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Evaluation of the anti-Thomsen–Friedenreich antibodies Nemod-TF1 and Nemod-TF2 as prognostic markers in breast cancer

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Abstract

Purpose

The TF (Thomsen–Friedenreich, CD176, Galβ1-3GalNAc) carbohydrate moiety is known as a specific oncofetal carbohydrate epitope present in fetal and neoplastic tissue as well as in stem cells. TF was demonstrated to mediate tumor-promoting features and to be highly immunogenic. The current study aimed to evaluate whether presence of the TF antigen is associated with clinico-pathological parameters and prognosis of early breast cancer (BC).

Methods

Primary BC tissue (n = 226) was stained for TF using two monoclonal anti-TF antibodies (Nemod-TF1, Nemod-TF2). Staining results were correlated to clinical data including survival.

Results

Nemod-TF1 staining was positively correlated to lymph node metastasis (p = 0.03) and the presence of tumor-associated MUC1 (TA-MUC1; p = 0.003). Further, the presence of the Nemod-TF1 epitope predicted worse prognosis in TA-MUC1 positive (overall survival: p = 0.026) as well as in triple negative (overall survival: p = 0.002; distant metastasis-free survival: p = 0.012) BC.

Conclusions

The data presented here further support a role of TF in BC tumor biology. Whether anti-TF directed treatment approaches may gain clinical relevance in those cases determined as triple negative or TA-MUC1 positive remains to be determined.

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Abbreviations

BC:

Breast cancer

CIS:

Carcinoma in situ

CSC:

Cancer stem cell

DFS:

Disease-free survival

DMFS:

Distant metastasis-free survival

ER:

Estrogen receptor

FFPE:

Formalin-fixed, paraffin-embedded

IRS:

Immunoreactive score

LRFS:

Local recurrence-free survival

OS:

Overall survival

PR:

Progesterone receptor

REMARK:

REporting recommendations for tumor MARKer prognostic studies

TA-MUC1:

Tumor-associated mucin-1

TF:

Thomsen–Friedenreich, CD176, Galβ1-3GalNAc

TNBC:

Triple negative BC

WHO:

World Health Organization

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Acknowledgements

The authors thank Irmgard Wiest and Susanne Kunze (†) for their excellent technical assistance. We would further like to acknowledge Jutta Engel and Max Wiedemann for their help with collecting follow-up data from the Munich Cancer Registry. Parts of this work are part of ME’s MD thesis obtained at the Department of Obstetrics and Gynecology, LMU Munich, Germany [36].

Funding

The study was financed by the Department of Obstetrics and Gynecology, Ludwig-Maximilians-University of Munich, Munich, Germany. No special funding was received. 

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Authors and Affiliations

  1. Department of Obstetrics and Gynecology, Heidelberg University Hospital, Heidelberg, Germany

    Sabine Heublein & Florian Schuetz

  2. Department of Obstetrics and Gynecology, Ludwig-Maximilians-University of Munich, Munich, Germany

    Sabine Heublein, Markus Egger, Junyan Zhu, Luisa Berger, Christina Kuhn, Simone S. Hofmann, Udo Jeschke & Nina Ditsch

  3. St. Anna Kinderspital, Vienna, Austria

    Markus Egger

  4. Department of Pathology, Ludwig-Maximilians-University of Munich, Munich, Germany

    Doris Mayr

  5. Klinikum Traunstein, Traunstein, Germany

    Christian Schindlbeck

  6. Department of Obstetrics and Gynecology, University Hospital Augsburg, Augsburg, Germany

    Udo Jeschke & Nina Ditsch

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  1. Sabine Heublein
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Contributions

SH participated in design and coordination of the study, significantly participated in experimental assays and analysis as well as in statistical analysis, and wrote the manuscript. ME participated in IHC assays and analysis and helped to draft the manuscript. LB participated in data collection and analysis. JZ participated in experimental assays. DM supervised IHC as a gynecologic pathologist and participated in IHC analysis. CS participated in the design of the study and carefully reviewed the manuscript for important intellectual content. CK optimized Nemod-TF staining and participated in immunohistochemistry. SSH participated in cell culture experiment design and conduction. FS helped to draft the manuscript and revised the manuscript for important intellectual content. UJ conceived and coordinated the study, participated in study design, and approved the final version of the manuscript. ND conceived and coordinated the study, participated in study design, and approved the final version of the manuscript.

Corresponding author

Correspondence to Sabine Heublein.

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Conflict of interest

All authors read the manuscript and agree to the publication of the manuscript. SH reports grants from the following organizations/companies: FöFoLe LMU Munich Medical Faculty, FERRING, Novartis Oncology, Astra Zeneca, Apceth, Heuer Stiftung, Deutsche Forschungsgemeinschaft. She further reports personal fees from Roche and non-financial support from Addex. ND held honorary speeches for Roche, AstraZeneca, Mentor, Omniamed, TEVA, and MSD. All the support listed here has been received outside the submitted work. Remaining authors have no competing interests to declare.

Ethical approval

This article does not contain any studies with animals performed by any of the authors. 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. The tissue samples were retrieved from the archive of Obstetrics and Gynecology, Ludwig-Maximilians-University, Munich, Germany. The study was approved by the Ethics Committee of the Ludwig-Maximilians-University, Munich, Germany (approval number: 18-166). The tumor tissue re-used for our analysis had initially been collected for histopathological diagnostics. At the time, the tissue was examined for the current study all diagnostic procedures had already been fully completed and thus the tissue was classified as “left-over” material. Clinical data and tissue specimens were irreversibly anonymized prior to inclusion in the study. Researchers were blinded from patient data during experimental and statistical analysis.

Informed consent

As per declaration of our ethics committee [Ludwig-Maximilians-University, Munich, Germany (approval number: 18-166)], no written informed consent of the participants is needed given the circumstances described above.

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10549_2019_5503_MOESM1_ESM.jpg

Supplementary material 1 (JPEG 63 kb) Supplementary Figure 1: Disease-free survival (DFS) of Nemod-TF1-positive vs. -negativepatients in the subgroup of TA-MUC1mem-positive (A) and triple negative (B) breast cancers

10549_2019_5503_MOESM2_ESM.jpg

Supplementary material 2 (JPEG 28 kb) Supplementary Figure 2: Distant metastasis-free survival (DMFS) of Nemod-TF1-positive vs. -negative patients in the subgroup of Her2-positive breast cancers

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Heublein, S., Egger, M., Zhu, J. et al. Evaluation of the anti-Thomsen–Friedenreich antibodies Nemod-TF1 and Nemod-TF2 as prognostic markers in breast cancer. Breast Cancer Res Treat 179, 643–652 (2020). https://doi.org/10.1007/s10549-019-05503-6

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