Skip to main content

Advertisement

SpringerLink
Log in

The spectrum of HER2 expression in breast cancer: linking immunohistochemistry quantification with in situ hybridization assay

  • Original Article
  • Published:
Virchows Archiv Aims and scope Submit manuscript

A Letter to the Editor to this article was published on 14 June 2022

Abstract

We aimed to document the pathological characteristics of breast cancer (BC) cases with different scores of HER2 by immunohistochemistry (IHC), as well as to establish a relationship between HER2 expression and HER2 amplification by in situ hybridization (ISH). A cohort of 258 primary BC cases was evaluated for HER2 gene amplification with bright-field ISH. All HER2-negative and HER2-positive cases by IHC were concordant with the ISH classification. BC cases with score of 0 had lower average of HER2 copy number compared to cases with score of 1 + . HER2-equivocal cases by IHC had intermediate pathological characteristics between HER2-negative and HER2-positive cases. About 12% of HER2-equivocal cases were classified as ISH-positive. HER2-equivocal cases with HER2 gene amplification had proliferation index, HER2/CEP17 ratio, and average of HER2 copy number between HER2-equivocal cases without HER2 gene amplification and HER2-positive cases by IHC. Additionally, HER2-equivocal cases with HER2 amplification had score of 2 + in at least 50% of the total tumor area, with a proportion of ISH-positive cases increasing with the amount of score of 2 + present in the tumor. The quantification of score of 2 + in the tumor predicted the ISH classification with an AUC of 0.902. A logistic regression model using the same HER2 quantification and the nuclear score was able to increase the abovementioned prediction to an AUC of 0.929. As such, we were able to link HER2 quantification by IHC and morphological analysis with HER2 amplification by ISH.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

Data availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

References

  1. Wolff AC, Hammond MEH, Allison KH, Harvey BE, Mangu PB, Bartlett JMS, Bilous M, Ellis IO, Fitzgibbons P, Hanna W, Jenkins RB, Press MF, Spears PA, Vance GH, Viale G, McShane LM, Dowsett M (2018) Human epidermal growth factor receptor 2 testing in breast cancer: American Society of Clinical Oncology/College of American Pathologists clinical practice guideline focused update. Arch Pathol Lab Med 142(11):1364–1382

    Article  Google Scholar 

  2. Allison KH, Hammond MEH, Dowsett M, McKernin SE, Carey LA, Fitzgibbons PL, Hayes DF, Lakhani SR, Chavez-MacGregor M, Perlmutter J, Perou CM, Regan MM, Rimm DL, Symmans WF, Torlakovic EE, Varella L, Viale G, Weisberg TF, McShane LM, Wolff AC (2020) Estrogen and progesterone receptor testing in breast cancer: American Society of Clinical Oncology/College of American Pathologists guideline update. Arch Pathol Lab Med 144(5):545–563

    Article  CAS  Google Scholar 

  3. Slamon DJ, Clark GM, Wong SG, Levin WJ, Ullrich A, McGuire WL (1987) Human breast cancer: correlation of relapse and survival with amplification of the HER-2/neu oncogene. Science 235(4785):177–182

    Article  CAS  Google Scholar 

  4. Press MF, Pike MC, Chazin VR, Hung G, Udove JA, Markowicz M, Danyluk J, Godolphin W, Sliwkowski M, Akita R et al (1993) Her-2/neu expression in node-negative breast cancer: direct tissue quantitation by computerized image analysis and association of overexpression with increased risk of recurrent disease. Cancer Res 53(20):4960–4970

    CAS  PubMed  Google Scholar 

  5. Press MF, Bernstein L, Thomas PA, Meisner LF, Zhou JY, Ma Y, Hung G, Robinson RA, Harris C, El-Naggar A, Slamon DJ, Phillips RN, Ross JS, Wolman SR, Flom KJ (1997) HER-2/neu gene amplification characterized by fluorescence in situ hybridization: poor prognosis in node-negative breast carcinomas. J Clin Oncol 15(8):2894–2904

    Article  CAS  Google Scholar 

  6. Slamon DJ, Leyland-Jones B, Shak S, Fuchs H, Paton V, Bajamonde A, Fleming T, Eiermann W, Wolter J, Pegram M, Baselga J, Norton L (2001) Use of chemotherapy plus a monoclonal antibody against HER2 for metastatic breast cancer that overexpresses HER2. N Engl J Med 344(11):783–792

    Article  CAS  Google Scholar 

  7. Cobleigh MA, Vogel CL, Tripathy D, Robert NJ, Scholl S, Fehrenbacher L, Wolter JM, Paton V, Shak S, Lieberman G, Slamon DJ (1999) Multinational study of the efficacy and safety of humanized anti-HER2 monoclonal antibody in women who have HER2-overexpressing metastatic breast cancer that has progressed after chemotherapy for metastatic disease. J Clin Oncol 17(9):2639–2648

    Article  CAS  Google Scholar 

  8. Piccart-Gebhart MJ, Procter M, Leyland-Jones B, Goldhirsch A, Untch M, Smith I, Gianni L, Baselga J, Bell R, Jackisch C, Cameron D, Dowsett M, Barrios CH, Steger G, Huang CS, Andersson M, Inbar M, Lichinitser M, Lang I, Nitz U, Iwata H, Thomssen C, Lohrisch C, Suter TM, Ruschoff J, Suto T, Greatorex V, Ward C, Straehle C, McFadden E, Dolci MS, Gelber RD (2005) Trastuzumab after adjuvant chemotherapy in HER2-positive breast cancer. N Engl J Med 353(16):1659–1672

    Article  CAS  Google Scholar 

  9. Vogel CL, Cobleigh MA, Tripathy D, Gutheil JC, Harris LN, Fehrenbacher L, Slamon DJ, Murphy M, Novotny WF, Burchmore M, Shak S, Stewart SJ, Press M (2002) Efficacy and safety of trastuzumab as a single agent in first-line treatment of HER2-overexpressing metastatic breast cancer. J Clin Oncol 20(3):719–726

    Article  CAS  Google Scholar 

  10. Tsai YF, Tseng LM, Lien PJ, Hsu CY, Lin YS, King KL, Wang YL, Chao TC, Liu CY, Chiu JH, Yang MH (2019) HER2 immunohistochemical scores provide prognostic information for patients with HER2-type invasive breast cancer. Histopathology 74(4):578–586

    Article  Google Scholar 

  11. Zhao J, Krishnamurti U, Zhang C, Meisel J, Wei Z, Suo A, Aneja R, Li Z, Li X (2020) HER2 immunohistochemistry staining positivity is strongly predictive of tumor response to neoadjuvant chemotherapy in HER2 positive breast cancer. Pathol Res Pract 216(11):153155

    Article  CAS  Google Scholar 

  12. Marchio C, Annaratone L, Marques A, Casorzo L, Berrino E, Sapino A (2021) Evolving concepts in HER2 evaluation in breast cancer: Heterogeneity, HER2-low carcinomas and beyond. Semin Cancer Biol 72:123–135

    Article  CAS  Google Scholar 

  13. Tarantino P, Hamilton E, Tolaney SM, Cortes J, Morganti S, Ferraro E, Marra A, Viale G, Trapani D, Cardoso F, Penault-Llorca F, Viale G, Andre F, Curigliano G (2020) HER2-low breast cancer: pathological and clinical landscape. J Clin Oncol 38(17):1951–1962

    Article  CAS  Google Scholar 

  14. Banerji U, van Herpen CML, Saura C, Thistlethwaite F, Lord S, Moreno V, Macpherson IR, Boni V, Rolfo C, de Vries EGE, Rottey S, Geenen J, Eskens F, Gil-Martin M, Mommers EC, Koper NP, Aftimos P (2019) Trastuzumab duocarmazine in locally advanced and metastatic solid tumours and HER2-expressing breast cancer: a phase 1 dose-escalation and dose-expansion study. Lancet Oncol 20(8):1124–1135

    Article  CAS  Google Scholar 

  15. Modi S, Park H, Murthy RK, Iwata H, Tamura K, Tsurutani J, Moreno-Aspitia A, Doi T, Sagara Y, Redfern C, Krop IE, Lee C, Fujisaki Y, Sugihara M, Zhang L, Shahidi J, Takahashi S (2020) Antitumor activity and safety of trastuzumab deruxtecan in patients with HER2-Low-expressing advanced breast cancer: results from a phase Ib study. J Clin Oncol 38(17):1887–1896

    Article  CAS  Google Scholar 

  16. Press MF, Villalobos I, Santiago A, Guzman R, Cervantes M, Gasparyan A, Campeau A, Ma Y, Tsao-Wei DD, Groshen S (2016) Assessing the new American Society of Clinical Oncology/College of American Pathologists guidelines for HER2 testing by fluorescence in situ hybridization: experience of an academic consultation practice. Arch Pathol Lab Med 140(11):1250–1258

    Article  Google Scholar 

  17. Press MF, Sauter G, Buyse M, Fourmanoir H, Quinaux E, Tsao-Wei DD, Eiermann W, Robert N, Pienkowski T, Crown J, Martin M, Valero V, Mackey JR, Bee V, Ma Y, Villalobos I, Campeau A, Mirlacher M, Lindsay MA, Slamon DJ (2016) HER2 gene amplification testing by fluorescent in situ hybridization (FISH): comparison of the ASCO-College of American Pathologists guidelines with FISH scores used for enrollment in Breast Cancer International Research Group clinical trials. J Clin Oncol 34(29):3518–3528

    Article  CAS  Google Scholar 

  18. Hendriks BS, Klinz SG, Reynolds JG, Espelin CW, Gaddy DF, Wickham TJ (2013) Impact of tumor HER2/ERBB2 expression level on HER2-targeted liposomal doxorubicin-mediated drug delivery: multiple low-affinity interactions lead to a threshold effect. Mol Cancer Ther 12(9):1816–1828

    Article  CAS  Google Scholar 

  19. Onsum MD, Geretti E, Paragas V, Kudla AJ, Moulis SP, Luus L, Wickham TJ, McDonagh CF, MacBeath G, Hendriks BS (2013) Single-cell quantitative HER2 measurement identifies heterogeneity and distinct subgroups within traditionally defined HER2-positive patients. Am J Pathol 183(5):1446–1460

    Article  CAS  Google Scholar 

  20. Curado M, Caramelo AS, Eloy C, Polonia A (2019) What to expect from the 2018 ASCO/CAP HER2 guideline in the reflex in situ hybridization test of immunohistochemically equivocal 2+ cases? Virchows Arch 475(3):303–311

    Article  CAS  Google Scholar 

  21. Polonia A, Caramelo A (2021) HER2 in situ hybridization test in breast cancer: quantifying margins of error and genetic heterogeneity. Mod Pathol 34(8):1478–1486

    Article  CAS  Google Scholar 

  22. Polonia A, Leitao D, Schmitt F (2016) Application of the 2013 ASCO/CAP guideline and the SISH technique for HER2 testing of breast cancer selects more patients for anti-HER2 treatment. Virchows Arch 468(4):417–423

    Article  CAS  Google Scholar 

  23. Polonia A, Oliveira G, Schmitt F (2017) Characterization of HER2 gene amplification heterogeneity in invasive and in situ breast cancer using bright-field in situ hybridization. Virchows Arch 471(5):589–598

    Article  CAS  Google Scholar 

  24. Murray C, D’Arcy C, Gullo G, Flanagan L, Quinn CM, Quinn CM (2018) Human epidermal growth factor receptor 2 testing by fluorescent in situ hybridization: positive or negative? ASCO/College of American Pathologists Guidelines 2007, 2013, and 2018. J Clin Oncol 36(35):3522–3523

    Article  CAS  Google Scholar 

  25. Xu B, Shen J, Guo W, Zhao W, Zhuang Y, Wang L (2019) Impact of the 2018 ASCO/CAP HER2 guidelines update for HER2 testing by FISH in breast cancer. Pathol Res Pract 215(2):251–255

    Article  CAS  Google Scholar 

  26. Krystel-Whittemore M, Xu J, Brogi E, Ventura K, Patil S, Ross DS, Dang C, Robson M, Norton L, Morrow M, Wen HY (2019) Pathologic complete response rate according to HER2 detection methods in HER2-positive breast cancer treated with neoadjuvant systemic therapy. Breast Cancer Res Treat 177(1):61–66

    Article  CAS  Google Scholar 

  27. Wu Z, Xu S, Zhou L, Yin W, Lin Y, Du Y, Wang Y, Jiang Y, Yin K, Zhang J, Lu J (2018) Clinical significance of quantitative HER2 gene amplification as related to its predictive value in breast cancer patients in neoadjuvant setting. Onco Targets Ther 11:801–808

    Article  Google Scholar 

  28. Hurvitz SA, Caswell-Jin JL, McNamara KL, Zoeller JJ, Bean GR, Dichmann R, Perez A, Patel R, Zehngebot L, Allen H, Bosserman L, DiCarlo B, Kennedy A, Giuliano A, Calfa C, Molthrop D, Mani A, Chen HW, Dering J, Adams B, Kotler E, Press MF, Brugge JS, Curtis C, Slamon DJ (2020) Pathologic and molecular responses to neoadjuvant trastuzumab and/or lapatinib from a phase II randomized trial in HER2-positive breast cancer (TRIO-US B07). Nat Commun 11(1):5824

    Article  CAS  Google Scholar 

  29. Katayama A, Miligy IM, Shiino S, Toss MS, Eldib K, Kurozumi S, Quinn CM, Badr N, Murray C, Provenzano E, Callagy G, Martyn C, Millican-Slater R, Purdie C, Purnell D, Pinder SE, Oyama T, Shaaban AM, Ellis I, Lee AHS, Rakha EA (2021) Predictors of pathological complete response to neoadjuvant treatment and changes to post-neoadjuvant HER2 status in HER2-positive invasive breast cancer. Mod Pathol 34(7):1271–1281

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Pathology, Ipatimup Diagnostics, Institute of Molecular Pathology and Immunology, University of Porto, Rua Júlio Amaral de Carvalho, 45, 4200-135, Porto, Portugal

    António Polónia, Carolina Canelas & Ana Caramelo

  2. I3S – Instituto de Investigação e Inovação em Saúde, University of Porto, Porto, Portugal

    António Polónia, Carolina Canelas & Ana Caramelo

Authors
  1. António Polónia
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Carolina Canelas
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ana Caramelo
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

AP and AC: designed the research study. AC and CC: responsible for the execution of the IHC and ISH technique. AP: retrieved and analyzed the data, wrote the first draft of the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to António Polónia.

Ethics declarations

Ethics approval and consent to participate

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. For this type of study, formal consent is not required.

Conflict of interest

The authors declare no competing interests.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 (DOCX 22 KB)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Polónia, A., Canelas, C. & Caramelo, A. The spectrum of HER2 expression in breast cancer: linking immunohistochemistry quantification with in situ hybridization assay. Virchows Arch 480, 1171–1179 (2022). https://doi.org/10.1007/s00428-022-03290-y

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00428-022-03290-y

Keywords

Search

Navigation