 1, the sensitivities of detection were similar to that of the ews control, ie, the. We examined gene expression profiles in cells expressing ewswt1. The chimeric transcript ewswt1 was only described in desmoplastic small round cell tumor 48, 49.
A chromosomal translocation fusion gene product ews wt1 is the defining genetic event in desmoplastic small round cell tumor dsrct, a rare but aggressive tumor with a high rate of mortality. Fusion of the ews gene to chn, a member of the steroidthyroid receptor gene superfamily, in a human myxoid chondrosarcoma. Ewswt1 cooperates with the loss of p53 to exert its. This region of chromosome 22 encodes the nterminal transactivation domain of the ews protein and that region may become joined to one of several other chromosomes which encode various transcription factors, see the expression of a chimeric protein with the ews. The oncogenic properties of ewswt1 of desmoplastic small. Mscs were cotransfected with the 1682pgl3 construct together with either an empty pmscv or a pmscv construct containing sequences encoding the ewsfli1, ewserg or fuserg fusion. The ewsr1 gene provides instructions for making the ews protein, whose function is not completely understood. Molecularly, ewing sarcoma is characterized by a translocation between chromosome 11 and 22, leading to a fusion protein of ewing sarcoma breakpoint region 1 ewsr1 with friend leukemia integration 1 transcription factor fli1. The engineering workstation ews labs currently include over 1,200 workstations and serve over 19,000 students taking engineering courses. Although one wt1 splicing variant encodes a wellcharacterized zinc finger transcription factor, little is known about the function of the most prevalent wt1 isoform, whose dna binding domain is disrupted by a threeamino acid kts insertion.
This chromosomal translocation generates a chimeric transcript that is formed by fusion of the 5. After the alignment steps, the software can generate final gene fusion candidates and integrating our software into the pipeline will come very easily to you. Foxo1 is a direct target of ewsfli1 oncogenic fusion protein. A genomic case study of desmoplastic small round cell tumor. The ews protein has two regions that contribute to its function. Ladanyi, characterization of the genomic breakpoint and chimeric transcripts in the ewswt1 gene fusion of desmoplastic small round cell tumor, proceedings of the national academy of sciences of the united states of america, vol. Ewswt1 oncoprotein activates neuronal reprogramming factor. The cell line expresses different variants of ewswt1, some already identified. Primers designed to amplify cdna spanning the junction region of potential transcripts from this ews wt1 gene fusion were usedto analyze rnafromsix dsrctbyrt. Characterization of the genomic breakpoint and chimeric transcripts in the ews wt1 gene fusion of desmoplastic small round cell tumor.
Fusion of the ews and wt1 genes in the desmoplastic small. Our study replicates previously reported susceptibility loci at 1p36. Desmoplastic small round cell tumor is dependent on the ewswt1. The oncogenic fusion gene ewswt1 is the defining chromosomal. Esft are dependent on insulinlike growth factor1 igf1 for growth and survival and recent evidence. Molecular variants of the ewswt1 gene fusion in desmoplastic small round cell tumor. An openlabel phase ii study was conducted to evaluate the efficacy and safety of ganitumab monotherapy in patients with metastatic ewing family tumors eft or desmoplastic small round cell tumors dsrct. Ewswt1 oncogene activates a neuronal reprogramming. Tamoxifen, 4hydroxytamoxifen 4ht and g418 sigma, and n2 supplement invitrogen were purchased.
Detection of the ews wt1 gene fusion or chimeric rna is diagnostic for dsrct. Jun 16, 2014 the oncogenic fusion gene ews wt1 is the defining chromosomal translocation in desmoplastic small round cell tumors dsrct, a rare but aggressive soft tissue sarcoma with a high rate of mortality. We show a clear dependence of the tumor on ewswt1 in two different cell lines. Surprisingly, there is striking overlap between the ews wt1 and ews fli1 gene signatures, despite the fact that the dnabinding domain of the fusion proteins, wt1 and fli1, is structurally unique. Sequence analysis of both cases revealed fusion between exon 7 of the ews gene and exon 8 of the wt1 gene. The ewswt1 fusion associated with desmoplastic small round cell tumors dsrct, and a truncated protein composed of the first 262 amino acids of fus with a. Ewswt1 oncoprotein activates neuronal reprogramming. The simultaneous presence of two major transcripts emerged, the first characterized by the fusion of exons 17 and 10 of ews gene to exons 810 of wt1, marked in fig. Houghton1,2,3, yidong chen1,2,4 and yuzuru shiio1,2,5 1 greehey childrens cancer research institute, the university of texas health science center, san antonio, texas, usa. Dsrct thus represents the third primitive sarcoma in which the ews gene is involved and the first instance of recurrent rearrangement of a tumor suppressor gene, wt1, in a specific tumor type. Upon treatment of these es cells with doxycycline, expression of the ews atf1 fusion transcript was detected by rtpcr figure 1b. Incidence and outcomes of desmoplastic small round cell tumor. Ewing sarcoma ews is a pediatric cancer characterized by the ewsr1fli1 fusion. Adenosine transporter ent4 is a direct target of ewswt1.
Cellcycle analysis was performed on modfit lt analytical software verity. The wt1 and ews genes encode dna and rna binding proteins involved in wilms tumor and ewing sarcoma pathogenesis, respectively. Mechanism of action of trabectedin in desmoplastic small round cell. This biomarker has been used in four posttreatment blood samples, 3 years after surgery, and no trace of ews wt1 gene fusion was detected, in. A multiplex realtime pcr assay for the detection of gene. The ews wt1 gene fusion in desmoplastic small round cell tumor. Development and evaluation of a pansarcoma fusion gene. Exon 7 of ews is the most frequent site of fusion in ewings sarcoma and related primitive neuroectodermal tumors, in which the ews gene is fused to the part encoding the dna. Northern blotting showed aberrant ews and wt1 transcripts of the same size, suggesting the presence of a chimeric messenger rna.
Reverse transcriptasepolymerase chain reaction as an. Molecular detection and targeting of ewsr1 fusion transcripts. We have recently shown that this translocation represents a rearrangement between the ews and wt1 genes, normally located at 22q12 and 11p, respectively, generating a fusion gene which encodes a chimeric rna resulting from an inframe junction of ews exon 7 to wt1 exon 8. Desmoplastic small round cell tumor of the middle ear. One region, the transcriptional activation domain, allows the ews protein to turn on activate the first step in the production of proteins from genes transcription. Rumsey memorial foundation, and the pew scholars program in the. We performed a genomewide association study of 733 ews cases and 46 unaffected individuals of european ancestry. Ews windows lab software engineering it shared services.
Apr 28, 2020 ews wt1 loss alters the transcriptome of dsrct cells. The role of fli1ews, a fusion gene reciprocal to ewsfli1. Jan 18, 20 wt1 gene, deleted in individuals with wilms tumor, encodes a zinc finger dnabinding protein that acts as a transcriptional activator or repressor, mainly regulating normal formation in the genitourinary system and mesothelial tissues. Activation of igfir promoter activity by the ewswt1 fusion protein in.
Ews wt1 functions as an aberrant transcription factor that drives tumorigenesis, but the mechanistic basis for its pathogenic activity is not well understood. This research was supported by the intramural research program of. Ewswt1 functions as an aberrant transcription factor that drives tumorigenesis, but the mechanistic basis for its pathogenic activity is not well understood. A novel ewswt1 gene fusion product in desmoplastic small round. Ent4 is transcriptionally activated by both isoforms of ewswt1 as. Consistent comigration of the rearranged ews and wt1 bands in multiple enzyme digests indicated fusion of the genomic sequences, presumably due to the translocation t11. Consistenttranslocations involving the wt1 genehavenotbeendescribed. The fusion of these two genes in dsrct results in the production of a putatively oncogenic protein composed of the zinc finger dna binding domains of wt1 linked to potential transcriptional regulatory domains of ews. The different translocation partners of the ews gene, all of which are putative or definite transcription factor genes, may be responsible for the.
A novel type of ewschop fusion gene in myxoid liposarcoma. Molecular genetic characterization of the ewsatf1 fusion. Ewswt1 loss alters the transcriptome of dsrct cells. Knockdown of ewsr1fli1 expression alters the transcriptome. Request pdf a novel ewswt1 gene fusion product in desmoplastic small round cell tumor is a potent transactivator of the insulinlike growth factori receptor.
Molecular variants of the ewswt1 gene fusion in desmoplastic. Axiovert 200m and analyzed with the lsm 510 confocal software zeiss. Desmoplastic small round cell tumor dsrct is a primitive sarcoma with a consistent cytogenetic abnormality, t11. Ewing sarcoma is characterized by chromosomal rearrangements between the gene ewsr1 and gene members of the ets. Desmoplastic small round cell tumor is dependent on the. Purpose ganitumab is a fully human monoclonal antibody against type1 insulinlike growth factor receptor igf1r. A novel ewswt1 gene fusion product in desmoplastic small round cell tumor is a potent transactivator of the insulinlike growth factori receptor igfir gene. The ews wt1 gene fusion protein serves as a diseasespecific marker and yields a definitive diagnosis of dsrct. Ews provides high performance, highly reliable instructional computing environment for the students and faculty of the college of engineering. Other previously identified targets of the fusion protein, such as pdgfa.
The ewswt1 gene fusion in desmoplastic small round cell tumor. The igfi receptor gene promoter is a molecular target for the. The oncogenic fusion gene ewswt1 is the defining chromosomal translocation in desmoplastic small round cell tumors dsrct, a rare but aggressive soft tissue sarcoma with a. Dnabinding segment of wt1, the wilms tumor suppressor gene. A novel ewswt1 gene fusion product in desmoplastic small round cell tumor is a potent transactivator of the insulinlike growth factori receptor igfir gene haim werner, gila idelman, moran rubinstein, patrick pattee, srinivasa r. Wt1 encodes a tumor suppressor first identified by its inactivation in wilms tumor. Ews wt1 oncogene acts as an aberrant transcription factor that drives tumorigenesis, but the mechanism by which ews wt1 causes tumorigenesis is not well understood. A novel ewswt1 gene fusion product in desmoplastic small. However, by performing effective filtering, sensitive and accurate gene fusion detection is possible. Icc analysis icc analysis was carried out on all 51 cases on cell blocks or on representative smears or both using a panel of antibodies against cytokeratin, vimentin, desmin, nse, cd99, and leukocyte common antigen.1555 978 1111 668 1361 449 1238 1398 628 276 1187 1516 603 1361 843 1224 1272 1089 1630 741 679 749 1112 380 1282 1010 1450 174 338 59 390 679 216 1289 1233 1436 707 379 837 1347 721 29 220 1334 145 466