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Role of high-throughput sequencing in the diagnosis of cutaneous T-cell lymphoma
  1. Bryan Rea1,
  2. Paul Haun1,
  3. Ryan Emerson2,
  4. Marissa Vignali2,
  5. Midhat Farooqi1,
  6. Sara Samimi1,
  7. Rosalie Elenitsas1,
  8. Ilan Kirsch2,
  9. Adam Bagg1
  1. 1Hospital of the University of Pennsylvania, Philadelphia, Pennsylvania, USA
  2. 2Adaptive Biotechnologies, Seattle, Washington, USA
  1. Correspondence to Dr Bryan Rea, Hospital of the University of Pennsylvania, Philadelphia, PA 19104, USA; bryan.rea{at}uphs.upenn.edu

Abstract

Aims Substantial clinicopathological overlap exists between cutaneous T-cell lymphoma (CTCL) and benign conditions, leading to diagnostic difficulties. We sought to delineate the utility of high-throughput sequencing (HTS) across a spectrum of histological findings in CTCL and reactive mimics.

Methods One hundred skin biopsies obtained for clinical concern for CTCL were identified, comprising 25 cases each from four histological categories: ‘definitive CTCL’, ‘atypical lymphoid infiltrate, concerning for CTCL’, ‘atypical lymphoid infiltrate, favour reactive’ or ‘reactive lymphoid infiltrate’. T-cell receptor gamma chain gene (TRG) PCR and T-cell receptor beta chain gene HTS were performed on both skin biopsy and concurrently collected peripheral blood; most peripheral blood samples were also analysed by flow cytometry.

Results Histologically defined CTCL specimens had significantly higher clonality scores and T-cell fractions via HTS than all other groups (all P<0.002 and P<0.03, respectively). HTS was more diagnostically specific than TRG PCR in skin (100% vs 88%), while diagnostic sensitivity (68% vs 72%) and accuracy (84% vs 80%) were similar. TRG PCR and flow cytometry performed on blood were the least diagnostically useful assays. Some identically sized peaks detected by TRG PCR in concurrent skin and peripheral blood specimens were non-identical by HTS analysis.

Conclusions HTS, by assessing both clonality and T-cell fractions in skin biopsies, is a powerful tool to aid in the diagnosis of CTCL. It is more specific than TRG PCR in distinguishing definitive CTCL from reactive and indeterminate histology. Identically sized peaks by TRG PCR, typically interpreted to be clonally related, are not always clonally identical by sequencing.

  • molecular pathology
  • dermatopathology
  • haematopathology
  • lymphoma

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Footnotes

  • Handling editor Mary Frances McMullin.

  • Contributors BR extracted and analysed all the data, and wrote the manuscript. PH identified patients, reviewed the pathology and helped write the manuscript. RyE and MV performed and analysed the next-generation sequencing data. MF contributed essential samples and interpreted PCR studies. SS provided clinical context. RoE helped identify patients and write the manuscript. IK helped write the manuscript. AB conceived and designed the study and wrote the manuscript. All authors read and approved the final manuscript.

  • Funding The authors have not declared a specific grant for this research from any funding agency in the public, commercial or not-for-profit sectors.

  • Competing interests RyE, MV and IK disclose employment and equity with Adaptive Biotechnologies, Seattle, WA.

  • Patient consent Not required.

  • Ethics approval UPHS Institutional Review Board.

  • Provenance and peer review Not commissioned; externally peer reviewed.

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