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Quality assurance trials for Ki67 assessment in pathology

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Abstract

Ki67 is a broadly used proliferation marker in surgical pathology with an obvious need for standardization to improve reproducibility of assessment. Here, we present results of the so far only existing round robin tests on Ki67, organized annually in Germany, Austria, and Switzerland from 2010 to 2015 with up to 160 participating laboratories (QuIP). In each quality assessment trial, eight probes from each breast cancer, neuroendocrine tumor, and malignant lymphoma were compiled on a tissue microarray (TMA). TMAs were stained in the participants’ laboratories with antibodies and procedures also applied in their daily routine. Participating pathologists were expected to assign Ki67 values to one of four different categories for each tumor type. All local stainings and evaluations were reassessed by the organizing panel and compared to a preset standard. On average, 95% of participants reached the benchmark of over 80% concordance rates with the Ki67 category pre-established by the panel. Automatization and type of antibody did not affect the success rate. Concordance rates differed between tumor entities being highest in each tumor type with either very high or very low labeling indices. Lower rates were seen for intermediate Ki67 levels. Staining quality improved during the observation period as did inter-observer concordance with 85% of participants achieving excellent agreement (kappa > 0.8) in the first year and over 95% in 2015. In conclusion, regular external quality assurance trials have been established as a tool to improve the reproducibility and reliability of the prognostic and predictive proliferation marker Ki67.

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References

  1. Gerdes J, Lemke H, Baisch H, Wacker HH, Schwab U, Stein H (1984) Cell cycle analysis of a cell proliferation-associated human nuclear antigen defined by the monoclonal antibody Ki-67. J Immunol 133:1710–1715

    CAS  PubMed  Google Scholar 

  2. Mengel M, von Wasielewski R, Wiese B, Rudiger T, Muller-Hermelink HK, Kreipe H (2002) Inter-laboratory and inter-observer reproducibility of immunohistochemical assessment of the Ki-67 labelling index in a large multi-centre trial. J Pathol 198:292–299

    Article  PubMed  Google Scholar 

  3. Gluz O, Nitz UA, Christgen M, Kates RE, Shak S, Clemens M, Kraemer S, Aktas B, Kuemmel S, Reimer T, Kusche M, Heyl V, Lorenz-Salehi F, Just M, Hofmann D, Degenhardt T, Liedtke C, Svedman C, Wuerstlein R, Kreipe HH, Harbeck N (2016) West German Study Group Phase III Plan B Trial: first prospective outcome data for the 21-gene recurrence score assay and concordance of prognostic markers by central and local pathology assessment. J Clin Oncol 34:2341–2349

    Article  PubMed  Google Scholar 

  4. Dowsett M, Smith IE, Ebbs SR, Dixon JM, Skene A, A’Hern R, Salter J, Detre S, Hills M, Walsh G, IMPACT Trialists Group (2007) Prognostic value of Ki67 expression after short-term presurgical endocrine therapy for primary breast cancer. J Natl Cancer Inst 99:167–170

    Article  CAS  PubMed  Google Scholar 

  5. Viale G, Giobbie-Hurder A, Regan MM, Coates AS, Mastropasqua MG, Dell’Orto P, Maiorano E, MacGrogan G, Braye SG, Ohlschlegel C, Neven P, Orosz Z, Olszewski WP, Knox F, Thurlimann B, Price KN, Castiglione-Gertsch M, Gelber RD, Gusterson BA, Goldhirsch A, Breast International Group Trial 1-98 (2008) Prognostic and predictive value of centrally reviewed Ki-67 labeling index in postmenopausal women with endocrine-responsive breast cancer: results from Breast International Group Trial 1-98 comparing adjuvant tamoxifen with letrozole. J Clin Oncol 26:5569–5575

    Article  PubMed  PubMed Central  Google Scholar 

  6. Niikura N, Iwamoto T, Masuda S, Kumaki N, Xiaoyan T, Shirane M, Mori K, Tsuda B, Okamura T, Saito Y, Suzuki Y, Tokuda Y (2012) Immunohistochemical Ki67 labeling index has similar proliferation predictive power to various gene signatures in breast cancer. Cancer Sci 103:1508–1512

    Article  CAS  PubMed  Google Scholar 

  7. von Minckwitz G, Schmitt WD, Loibl S, Muller BM, Blohmer JU, Sinn BV, Eidtmann H, Eiermann W, Gerber B, Tesch H, Hilfrich J, Huober J, Fehm T, Barinoff J, Rudiger T, Erbstoesser E, Fasching PA, Karn T, Muller V, Jackisch C, Denkert C (2013) Ki67 measured after neoadjuvant chemotherapy for primary breast cancer. Clin Cancer Res 19:4521–4531

    Article  Google Scholar 

  8. Nitz U, Gluz O, Huober J, Kreipe HH, Kates RE, Hartmann A, Erber R, Scholz M, Lisboa B, Mohrmann S, Mobus V, Augustin D, Hoffmann G, Weiss E, Bohmer S, Kreienberg R, Du Bois A, Sattler D, Thomssen C, Kiechle M, Janicke F, Wallwiener D, Harbeck N, Kuhn W (2014) Final analysis of the prospective WSG-AGO EC-Doc versus FEC phase III trial in intermediate-risk (pN1) early breast cancer: efficacy and predictive value of Ki67 expression. Ann Oncol 25:1551–1557

    Article  CAS  PubMed  Google Scholar 

  9. Denkert C, Loibl S, Muller BM, Eidtmann H, Schmitt WD, Eiermann W, Gerber B, Tesch H, Hilfrich J, Huober J, Fehm T, Barinoff J, Jackisch C, Prinzler J, Rudiger T, Erbstosser E, Blohmer JU, Budczies J, Mehta KM, von Minckwitz G (2013) Ki67 levels as predictive and prognostic parameters in pretherapeutic breast cancer core biopsies: a translational investigation in the neoadjuvant GeparTrio trial. Ann Oncol 24:2786–2793

    Article  CAS  PubMed  Google Scholar 

  10. Tang LH, Gonen M, Hedvat C, Modlin IM, Klimstra DS (2012) Objective quantification of the Ki67 proliferative index in neuroendocrine tumors of the gastroenteropancreatic system: a comparison of digital image analysis with manual methods. Am J Surg Pathol 36:1761–1770

    Article  PubMed  Google Scholar 

  11. Bosman F, Carneiro F, Hruban R, Theise N (2010) WHO classification of tumours of the digestive system. International Agency for Research on Cancer, Lyon

    Google Scholar 

  12. Polley MY, Leung SC, LM MS, Gao D, Hugh JC, Mastropasqua MG, Viale G, Zabaglo LA, Penault-Llorca F, Bartlett JM, Gown AM, Symmans WF, Piper T, Mehl E, Enos RA, Hayes DF, Dowsett M, Nielsen TO, International Ki67 in Breast Cancer Working Group of the Breast International Group and North American Breast Cancer Group (2013) An international Ki67 reproducibility study. J Natl Cancer Inst 105:1897–1906

    Article  PubMed  PubMed Central  Google Scholar 

  13. Polley MY, Leung SC, Gao D, Mastropasqua MG, Zabaglo LA, Bartlett JM, McShane LM, Enos RA, Badve SS, Bane AL, Borgquist S, Fineberg S, Lin MG, Gown AM, Grabau D, Gutierrez C, Hugh JC, Moriya T, Ohi Y, Osborne CK, Penault-Llorca FM, Piper T, Porter PL, Sakatani T, Salgado R, Starczynski J, Laenkholm AV, Viale G, Dowsett M, Hayes DF, Nielsen TO (2015) An international study to increase concordance in Ki67 scoring. Mod Pathol 28:778–786

    Article  PubMed  Google Scholar 

  14. Zabaglo L, Salter J, Anderson H, Quinn E, Hills M, Detre S, A’Hern R, Dowsett M (2010) Comparative validation of the SP6 antibody to Ki67 in breast cancer. J Clin Pathol 63:800–804

    Article  PubMed  Google Scholar 

  15. Christgen M, von Ahsen S, Christgen H, Langer F, Kreipe H (2015) The region-of-interest size impacts on Ki67 quantification by computer-assisted image analysis in breast cancer. Hum Pathol 46:1341–1349

    Article  PubMed  Google Scholar 

  16. Cserni G, Voros A, Liepniece-Karele I, Bianchi S, Vezzosi V, Grabau D, Sapino A, Castellano I, Regitnig P, Foschini MP, Zolota V, Varga Z, Figueiredo P, Decker T, Focke C, Kulka J, Kaya H, Reiner-Concin A, Amendoeira I, Callagy G, Caffrey E, Wesseling J, Wells C (2014) Distribution pattern of the Ki67 labelling index in breast cancer and its implications for choosing cut-off values. Breast 23:259–263

    Article  PubMed  Google Scholar 

  17. Wasielewski R, Hasselmann S, Ruschoff J, Fisseler-Eckhoff A, Kreipe H (2008) Proficiency testing of immunohistochemical biomarker assays in breast cancer. Virchows Arch 453:537–543

    Article  PubMed  Google Scholar 

  18. Broyde A, Boycov O, Strenov Y, Okon E, Shpilberg O, Bairey O (2009) Role and prognostic significance of the Ki-67 index in non-Hodgkin’s lymphoma. Am J Hematol 84:338–343

    Article  PubMed  Google Scholar 

  19. Inwald EC, Klinkhammer-Schalke M, Hofstadter F, Zeman F, Koller M, Gerstenhauer M, Ortmann O (2013) Ki-67 is a prognostic parameter in breast cancer patients: results of a large population-based cohort of a cancer registry. Breast Cancer Res Treat 139:539–552

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Cheang MC, Chia SK, Voduc D, Gao D, Leung S, Snider J, Watson M, Davies S, Bernard PS, Parker JS, Perou CM, Ellis MJ, Nielsen TO (2009) Ki67 index, HER2 status, and prognosis of patients with luminal B breast cancer. J Natl Cancer Inst 101:736–750

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Laurinavicius A, Plancoulaine B, Rasmusson A, Besusparis J, Augulis R, Meskauskas R, Herlin P, Laurinaviciene A, Abdelhadi Muftah AA, Miligy I, Aleskandarany M, Rakha EA, Green AR, Ellis IO (2016) Bimodality of intratumor Ki67 expression is an independent prognostic factor of overall survival in patients with invasive breast carcinoma. Virchows Arch 468:493–502

    Article  CAS  PubMed  Google Scholar 

  22. Penault-Llorca F, Andre F, Sagan C, Lacroix-Triki M, Denoux Y, Verriele V, Jacquemier J, Baranzelli MC, Bibeau F, Antoine M, Lagarde N, Martin AL, Asselain B, Roche H (2009) Ki67 expression and docetaxel efficacy in patients with estrogen receptor-positive breast cancer. J Clin Oncol 27:2809–2815

    Article  CAS  PubMed  Google Scholar 

  23. Viale G, Regan MM, Mastropasqua MG, Maffini F, Maiorano E, Colleoni M, Price KN, Golouh R, Perin T, Brown RW, Kovacs A, Pillay K, Ohlschlegel C, Gusterson BA, Castiglione-Gertsch M, Gelber RD, Goldhirsch A, Coates AS, International Breast Cancer Study Group (2008) Predictive value of tumor Ki-67 expression in two randomized trials of adjuvant chemoendocrine therapy for node-negative breast cancer. J Natl Cancer Inst 100:207–212

    Article  CAS  PubMed  Google Scholar 

  24. Wiesner FG, Magener A, Fasching PA, Wesse J, Bani MR, Rauh C, Jud S, Schrauder M, Loehberg CR, Beckmann MW, Hartmann A, Lux MP (2009) Ki-67 as a prognostic molecular marker in routine clinical use in breast cancer patients. Breast 18:135–141

    Article  PubMed  Google Scholar 

  25. Dowsett M, Nielsen TO, A’Hern R, Bartlett J, Coombes RC, Cuzick J, Ellis M, Henry NL, Hugh JC, Lively T, McShane L, Paik S, Penault-Llorca F, Prudkin L, Regan M, Salter J, Sotiriou C, Smith IE, Viale G, Zujewski JA, Hayes DF, International Ki-67 in Breast Cancer Working Group (2011) Assessment of Ki67 in breast cancer: recommendations from the international Ki67 in breast cancer working group. J Natl Cancer Inst 103:1656–1664

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Coates AS, Winer EP, Goldhirsch A, Gelber RD, Gnant M, Piccart-Gebhart M, Thurlimann B, Senn HJ, Panel Members (2015) Tailoring therapies—improving the management of early breast cancer: St Gallen International Expert Consensus on the primary therapy of early breast cancer 2015. Ann Oncol 26:1533–1546

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Endris V, Stenzinger A, Pfarr N, Penzel R, Möbs M, Lenze D, Darb-Esfahani S, Hummel M, Sabine-Merkelbach-Bruse JA, Lehmann U, Kreipe H, Kirchner T, Büttner R, Jochum W, Höfler G, Dietel M, Weichert W, Schirmacher P (2016) NGS-based BRCA1/2 mutation testing of high-grade serous ovarian cancer tissue: results and conclusions of the first international round robin trial. Virchows Arch 468:697–705. doi:10.1007/s00428-016-1919-8

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

This study was supported by the Deutsche Krebsgesellschaft. The authors gratefully acknowledge Gillian Teicke for her support in preparing the manuscript.

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Author notes

  1. M. Raap and S. Ließem contributed equally to this work.

Authors and Affiliations

  1. Institute of Pathology, Hannover Medical School, Carl-Neuberg-Str. 1, 30625, Hannover, Germany

    M. Raap, S. Ließem & H. Kreipe

  2. Institute of Pathology Nordhessen, Kassel, Germany

    J. Rüschoff

  3. Institute of Pathology and Cytology, Wiesbaden, Germany

    A. Fisseler-Eckhoff

  4. Department of Pathology and Bacteriology, Danube Hospital, Vienna, Austria

    A. Reiner

  5. Institute of Pathology, University Hospital of Basel, Basel, Switzerland

    S. Dirnhofer

  6. Institute of Pathology, Nordstadt Hospital, Hannover, Germany

    R. von Wasielewski

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  1. M. Raap
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Contributions

Authorship was based on the ICMJE recommendation for the qualification of authorship.

M. Raap, S. Ließem, and H. Kreipe designed the study. All authors made substantial contributions to the acquisition, analysis, and interpretation of data; revised the manuscript for important intellectual content; approved the final version; and agreed to be accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved.

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Correspondence to H. Kreipe.

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Conflict of interest

The authors declare that they have no conflict of interest.

Disclosure statement

R. von Wasielewski is the founder and partial owner of “multiblock GmbH” which served as a logistics center in the trial.

Research did not involve human participants or animals.

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Raap, M., Ließem, S., Rüschoff, J. et al. Quality assurance trials for Ki67 assessment in pathology. Virchows Arch 471, 501–508 (2017). https://doi.org/10.1007/s00428-017-2142-y

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  • DOI: https://doi.org/10.1007/s00428-017-2142-y

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