A novel t(4;22)(q31;q12) produces an EWSR1-SMARCA5 fusion in extraskeletal Ewing sarcoma/primitive neuroectodermal tumor

Janos Sumegi, Jun Nishio, Marilu Nelson, Robert W. Frayer, Deborah Perry, Julia A. Bridge

Research output: Contribution to journalArticle

63 Citations (Scopus)

Abstract

Over 90% of Ewing sarcoma/primitive neuroectodermal tumors (PNETs) feature an 11;22 translocation leading to an EWSR1-FLI1 fusion. Less commonly, a member of the ETS-transcription factor family other than FLI1 is fused with EWSR1. In this study, cytogenetic analysis of an extraskeletal Ewing sarcoma/PNET revealed a novel chromosomal translocation t(4;22)(q31;q12) as the sole anomaly. Following confirmation of an EWSR1 rearrangement by the use of EWSR1 breakpoint flanking probes, a fluorescence in situ hybridization positional cloning strategy was used to further narrow the 4q31 breakpoint. These analyses identified the breakpoint within RP11-481K16, a bacterial artificial chromosome (BAC) clone containing two gene candidates FREM and SMARCA5. Subsequent RACE, RT-PCR, and sequencing studies were conducted to further characterize the fusion transcript. An in-frame fusion of the first 7 exons of EWSR1 to the last 19 exons of SMARCA5 was identified. SMARCA5 encodes for hSNF2H, a chromatin-remodeling protein. Analogous to EWSR1-ETS-expressing NIH3T3 cells, NIH3T3 cells expressing EWSR1-hSNF2H exhibited anchorage-independent growth and formed colonies in soft agar, indicating chimeric protein tumorigenic potential. Conversely, expression of EWSR1-hSNF2H in NIH3T3 cells, unlike EWSR1-ETS fusions, did not induce EAT-2 expression. Mapping analysis demonstrated that deletion of the C-terminus (SLIDE or SANT motives) of hSNF2H impaired, and deletion of the SNF2-N domain fully abrogated NIH3T3 cell transformation by EWSR1-SMARCA5. It is proposed that EWSR1-hSNF2H may act as an oncogenic chromatin-remodeling factor and that its expression contributes to Ewing sarcoma/primitive neuroectodermal tumorigenesis. To the best of our knowledge, this is the first description of a fusion between EWSR1 and a chromatin-reorganizing gene in Ewing sarcoma/PNET and thus expands the EWSR1 functional partnership beyond transcription factor and zinc-finger gene families.

Original languageEnglish (US)
Pages (from-to)333-342
Number of pages10
JournalModern Pathology
Volume24
Issue number3
DOIs
StatePublished - Mar 1 2011

Fingerprint

Primitive Neuroectodermal Tumors
Ewing's Sarcoma
Chromatin Assembly and Disassembly
Exons
Transcription Factors
Genes
Bacterial Artificial Chromosomes
Genetic Translocation
Cytogenetic Analysis
Zinc Fingers
Fluorescence In Situ Hybridization
Agar
Chromatin
Organism Cloning
Carcinogenesis
Proteins
Clone Cells
Polymerase Chain Reaction
Growth

Keywords

  • EWSR1
  • Ewing sarcoma
  • SMARCA5
  • chromatin remodeling
  • fusion gene
  • translocation

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

Cite this

A novel t(4;22)(q31;q12) produces an EWSR1-SMARCA5 fusion in extraskeletal Ewing sarcoma/primitive neuroectodermal tumor. / Sumegi, Janos; Nishio, Jun; Nelson, Marilu; Frayer, Robert W.; Perry, Deborah; Bridge, Julia A.

In: Modern Pathology, Vol. 24, No. 3, 01.03.2011, p. 333-342.

Research output: Contribution to journalArticle

Sumegi, Janos ; Nishio, Jun ; Nelson, Marilu ; Frayer, Robert W. ; Perry, Deborah ; Bridge, Julia A. / A novel t(4;22)(q31;q12) produces an EWSR1-SMARCA5 fusion in extraskeletal Ewing sarcoma/primitive neuroectodermal tumor. In: Modern Pathology. 2011 ; Vol. 24, No. 3. pp. 333-342.
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