RNA-based determination of ESR1 and HER2 expression and response to neoadjuvant chemotherapy.

BACKGROUND Hormone and human epidermal growth factor receptor 2 (HER2) receptors are the most important breast cancer biomarkers, and additional objective and quantitative test methods such as messenger RNA (mRNA)-based quantitative analysis are urgently needed. In this study, we investigated the clinical validity of RT-PCR-based evaluation of estrogen receptor (ESR1) and HER2 mRNA expression. PATIENTS AND METHODS A total of 1050 core biopsies from two retrospective (GeparTrio, GeparQuattro) and one prospective (PREDICT) neoadjuvant studies were evaluated by quantitative RT-PCR for ESR1 and HER2. RESULTS ESR1 mRNA was significantly predictive for reduced response to neoadjuvant chemotherapy in univariate and multivariate analysis in all three cohorts. The complete pathologically documented response (pathological complete response, pCR) rate for ESR1+/HER2- tumors was 7.3%, 8.0% and 8.6%; for ESR1-/HER2- tumors it was 34.4%, 33.7% and 37.3% in GeparTrio, GeparQuattro and PREDICT, respectively (P < 0.001 in each cohort). In the Kaplan-Meier analysis in GeparTrio patients with ESR1+/HER2- tumors had the best prognosis, compared with ESR1-/HER2- and ESR1-/HER2+ tumors [disease-free survival (DFS): P < 0.0005, overall survival (OS): P < 0.0005]. CONCLUSIONS Our results suggest that mRNA levels of ESR1 and HER2 predict response to neoadjuvant chemotherapy and are significantly associated with long-term outcome. As an additional option to standard immunohistochemistry and gene-array-based analysis, quantitative RT-PCR analysis might be useful for determination of the receptor status in breast cancer.

[1]  H. Höfler,et al.  Decentral gene expression analysis for ER+/Her2− breast cancer: results of a proficiency testing program for the EndoPredict assay , 2012, Virchows Archiv.

[2]  M. Rezai,et al.  Lapatinib versus trastuzumab in combination with neoadjuvant anthracycline-taxane-based chemotherapy (GeparQuinto, GBG 44): a randomised phase 3 trial. , 2012, The Lancet. Oncology.

[3]  Tanja Fehm,et al.  Neoadjuvant chemotherapy and bevacizumab for HER2-negative breast cancer. , 2012, The New England journal of medicine.

[4]  D. Dabbs,et al.  High false-negative rate of HER2 quantitative reverse transcription polymerase chain reaction of the Oncotype DX test: an independent quality assurance study. , 2011, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[5]  L. Pusztai,et al.  Gene expression profiling in breast cancer: classification, prognostication, and prediction , 2011, The Lancet.

[6]  R Peto,et al.  Relevance of breast cancer hormone receptors and other factors to the efficacy of adjuvant tamoxifen: patient-level meta-analysis of randomised trials , 2011, The Lancet.

[7]  M. Dowsett,et al.  Estrogen receptor: a never ending story? , 2011, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[8]  R. Greil,et al.  A New Molecular Predictor of Distant Recurrence in ER-Positive, HER2-Negative Breast Cancer Adds Independent Information to Conventional Clinical Risk Factors , 2011, Clinical Cancer Research.

[9]  Carsten Denkert,et al.  Quantitative Determination of Estrogen Receptor, Progesterone Receptor, and HER2 mRNA in Formalin-fixed Paraffin-embedded Tissue—A New Option for Predictive Biomarker Assessment in Breast Cancer , 2011, Diagnostic molecular pathology : the American journal of surgical pathology, part B.

[10]  T. Fehm,et al.  Integrating bevacizumab, everolimus, and lapatinib into current neoadjuvant chemotherapy regimen for primary breast cancer. Safety results of the GeparQuinto trial. , 2011, Annals of oncology : official journal of the European Society for Medical Oncology.

[11]  J. Cuzick,et al.  ER and HER2 expression are positively correlated in HER2 non-overexpressing breast cancer , 2009, Breast Cancer Research.

[12]  C. Denkert,et al.  Comparison of different approaches for assessment of HER2 expression on protein and mRNA level: prediction of chemotherapy response in the neoadjuvant GeparTrio trial (NCT00544765) , 2011, Breast Cancer Research and Treatment.

[13]  Mitch Dowsett,et al.  Current and emerging biomarkers in breast cancer: prognosis and prediction. , 2010, Endocrine-related cancer.

[14]  Hiltrud Brauch,et al.  Automated extraction of DNA and RNA from a single formalin-fixed paraffin-embedded tissue section for analysis of both single-nucleotide polymorphisms and mRNA expression. , 2010, Clinical chemistry.

[15]  L. Goldstein,et al.  Human epidermal growth factor receptor 2 assessment in a case-control study: comparison of fluorescence in situ hybridization and quantitative reverse transcription polymerase chain reaction performed by central laboratories. , 2010, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[16]  C. Denkert,et al.  Effect of neoadjuvant anthracycline–taxane-based chemotherapy in different biological breast cancer phenotypes: overall results from the GeparTrio study , 2010, Breast Cancer Research and Treatment.

[17]  Anthony Rhodes,et al.  American Society of Clinical Oncology/College of American Pathologists guideline recommendations for immunohistochemical testing of estrogen and progesterone receptors in breast cancer. , 2010, Archives of pathology & laboratory medicine.

[18]  M. Dowsett,et al.  Feasibility of using tissue microarray cores of paraffin-embedded breast cancer tissue for measurement of gene expression: a proof-of-concept study , 2010, Journal of Clinical Pathology.

[19]  B. Thürlimann,et al.  Trastuzumab (T) treatment beyond progression in metastatic breast cancer (MBC): Patterns of care in Swiss clinical practice. , 2010 .

[20]  M. Rezai,et al.  Capecitabine in addition to anthracycline- and taxane-based neoadjuvant treatment in patients with primary breast cancer: phase III GeparQuattro study. , 2010, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[21]  Carsten Denkert,et al.  Tumor-associated lymphocytes as an independent predictor of response to neoadjuvant chemotherapy in breast cancer. , 2010, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[22]  A. Goldhirsch,et al.  Lessons on responsiveness to adjuvant systemic therapies learned from the neoadjuvant setting. , 2009, Breast.

[23]  W. Weichert,et al.  Identification of biology-based breast cancer types with distinct predictive and prognostic features: role of steroid hormone and HER2 receptor expression in patients treated with neoadjuvant anthracycline/taxane-based chemotherapy , 2009, Breast Cancer Research.

[24]  Benjamin Haibe-Kains,et al.  Improvement of the clinical applicability of the Genomic Grade Index through a qRT-PCR test performed on frozen and formalin-fixed paraffin-embedded tissues , 2009, BMC Genomics.

[25]  Uwe Rogel,et al.  RNA extraction from archival formalin-fixed paraffin-embedded tissue: a comparison of manual, semiautomated, and fully automated purification methods. , 2009, Clinical chemistry.

[26]  A. Tutt,et al.  A single-tube quantitative assay for mRNA levels of hormonal and growth factor receptors in breast cancer specimens. , 2009, The Journal of molecular diagnostics : JMD.

[27]  N. Pavlidis,et al.  Gene expression of estrogen receptor, progesterone receptor and microtubule-associated protein Tau in high-risk early breast cancer: a quest for molecular predictors of treatment benefit in the context of a Hellenic Cooperative Oncology Group trial , 2009, Breast Cancer Research and Treatment.

[28]  A. Luini,et al.  Increasing steroid hormone receptors expression defines breast cancer subtypes non responsive to preoperative chemotherapy , 2009, Breast Cancer Research and Treatment.

[29]  Edith A Perez,et al.  Estrogen- and progesterone-receptor status in ECOG 2197: comparison of immunohistochemistry by local and central laboratories and quantitative reverse transcription polymerase chain reaction by central laboratory. , 2008, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[30]  S. Loibl,et al.  Neoadjuvant vinorelbine-capecitabine versus docetaxel-doxorubicin-cyclophosphamide in early nonresponsive breast cancer: phase III randomized GeparTrio trial. , 2008, Journal of the National Cancer Institute.

[31]  S. Loibl,et al.  Intensified neoadjuvant chemotherapy in early-responding breast cancer: phase III randomized GeparTrio study. , 2008, Journal of the National Cancer Institute.

[32]  C. Perou,et al.  The Triple Negative Paradox: Primary Tumor Chemosensitivity of Breast Cancer Subtypes , 2007, Clinical Cancer Research.

[33]  Anthony Rhodes,et al.  American Society of Clinical Oncology/College of American Pathologists guideline recommendations for human epidermal growth factor receptor 2 testing in breast cancer. , 2006, Archives of pathology & laboratory medicine.

[34]  Christine Desmedt,et al.  Proliferation: the Most Prominent Predictor of Clinical Outcome in Breast Cancer , 2006, Cell cycle.

[35]  R. Schiff,et al.  Advanced concepts in estrogen receptor biology and breast cancer endocrine resistance: implicated role of growth factor signaling and estrogen receptor coregulators , 2005, Cancer Chemotherapy and Pharmacology.

[36]  R. Schiff,et al.  Crosstalk between estrogen receptor and growth factor receptor pathways as a cause for endocrine therapy resistance in breast cancer. , 2005, Clinical Cancer Research.

[37]  I. Zuna,et al.  In vivo chemosensitivity-adapted preoperative chemotherapy in patients with early-stage breast cancer: the GEPARTRIO pilot study. , 2005, Annals of oncology : official journal of the European Society for Medical Oncology.

[38]  Joon-Oh Park,et al.  Mitomycin-C and capecitabine as third-line chemotherapy in patients with advanced colorectal cancer: a phase II study , 2005, Cancer Chemotherapy and Pharmacology.

[39]  Claus Lindbjerg Andersen,et al.  Normalization of Real-Time Quantitative Reverse Transcription-PCR Data: A Model-Based Variance Estimation Approach to Identify Genes Suited for Normalization, Applied to Bladder and Colon Cancer Data Sets , 2004, Cancer Research.

[40]  P. V. van Diest,et al.  Comparison of the prognostic value of four methods to assess mitotic activity in 186 invasive breast cancer patients: classical and random mitotic activity assessments with correction for volume percentage of epithelium. , 1995, Human pathology.