Long read sequencing identifies complex structural variant landscape and recurrent TERT rearrangements in mucoepidermoid carcinoma

Oral Oncol. 2024 Dec:159:107108. doi: 10.1016/j.oraloncology.2024.107108. Epub 2024 Nov 15.

Abstract

Mucoepidermoid Carcinoma (MEC) is a common salivary malignant neoplasm. Approximately 60 % of MECs harbor translocations between CRTC1 or CRTC3 and MAML2, which are thought to drive disease pathogenesis. However, the precise structural mechanism driving this rearrangement remains uncharacterized. Here, we performed multi-omic and long read genomic sequencing, discovering a chain of alterations that created the CRTC1::MAML2 fusion, but also an unexpected MAML2 to MYBL1 rearrangement, suggesting that MYBL1 may play a larger role in salivary gland cancers than previously recognized. Furthermore, we discovered and validated recurrent TERT rearrangements and amplifications in MEC models. 5/5 MEC cell lines and 36/39 (92 %) primary MEC tumors harbored a TERT rearrangement or copy number amplification. Custom sequencing of the TERT locus confirmed translocation breakpoints in 13/33 (39 %) MECs, while exome sequencing confirmed frequent TERT amplifications. Critically, TERT knockdown in NCI-H292, a cell line with TERT promoter rearrangement, reduced clonogenic cell survival, supporting a critical role of this gene in MEC tumorigenesis. Overall, our data suggest that complex chromothripsis rearrangement mechanisms drive the formation of structural variation in CRTC1::MAML2 fusion positive and negative tumors and reveal highly recurrent structural variation driving TERT rearrangement in MEC.

Keywords: CRTC1; MAML2; MEC; NOTCH2; Structural variation; TERT; Translocation.

MeSH terms

  • Carcinoma, Mucoepidermoid* / genetics
  • Carcinoma, Mucoepidermoid* / pathology
  • Cell Line, Tumor
  • DNA-Binding Proteins / genetics
  • Female
  • Gene Rearrangement*
  • Humans
  • Male
  • Oncogene Proteins, Fusion / genetics
  • Proto-Oncogene Proteins
  • Salivary Gland Neoplasms / genetics
  • Salivary Gland Neoplasms / pathology
  • Telomerase* / genetics
  • Trans-Activators / genetics
  • Transcription Factors / genetics
  • Translocation, Genetic / genetics

Substances

  • Telomerase
  • TERT protein, human
  • MAML2 protein, human
  • Transcription Factors
  • CRTC1 protein, human
  • Trans-Activators
  • DNA-Binding Proteins
  • Oncogene Proteins, Fusion
  • MYBL1 protein, human
  • Proto-Oncogene Proteins