Improving sarcoma classification by using RNA hybridisation capture sequencing in sarcomas of uncertain histogenesis of young individuals

Pathology. 2023 Jun;55(4):478-485. doi: 10.1016/j.pathol.2022.11.012. Epub 2023 Feb 14.


Our aim was to utilise a 241-gene RNA hybridisation capture sequencing (CaptureSeq) gene panel to identify unexpected fusions in undifferentiated, unclassified or partly classified sarcomas of young individuals (<40 years). The purpose was to determine the utility and yield of a large, targeted fusion panel as a tool for classifying tumours that do not fit typical diagnostic entities at the time of the original diagnosis. RNA hybridisation capture sequencing was performed on 21 archival resection specimens. Successful sequencing was obtained in 12 of 21 samples (57%), two of which (16.6%) harboured translocations. A novel NEAT1::GLI1 fusion, not previously reported in the literature, presented in a young patient with a tumour in the retroperitoneum, which displayed low grade epithelioid cells. The second case, a localised lung metastasis in a young male, demonstrated a EWSR1::NFATC2 translocation. No targeted fusions were identified in the remaining 83.4% (n=10) of cases. Forty-three per cent of the samples failed sequencing as a result of RNA degradation. RNA-based sequencing is an important tool, which helps to redefine the classification of unclassified or partly classified sarcomas of young adults by identifying pathogenic gene fusions in up to 16.6% of the cases. Unfortunately, 43% of the samples underwent significant RNA degradation, falling below the sequencing threshold. As CaptureSeq is not yet available in routine pathology practice, increasing awareness of the yield, failure rate and possible aetiological factors for RNA degradation is fundamental to maximise laboratory procedures to improve RNA integrity, allowing the potential identification of significant gene alterations in solid tumours.

Keywords: GLI1; NFATC2; RNA sequencing; Undifferentiated; gene fusion; sarcoma.

MeSH terms

  • Gene Fusion
  • Gene Rearrangement
  • Humans
  • Male
  • Oncogene Proteins, Fusion / genetics
  • Sarcoma* / diagnosis
  • Sarcoma* / genetics
  • Sarcoma* / pathology
  • Soft Tissue Neoplasms* / diagnosis
  • Soft Tissue Neoplasms* / genetics
  • Soft Tissue Neoplasms* / pathology
  • Transcription Factors / genetics
  • Young Adult


  • Transcription Factors
  • Oncogene Proteins, Fusion