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Identification of tumor biomarkers for pathological complete response to neoadjuvant treatment in locally advanced breast cancer

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Abstract

Background

Therapeutic response predictors like age, nodal status, and tumor grade and markers, like ER/PR, HER2, and Ki67, are not reliable in predicting the response to a specific form of chemotherapy. The current study aims to identify and validate reliable markers that can predict pathological complete response (pCR) in fluorouracil, epirubicin, and cyclophosphamide (FEC)-based neoadjuvant therapy with (NACT/RT) and without concurrent radiation (NACT).

Materials and methods

Tandem mass tag (TMT) quantitative liquid chromatography-tandem mass spectrometry (LC–MS/MS) was used to identify differentially expressed proteins from core needle breast biopsy between pCR (n = 4) and no-pCR (n = 4). Immunoblotting of shortlisted proteins with the tissue lysates confirmed the differential expression of the markers. Further, immunohistochemistry (IHC) was performed on formalin-fixed paraffin-embedded sections of treatment-naive core needle biopsies. In the NACT, 29 pCR and 130 no-pCR and in NACT/RT, 32 pCR and 71 no-pCR were used.

Results

733 and 807 proteins were identified in NACT and NACT/RT groups, respectively. Ten proteins were shortlisted for validation as potential pCR-predictive markers. THBS1, TNC, and DCN were significantly overexpressed in no-pCR in both the groups. In NACT, CPA3 was significantly upregulated in the no-pCR. In NACT/RT, HnRNPAB was significantly upregulated and HMGB1 significantly downregulated in the no-pCR. HMGB1 was the only marker to show prognostic significance.

Conclusion

Quantitative proteomics followed by IHC identified and validated potential biomarkers for predicting patient response to therapy. These markers can be used, following larger-scale validation, in combination with routine histological analysis providing vital indications of treatment response.

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Data availability

The authors confirm that the data supporting the findings of this study are available within the article [and/or] its supplementary materials.

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Acknowledgements

We acknowledge the Department of Science, Government of India for the funding of the project (No. SERB/F/2369/2014-15, dt. 29-06-2014). We also thank the Indian Council of Medical Research for the financial support of Ms. Prarthana Gopinath, through their Senior Research Fellowship Program (5/3/8/37/ITR-F/2018-ITR dt. 07-06-2018).

Funding

We acknowledge the Department of Science, Government of India for the funding of the project (No. SERB/F/2369/2014–15, dt. 29–06-2014). We also thank the Indian Council of Medical Research for the financial support of Ms. Prarthana Gopinath, through their Senior Research Fellowship program (5/3/8/37/ITR-F/2018-ITR dt. 07–06-2018).

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Authors and Affiliations

  1. Department of Molecular Oncology, Cancer Institute WIA, Chennai, 600036, Tamil Nadu, India

    Prarthana Gopinath, Gopal Gopisetty & Rajkumar Thangarajan

  2. Department of Surgical Oncology, Cancer Institute WIA, Chennai, 600036, Tamil Nadu, India

    Sridevi Veluswami

  3. Department of Oncopatholology, Cancer Institute WIA, Chennai, Tamil Nadu, India

    Shirley Sundersingh

  4. Department of Epidemiology, Biostatistics and Cancer Registry, Cancer Institute WIA, Chennai, Tamil Nadu, India

    Swaminathan Rajaraman

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  1. Prarthana Gopinath
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  2. Sridevi Veluswami
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  3. Gopal Gopisetty
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  4. Shirley Sundersingh
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  5. Swaminathan Rajaraman
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  6. Rajkumar Thangarajan
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Contributions

All authors contributed to the study conception and design. PG, SV, SS, and RS: contributed to resource and sample collection. PG, GG, and TR: contributed in the methodology, data curation, formal analysis, investigation, and validation. Visualization of the histology results was performed by SS. The first draft of the manuscript was written by PG and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Sridevi Veluswami or Gopal Gopisetty.

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The study was approved by the Institutional Ethical Committee. Informed consent was obtained from all individual participants included in the study.

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Supplementary Information

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Supplementary file1 (PDF 235 KB)

10549_2022_6617_MOESM2_ESM.jpg

Supplementary file2 (JPG 188 KB) Prognostic significance of the candidate markers Kaplan–Meier plots for DFS and OS for low (green) vs. high protein expression (red). p-value ≤ 0.05 was considered statistically significant. Graphs were generated using GraphPad Prism v5.

Supplementary file3 (XLSX 245 KB)

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Gopinath, P., Veluswami, S., Gopisetty, G. et al. Identification of tumor biomarkers for pathological complete response to neoadjuvant treatment in locally advanced breast cancer. Breast Cancer Res Treat 194, 207–220 (2022). https://doi.org/10.1007/s10549-022-06617-0

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