Research Article

Blood, Lymphatics, Immune System and Stem Cells

Disruption of direct 3D telomere–TRF2 interaction through two molecularly disparate mechanisms is a hallmark of primary Hodgkin and Reed–Sternberg cells

  • Laboratory Investigation volume 97, pages 772781 (2017)
  • doi:10.1038/labinvest.2017.33
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Abstract

In classical Hodgkin’s lymphoma (cHL), specific changes in the 3D telomere organization cause progression from mononuclear Hodgkin cells (H) to multinucleated Reed–Sternberg cells (RS). In a post-germinal center B-cell in vitro model, permanent latent membrane protein 1 (LMP1) expression, as observed in Epstein–Barr virus (EBV)-associated cHL, results in multinuclearity and complex chromosomal aberrations through downregulation of key element of the shelterin complex, the telomere repeat binding factor 2 (TRF2). Thus, we hypothesized that the three-dimensional (3D) telomere–TRF2 interaction was progressively disturbed during transition from H to RS cells. To this end, we developed and applied for the first time a combined quantitative 3D TRF2-telomere immune fluorescent in situ hybridization (3D TRF2/Telo-Q-FISH) technique to monolayers of primary H and RS cells, and adjacent benign internal control lymphocytes of lymph node biopsy suspensions from diagnostic lymph node biopsies of 14 patients with cHL. We show that H and RS cells are characterized by two distinct patterns of disruption of 3D telomere–TRF2 interaction. Disruption pattern A is defined by massive attrition of telomere signals and a considerable increase of TRF2 signals not associated with telomeres. This pattern is restricted to EBV-negative cHL. Disruption pattern B is defined by telomere de-protection due to an impressive loss of TRF2 signals, physically linked to telomeres. This pattern is typical of, but is not restricted to, LMP1+EBV-associated cHL. In the disruption pattern B group, so-called 'ghost' end-stage RS cells, void of both TRF2 and telomere signals, were identified, whether or not associated with EBV. Our findings demonstrate that two molecularly disparate mechanisms converge on the level of 3D telomere–TRF2 interaction in the formation of RS cells.

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Acknowledgements

We thank Mary Cheang, Biostatistician, for statistical analysis of data. The 3D Telomeres Technology platform (3DTT) and the TeloView are property to 3D Signatures and were used with the company's permission. We are grateful to receive research support from the Cole Foundation (NJ) and the Canadian Institutes of Health Research (SM; Grant MOP110982).

Author information

Affiliations

  1. Division of Hematology, Department of Medicine, Jewish General Hospital, McGill University, Montréal, QC, Canada

    • Hans Knecht
    • , Nathalie A Johnson
    •  & Tina Haliotis
  2. Department of Pathology, Jewish General Hospital, McGill University, Montréal, QC, Canada

    • Tina Haliotis
  3. MICB, University of Manitoba, Winnipeg, MB, Canada

    • Daniel Lichtensztejn
    •  & Sabine Mai

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Competing interests

SM is a founder of and currently a director, officer (CSO) and shareholder of 3D Signatures. HK is a member of the Scientific Advisory Board and shareholder of 3D Signatures. The remaining authors declare no conflict of interest.

Corresponding author

Correspondence to Hans Knecht.

Supplementary information

Supplementary Information accompanies the paper on the Laboratory Investigation website (http://www.laboratoryinvestigation.org)