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Comprehensive immunohistochemical analysis of RET, BCAR1, and BCAR3 expression in patients with Luminal A and B breast cancer subtypes

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Abstract

Purpose

Breast cancer (BC) is considered a heterogeneous disease composed of distinct subtypes with diverse clinical outcomes. Luminal subtype tumors have the best prognosis, and patients benefit from endocrine therapy. However, resistance to endocrine therapies in BC is an obstacle to successful treatment, and novel biomarkers are needed to understand and overcome this mechanism. The RET, BCAR1, and BCAR3 genes may be associated with BC progression and endocrine resistance.

Methods

Aiming to evaluate the expression profile and prognostic value of RET, BCAR1, and BCAR3, we performed immunohistochemistry on tissue microarrays (TMAs) containing a cohort of 361 Luminal subtype BC.

Results

Low expression levels of these three proteins were predominantly observed. BCAR1 expression was correlated with nuclear grade (p = 0.057), and BCAR3 expression was correlated with lymph node status (p = 0.011) and response to hormonal therapy (p = 0.021). Further, low expression of either BCAR1 or BCAR3 was significantly associated with poor prognosis (p = 0.005; p = 0.042). Pairwise analysis showed that patients with tumors with low BCAR1/low BCAR3 expression had a poorer overall survival (p = 0.013), and the low BCAR3 expression had the worst prognosis with RET high expression stratifying these patients into two different groups. Regarding the response to hormonal therapy, non-responder patients presented lower expression of RET in comparison to the responder group (p = 0.035). Additionally, the low BCAR1 expression patients had poorer outcomes than BCAR1 high (p = 0.015).

Conclusion

Our findings suggest RET, BCAR1, and BCAR3 as potential candidate markers for endocrine therapy resistance in Luminal BC.

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Funding

This work was supported by the Fundação de Amparo a Pesquisa do Estadode São Paulo (FAPESP; 2019/05252-4) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq; 309524/2017-2).

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Authors and Affiliations

  1. Discipline of Oncology, Department of Radiology and Oncology, Faculty of Medicine, University of São Paulo, São Paulo, 01246-903, Brazil

    Ana Carolina Pavanelli, Flavia Rotea Mangone, Simone Aparecida Bessa & Maria Aparecida Nagai

  2. Laboratory of Molecular Genetics, Center for Translational Research in Oncology, Cancer Institute of Sao Paulo, São Paulo, 01246-000, Brazil

    Ana Carolina Pavanelli, Flavia Rotea Mangone, Simone Aparecida Bessa & Maria Aparecida Nagai

  3. Department of Pathology and Molecular Medicine, Cancer Research Institute, Queen’s University Kingston, 18 Stuart Street, Kingston, ON, K7L 3N6, Canada

    Piriya Yoganathan & Lois M. Mulligan

  4. Department of Pathological Anatomy, A. C. Camargo Cancer Center, São Paulo, 01509-020, Brazil

    Suely Nonogaki, Cynthia A. B. de Toledo Osório & Victor Piana de Andrade

  5. Department of Pathology, Cancer Institute of Sao Paulo, Hospital das Clinicas, Faculty of Medicine, University of São Paulo, HCFMUSP, São Paulo, 01246-903, Brazil

    Iberê Cauduro Soares & Evandro Sobrosa de Mello

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  1. Ana Carolina Pavanelli
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Correspondence to Maria Aparecida Nagai.

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This study was conducted according to the guidelines laid down in the Declaration of Helsinki and was approved by the Institutional Review Boards of the Medical School University of São Paulo, São Paulo, Brasil (CAAE: 35295514.1.0000.0065).

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10549_2021_6452_MOESM1_ESM.tiff

Supplementary file1—Supplementary Figure 1 - Kaplan-Meier curves of disease-free survival in Luminal breast cancer patients stratified according to RET (a), BCAR1 (b), and BCAR3 (c) protein expression. Tumors were classified as low (negative or weak) or high (moderate or intense) according to RET, BCAR1, or BCAR3 protein immunostaining. Log-rank test was performed for curve comparisons. HR - hazard ratio. (TIFF 109 kb)

10549_2021_6452_MOESM2_ESM.tiff

Supplementary file2—Supplementary Figure 2 - Prognostic association of the pairwise combinations of RET, BCAR1 and BCAR3 expression. Kaplan-Meier curves of disease-free survival in luminal breast cancer patients stratified according to the combination of RET and BCAR3 (a), RET and BCAR1 (b) and, BCAR3 and BCAR1 (c) protein expression. Tumors were classified as low (negative or weak) or high (moderate or intense) according to RET, BCAR3, or BCAR1 protein immunostaining. Log-rank test was performed for curve comparisons. NA – not analyzed because 100% of the cases was censored cases. HR -hazard ratio. (TIFF 331 kb)

10549_2021_6452_MOESM3_ESM.tiff

Supplementary file3—Supplementary Figure 3 - Prognostic association between RET, BCAR1, and BCAR3 expression and response to endocrine therapy. Kaplan-Meier curves of disease-free survival in Luminal breast cancer patients stratified according to RET (a), BCAR1 (b), and BCAR3 (c) protein expression in combination with response to endocrine therapy. Tumors were classified as low (negative or weak) or high (moderate or intense) according to RET, BCAR1, or BCAR3 protein immunostaining and response to hormonal therapy was considered as absence of disease recurrence. Log-rank test was performed for curve comparisons. The p-values obtained for comparison between responder and/or non responder curves are presented inside the figures. NA – not analyzed because 100% of the cases was censored cases. HR - Hazard ratio. (TIFF 127 kb)

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Pavanelli, A.C., Mangone, F.R., Yoganathan, P. et al. Comprehensive immunohistochemical analysis of RET, BCAR1, and BCAR3 expression in patients with Luminal A and B breast cancer subtypes. Breast Cancer Res Treat 192, 43–52 (2022). https://doi.org/10.1007/s10549-021-06452-9

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