- Case Report
- Open Access
Negative genic switch of HER-2 in the primary tumor instead of the synchronous metastatic nodal lesions after neoadjuvant chemotherapy in a patient with primary HER2-positive breast cancer
- Hao-ran Chen†1,
- Yu-tuan Wu†1,
- Qiu-bo Yu†2,
- Ya-ying Yang†3,
- Yu-xian Wei1,
- Hong-yuan Li1,
- Kai-nan Wu1 and
- Ling-quan Kong1Email author
© The Author(s). 2017
Received: 1 February 2017
Accepted: 9 October 2017
Published: 19 October 2017
A few retrospective studies have indicated that neoadjuvant chemotherapy (NAC) in breast cancer may change biomarker profiles of the primary tumor. Little is known about the status of HER-2 gene of the synchronous nodal metastases when that of the residual tumor undergoes negative conversion in a neoadjuvant setting.
We describe a female patient with left breast cancer (T2N2M0) who underwent negative conversion of HER-2 in the primary tumor instead of the synchronous nodal lesions after NAC. Core needle biopsy showed invasive ductal carcinoma with HER2 immunohistochemistry (IHC) (2+) and amplified HER-2 gene determined by fluorescence in situ hybridization (FISH). Then, the patient underwent 4 cycles of anthracycline- and taxane-based NAC and subsequent left modified radical mastectomy. Postoperative pathology showed invasive ductal carcinoma involving 4 of 12 surgically excised axillary lymph nodes with HER2 IHC (1+) and FISH negative (HER2 gene not amplified) in the residual tumor of the breast specimen. Due to the negative genic switch of HER2 after NAC, the patient rejected to accept trastuzumab. Under the patient’s consent, the synchronous nodal lesions were further investigated and showed HER2 IHC(−) but FISH positive (HER-2 gene amplified). Therefore, the patient agreed to accept adjuvant trastuzumab treatment every 3 weeks for 1 year.
We propose further assessment of HER2 gene in the synchronous nodal metastases, especially when negative genic switch of HER-2 occurs in the primary tumor after NAC in order to tailor the systemic regimens for breast cancer patients.
Neoadjuvant chemotherapy (NAC) is utilized in the context of locally advanced breast cancer to downstage tumors, improve operability, and increase the chance of breast-conserving surgery . Patients receiving NAC shared equivalent disease progression and overall survival with those only receiving post-operative chemotherapy . In early breast cancer, anthracycline and/or taxane-based regimens in a neoadjuvant setting provides information about the tumor’s sensitivity to chemotherapy and clinical outcomes after post-operative systemic therapy [3–5]. Approximately 15% of patients have achieved complete remission of the primary tumor and acquired better clinical outcomes by NAC [6, 7].
Recently, a trend has emerged in distinguishing prognostic factors by studying alterations of biomarkers in residual tumoral lesions. A few retrospective studies [8–11] have suggested that NAC in breast cancer may change biomarker profiles of the primary tumor. But, little is known about the status of the HER-2 gene of the synchronous nodal metastases after NAC when that of the residual tumor undergoes negative conversion , which poses a challenge to the inclusion of trastuzumab in the systemic therapy regimens. This issue has been significant in recent years due to the common usage of trastuzumab in HER-2 positive tumor, and decreased HER2 expression in invasive breast cancer after NAC has been noted [11, 13]. Given that sentinel lymph node biopsy or axillary lymph node dissection are performed in standard surgical practice and routine pathological assessment of nodes is carried out to evaluate the axillary surgical staging, this case report shows that there may be additional benefit to perform molecular testing on nodal metastases, especially when both HER-2 gene and its oncogenic receptor of the primary tumor underwent negative conversion after NAC.
Here, this case study reported on the loss of HER-2 gene in the primary focus of breast cancer after NAC and meanwhile, we found by FISH that HER-2 gene was still amplified in the synchronous metastatic axillary lymph nodes. Then, the patient agreed to accept trastuzumab every 3 weeks for 1 year.
In this patient, HER2 status of the primary tumor converted from positivity to negativity after NAC, which is in accordance with the results of one preclinical study  showing that paclitaxel downregulated HER2 expression in MDA-MB 453 cells and one retrospective study by Niikura et al. , which showed that 21.4% of HER-2 positive primary tumor had lost HER-2 expression in a neoadjuvant setting, and strongly supported the necessity to retest HER-2 status of residual tumor after neoadjuvant therapy in order to accurately determine appropriate utilization of anti-HER-2 therapy. The primary tumor of the patient after NAC was proved to be HER-2 negative by IHC and FISH. And then, the patient rejected to accept trastuzumab. Similarly, in the study of Niikura et al. , 28% of patients whose cancer lost HER-2 expression in a neoadjuvant setting did not receive trastuzumab treatment. Indeed, HER-2 status has been widely and successfully established in certified diagnostic pathology laboratories; potential diagnostic pitfalls of this “simple” marker can often occur in a neoadjuvant setting due to the modulatory effect of NAC on HER-2 expression of the primary tumor [8–12] and intratumoral heterogeneity [10, 15]. We appreciate the productive work of Niikura et al.  in throwing light upon the phenomenon of negative conversion of HER2 status in residual tumors after NAC and its potential influence on the inclusion of trastuzumab in systemic therapy regimens. In the absence of synchronous nodal metastases, their study bears much significance because negative genic switch of HER2 in residual tumors after NAC poses a challenge to the utilization of targeted therapy with a proportion of patients whose cancer lost HER2 expression after NAC in their study not receiving trastuzumab. But in the presence of synchronous nodal metastases, the situation becomes more complicated because metastatic tumor cells that originate from the HER-2 positive primary tumor prior to NAC may still harbor amplified HER2 gene. Various studies [16–18] have revealed certain discrepancies of HER2 expression between primary tumors and synchronous nodal metastases in the absence of chemotherapy, ranging from 0 to 9%. Little is known about the status of HER-2 gene of synchronous nodal metastases in a neoadjuvant setting, as confirmation of this oncogene in nodal lesions is crucial under the condition in which HER-2 gene status of the primary tumor converted from amplification to non-amplification after NAC . So, we shifted our attention to the pathologically confirmed metastatic lesions within axillary lymph nodes (4/12). In order to optimize the patient’s clinical outcomes, the synchronous nodal metastases were submitted for IHC, only to find the nodal lesions shared the same negative HER-2 status with the primary tumor, which indicated that HER-2 protein expression of nodal lesions was also suppressed by NAC, as Oldham et al.  demonstrated that paclitaxel downregulated HER2 expression in MDA-MB 453 cells. Finally, FISH was performed and confirmed amplification of HER-2 gene of the synchronous nodal metastases. Then, the patient agreed to receive trastuzumab. This discrepancy between HER-2 gene and its oncogenic receptor as revealed by IHC and FISH, respectively, was in accordance with one study  indicating that the HER-2 expression at the transcriptomic level is not always parallel to that at the proteomic level based on ERBB2 mRNA analysis after neoadjuvant systemic therapy, thus emphasizing that conclusions drawn from proteomic level should be further investigated in genomic and transcriptomic level. We speculated that suppression of HER2 protein expression of the nodal lesions in a neoadjuvant setting in this patient was possibly contemporary and the oncogenic receptor would soon be regained following a cessation of chemotherapy, due to the persistent amplification of HER-2 gene in nodal lesions.
Aitken et al.  confirmed by breast tissue microarray (TMA) and matched node TMA that biomarkers’ status of metastatic nodal lesions could be a more accurate measurement for guiding adjuvant therapy. It is postulated that biomarker profiles expressed by locoregional nodal lesions are more indicative of the biological behaviors of distant micro-metastases and circulating tumor cells (CTCs) that are more invasive and closely correlated with the potential clinical outcomes [21, 22] and vice versa. Onsten et al.  elaborated two cases in which gene expression profiles of CTCs bore better resemblance to that of the nodal metastases than the primary tumor when molecular discordance between CTCs and the primary tumor were investigated by reverse transcription quantitative PCR. Aktas et al.  demonstrated that detection and evaluation of HER-2 status in CTCs by liquid biopsy was able to make relatively accurate prediction about HER-2 gene and its oncogenic receptor status on metastases in a prospective open non-randomized study. Investigation of the biology of CTCs is significant, as a classical genetic model of human cancer progression is provided by the analysis of CTCs development . White et al.  demonstrated that disseminated tumor cells isolated at the time of diagnosis shared the same subclonal cell DNA copy number aberration with relatively large proportions of breast cancer cells in the lymph node metastases.
While the necessity to retest the HER2 status of residual tumors after NAC to tailor the utilization of anti-HER2 therapy is being emphasized , the possibility always lost sight that synchronous nodal metastases, distant micrometastases, and even CTCs, if any, which originate from the HER2 positive primary tumor, may still harbor tumor cells that are HER2-gene amplified.
This case report highlights, in a neoadjuvant setting, the significance of further assessment of HER-2 gene in the synchronous nodal metastases, especially when negative genic switch of HER-2 occurs in the primary tumor in order to formulate a more tailored adjuvant therapy, thus improving the patient’s outcome. Even if HER-2 gene status of synchronous nodal metastases undergoes negative conversion, whether to adopt the protocol of trastuzumab-containing adjuvant therapy still remains uncertain, because we are still not sure whether it can completely represent the characteristics of the distant metastatic cancer cells or CTCs. Prospective studies of HER-2 gene analyses among various matched lesions (i.e., primary tumors, synchronous nodal metastases, distant micro-metastases, and CTCs) in a neoadjuvant setting are needed for further study. Furthermore, a randomized controlled trial on whether to adopt the protocol of trastuzumab-containing adjuvant therapy among patients whose cancer lose HER2 expression not only in the primary tumor but also in the synchronous metastatic nodal lesions after NAC should be carried out.
The case report was supported by the National Natural Science Foundation of China (NSFC) (81372851) and through provision of data by the Breast Cancer Center of Chongqing, The First Affiliated Hospital of Chongqing Medical University, People’s Republic of China.
Availability of data and materials
The data and figures supporting the conclusions of this article are included within the article.
K-LQ and C-HR conceived of the study. C-HR and W-YT drafted this manuscript; Y-QB and Y-YY were involved in preparing the figures. L-HY and W-KN carried out the literature searching. K-LQ revised the manuscript. All authors read and approved the final manuscript.
Ethics approval and consent to participate
The analysis was performed in accordance with the ethical standards of the First Affiliated Hospital of Chongqing Medical University.
Consent for publication
The authors declare that they have no competing interests.
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- Kaufmann M, von Minckwitz G, Smith R, et al. International expert panel on the use of primary (preoperative) systemic treatment of operable breast cancer: review and recommendations. J Clin Oncol. 2003;21:2600–8.View ArticlePubMedGoogle Scholar
- Mauri D, Pavlidis N, Ioannidis JP. Neoadjuvant versus adjuvant systemic treatment in breast cancer: a meta-analysis. J Natl Cancer Inst. 2005;97:188–94.View ArticlePubMedGoogle Scholar
- Fisher B, Brown A, Mamounas E, et al. Effect of preoperative chemotherapy on local-regional disease in women with operable breast cancer: findings from National Surgical Adjuvant Breast and Bowel Project B-18. J Clin Oncol. 1997;15:2483–93.View ArticlePubMedGoogle Scholar
- Bear HD, Anderson S, Smith RE, et al. Sequential preoperative or postoperative docetaxel added to preoperative doxorubicin plus cyclophosphamide for operable breast cancer: National Surgical Adjuvant Breast and Bowel Project Protocol B-27. J Clin Oncol. 2006;24:2019–27.View ArticlePubMedGoogle Scholar
- Wolff AC, Berry D, Carey LA, et al. Research issues affecting preoperative systemic therapy for operable breast cancer. J Clin Oncol. 2008;26:806–13.View ArticlePubMedGoogle Scholar
- Pierga JY, Mouret E, Laurence V, et al. Prognostic factors for survival after neoadjuvant chemotherapy in operable breast cancer. The role of clinical response. Eur J Cancer. 2003;39:1089–96.View ArticlePubMedGoogle Scholar
- Wolmark N, Wang J, Mamounas E et al. Preoperative chemotherapy in patients with operable breast cancer: nine-year results from National Surgical Adjuvant Breast and Bowel Project B-18. J Natl Cancer Inst Monogr. 2001;(30):96–102.Google Scholar
- Lorgis V, Algros MP, Villanueva C, et al. Discordance in early breast cancer for tumour grade, estrogen receptor, progesteron receptors and human epidermal receptor-2 status between core needle biopsy and surgical excisional primary tumour. Breast. 2011;20:284–7.View ArticlePubMedGoogle Scholar
- Kumaki N, Umemura S, Tang X, et al. Alteration of immunohistochemical biomarkers between pre- and post-chemotherapy: hormone receptors, HER2 and Ki-67. Breast Cancer. 2011;18:98–102.View ArticlePubMedGoogle Scholar
- Zhou X, Zhang J, Yun H, et al. Alterations of biomarker profiles after neoadjuvant chemotherapy in breast cancer: tumor heterogeneity should be taken into consideration. Oncotarget. 2015;6:36894–902.PubMedPubMed CentralGoogle Scholar
- Li P, Liu T, Wang Y, et al. Influence of neoadjuvant chemotherapy on HER2/neu status in invasive breast cancer. Clin Breast Cancer. 2013;13:53–60.View ArticlePubMedGoogle Scholar
- Cockburn A, Yan J, Rahardja D, et al. Modulatory effect of neoadjuvant chemotherapy on biomarkers expression; assessment by digital image analysis and relationship to residual cancer burden in patients with invasive breast cancer. Hum Pathol. 2014;45:249–58.View ArticlePubMedGoogle Scholar
- Oldham EA, Li C, Ke S, et al. Comparison of action of paclitaxel and poly(L-glutamic acid)-paclitaxel conjugate in human breast cancer cells. Int J Oncol. 2000;16:125–32.PubMedGoogle Scholar
- Niikura N, Tomotaki A, Miyata H, et al. Changes in tumor expression of HER2 and hormone receptors status after neoadjuvant chemotherapy in 21,755 patients from the Japanese breast cancer registry. Ann Oncol. 2016;27:480–7.View ArticlePubMedGoogle Scholar
- Hanna WM, Ruschoff J, Bilous M, et al. HER2 in situ hybridization in breast cancer: clinical implications of polysomy 17 and genetic heterogeneity. Mod Pathol. 2014;27:4–18.View ArticlePubMedGoogle Scholar
- Ataseven B, Gologan D, Gunesch A, et al. HER2/neu, topoisomerase 2a, estrogen and progesterone receptors: discordance between primary breast cancer and metastatic axillary lymph node in expression and amplification characteristics. Breast Care (Basel). 2012;7:465–70.View ArticleGoogle Scholar
- Jensen JD, Knoop A, Ewertz M, Laenkholm AV. ER, HER2, and TOP2A expression in primary tumor, synchronous axillary nodes, and asynchronous metastases in breast cancer. Breast Cancer Res Treat. 2012;132:511–21.View ArticlePubMedGoogle Scholar
- Santinelli A, Pisa E, Stramazzotti D, Fabris G. HER-2 status discrepancy between primary breast cancer and metastatic sites. Impact on target therapy. Int J Cancer. 2008;122:999–1004.View ArticlePubMedGoogle Scholar
- Gonzalez-Angulo AM, Iwamoto T, Liu S, et al. Gene expression, molecular class changes, and pathway analysis after neoadjuvant systemic therapy for breast cancer. Clin Cancer Res. 2012;18:1109–19.View ArticlePubMedPubMed CentralGoogle Scholar
- Aitken SJ, Thomas JS, Langdon SP, et al. Quantitative analysis of changes in ER, PR and HER2 expression in primary breast cancer and paired nodal metastases. Ann Oncol. 2010;21:1254–61.View ArticlePubMedGoogle Scholar
- Kroigard AB, Larsen MJ, Thomassen M, Kruse TA. Molecular concordance between primary breast cancer and matched metastases. Breast J. 2016;22:420–30.View ArticlePubMedGoogle Scholar
- Yao ZX, LJ L, Wang RJ, et al. Discordance and clinical significance of ER, PR, and HER2 status between primary breast cancer and synchronous axillary lymph node metastasis. Med Oncol. 2014;31:798.View ArticlePubMedGoogle Scholar
- Onstenk W, Sieuwerts AM, Weekhout M, et al. Gene expression profiles of circulating tumor cells versus primary tumors in metastatic breast cancer. Cancer Lett. 2015;362:36–44.View ArticlePubMedGoogle Scholar
- Aktas B, Kasimir-Bauer S, Muller V, et al. Comparison of the HER2, estrogen and progesterone receptor expression profile of primary tumor, metastases and circulating tumor cells in metastatic breast cancer patients. BMC Cancer. 2016;16:522.View ArticlePubMedPubMed CentralGoogle Scholar
- Vanharanta S, Massague J. Origins of metastatic traits. Cancer Cell. 2013;24:410–21.View ArticlePubMedPubMed CentralGoogle Scholar
- Demeulemeester J, Kumar P, Moller EK, et al. Tracing the origin of disseminated tumor cells in breast cancer using single-cell sequencing. Genome Biol. 2016;17:250.View ArticlePubMedPubMed CentralGoogle Scholar
- Wolff AC, Hammond ME, Hicks DG, et al. Recommendations for human epidermal growth factor receptor 2 testing in breast cancer: American Society of Clinical Oncology/College of American Pathologists clinical practice guideline update. J Clin Oncol. 2013;31:3997–4013.View ArticlePubMedGoogle Scholar