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Reproducibility of histologic prognostic parameters for mantle cell lymphoma: cytology, Ki67, p53 and SOX11

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Abstract

Mantle cell lymphoma (MCL) shows a clinical aggressiveness that varies from patient to patient. Despite major advances in outcomes with current immunochemotherapy, the future development of therapies requires risk stratification to tailor therapy intensity. Within the group of reference pathologists for the ongoing trials of the European MCL Network, we performed a round robin test on a tissue microarray to evaluate the reproducibility in assessing the biomarkers of outcome in MCL. Cytological subtype, Ki67-index and expression of p53 and SOX11 were evaluated on 20 diagnostic tumour samples by eight participating labs independently. We demonstrate that the assessment of the proliferation index by counting the Ki67 positive cells as well as assessment of SOX11 and p53 expression status is reproducible between labs. For the most established prognostic biomarker, Ki67, the intra-class correlation coefficient was very good when assessed as a continuous parameter (0.87). The agreement was lower when the values were analysed in a dichotomized way applying the commonly used cutoff of 30% (kappa = 0.65, complete concordance of all labs in 13/20 (65%)). Cases with discrepant results between labs in the dichotomized analysis showed mean values close to the cutoff of 30%. Centralised scoring and digital image analysis revealed results in line with the scores from individual labs. All cases in our cohort were additionally assessed for gene expression signatures and of TP53 gene alterations. Given the good reproducibility when guidelines of assessment are applied, the biomarker studied in this inter-laboratory test presents potential candidates to be enhanced for risk-stratification in the future clinical trials.

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Acknowledgements

The authors thank Dana Germer, Charlotte Botz von Drathen and Reine Zühlke-Jenisch for excellent technical support.

Contributions

All authors contributed to the study conception and design. Giorgio A. Croci and Wolfram Klapper designed the research; Giorgio A. Croci, Wolfram Klapper, José Cabeçadas, Elias Campo, Luis Veloza, Erik Clasen-Linde, Rashmi S. Goswami, Lars Helgeland, Stefano Pileri and Grzegorz Rymkiewicz performed histopathological assessment; Giorgio A. Croci and Sarah Reinke performed image analysis; Sílvia Beà, David W. Scott, Guillem Clot and Anna Enjuanes performed molecular and bioinformatic analyses; Eva Hoster planned and performed statistical analyses; Martin Dreyling provided medical support; Giorgio A. Croci, Wolfram Klapper, Sílvia Beà and Eva Hoster wrote the manuscript; all authors approved the final version of the manuscript.

Funding

The work was supported in part by the NIH, grant number 1P01CA229100.

Author information

Authors and Affiliations

  1. Hematopathology Section, Institute of Pathology and Lymph Node Registry, Universitätsklinikum Schleswig-Holstein, Campus Kiel, Kiel, Germany

    Giorgio A. Croci, Sarah Reinke & Wolfram Klapper

  2. Pathology Unit, Department of Pathophysiology and Transplantation, University of Milan, Milan, Italy

    Giorgio A. Croci

  3. UOC di Anatomia Patologica, IRCCS Fondazione Ca’ Granda – Ospedale Maggiore Policlinico, Via Francesco Sforza, 35 - 20122, Milano, Italy

    Giorgio A. Croci

  4. Institute for Medical Information Processing, Biometry and Epidemiology (IBE), Ludwig-Maximilians-Universität Munich, Munich, Germany

    Eva Hoster

  5. Department of Medicine III, University Hospital of the Ludwig Maximilians University Munich, Munich, Germany

    Eva Hoster & Martin Dreyling

  6. Institute for Biomedical Research August Pi i Sunyer (IDIBAPS), Barcelona, Spain

    Sílvia Beà, Guillem Clot, Anna Enjuanes & Elias Campo

  7. Centro de Investigación Biomédica en Red de Cáncer (CIBERONC), Barcelona, Spain

    Sílvia Beà, Guillem Clot, Anna Enjuanes & Elias Campo

  8. Centre for Lymphoid Cancer, British Columbia Cancer, Vancouver, Canada

    David W. Scott

  9. Department of Pathology, Instituto Português de Oncologia de Lisboa Francisco Gentil, E.P.E, Lisbon, Portugal

    José Cabeçadas

  10. Hematopathology Unit-Laboratory of Pathology, Hospital Clínic of Barcelona, University of Barcelona, Barcelona, Spain

    Luis Veloza & Elias Campo

  11. Department of Pathology, Rigshospitalet, Copenhagen, Denmark

    Erik Clasen-Linde

  12. Sunnybrook Health Sciences Centre, Dept. of Laboratory Medicine and Molecular Diagnostics, and Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Canada

    Rashmi S. Goswami

  13. Department of Pathology, Haukeland University Hospital, Bergen, Norway

    Lars Helgeland

  14. Division of Diagnostic Haematopathology, European Institute of Oncology, IRCCS, Milan, Italy

    Stefano Pileri

  15. Department of Pathology and Laboratory Diagnostics, Maria Sklodowska-Curie Institute-Oncology Centre, Warsaw, Poland

    Grzegorz Rymkiewicz

Authors
  1. Giorgio A. Croci
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  2. Eva Hoster
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Correspondence to Giorgio A. Croci.

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Supplementary figure 1

Cytology scored in individual labs (A-H) for all cases (1–20) according to the 4-tiered system (left panel) and dichotomized as low grade vs high grade (right panel). NA = not available. (PNG 233 kb)

High Resolution (TIF 189 kb)

Supplementary figure 2

Bland-Altman-Plot of mean and difference to Ki67 value using reference value by Kiel lab: each lab is depicted by a colour (reference = black), the sample is indicated by a small case letter. (PNG 265 kb)

High Resolution (TIF 31 kb)

Supplementary figure 3

p53 scored in individual labs (A-H) for all cases (1-20) according to the 4-tiered system (left panel) and dichotomized as non-high and high (right panel). NA = not available. (PNG 298 kb)

High Resolution (TIF 184 kb)

Supplementary figure 4

SOX11 scored in individual labs (A-H) for all cases (1-20) according to the 3-tiered system (left panel) and dichotomized as non-positive and positive (right panel). (PNG 211 kb)

High Resolution (TIF 177 kb)

Supplementary figure 5

Guideline for Ki67 assessment. In this MCL case with mantle zone pattern, Ki67 count should be performed ruling out non-neoplastic proliferative foci, such as residual germinal centres (square area) and T-cell reactive component at the periphery; b) once the region of interest (square) has been selected, Ki67 positive cells should be counted as the percentage over 100 cells. (PNG 2636 kb)

High Resolution (TIF 1007 kb)

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Croci, G.A., Hoster, E., Beà, S. et al. Reproducibility of histologic prognostic parameters for mantle cell lymphoma: cytology, Ki67, p53 and SOX11. Virchows Arch 477, 259–267 (2020). https://doi.org/10.1007/s00428-020-02750-7

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