RET Fluorescence In Situ Hybridization Analysis Is a Sensitive but Highly Unspecific Screening Method for RET Fusions in Lung Cancer

T. Radonic*, W.R.R. Geurts-Giele, K.G. Samsom, G.M.J.M. Roemen, J.H. von der Thusen, E. Thunnissen, I.C. Meijssen, H.F.B.M. Sleddens, W.N.M. Dinjens, M.C. Boelens, K. Weijers, E.J.M. Speel, S.P. Finn, C. O'Brien, T. van Wezel, D. Cohen, K. Monkhorst, P. Roepman, H.J. Dubbink

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Introduction: RET gene fusions are established oncogenic drivers in 1% of NSCLC. Accurate detection of advanced patients with RET fusions is essential to ensure optimal therapy choice. We investigated the performance of fluorescence in situ hybridization (FISH) as a diagnostic test for detecting functional RET fusions.Methods: Between January 2016 and November 2019, a total of 4873 patients with NSCLC were routinely screened for RET fusions using either FISH (n = 2858) or targeted RNA next-generation sequencing (NGS) (n = 2015). If sufficient material was available, positive cases were analyzed by both methods (n = 39) and multiple FISH assays (n = 17). In an independent cohort of 520 patients with NSCLC, whole-genome sequencing data were investigated for disruptive structural variations and functional fusions in the RET and compared with ALK and ROS1 loci.Results: FISH analysis revealed RET rearrangement in 48 of 2858 cases; of 30 rearranged cases double tested with NGS, only nine had a functional RET fusion. RNA NGS yielded RET fusions in 14 of 2015 cases; all nine cases double tested by FISH had RET locus rearrangement. Of these 18 verified RET fusion cases, 16 had a split signal and two a complex rearrangement by FISH. By whole-genome sequencing, the prevalence of functional fusions compared with all disruptive events was lower in the RET (4 of 9, 44%) than the ALK (27 of 34, 79%) and ROS1 (9 of 12, 75%) loci.Conclusions: FISH is a sensitive but unspecific technique for RET screening, always requiring a confirmation using an orthogonal technique, owing to frequently occurring RET rearrangements not resulting in functional fusions in NSCLC. (C) 2021 International Association for the Study of Lung Cancer. Published by Elsevier Inc.
Original languageEnglish
Pages (from-to)798-806
Number of pages9
JournalJournal of Thoracic Oncology
Volume16
Issue number5
DOIs
Publication statusPublished - 1 May 2021

Keywords

  • Non-small cell carcinoma
  • RET
  • FISH
  • RNA NGS
  • ROS1 REARRANGEMENT
  • ALK
  • MULTICENTER

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