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Comparison of genetic, antigenic and clinical features of extra-osseous Ewing Sarcoma (EO-EWS) and osseous Ewing sarcoma (O-EWS). Amanda Rivera-Begeman; Carrye Cost; Stephen Lessnick; Richard Smith; Charles Timmons; Patrick Leavey Annual CTOS Meeting; Seattle, Washington, November 2007.

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slide1

Comparison of genetic, antigenic and clinical features of extra-osseous Ewing Sarcoma (EO-EWS) and osseous Ewing sarcoma (O-EWS)

Amanda Rivera-Begeman; Carrye Cost;

Stephen Lessnick; Richard Smith; Charles Timmons; Patrick Leavey

Annual CTOS Meeting; Seattle, Washington, November 2007

background
Background
  • Ewing\'s sarcoma family of tumors (EFT)
    • Osseous EWS (O-EWS), Extraosseous ES (EO-EWS), Peripheral primitive neuroectodermal tumor (pPNET) and Askin\'s tumor of chest wall (Carvajal,R. et al; Hematol Oncol Clin North Am 2005)
  • Cell surface protein MIC-2 (CD99) expressed on both O-EWS and EO-EWS (Ambros, IM; Cancer; 1991)
  • FLI-1 nuclear immunostain + ve in 70% of EWS and PNET cases (Folpe et al; Am J Surg Pathol; 2000)
background1
Background
  • EWS/FLI-1 is seen in patients with EFT (O’Sullivan,MJ et al; Hum Pathol; 2001)
  • Prior reports of treatment for EO-EWS demonstrated no advantage to the addition of Doxorubicin (Raney RB et al.; J Clin Oncol; 1997)
  • EO-EWS should be treated with strategies used for O-EWS vs. malignant mesenchymal tumors (Castex,M.P.; J Clin Oncol; 2007)
objective
Objective
  • To describe genetic, antigenic, and clinical features of patients with EO-EWS, primarily those of intra-abdominal origin
  • To compare genetic and antigenic features of EO-EWS to those of randomly chosen patients with O-EWS
patients
Patients
  • Eligibility criteria
    • EFT treated at Children’s Medical Center Dallas (1995 – 2005; n=52)
    • Availability of archival diagnostic material and clinical data
methods
Methods
  • All immunostains were independently reviewed by 2 pathologists (CT, ARB)
  • Interpretation was subjective +ve vs. –ve
    • No grading was attempted
slide8

Fli-1

weak positive

Fli-1

strong positive

slide9

CD99 (O13)

Weak positive

CD99 (O13)

strong positive

slide12

Cytokeratin Cam 5.2

positive

Carcinoembryonic antigen (CEA)

positive

methods1
Methods
  • RT-PCR
    • RNA extracted from formalin fixed, paraffin embedded tumor (Roche high pure RNA paraffin kit)
    • PCR was performed for EWS/FLI-1
      • EWS/FLI-1 fusion type identified by melt curve analysis
      • EWS-FLI-1 fusion type confirmed by agarose gel electrophoresis
    • Alternate partners were examined as necessary (ews/fev, etv1, etv4 and erg)
results ews fli 1 type
Results – EWS-FLI-1 type
  • Not type 1 or 2
    • EO-EWS: confirmed t(11:22)(q24;q12) but no translocation products amplified
    • O-EWS: PCR product melting curve between types 1 and 2 but failed sequencing
summary
Summary
  • Negative FLI-1 nuclear staining does not exclude EWS-FLI-1 translocation positive EFT
  • While CEA +ve staining can be seen in EO-EWS it does not differentiate this from O-EWS
  • More type 2 fusions noted in patients with EO-EWS
conclusion
Conclusion
  • Variability may occur in immunostaining and genotype analysis of patients with extra-osseous Ewing sarcoma vs. osseous Ewing sarcoma.
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