Desmoplastic small round cell tumor dsrct is a primitive sarcoma with a consistent cytogenetic abnormality, t11. 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. 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 and wt1 genes in the desmoplastic small. Mechanism of action of trabectedin in desmoplastic small. Axiovert 200m and analyzed with the lsm 510 confocal software zeiss. The engineering workstation ews labs currently include over 1,200 workstations and serve over 19,000 students taking engineering courses. 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. Rumsey memorial foundation, and the pew scholars program in the.
Molecular variants of the ewswt1 gene fusion in desmoplastic small round cell tumor. Ewswt1 loss alters the transcriptome of dsrct cells. Molecular genetic characterization of the ewsatf1 fusion. Mechanism of action of trabectedin in desmoplastic small round cell. Dnabinding segment of wt1, the wilms tumor suppressor gene. This research was supported by the intramural research program of.
A novel ewswt1 gene fusion product in desmoplastic small round cell tumor is a potent transactivator of the insulinlike growth factori receptor igfir gene. 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. Ewswt1 oncoprotein activates neuronal reprogramming. We performed a genomewide association study of 733 ews cases and 46 unaffected individuals of european ancestry. We examined gene expression profiles in cells expressing ews wt1. Molecular detection and targeting of ewsr1 fusion transcripts. Primers designed to amplify cdna spanning the junction region of potential transcripts from this ews wt1 gene fusion were usedto analyze rnafromsix dsrctbyrt. 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. 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. Generation of ewswt1 knockin mice and animal care to generate a conditional ewswt1 mouse, we followed. 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.
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. 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. Pdf ewswt1 oncoprotein activates neuronal reprogramming. Ent4 is transcriptionally activated by both isoforms of ewswt1 as. We examined gene expression profiles in cells expressing ewswt1. The igfi receptor gene promoter is a molecular target for the. Tamoxifen, 4hydroxytamoxifen 4ht and g418 sigma, and n2 supplement invitrogen were purchased. Ewing sarcoma is a rare malignancy arising in the bone and softtissue and most frequently occurs in children and adolescence. Molecular heterogeneity and function of ewswt1 fusion. The ewsr1 gene provides instructions for making the ews protein, whose function is not completely understood. 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. However, by performing effective filtering, sensitive and accurate gene fusion detection is possible. Ews windows lab software engineering it shared services.
Ews wt1 oncoprotein activates neuronal reprogramming factor. Ews wt1 functions as an aberrant transcription factor that drives tumorigenesis, but the mechanistic basis for its pathogenic activity is not well understood. Desmoplastic small round cell tumor is dependent on the ewswt1. Development and evaluation of a pansarcoma fusion gene. Ewswt1 functions as an aberrant transcription factor that drives tumorigenesis, but the mechanistic basis for its pathogenic activity is not well understood. 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. The ewswt1 gene fusion in desmoplastic small round cell.
Molecular variants of the ewswt1 gene fusion in desmoplastic. Desmoplastic small round cell tumor dsrct is characterized by the presence of a fusion protein ews wt1, arising from the t 11. Rnabinding protein ews is a protein that in humans is encoded by the ewsr1 gene on human chromosome 22, specifically 22q12. A novel ewswt1 gene fusion product in desmoplastic small round. Consistenttranslocations involving the wt1 genehavenotbeendescribed.
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. 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. Foxo1 is a direct target of ewsfli1 oncogenic fusion protein. We show a clear dependence of the tumor on ewswt1 in two different cell lines.
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. 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. 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. Dsrct expressing the ewswt1 fusion protein, we investigated the ability.
The ews wt1 gene fusion in desmoplastic small round cell tumor. Apr 28, 2020 ews wt1 loss alters the transcriptome of dsrct cells. Ewswt1 cooperates with the loss of p53 to exert its. This chromosomal translocation generates a chimeric transcript that is formed by fusion of the 5. Reverse transcriptasepolymerase chain reaction as an. Characterization of the genomic breakpoint and chimeric pnas.
The ewswt1 gene fusion in desmoplastic small round cell tumor. Although a number of individual targets of ewswt1 have been reported in the literature, the gene signature of the fusion protein and a. Desmoplastic small round cell tumor of the middle ear. Activation of igfir promoter activity by the ewswt1 fusion protein in. Desmoplastic small round cell tumor is dependent on the. Our study replicates previously reported susceptibility loci at 1p36. The ews wt1 gene fusion protein serves as a diseasespecific marker and yields a definitive diagnosis of dsrct.
Here, we present a comprehensive genomic profiling of one dsrct affecting a 26yearold male, who achieved an excellent outcome. Characterization of the genomic breakpoint and chimeric transcripts in the ews wt1 gene fusion of desmoplastic small round cell tumor. A multiplex realtime pcr assay for the detection of gene. The oncogenic properties of ewswt1 of desmoplastic small round. 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. 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. Purpose ganitumab is a fully human monoclonal antibody against type1 insulinlike growth factor receptor igf1r. The wt1 and ews genes encode dna and rna binding proteins involved in wilms tumor and ewing sarcoma pathogenesis, respectively. A practical approach to the clinical diagnosis of ewings sarcoma primitive neuroectodermal tumour and other small round cell tumours sharing ews rearrangement using new fluorescence in situ hybridisation probes for ewsr1.
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. A novel type of ewschop fusion gene in myxoid liposarcoma. 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. 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. Ewing sarcoma is characterized by chromosomal rearrangements between the gene ewsr1 and gene members of the ets. Adenosine transporter ent4 is a direct target of ewswt1. A novel ewswt1 gene fusion product in desmoplastic small. Pdf the ewswt1 translocation product induces pdgfa in. Northern blotting showed aberrant ews and wt1 transcripts of the same size, suggesting the presence of a chimeric messenger rna. Esft are dependent on insulinlike growth factor1 igf1 for growth and survival and recent evidence. Desmoplastic small round cell tumor is dependent on the ews. In addition to the replacement of transcriptional regulatory domain of wt1 with that of ews, the nterminal first zinc finger that contributes to the binding specificity of wt1 is deleted during the generation of the ews wt1 fusion protein. 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. Cellcycle analysis was performed on modfit lt analytical software verity.
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. Request pdf a novel ewswt1 gene fusion product in desmoplastic small round cell tumor is a potent transactivator of the insulinlike growth factori receptor. Phase ii study of ganitumab, a fully human antitype1. Ews and wt1 gene fusion in desmoplastic small round cell. 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. The translocation fuses exon 7 of the ews gene to exon 8 of the wt1 gene ews wt1. The different translocation partners of the ews gene, all of which are putative or definite transcription factor genes, may be responsible for the. 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. 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. 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.
The ews wt1 protein can attenuate tumor suppressor function of ews gene and promote the expression of growth factor which is inhibited by normal wt1 gene. The cell line expresses different variants of ewswt1, some already identified. Sequence analysis of both cases revealed fusion between exon 7 of the ews gene and exon 8 of the wt1 gene. Ewswt1 oncoprotein activates neuronal reprogramming factor. Upon treatment of these es cells with doxycycline, expression of the ews atf1 fusion transcript was detected by rtpcr figure 1b. The chimeric transcript ewswt1 was only described in desmoplastic small round cell tumor 48, 49. The oncogenic fusion gene ews wt1 is the defining chromosomal translocation in desmoplastic small roundcell tumors dsrct, a rare but aggressive soft tissue sarcoma with a high rate of mortality. 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. Wt1 encodes a tumor suppressor first identified by its inactivation in wilms tumor. The ews protein has two regions that contribute to its function. 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. Igf1 is a common target gene of ewings sarcoma fusion.
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. Detection of the ews wt1 gene fusion or chimeric rna is diagnostic for dsrct. Incidence and outcomes of desmoplastic small round cell tumor. The role of fli1ews, a fusion gene reciprocal to ewsfli1. Ewing sarcoma ews is a pediatric cancer characterized by the ewsr1fli1 fusion.
Other previously identified targets of the fusion protein, such as pdgfa. Knockdown of ewsr1fli1 expression alters the transcriptome. The oncogenic properties of ewswt1 of desmoplastic small. The oncogenic fusion gene ewswt1 is the defining chromosomal. Ewswt1 oncogene activates a neuronal reprogramming.
Ews provides high performance, highly reliable instructional computing environment for the students and faculty of the college of engineering. Jci ewsatf1 expression induces sarcomas from neural crest. Fusion of the ews gene to chn, a member of the steroidthyroid receptor gene superfamily, in a human myxoid chondrosarcoma. A genomic case study of desmoplastic small round cell tumor. Although a number of individual targets of ews wt1 have been reported in the literature, the gene signature of the fusion protein and a.