Characterization of the genomic breakpoint and chimeric transcripts in the EWS-WT1 gene fusion of desmoplastic small round cell tumor (original) (raw)

Abstract

Desmoplastic small round cell tumor is a recently recognized distinctive tumor shown to be associated with a recurrent translocation, t(11;22)(p13;q12), and rearrangement of the genes for Ewing sarcoma (EWS) and Wilms tumor (WT1). A genomic DNA fragment containing the EWS-WT1 gene fusion has been isolated from a desmoplastic small round cell tumor, and the breakpoint has been characterized. The breakpoints involve the intron between EWS exons 7 and 8 and the intron between WT1 exons 7 and 8. Chimeric transcripts corresponding to the fusion gene were detected in four of six cases studied. Analysis of these transcripts show an in-frame fusion of RNA encoding the amino-terminal domain of EWS to both alternatively spliced forms of the last three zinc fingers of the DNA-binding domain of WT1. Desmoplastic small round cell tumor represents the third tumor type associated with translocation of EWS and the first tumor associated with consistent translocation of WT1. The chimeric products are predicted to modulate transcription at WT1 target sites and contribute to development of this unique tumor.

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  1. Bickmore W. A., Oghene K., Little M. H., Seawright A., van Heyningen V., Hastie N. D. Modulation of DNA binding specificity by alternative splicing of the Wilms tumor wt1 gene transcript. Science. 1992 Jul 10;257(5067):235–237. doi: 10.1126/science.1321494. [DOI] [PubMed] [Google Scholar]
  2. Biegel J. A., Conard K., Brooks J. J. Translocation (11;22)(p13;q12): primary change in intra-abdominal desmoplastic small round cell tumor. Genes Chromosomes Cancer. 1993 Jun;7(2):119–121. doi: 10.1002/gcc.2870070210. [DOI] [PubMed] [Google Scholar]
  3. Bonetta L., Kuehn S. E., Huang A., Law D. J., Kalikin L. M., Koi M., Reeve A. E., Brownstein B. H., Yeger H., Williams B. R. Wilms tumor locus on 11p13 defined by multiple CpG island-associated transcripts. Science. 1990 Nov 16;250(4983):994–997. doi: 10.1126/science.2173146. [DOI] [PubMed] [Google Scholar]
  4. Bonin G., Scamps C., Turc-Carel C., Lipinski M. Chimeric EWS-FLI1 transcript in a Ewing cell line with a complex t(11;22;14) translocation. Cancer Res. 1993 Aug 15;53(16):3655–3657. [PubMed] [Google Scholar]
  5. Brenner B., Wildhardt G., Schneider S., Royer-Pokora B. RNA polymerase chain reaction detects different levels of four alternatively spliced WT1 transcripts in Wilms' tumors. Oncogene. 1992 Jul;7(7):1431–1433. [PubMed] [Google Scholar]
  6. Call K. M., Glaser T., Ito C. Y., Buckler A. J., Pelletier J., Haber D. A., Rose E. A., Kral A., Yeger H., Lewis W. H. Isolation and characterization of a zinc finger polypeptide gene at the human chromosome 11 Wilms' tumor locus. Cell. 1990 Feb 9;60(3):509–520. doi: 10.1016/0092-8674(90)90601-a. [DOI] [PubMed] [Google Scholar]
  7. Crozat A., Aman P., Mandahl N., Ron D. Fusion of CHOP to a novel RNA-binding protein in human myxoid liposarcoma. Nature. 1993 Jun 17;363(6430):640–644. doi: 10.1038/363640a0. [DOI] [PubMed] [Google Scholar]
  8. Delattre O., Zucman J., Plougastel B., Desmaze C., Melot T., Peter M., Kovar H., Joubert I., de Jong P., Rouleau G. Gene fusion with an ETS DNA-binding domain caused by chromosome translocation in human tumours. Nature. 1992 Sep 10;359(6391):162–165. doi: 10.1038/359162a0. [DOI] [PubMed] [Google Scholar]
  9. Douglass E. C., Valentine M., Etcubanas E., Parham D., Webber B. L., Houghton P. J., Houghton J. A., Green A. A. A specific chromosomal abnormality in rhabdomyosarcoma. Cytogenet Cell Genet. 1987;45(3-4):148–155. doi: 10.1159/000132446. [DOI] [PubMed] [Google Scholar]
  10. Drummond I. A., Madden S. L., Rohwer-Nutter P., Bell G. I., Sukhatme V. P., Rauscher F. J., 3rd Repression of the insulin-like growth factor II gene by the Wilms tumor suppressor WT1. Science. 1992 Jul 31;257(5070):674–678. doi: 10.1126/science.1323141. [DOI] [PubMed] [Google Scholar]
  11. Drummond I. A., Rupprecht H. D., Rohwer-Nutter P., Lopez-Guisa J. M., Madden S. L., Rauscher F. J., 3rd, Sukhatme V. P. DNA recognition by splicing variants of the Wilms' tumor suppressor, WT1. Mol Cell Biol. 1994 Jun;14(6):3800–3809. doi: 10.1128/mcb.14.6.3800. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Galili N., Davis R. J., Fredericks W. J., Mukhopadhyay S., Rauscher F. J., 3rd, Emanuel B. S., Rovera G., Barr F. G. Fusion of a fork head domain gene to PAX3 in the solid tumour alveolar rhabdomyosarcoma. Nat Genet. 1993 Nov;5(3):230–235. doi: 10.1038/ng1193-230. [DOI] [PubMed] [Google Scholar]
  13. Gerald W. L., Chao J., Chao L. Immunological identification of rat tissue kallikrein cDNA and characterization of the kallikrein gene family. Biochim Biophys Acta. 1986 Feb 24;866(1):1–14. doi: 10.1016/0167-4781(86)90093-x. [DOI] [PubMed] [Google Scholar]
  14. Gerald W. L., Miller H. K., Battifora H., Miettinen M., Silva E. G., Rosai J. Intra-abdominal desmoplastic small round-cell tumor. Report of 19 cases of a distinctive type of high-grade polyphenotypic malignancy affecting young individuals. Am J Surg Pathol. 1991 Jun;15(6):499–513. [PubMed] [Google Scholar]
  15. Gerald W. L., Rosai J. Desmoplastic small cell tumor with multi-phenotypic differentiation. Zentralbl Pathol. 1993 Jun;139(2):141–151. [PubMed] [Google Scholar]
  16. Gessler M., Poustka A., Cavenee W., Neve R. L., Orkin S. H., Bruns G. A. Homozygous deletion in Wilms tumours of a zinc-finger gene identified by chromosome jumping. Nature. 1990 Feb 22;343(6260):774–778. doi: 10.1038/343774a0. [DOI] [PubMed] [Google Scholar]
  17. Haber D. A., Buckler A. J. WT1: a novel tumor suppressor gene inactivated in Wilms' tumor. New Biol. 1992 Feb;4(2):97–106. [PubMed] [Google Scholar]
  18. Haber D. A., Housman D. E. Role of the WT1 gene in Wilms' tumour. Cancer Surv. 1992;12:105–117. [PubMed] [Google Scholar]
  19. Haber D. A., Park S., Maheswaran S., Englert C., Re G. G., Hazen-Martin D. J., Sens D. A., Garvin A. J. WT1-mediated growth suppression of Wilms tumor cells expressing a WT1 splicing variant. Science. 1993 Dec 24;262(5142):2057–2059. doi: 10.1126/science.8266105. [DOI] [PubMed] [Google Scholar]
  20. Haber D. A., Sohn R. L., Buckler A. J., Pelletier J., Call K. M., Housman D. E. Alternative splicing and genomic structure of the Wilms tumor gene WT1. Proc Natl Acad Sci U S A. 1991 Nov 1;88(21):9618–9622. doi: 10.1073/pnas.88.21.9618. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Haber D. A., Timmers H. T., Pelletier J., Sharp P. A., Housman D. E. A dominant mutation in the Wilms tumor gene WT1 cooperates with the viral oncogene E1A in transformation of primary kidney cells. Proc Natl Acad Sci U S A. 1992 Jul 1;89(13):6010–6014. doi: 10.1073/pnas.89.13.6010. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Ladanyi M., Gerald W. Fusion of the EWS and WT1 genes in the desmoplastic small round cell tumor. Cancer Res. 1994 Jun 1;54(11):2837–2840. [PubMed] [Google Scholar]
  23. Madden S. L., Cook D. M., Morris J. F., Gashler A., Sukhatme V. P., Rauscher F. J., 3rd Transcriptional repression mediated by the WT1 Wilms tumor gene product. Science. 1991 Sep 27;253(5027):1550–1553. doi: 10.1126/science.1654597. [DOI] [PubMed] [Google Scholar]
  24. Maheswaran S., Park S., Bernard A., Morris J. F., Rauscher F. J., 3rd, Hill D. E., Haber D. A. Physical and functional interaction between WT1 and p53 proteins. Proc Natl Acad Sci U S A. 1993 Jun 1;90(11):5100–5104. doi: 10.1073/pnas.90.11.5100. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. May W. A., Gishizky M. L., Lessnick S. L., Lunsford L. B., Lewis B. C., Delattre O., Zucman J., Thomas G., Denny C. T. Ewing sarcoma 11;22 translocation produces a chimeric transcription factor that requires the DNA-binding domain encoded by FLI1 for transformation. Proc Natl Acad Sci U S A. 1993 Jun 15;90(12):5752–5756. doi: 10.1073/pnas.90.12.5752. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. May W. A., Lessnick S. L., Braun B. S., Klemsz M., Lewis B. C., Lunsford L. B., Hromas R., Denny C. T. The Ewing's sarcoma EWS/FLI-1 fusion gene encodes a more potent transcriptional activator and is a more powerful transforming gene than FLI-1. Mol Cell Biol. 1993 Dec;13(12):7393–7398. doi: 10.1128/mcb.13.12.7393. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Ohno T., Rao V. N., Reddy E. S. EWS/Fli-1 chimeric protein is a transcriptional activator. Cancer Res. 1993 Dec 15;53(24):5859–5863. [PubMed] [Google Scholar]
  28. Park S., Schalling M., Bernard A., Maheswaran S., Shipley G. C., Roberts D., Fletcher J., Shipman R., Rheinwald J., Demetri G. The Wilms tumour gene WT1 is expressed in murine mesoderm-derived tissues and mutated in a human mesothelioma. Nat Genet. 1993 Aug;4(4):415–420. doi: 10.1038/ng0893-415. [DOI] [PubMed] [Google Scholar]
  29. Park S., Tomlinson G., Nisen P., Haber D. A. Altered trans-activational properties of a mutated WT1 gene product in a WAGR-associated Wilms' tumor. Cancer Res. 1993 Oct 15;53(20):4757–4760. [PubMed] [Google Scholar]
  30. Plougastel B., Zucman J., Peter M., Thomas G., Delattre O. Genomic structure of the EWS gene and its relationship to EWSR1, a site of tumor-associated chromosome translocation. Genomics. 1993 Dec;18(3):609–615. doi: 10.1016/s0888-7543(05)80363-5. [DOI] [PubMed] [Google Scholar]
  31. Pritchard-Jones K., Fleming S. Cell types expressing the Wilms' tumour gene (WT1) in Wilms' tumours: implications for tumour histogenesis. Oncogene. 1991 Dec;6(12):2211–2220. [PubMed] [Google Scholar]
  32. Pritchard-Jones K., Fleming S., Davidson D., Bickmore W., Porteous D., Gosden C., Bard J., Buckler A., Pelletier J., Housman D. The candidate Wilms' tumour gene is involved in genitourinary development. Nature. 1990 Jul 12;346(6280):194–197. doi: 10.1038/346194a0. [DOI] [PubMed] [Google Scholar]
  33. Rauscher F. J., 3rd, Morris J. F., Tournay O. E., Cook D. M., Curran T. Binding of the Wilms' tumor locus zinc finger protein to the EGR-1 consensus sequence. Science. 1990 Nov 30;250(4985):1259–1262. doi: 10.1126/science.2244209. [DOI] [PubMed] [Google Scholar]
  34. Rodriguez E., Sreekantaiah C., Gerald W., Reuter V. E., Motzer R. J., Chaganti R. S. A recurring translocation, t(11;22)(p13;q11.2), characterizes intra-abdominal desmoplastic small round-cell tumors. Cancer Genet Cytogenet. 1993 Aug;69(1):17–21. doi: 10.1016/0165-4608(93)90105-u. [DOI] [PubMed] [Google Scholar]
  35. Sawyer J. R., Tryka A. F., Lewis J. M. A novel reciprocal chromosome translocation t(11;22)(p13;q12) in an intraabdominal desmoplastic small round-cell tumor. Am J Surg Pathol. 1992 Apr;16(4):411–416. doi: 10.1097/00000478-199204000-00010. [DOI] [PubMed] [Google Scholar]
  36. Sharma P. M., Yang X., Bowman M., Roberts V., Sukumar S. Molecular cloning of rat Wilms' tumor complementary DNA and a study of messenger RNA expression in the urogenital system and the brain. Cancer Res. 1992 Nov 15;52(22):6407–6412. [PubMed] [Google Scholar]
  37. Shen W. P., Towne B., Zadeh T. M. Cytogenetic abnormalities in an intraabdominal desmoplastic small cell tumor. Cancer Genet Cytogenet. 1992 Dec;64(2):189–191. doi: 10.1016/0165-4608(92)90355-c. [DOI] [PubMed] [Google Scholar]
  38. Sorensen P. H., Lessnick S. L., Lopez-Terrada D., Liu X. F., Triche T. J., Denny C. T. A second Ewing's sarcoma translocation, t(21;22), fuses the EWS gene to another ETS-family transcription factor, ERG. Nat Genet. 1994 Feb;6(2):146–151. doi: 10.1038/ng0294-146. [DOI] [PubMed] [Google Scholar]
  39. Tadokoro K., Oki N., Fujii H., Ohshima A., Inoue T., Yamada M. Genomic organization of the human WT1 gene. Jpn J Cancer Res. 1992 Nov;83(11):1198–1203. doi: 10.1111/j.1349-7006.1992.tb02745.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Turc-Carel C., Aurias A., Mugneret F., Lizard S., Sidaner I., Volk C., Thiery J. P., Olschwang S., Philip I., Berger M. P. Chromosomes in Ewing's sarcoma. I. An evaluation of 85 cases of remarkable consistency of t(11;22)(q24;q12). Cancer Genet Cytogenet. 1988 Jun;32(2):229–238. doi: 10.1016/0165-4608(88)90285-3. [DOI] [PubMed] [Google Scholar]
  41. Turc-Carel C., Lizard-Nacol S., Justrabo E., Favrot M., Philip T., Tabone E. Consistent chromosomal translocation in alveolar rhabdomyosarcoma. Cancer Genet Cytogenet. 1986 Jan 15;19(3-4):361–362. doi: 10.1016/0165-4608(86)90069-5. [DOI] [PubMed] [Google Scholar]
  42. Wang-Wuu S., Soukup S., Ballard E., Gotwals B., Lampkin B. Chromosomal analysis of sixteen human rhabdomyosarcomas. Cancer Res. 1988 Feb 15;48(4):983–987. [PubMed] [Google Scholar]
  43. Whang-Peng J., Triche T. J., Knutsen T., Miser J., Douglass E. C., Israel M. A. Chromosome translocation in peripheral neuroepithelioma. N Engl J Med. 1984 Aug 30;311(9):584–585. doi: 10.1056/NEJM198408303110907. [DOI] [PubMed] [Google Scholar]
  44. Zucman J., Delattre O., Desmaze C., Epstein A. L., Stenman G., Speleman F., Fletchers C. D., Aurias A., Thomas G. EWS and ATF-1 gene fusion induced by t(12;22) translocation in malignant melanoma of soft parts. Nat Genet. 1993 Aug;4(4):341–345. doi: 10.1038/ng0893-341. [DOI] [PubMed] [Google Scholar]
  45. Zucman J., Delattre O., Desmaze C., Plougastel B., Joubert I., Melot T., Peter M., De Jong P., Rouleau G., Aurias A. Cloning and characterization of the Ewing's sarcoma and peripheral neuroepithelioma t(11;22) translocation breakpoints. Genes Chromosomes Cancer. 1992 Nov;5(4):271–277. doi: 10.1002/gcc.2870050402. [DOI] [PubMed] [Google Scholar]
  46. Zucman J., Melot T., Desmaze C., Ghysdael J., Plougastel B., Peter M., Zucker J. M., Triche T. J., Sheer D., Turc-Carel C. Combinatorial generation of variable fusion proteins in the Ewing family of tumours. EMBO J. 1993 Dec;12(12):4481–4487. doi: 10.1002/j.1460-2075.1993.tb06137.x. [DOI] [PMC free article] [PubMed] [Google Scholar]