PMID- 36636421
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20230115
IS  - 2218-6751 (Print)
IS  - 2226-4477 (Electronic)
IS  - 2218-6751 (Linking)
VI  - 11
IP  - 12
DP  - 2022 Dec
TI  - Comparison of ROS1-rearrangement detection methods in a cohort of surgically 
      resected non-small cell lung carcinomas.
PG  - 2477-2494
LID - 10.21037/tlcr-22-504 [doi]
AB  - BACKGROUND: Patients with non-small cell lung cancer (NSCLC) harboring a ROS 
      proto-oncogene 1 (ROS1)-rearrangement respond to treatment with ROS1 inhibitors. 
      To distinguish these rare cases, screening with immunohistochemistry (IHC) for 
      ROS1 protein expression has been suggested. However, the reliability of such an 
      assay and the comparability of the antibody clones has been debated. Therefore we 
      evaluated the diagnostic performance of current detection strategies for 
      ROS1-rearrangement in two NSCLC-patient cohorts. METHODS: Resected tissue 
      samples, retrospectively collected from consecutive NSCLC-patients surgically 
      treated at Uppsala University Hospital were incorporated into tissue microarrays 
      [all n=676, adenocarcinomas (AC) n=401, squamous cell carcinomas (SCC) n=213, 
      other NSCLC n=62]. ROS1-rearrangements were detected using fluorescence in situ 
      hybridization (FISH) (Abbott Molecular; ZytoVision). In parallel, ROS1 protein 
      expression was detected using IHC with three antibody clones (D4D6, SP384, 
      EPMGHR2) and accuracy, sensitivity, and specificity were determined. Gene 
      expression microarray data (Affymetrix) and RNA-sequencing data were available 
      for a subset of patients. NanoString analyses were performed for samples with 
      positive or ambiguous results (n=21). RESULTS: Using FISH, 2/630 (0.3% all NSCLC; 
      0.5% non-squamous NSCLC) cases were positive for ROS1 fusion. Additionally, nine 
      cases demonstrated ambiguous FISH results. Using IHC, ROS1 protein expression was 
      detected in 24/665 (3.6% all NSCLC; 5.1% non-squamous NSCLC) cases with clone 
      D4D6, in 18/639 (2.8% all NSCLC; 3.9% non-squamous NSCLC) cases with clone SP384, 
      and in 1/593 (0.2% all NSCLC; 0.3% non-squamous NSCLC) case with clone EPMGHR2. 
      Elevated RNA-levels were seen in 19/369 (5.1%) cases (Affymetrix and 
      RNA-sequencing combined). The overlap of positive results between the assays was 
      poor. Only one of the FISH-positive cases was positive with all antibodies and 
      demonstrated high RNA-expression. This rearrangement was confirmed in the 
      NanoString-assay and also in the RNA-sequencing data. Other cases with high 
      protein/RNA-expression or ambiguous FISH were negative in the NanoString-assay. 
      CONCLUSIONS: The occurrence of ROS1 fusions is low in our cohorts. The IHC assays 
      detected the fusions, but the accuracy varied depending on the clone. The 
      presumably false-positive and uncertain FISH results questions this method for 
      detection of ROS1-rearrangements. Thus, when IHC is used for screening, 
      transcript-based assays are preferable for validation in clinical diagnostics.
CI  - 2022 Translational Lung Cancer Research. All rights reserved.
FAU - Thurfjell, Viktoria
AU  - Thurfjell V
AD  - Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, 
      Sweden.
FAU - Micke, Patrick
AU  - Micke P
AD  - Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, 
      Sweden.
FAU - Yu, Hui
AU  - Yu H
AD  - Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, 
      Sweden.
FAU - Krupar, Rosemarie
AU  - Krupar R
AD  - Division of Pathology, Research Center Borstel, Leibniz Lung Center, Borstel, 
      Germany.
AD  - Institute of Pathology, University Hospital Schleswig-Holstein, Campus Lubeck, 
      Lubeck, Germany.
FAU - Svensson, Maria A
AU  - Svensson MA
AD  - Clinical Research Center, Faculty of Medicine and Health, Orebro University, 
      Orebro, Sweden.
FAU - Brunnstrom, Hans
AU  - Brunnstrom H
AD  - Division of Pathology, Lund University and Laboratory Medicine Region Skane, 
      Lund, Sweden.
FAU - Lamberg, Kristina
AU  - Lamberg K
AD  - Department of Pulmonary and Allergic Diseases, Uppsala University Hospital, 
      Uppsala, Sweden.
FAU - Moens, Lotte N J
AU  - Moens LNJ
AD  - Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, 
      Sweden.
AD  - Clinical Genomics Uppsala, Science for Life Laboratory, Uppsala, Sweden.
FAU - Strell, Carina
AU  - Strell C
AD  - Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, 
      Sweden.
FAU - Gulyas, Miklos
AU  - Gulyas M
AD  - Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, 
      Sweden.
FAU - Helenius, Gisela
AU  - Helenius G
AD  - Department of Laboratory Medicine, Faculty of Medicine and Health, Orebro 
      University, Orebro, Sweden.
FAU - Yoshida, Akihiko
AU  - Yoshida A
AD  - Department of Diagnostic Pathology, National Cancer Center Hospital, Tokyo, 
      Japan.
FAU - Goldmann, Torsten
AU  - Goldmann T
AD  - Division of Pathology, Research Center Borstel, Leibniz Lung Center, Borstel, 
      Germany.
AD  - Airway Research Center North (ARCN), Member of the German Center for Lung 
      Research (DZL), Grosshansdorf, Germany.
FAU - Mattsson, Johanna Sofia Margareta
AU  - Mattsson JSM
AD  - Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, 
      Sweden.
LA  - eng
PT  - Journal Article
PL  - China
TA  - Transl Lung Cancer Res
JT  - Translational lung cancer research
JID - 101646875
PMC - PMC9830269
OTO - NOTNLM
OT  - Crizotinib
OT  - ROS proto-oncogene 1 (ROS1)
OT  - fusion gene detection
OT  - molecular pathology
OT  - targeted therapy
COIS- Conflicts of Interest: All authors have completed the ICMJE uniform disclosure 
      form (available at 
      https://tlcr.amegroups.com/article/view/10.21037/tlcr-22-504/coif). VT reports 
      support for study materials from the Lions Cancer Foundation, Uppsala, Sweden. PM 
      reports support for the present manuscript from the Swedish Cancer Society, 
      Vetenskapsradet, Uppsala-Orebro Regional Research Council, and the Sjoberg 
      Foundation. RK reports consulting fees from Aignostics GmbH. CS reports grants 
      from the Swedish Cancer Society, Swedish Cancer Society Radiotherapy Fellowship, 
      and Cancer- och Allergifonden Research. GH reports grants from Roche, honoraria 
      from Bristol Meyer Squibb, payment for expert testimony from Astra Zeneca and 
      Pfizer, and support for attending meetings and/or travel from Astra Zeneca. JSMM 
      reports support for study materials from Selanders Stiftelse, Uppsala, Sweden, 
      support for attending meetings from the Swedish Cancer Society, and receipt of 
      materials from Zytomed Systems (anti-ROS1 mAb EPMGHR2). The other authors have no 
      conflicts of interest to declare.
EDAT- 2023/01/14 06:00
MHDA- 2023/01/14 06:01
PMCR- 2022/12/01
CRDT- 2023/01/13 02:01
PHST- 2022/07/04 00:00 [received]
PHST- 2022/11/06 00:00 [accepted]
PHST- 2023/01/13 02:01 [entrez]
PHST- 2023/01/14 06:00 [pubmed]
PHST- 2023/01/14 06:01 [medline]
PHST- 2022/12/01 00:00 [pmc-release]
AID - tlcr-11-12-2477 [pii]
AID - 10.21037/tlcr-22-504 [doi]
PST - ppublish
SO  - Transl Lung Cancer Res. 2022 Dec;11(12):2477-2494. doi: 10.21037/tlcr-22-504.