RNA sequencing of esophageal adenocarcinomas identifies novel fusion transcripts, including NPC1-MELK, arising from a complex chromosomal rearrangement

Cancer. 2017 Oct 15;123(20):3916-3924. doi: 10.1002/cncr.30837. Epub 2017 Jun 22.

Abstract

Background: Studies of chromosomal rearrangements and fusion transcripts have elucidated mechanisms of tumorigenesis and led to targeted cancer therapies. This study was aimed at identifying novel fusion transcripts in esophageal adenocarcinoma (EAC).

Methods: To identify new fusion transcripts associated with EAC, targeted RNA sequencing and polymerase chain reaction (PCR) verification were performed in 40 EACs and matched nonmalignant specimens from the same patients. Genomic PCR and Sanger sequencing were performed to find the breakpoint of fusion genes.

Results: Five novel in-frame fusion transcripts were identified and verified in 40 EACs and in a validation cohort of 15 additional EACs (55 patients in all): fibroblast growth factor receptor 2 (FGFR2)-GRB2-associated binding protein 2 (GAB2) in 2 of 55 or 3.6%, Niemann-Pick C1 (NPC1)-maternal embryonic leucine zipper kinase (MELK) in 2 of 55 or 3.6%, ubiquitin-specific peptidase 54 (USP54)-calcium/calmodulin dependent protein kinase II γ (CAMK2G) in 2 of 55 or 3.6%, megakaryoblastic leukemia (translocation) 1 (MKL1)-fibulin 1 (FBLN1) in 1 of 55 or 1.8%, and CCR4-NOT transcription complex subunit 2 (CNOT2)-chromosome 12 open reading frame 49 (C12orf49) in 1 of 55 or 1.8%. A genomic analysis indicated that NPC1-MELK arose from a complex interchromosomal translocation event involving chromosomes 18, 3, and 9 with 3 rearrangement points, and this was consistent with chromoplexy.

Conclusions: These data indicate that fusion transcripts occur at a stable frequency in EAC. Furthermore, our results indicate that chromoplexy is an underlying mechanism that generates fusion transcripts in EAC. These and other fusion transcripts merit further study as diagnostic markers and potential therapeutic targets in EAC. Cancer 2017;123:3916-24. © 2017 American Cancer Society.

Keywords: Niemann-Pick c (NPC) fusion; chromoplexy; esophageal adenocarcinomas; fibroblast growth factor receptor 2 (FGFR2) fusion; fusion transcript.

MeSH terms

  • Adaptor Proteins, Signal Transducing / genetics
  • Adenocarcinoma / genetics*
  • Adult
  • Aged
  • Aged, 80 and over
  • Calcium-Binding Proteins / genetics
  • Calcium-Calmodulin-Dependent Protein Kinase Type 2 / genetics
  • Carrier Proteins / genetics
  • Case-Control Studies
  • Cell Line, Tumor
  • Esophageal Neoplasms / genetics*
  • Female
  • Gene Rearrangement / genetics*
  • Humans
  • Intracellular Signaling Peptides and Proteins
  • Male
  • Membrane Glycoproteins / genetics
  • Middle Aged
  • Mutant Chimeric Proteins / genetics*
  • Niemann-Pick C1 Protein
  • Protein Serine-Threonine Kinases / genetics
  • RNA, Messenger / metabolism*
  • Receptor, Fibroblast Growth Factor, Type 2 / genetics
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sequence Analysis, RNA
  • Trans-Activators / genetics
  • Ubiquitin-Specific Proteases / genetics

Substances

  • Adaptor Proteins, Signal Transducing
  • Calcium-Binding Proteins
  • Carrier Proteins
  • GAB2 protein, human
  • Intracellular Signaling Peptides and Proteins
  • MRTFA protein, human
  • Membrane Glycoproteins
  • Mutant Chimeric Proteins
  • NPC1 protein, human
  • Niemann-Pick C1 Protein
  • RNA, Messenger
  • Trans-Activators
  • fibulin
  • MELK protein, human
  • FGFR2 protein, human
  • Receptor, Fibroblast Growth Factor, Type 2
  • Protein Serine-Threonine Kinases
  • CAMK2G protein, human
  • Calcium-Calmodulin-Dependent Protein Kinase Type 2
  • USP54 protein, human
  • Ubiquitin-Specific Proteases

Supplementary concepts

  • Adenocarcinoma Of Esophagus