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Next-generation sequencing is highly sensitive for the detection of beta-catenin mutations in desmoid-type fibromatoses


Desmoid-type fibromatoses are locally aggressive and frequently recurrent tumours, and an accurate diagnosis is essential for patient management. The majority of sporadic lesions harbour beta-catenin (CTNNB1) mutations. We used next-generation sequencing to detect CTNNB1 mutations and to compare the sensitivity and specificity of next-generation sequencing with currently employed mutation detection techniques: mutation-specific restriction enzyme digestion and polymerase chain reaction amplification. DNA was extracted from formalin-fixed paraffin-embedded needle biopsy or resection tissue sections from 144 patients with sporadic desmoid-type fibromatoses, four patients with syndrome-related desmoid-type fibromatoses and 11 morphological mimics. Two primer pairs were designed for CTNNB1 mutation hotspots. Using ≥10 ng of DNA, libraries were generated by Fluidigm and sequenced on the Ion Torrent Personal Genome Machine. Next-generation sequencing had a sensitivity of 92.36 % (133/144, 95 % CIs: 86.74 to 96.12 %) and a specificity of 100 % for the detection of CTNNB1 mutations in desmoid-type fibromatoses-like spindle cell lesions. All mutations detected by mutation-specific restriction enzyme digestion were identified by next-generation sequencing. Next-generation sequencing identified additional mutations in 11 tumours that were not detected by mutation-specific restriction enzyme digestion, two of which have not been previously described. Next-generation sequencing is highly sensitive for the detection of CTNNB1 mutations. This multiplex assay has the advantage of detecting additional mutations compared to those detected by mutation-specific restriction enzyme digestion (sensitivity 82.41 %). The technology requires minimal DNA and is time- and cost-efficient.

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This work was supported by Skeletal Cancer Action Trust (SCAT), UK. The material was obtained from the RNOH Musculoskeletal Research Programme and Biobank. Support was provided to AMF (UCL) by the National Institute for Health Research, UCLH Biomedical Research Centre and the UCL Experimental Cancer Centre. SJA is a NIHR-funded Academic Clinical Fellow. We are grateful to the patients for participating in the research and to the clinicians and support staff of the London Sarcoma Service.

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No conflicts of interest to declare.

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Correspondence to Adrienne M. Flanagan.

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Aitken, S.J., Presneau, N., Kalimuthu, S. et al. Next-generation sequencing is highly sensitive for the detection of beta-catenin mutations in desmoid-type fibromatoses. Virchows Arch 467, 203–210 (2015). https://doi.org/10.1007/s00428-015-1765-0

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  • Next-generation sequencing
  • Desmoid-type fibromatosis
  • Beta-catenin
  • CTNNB1
  • Molecular pathology