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Introduction Breast cancer diagnosed in young women who are 40 years (yrs) or younger is a relatively rare disease. However, it represents the most common cause of cancer-related deaths in this age-group. Furthermore, young age at diagnosis is associated with an increased risk of recurrence and worse survival. To date, general concepts concerning oncological cancer management should be driven by clinicopathological tumor characteristics and should adhere to standardized protocols for patients in general, but little is known about the oncological cancer treatment and outcome of this very young women in today's clinical practice. Patients and Methods The breast cancer in pregnancy registry study (BCP/GBG29/BIG 03-02) is a multicenter, international, observational study. BCP was established to investigate the oncological management and outcome of breast cancer in pregnancy. Since 2014 non-pregnant patients who are 40 yrs or younger are eligible if diagnosed with histological confirmed invasive breast cancer, independent of the type of treatment as control cohort. All patients received oncological treatment according to local standards. In this study the following endpoints will be analyzed descriptively for the young non-pregnant women cohort: breast cancer staging at diagnosis, biological characteristics of breast cancer at diagnosis, diagnostic procedures, treatment modalities, toxicity, pathological complete response after neoadjuvant chemotherapy, disease-free survival and overall survival. Results From February 2014 until June 2018, 969 non-pregnant patients ≤40 yrs have been registered. The median age at diagnosis was 35 yrs (range 19-40). Overall, 90.1% of patients had a stage T1-2 at diagnosis and 67.1% of patients had negative lymph nodes. 86.7% of tumors were invasive ductal carcinomas and 4.1% lobular carcinomas. Grading (G) 3 was reported in 55.5%. 26.6% of tumors were luminal A-like (ER- and/or PgR-positive, HER2-negative, G1-2), 40.0% luminal B-like (ER- and/or PgR-positive, HER2-negative, G3 or ER- and/or PgR-positive, HER2-positive, any G), 7.7% HER2 positive non-luminal-like, and 25.7% triple negative breast cancers. 3.8% of young non-pregnant patients had metastatic disease at primary diagnosis. Conclusion This registry comprises a large cohort of young non-pregnant patients with breast cancer diagnosed at the age of 40 yrs or younger and provides important data about a modern breast cancer treatment as well as oncological outcome in this setting of young women. Further results including oncological management, toxicity, and survival will be presented at the meeting. Citation Format: Seiler S, Schmatloch S, Reinisch M, Neunhöffer T, Schmidt M, Bechtner C, Marmé F, Wagner M, Möbus V, Reimer T, Kleine-Tebbe A, Sinn B, Stickeler E, Untch M, Janni W, Seither F, Loibl S. Cancer management and outcome of very young non-pregnant patients with breast cancer diagnosed at 40 years or younger– GBG 29 [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P1-17-07.

Background: The treatment of triple negative breast cancer (TNBC) is hampered by the lack of established therapeutic targets such as hormone receptors or HER-2. Chemotherapy and radiotherapy is the standard of care, yet survival rates in TNBC remain poor. Approaches tailored to the patient's individual tumor signature may lead to improved therapeutic outcome. We have set up a clinical workflow covering drug development (from target discovery to manufacturing) and drug release providing a custom-made investigational medicinal product (IMP) for each individual patient. Trial Design: A phase I/II trial assesses the feasibility, safety and biological efficacy of this personalized immunotherapy in three clinical sites in Germany and Sweden. TNBC patients (pT1cN0M0 – TxNxM0) after completion of initial standard of care therapy will be allocated to one of two study arms. Patients in ARM1 receive 8 vaccination cycles with a personalized combination of shared tumor-associated antigens, selected based on each patient tumor's antigen-expression profile out of a WAREHOUSE of pre-manufactured mRNA vaccine. Patients in ARM2 receive the personalized mRNA WAREHOUSE vaccine followed by 8 vaccination cycles of an on-demand manufactured mRNA MUTANOME vaccine encoding up to twenty unique neo-epitopes of the individual patient identified by next generation sequencing. The mRNAs are administered intravenously as a nanoparticulate lipoplex formulation, which protects RNA from degradation, activates innate immunity, transfects APCs and consequently induces highly potent antigen-specific T-cell responses. The treatment of 12 patients in ARM1 is completed and enrolment of patients for ARM2 has started. Preliminary data show that the RNA-WAREHOUSE approach is feasible and can be applied safely. Biomarker analysis is ongoing. This approach is promising as it addresses the heterogeneity of TNBC. The TNBC-MERIT trial was initially funded by the EU Commission's FP7 and led by BioNTech AG. Citation Format: Schmidt M, Bolte S, Frenzel K, Heesen L, Derhovanessian E, Bukur V, Diken M, Gruetzner J, Kreiter S, Klein A, Kuhn A, Langer D, Loewer M, Lindman H, Schneeweiss A, Tuereci O, Sahin U. Highly innovative personalized RNA-immunotherapy for patients with triple negative breast cancer [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr OT2-06-01.

Introduction: Currently, a multi-center, open-label, phase I/IIA, dose-escalation study with the fetal estrogen estetrol (E4) is ongoing in Germany in postmenopausal patients with advanced breast cancer (ABCE4 study). The objective of the study is to assess safety and tolerability of different doses of E4 (range 20-60 mg per day). In addition initial anti-tumor response will be determined. Study design: Patients are treated for 12 weeks with E4; 4 weeks in Phase I and thereafter 8 weeks in Phase IIA. Phase I of the study will follow the traditional 3 + 3 design to determine the optimal dose in patients with advanced breast cancer. Patients are treated in cohorts of three all receiving the same dose. Occurrence of dose limiting toxicity (DLT) at completion of phase I (4 weeks treatment) will determine escalation to the next higher dose in the study. After completion of Phase I, patients will continue treatment for 8 weeks to assess preliminary anti-tumor response in Phase IIA. Treatment may continue beyond 12 weeks based on evaluation of the patient and her treating physician. Results: Phase I of the first treatment cohort with 20 mg E4 per day has been completed. A total of six postmenopausal women with advanced breast cancer has been included in the first cohort. One patient withdrew consent before treatment with E4 started. She was replaced. Two patients discontinued the study before completion of Phase I for reasons other than DLTs. These patients were unevaluable for toxicity and were also replaced. Three patients completed Phase I. One patient discontinued the study during Phase IIA due to disease progression after 9,5 weeks of E4 treatment. One patient completed both the Phase I and IIA part of the study. She had stable disease at study completion and wanted to continue E4 treatment because of improved well-being. Tumor assessment after 24 weeks of E4 treatment showed again stable disease. One patient is presently in Phase IIA of the study. None of the patients experienced a DLT. The 20 mg E4 dose was well tolerated by all patients. In total 17 adverse events were reported. Adverse events were mainly of mild or moderate intensity. Five of 17 events fulfilled criteria of seriousness; none of these events were considered to be related to the E4 treatment. Four events were considered to be possibly related to the E4 treatment. These events were increased endometrial thickness, dry skin, pruritus and fatigue, all of mild intensity. A remarkable finding was that three of the five patients treated with E4 reported to the investigator to “feel better” than before the start of E4 treatment. This '”feeling better” was described by one of the patients as: '”feeling less down and exhausted; instead feeling much more optimistic, powerful and positive when taking E4”. So far anti-tumor response could be estimated in one patient. This patient, who started the study with progressive disease, had stable disease as shown by tumor assessments after 12 weeks and 24 weeks of E4 treatment. Conclusion: Based on these results, we conclude that a daily dose of 20 mg E4 is well tolerated. The majority of patients experienced favorable subjective effects on wellbeing. The data obtained with the 20 mg dose E4 allow dose escalation to the next higher dose of 40 mg E4 per day. Citation Format: Schmidt M, Hönig A, Verhoeven C, Almstedt K, Battista M, Lenhard HG, Krijgh J, Coelingh Bennink H. Estetrol for treatment of advanced ER+ breast cancer [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P4-13-12.

Background: In the pivotal BOLERO-2 trial, everolimus (EVE) + exemestane (EXE) more than doubled the median progression-free survival (PFS) vs EXE alone in hormone receptor positive (HR+), human epidermal growth factor-receptor 2-negative (HER2-) advanced breast cancer (ABC) recurring/progressing on/after prior non-steroidal aromatase inhibitors (NSAIs). BRAWO is a German non-interventional study conducted in patients (pts) with HR+, HER2–ABC receiving EVE + EXE, according to Summary of Product Characteristics (SmPC), in routine clinical practice. Here we report the final PFS and safety results. Methods: This multicenter study documented 2100 pts between October 2012 and December 2017 across 341 sites in Germany. Postmenopausal women with HR+, HER2– ABC with recurrence or progression after a NSAI were included. Primary observation parameters included the evaluation of the effectiveness of EVE + EXE used in routine care for the entire pt group. Results: In the final analysis, out of the 2100 documented pts, 2074 were included in the full analysis set. The median time since the primary diagnosis was 7.1 years and the median time from first sign of relapse (local recurrence or distant metastases) was 2.1 years. At baseline, 54.1% of pts presented with visceral metastases and 50.1% had an ECOG performance status of 0. Approximately, 63% of pts started with EVE 10 mg (median duration of exposure: 5.1 months; 95% CI, 4.6-5.4), while 34.1% started with EVE 5 mg (median duration of exposure: 4.6 months; 95% CI, 4.1-5.2). The distribution of treatment lines was as follows: first line, 28.7% (n=595); second line, 31.9% (n=662); third line, 18.1% (n=376); fourth line, 10.7% (n=221) and, fifth line and later, 10.6% (n=220). Treatment was discontinued by 55.7% of pts (n=1170) due to progressive disease and 26% of pts (n=546) due to adverse events. The Kaplan-Meier estimate of the median PFS was 6.6 months (95% CI, 6.2-7.0). The best overall responses, based on clinical routine, were complete response, 0.8% (n=17), partial response, 7.4% (n=150), and stable disease, 41.3% (n=842). The general safety profile was consistent with the previously reported safety findings. The most common adverse events were stomatitis (any grade: 42.6%, grade 3: 3.8%, grade 4: Conclusions: Data from BRAWO support EVE + EXE as a suitable treatment option with a reasonable safety profile for HR+, HER2− ABC recurring or progressing on/after prior NSAIs. Citation Format: Lüftner D, Schuetz F, Schneeweiss A, Grischke E-M, Bloch W, Decker T, Uleer C, Salat C, Förster F, Schmidt M, Mundhenke C, Tesch H, Jackisch C, Fischer T, Guderian G, Hanson S, Fasching P. Everolimus + exemestane for HR+ advanced breast cancer in routine clinical practice- Final results from the non-interventional trial, BRAWO [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P6-18-08.

Background Chemotherapy (CTx) is a cornerstone in HR+/HER2- advanced/metastatic breast cancer (a/mBC) after failure of endocrine treatment. In this indication, vinorelbine (VRL) is a well-established cytotoxic drug. There is a high medical need for new options that prolong the time between endocrine failure and intensive CTx, which is commonly associated with impaired quality of life and serious side effects. Metronomic CTx was shown to induce disease control in a/mBC with a favorable safety profile. This innovative approach involving continuous daily dosing of oral VRL, which could provide anti-angiogenic and immune-modulatory properties, has not been investigated so far in this indication. Trial Design VinoMetro is an open-label, single-arm, phase II study (Simon two-stage minimax) of metronomic daily oral VRL (30 mg/day) as first-line CTx. The study involves strict safety monitoring with an initial safety run-in. It is accompanied by a steering committee and supervised by an independent data safety and monitoring board (DSMB). The main objectives are to estimate efficacy in terms of clinical benefit rate after 24 weeks of treatment (primary endpoint) and the progression-free survival, amongst others, as well as the assessment of safety and quality of life. Patients with HR+/HER2- a/mBC having failed or being no candidate for endocrine therapy (targeted combinations allowed) and being naïve to palliative CTx are eligible, if they exhibit ECOG 0-1. The main exclusion criteria are prior vinca-alkaloids, aggressive disease requiring combination CTx and CNS involvement. Until 2017-05-31, 7 patients were enrolled. It is planned to include 45 (39 evaluable) patients at 8 German sites until 09/2018. Scheduled completion date is 09/2019. Two interim analyses are planned (first analysis: safety evaluation based on the 10 initial patients with predefined stopping rules). Depending on recruitment, it is planned to include the interim safety data in the congress presentation. VinoMetro is an investigator initiated trial (NCT03007992) sponsored by the University Medical Centre of Johannes Gutenberg-University Mainz, Germany, and supported by an unrestricted grant provided by Pierre Fabre Pharma GmbH (Freiburg, Germany). Citation Format: Schmidt M, Decker T, Fehm T, Fuxius S, Harbeck N, Juhasz-Böss I, Thomssen C, Seehase M, Schollenberger L, Ruckes C, Möbus V. A phase II study of metronomic daily oral vinorelbine as first-line chemotherapy in advanced/metastatic hormone receptor positive (HR+) / human epidermal growth factor receptor 2 negative (HER2-) breast cancer resistant to endocrine therapy - VinoMetro [abstract]. In: Proceedings of the 2017 San Antonio Breast Cancer Symposium; 2017 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2018;78(4 Suppl):Abstract nr OT2-05-02.

Background:Oncogene-induced senescence is considered as a barrier to tumor progression that arrests cells at risk for malignant transformation. Nevertheless, numerous findings demonstrate that senescent cells may also have the opposite function and promote tumor progression through the release of multiple factors called the senescence-associated secretory phenotype or senescence secretome. It is likely that the composition and the physiological consequences mediated by the senescence secretome are dependent on the oncogenes that trigger the senescence program. Breast cancer represents a heterogenous disease that can be divided into breast cancer subtypes caused by different subsets of genetic and epigenetic abnormalities. Therefore, tumor initiation and progression of breast cancer subtypes is triggered by variable oncogenic stimuli, and differences in the senescence secretomes within breast tumors might be responsible for tumor initiation, progression, metastasis and therapeutic response. Beside many studies concerning the role of senescence as a barrier to tumor progression using murine xenograft models very few investigations have been performed to elucidate how often senescent tumor cells appear within untreated human tumors, and if present whether these senescent tumor cells may play a role in disease progression, cancer immunosurveillance and therapy resistance. Methode: In the present study we analysed the appearance of senescent cells within invasive human breast cancers from 129 untreated patients. Cellular senescence was detected by the use of SAβ-gal staining and by immunohistochemical detection of p16, p21, p53, Ki67 and lamin B1. Results: Detection of cellular senescence by the use of SAβ-gal staining and detection of p16, p53, Ki67 and lamin B1 within invasive breast carcinomas indicate that senescent tumor cells varies strongly according to the breast cancer subtype. The highest percentages of senescent tumor cells exist within in the HER2-positive and luminal A breast carcinomas whereas no or very few senescent tumor cells were found in triple negative breast tumors. Based on these findings we suggest that the composition of secretomes released by senescent tumor cells from different breast cancer subtypes might be very distinct in respect to their ability to recruit immune cells, which can eliminate senescent tumor cells on one hand and regulate tumor growth, immune surveillance and therapy resistance on the other. Conclusion: Further characterization of senescent secretomes from HER2 and other breast cancer subtypes and their potential role in tumor progression, immune surveillance and therapy response might be warranted for the understanding of cancer biology as well as prognostic and therapeutic applications. Citation Format: Thaler S, Schad A, Kirkpatrick CJ, Sleeman JP, Springer E, Schmidt M, Cotarelo CL. Cellular senescence within HER2-amplified breast cancer: Potential implications for breast cancer immune surveillance and HER2-targeted therapy resistance [abstract]. In: Proceedings of the 2017 San Antonio Breast Cancer Symposium; 2017 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2018;78(4 Suppl):Abstract nr P5-10-01.

Introduction: The aim of shared decision making (SDM), defined as an interaction between patient and attending physician(s), is a treatment decision in which patients are meaningfully involved. Based on mutual agreement and active participation the awareness of a choice should be created and the choice respected. Many preference-sensitive decisions have to be made in breast cancer treatment. However, little is known about the implementation of SDM in German breast cancer care. We therefore investigated the process of SDM from the patients' perspective. Methods: All breast cancer patients who underwent surgery in one of four certified breast cancer centers in Germany between 07/2016 and 12/2016 were invited by mail to participate in the survey. The experienced decision-making process was assessed using the 9-item Shared Decision Making Questionnaire (SDM-Q-9). SDM-Q-9 items were rated on a 6-point scale ranging from "completely disagree" to "completely agree", added together and transformed into a scale ranging from 0 to 100. The higher the total score the higher the experienced degree of participation in the decision-making process. The survey also assessed patients' satisfaction with treatment, satisfaction with decisions and decisional control preferences, and included a range of demographic and clinical questions. For most items we asked the participants to separately rate decision-making consultations with their inpatient hospital doctors, outpatient gynecologists, outpatient oncologists and primary care providers (PCP). The project is still ongoing, data of approximately 300 patients will be presented at the meeting in December. Results: Of 289 patients approached by mail, 143 filled in the survey (response rate: 49%). Median age at the time of the survey was 62 years (36-89). 83% had breast conserving surgery, 17% mastectomy. 74% were treated with radiation, 31% received neo-/adjuvant chemotherapy. 14% were off-treatment at the time of survey participation, 67% still received antihormonal therapy, 9% anti HER2 treatment, 7% chemotherapy and 2% radiation. Inpatient hospital doctors achieved the highest SDM-Q-9 score (mean of 75, standard deviation of 22) indicating the highest degree of SDM. Oncologists, gynecologists and PCP were rated quite comparable with a mean score of 72 each and standard deviations (SD) of 27, 22 and 31. The mean score for all groups of doctors was 73. For items concerning satisfaction with quality and amount of doctors' information and participation in medical decisions patients showed a high degree of satisfaction, resulting in mean values of 3.6 and 3.5 with SD of 0.6 on a 4-point scale ranging from "1" "very unsatisfied" to "4" "very satisfied". Conclusions: A considerable number of patients took part in the survey. Overall, patients reported to have experienced SDM in many situations where treatment decisions were necessary. Patients were quite satisfied with the quality of information and their participation in medical decisions. However, we do not know whether non-respondents might have had different experiences regarding their treatment decision-making. Further research should include SDM expert observations of breast cancer treatment decisions to validate these findings. Citation Format: Feiten S, Scholl I, Schmidt M, Dünnebacke J, Franzen A, Ernst W, Weide R. Shared decision making (SDM) in routine care treatment of breast cancer patients – a survey of patients following surgery [abstract]. In: Proceedings of the 2017 San Antonio Breast Cancer Symposium; 2017 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2018;78(4 Suppl):Abstract nr P4-11-07.

Background Over 90% of male breast cancer (BC) patients (pts) have hormone receptor (HR) positive disease. Until today tamoxifen is considered to be the standard of care in male BC pts. Due to the low incidence of male BC, there is a lack of data regarding efficacy and safety. Therapeutic strategies are extrapolated from the standard of care of female BC. However, no prospectively randomized study in male BC pts has been conducted so far. The Male-GBG54 study is the first prospective, randomized, multicenter trial evaluating the efficacy and safety of different endocrine treatment options in male BC patients. Patients and Methods In the phase II Male-GBG54 trial (NCT01638247), pts were randomized to receive either tamoxifen 20 mg/day per os (p.o.) or tamoxifen 20mg/day p.o. + GnRHa subcutaneous (s.c.) q3m or exemestane 25 mg/day p.o. + GnRHa s.c. for 6 months as (neo)adjuvant or metastatic therapy. Further treatment was conducted as per local guidelines. Primary objective was the suppression of oestradiol in the three treatment arms after 3 months of therapy. Secondary objectives comprised oestradiol suppression after six months, compliance and safety of the three therapies, the level of different steroidal hormones (testosterone, dihydrotestosterone, SHBG, FSH, LH) and osteocalcin in the three arms. Quality of Life was assessed using the Aging Male Symptom Score, International Index of Erectile function and International Prostate Symptom Score. Tissue and blood was collected for translational research. Male BC pts with a Karnofsky Index ≥60%, normal blood lipids, and no history or evidence of prostate cancer were eligible. 14 pts per group were needed for the F-test to have 80% power to detect a difference in mean oestradiol decrease between the three therapeutic groups at the 5% significance level. The final sample size was calculated as 48 patients, as a non-parametric test (Kruskal-Wallis) was chosen in case the oestradiol levels are not normally distributed within the study population. Results Between October 2012 and May 2017, 55 pts were randomized within 24 centers in Germany. 52 pts were included in the adjuvant and 2 pts in the metastatic setting of whom 48 pts were fully evaluable and comprised the analysis set. The median age was 62 years (range 37-83 years). The baseline characteristics were well balanced between the 3 treatment groups. Karnofsky Index ranged from 60%-100%, 36.4% had received prior chemotherapy and 12.7% had HER2-positive BC. The majority had a pT2 (52%), pN0 (58.0%) BC. 3 serious adverse events were reported, none was related to the study treatment. Conclusion This is the first prospective randomized trial worldwide evaluating the impact of three different endocrine treatments in male breast cancer. The therapy was well tolerated without safety signals. At the meeting, final results of the primary and secondary endpoints will be present. Citation Format: Reinisch M, Seiler S, Hauzenberger T, Schmatloch S, Strittmatter HJ, Zahm DM, Thode C, Jackisch C, Furlanetto J, Strik D, Stickeler E, Marmé F, Janni W, Schmidt M, Kamischke A, Rudlowski C, Nekljudova V, von Minckwitz G, Loibl S. Male-GBG54: A prospective, randomised multi-centre phase II study evaluating endocrine treatment with either tamoxifen +/- gonadotropin releasing hormone analogue (GnRHa) or an aromatase inhibitor + GnRHa in male breast cancer patients [abstract]. In: Proceedings of the 2017 San Antonio Breast Cancer Symposium; 2017 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2018;78(4 Suppl):Abstract nr PD7-10.

Background A new subcutaneous (s.c.) formulation of trastuzumab became available in 2013 based on equivalent efficacy, pharmacokinetic (PK) profile and safety with the standard intravenous (i.v.) administration, where the s.c. trastuzumab was administered only into the thigh. As an s.c. injection into the abdominal wall (abdw) might be more convenient for patients (pts) and health care professionals, the PK profile of s.c. trastuzumab injected into the thigh vs the abdw in pts with HER2+ early BC needs to be evaluated. Methods GAIN-2 study compared two intense dose-dense (idd) anthracycline/taxane containing regimens. After completion of the anthracycline and i.v. trastuzumab given concurrently with taxanes, HER2+ BC pts were randomized in a 1:1 ratio to continue adjuvant s.c. trastuzumab 600mg fixed dose injected every 3 weeks either into the thigh or the abdw. Randomization was stratified according to CT arm [(iddEnPC) vs tailored dd CT (dtEC-dtD)] and age (≤50 vs >50). Pts in the EnPC arm received 14 and in the dtEC-dtD arm 15 cycles of s.c. trastuzumab. For the PK profile of s.c. trastuzumab serum samples collected before cycle 7, on days 2, 4, 8, 15 and 21 of cycle 7 are evaluated. With a total sample size of 30 (15 per group), the simulated 90% two-sided CI for the area under the plasma concentration (AUC0-last) will be (0.79-1.27) and for the peak drug concentration (Cmax) will be (0.77-1.30). Allowing for a dropout rate of 15%, 18 pts per group will be included in the PK analysis. The primary objective was to assess the PK profile of s.c. trastuzumab injected into the thigh vs the abdw. The secondary objectives included safety and tolerability. Results The per-protocol (pp) set consists of 30 pts (17 in the thigh group and 13 in the abdw group). Baseline characteristics were well balanced between the groups. The log-transformed Geometric Least Square Means (GLSM) for Cmax were 11.77 and 11.52 in the thigh and the abdw group, respectively. The geo-mean ratio (on the original scale) for Cmax was 1.29 (90% CI 1.05-1.58). The log-transformed GLSM for AUC0-last were 14.54 and 14.28 in the thigh and the abdw group, respectively. The geo-mean ratio for AUC0-last was 1.29 (90% CI 1.02-1.63). Overall 29 pts (96.7%) reported any grade and 5 pts (16.7%) high grade adverse events (AEs). The incidence of any grade AEs was similar between the two groups. The most common AEs were anemia (70.6% for the thigh vs 61.5% for the abdw group, p=0.705), leukopenia (80.0% for both groups, p=1.000) and fatigue (47.1% for the thigh vs 76.9% for the abdw group, p=0.141). 6 serious AEs were reported (2 in the thigh vs 4 in the abdw group). The final PK results of s.c. trastuzumab will be presented at the meeting. Conclusions Bioavailability of s.c. trastuzumab as reflected by peak and total exposure measured in cycle 7 was approx. 30% higher if administered into the thigh than into the abdw in pts with HER2+ primary BC treated after dose-dense CT plus i.v. trastuzumab. However, no increased toxicity was observed. Study limitations were that no cross-over design was used and number of pts satisfying criteria for pp-set were different in the arms. Citation Format: Möbus V, Mahlberg R, Janni W, Tomé O, Marmé F, Forstbauer H, Reimer T, von der Assen A, Reinisch M, Lorenz R, Schmatloch S, Schmidt M, Sinn B, Klutinus N, Stickeler E, Untch M, Seiler S, Burchardi N, von Minckwitz G, Loibl S. Pharmacokinetic results of a subcutaneous injection of trastuzumab into the thigh versus into the abdominal wall in patients with HER2-positive primary breast cancer (BC) treated within the neo-/adjuvant GAIN-2 study [abstract]. In: Proceedings of the 2017 San Antonio Breast Cancer Symposium; 2017 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2018;78(4 Suppl):Abstract nr P5-20-09.

Background - Fatigue is one of the most common and disabling complaints in patients with breast cancer and can effectively be reduced by physical exercise, with small to moderate effect sizes. To identify heterogeneity in responses to exercise and to further personalize exercise prescriptions, moderators of exercise effects on fatigue should be investigated. However, most randomized controlled trials (RCTs) are not adequately powered for such analyses. Therefore we conducted meta-analyses using the individual patient data of several exercise RCTs. The aim is to investigate the effect and moderators of physical exercise on cancer-related fatigue in patients with breast cancer. Methods - Within the Predicting OptimaL cAncer RehabIlitation and Supportive care (POLARIS) consortium, principal investigators of 34 exercise RCTs worldwide have shared their individual patient data. Twenty-two of these RCTs included patients with breast cancer with a total sample size of 3,061. Different questionnaires to assess level of fatigue were used, which was acknowledged by using z-scores in the analysis. A one-step individual patient data meta-analysis, using a linear mixed-effect model adjusted for baseline fatigue, with a random intercept on study (to account for study clustering) was undertaken to investigate effect of exercise on fatigue. The result, a between-group difference in z-scores, corresponds to a Cohen's d effect size. An interaction term was included in the model to assess potential moderators including demographic (age, marital status, education), clinical (body mass index, presence of distant metastasis), intervention-related (intervention timing, delivery mode and duration), and exercise-related (exercise type, frequency, intensity, duration) characteristics. Results – Exercise significantly reduced fatigue reported by women with breast cancer (β= -0.15, 95% CI -0.21;-0.09). This effect did not differ significantly between patients with different demographic and clinical characteristics (p-valuesinteraction >0.05). Also, neither timing (during or post-treatment) and duration of the intervention, nor exercise-related factors moderated intervention effects on fatigue. Supervised exercise had significantly larger effects on fatigue than unsupervised exercise (βdifference= -0.17, 95%CI -0.28;-0.05). Compared to the control group, supervised exercise significantly improved fatigue (β = -0.21, 95%CI = -0.28;-0.14), while unsupervised exercise did not (β = -0.04, 95%CI = -0.14;0.06). Conclusion – Exercise significantly reduces fatigue in patients with breast cancer across subgroups formed on the basis of age, marital status, education level, body mass index, and presence of distant metastasis. The effect of exercise is significantly larger when performed under supervision. Hence, exercise, and preferably supervised exercise, represents a viable intervention for the prevention and treatment of fatigue among patients with breast cancer. Citation Format: van Vulpen JK, Sweegers MG, Kalter J, Peeters PH, Courneya KS, Newton RU, Aaronson NK, Jacobsen PB, Steindorf K, Stuiver MM, Hayes S, Mesters I, Knoop H, Goedendorp M, Mutrie N, Thorsen L, Schmidt M, Sonke GS, Bohus M, James EL, Oldenburg HS, Velthuis MJ, Nollet F, Wenzel J, Wiskemann J, Galvão DA, Chinapaw MJ, Irwin ML, Griffith KA, van Weert E, Daley AJ, McConnachie A, Schulz K-H, Short CE, Plotnikoff RC, Potthoff K, van Beurden M, van Harten WH, Schmitz KH, Winters-Stone KM, Taaffe DR, van Mechelen W, Kersten M-J, Verdonck-de Leeuw IM, Brug J, Buffart LM, May AM. Effect and moderators of exercise on fatigue in patients with breast cancer: Meta-analysis of individual patient data [abstract]. In: Proceedings of the 2017 San Antonio Breast Cancer Symposium; 2017 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2018;78(4 Suppl):Abstract nr P6-12-06.

Background: The importance of the cellular as well as the humoral immune system is increasingly recognized in breast cancer. The transcription factor nuclear factor kappa B (NFKB1) plays an important role in immune cell development. We examined the prognostic significance of NFKB1 in node-negative breast cancer. Methods: Microarray based gene-expression data for NFKB1 (209239_at) were analysed in four previously published cohorts (Mainz, Rotterdam, Transbig, Yu) of node-negative breast cancer patients not treated with adjuvant therapy (n=824). A meta-analysis of previously published cohorts was performed using a random effects model. Intrinsic subtypes were determined using gene-expression data. Prognostic significance of NFKB1 for metastasis-free survival (MFS) was examined in the whole cohort, in estrogen receptor (ER) positive as well as ER negative patients, and in different intrinsic subtypes: luminal A, liminal B, basal-like, HER2-enriched (HER2-E), and normal-like. Independent prognostic relevance was analysed using multivariate Cox regression. Results: Higher RNA expression of NFKB1 was related to better MFS in a meta-analysis of the whole cohort (HR 0.48, 95% CI 0.34-0.67, P Conclusions: The transcription factor NFKB1 shows prognostic significance in node-negative breast cancer. Higher expression of NFKB1 is associated with improved outcome. The prognostic impact of NFKB1 differs between intrinsic subtypes and is most pronounced in luminal B breast cancer. Citation Format: Schmidt M, Madjar K, Heimes A-S, Battista M, Lebrecht A, Almsetdt K, Hasenburg A, Rahnenführer J, Hengstler JG. Prognostic significance of nuclear factor kappa B in node-negative breast cancer is most pronounced in luminal B breast cancer [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P2-05-22.

Background: Although endocrine therapy (ET) is recommended as first-line therapy for hormone receptor (HR)-positive, HER2-negative metastatic breast cancer (MBC) up to 50% of patients receive chemotherapy in this setting. Meanwhile new targeted treatment options for combination with ET have been developed and endocrine-based therapy with the CDK4/6 inhibitor Palbociclib (P) improves the progression free survival (PFS) of ET alone by about 50%. So far, there is no data comparing chemotherapy with or without maintenance ET and ET in combination with P as first-line therapy. Patients included in clinical trials are often criticized not to mirror the general breast cancer population and every-day clinical practice due to rigid inclusion and exclusion criteria, limited number of treatment options, strict monitoring intervals and study assessments. Methods: PADMA trial is a so called low intervention trial with no rigid inclusion and exclusion criteria, and study assessments.Patients with first-line HR+/HER2- MBC who are candidate for mono-chemotherapy will be eligible to receive either P plus ET per label or mono-chemotherapy per investigator´s choice with or without maintenance ET (1:1 randomization). Primary objective is to compare the time-to-treatment failure (TTF) for patients randomized to receive the mono-chemotherapy treatment strategy versus those randomized to receive P and ET. TTF is defined as time from randomization to discontinuation of treatment due to disease progression, treatment toxicity, patient's preference, or death. Main secondary objectives are progression free survival, overall survival at 36 months, amongst other time to event endpoints as well as toxicity and compliance. All patients receive a specific mobile device (PADMA-Phone) and a validated wearable device (ActiWatch) in order to collect data regarding sleep and activity levels, patient well-being and health care utilization (number and duration of phone calls, and patient visits to investigator site) for assessment of daily monitoring treatment impact (DMTI). Results: Overall, 360 patients will be accrued to show an improved TTF for P in combination with ET compared to mono-chemotherapy of investigator´s choice with or without maintenance ET. Recruitment will start in QIII/2017 and is planned for approximately 18 months in 100 sites in Germany, Spain, Poland, Italy, France, UK and Canada. Conclusions: The aim of PADMA is to demonstrate that an endocrine-based strategy consisting of ET plus P is superior to a chemotherapy-based strategy as first-line therapy in women with HR+/HER2- MBC in a real world setting. Assessment of patient-reported outcome, health care utilization, and sleep and activity levels will deliver important information on the differences between endocrine-based and chemotherapy-based treatment. Citation Format: Loibl S, Barinoff J, Seiler S, Decker T, Denkert C, Hardy-Bessard A-C, Senkus-Konefka E, Cognetti F, Palmieri C, Gelmon K, Luebbe K, Furlanetto J, Mueller V, Mundhenke C, Schmidt M, von Minckwitz G, Uhlig M, Burchardi N, Thill M. A randomized, open-label, multi-center phase IV study evaluating palbociclib plus endocrine treatment versus a chemotherapy-based treatment strategy in patients with hormone receptor-positive, HER2-negative metastatic breast cancer in a real world setting (PADMA) [abstract]. In: Proceedings of the 2017 San Antonio Breast Cancer Symposium; 2017 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2018;78(4 Suppl):Abstract nr OT3-05-04.

Background: The incidence of brain metastases (BM) in breast cancer patients is rising and has become a major clinical challenge. So far, limited therapeutic options and insights into the biology of BM exist since only a few studies analyzed exclusively data of breast cancer patients. In order to improve this situation, our multicenter registry was initiated in 2014: Brain Metastases in Breast Cancer Network Germany (BMBC, GBG79). Materials and Methods: Patients with BM diagnosed since 2000, a history of breast cancer and no history of other malignant or neurologic disease can be included. Registration is allowed retrospectively as well as prospectively into a web–based database ("MedCodes"). Characteristics of the primary tumor, metastatic disease and BM as well as treatment details are documented. For this first analysis, 548 patients from 39 German centers were included. Results: Median age at first diagnosis of BM was 55 years (25 – 90 years). 43% of patients (233/548) were HER2 positive, 19% (n=105) were triple–negative and 25% (n= 138) had luminal primary tumors indicating a selection of patients with specific tumor biology who develop BM. 54 % of the patients (n=267) had up to three BM whereas 45% (n=223) had more than three BM. 19% of patients (n=106) had BM without evidence of extracranial disease. 27% of the patients (n=146) underwent surgery of the BM. Of these patients, 61% (n= 89) were treated with whole brain radiotherapy and 16% (n=23) with stereotactic radiotherapy. In patients without surgery (n=397), 73% (n=289) received whole brain radiotherapy and 7% (n=28) stereotactic radiotherapy. Median time from diagnosis of primary breast cancer to BM was 38.5 month for the entire cohort (CI95% 35.4 – 43.3). The time from first diagnosis to BM was shorter for triple–negative patients (20.9 month, CI95% 15.5 – 25.9) compared with patients with HER2–positive (37.0 month, CI95% 30.5 – 42.0) or luminal tumors (48.3 month, CI95% 38.2 – 54.0) (p Conclusion: This is so far the largest analysis of breast cancer patients with BM treated in Germany. In this cohort, triple–negative subtype or more than three BM were associated with shorter survival from the diagnosis of BM. HER2 positive patients with no HER2 directed therapy after the diagnosis of BM showed a shorter survival. The recruitment of the registry is ongoing and we aim to include more than 1000 patients by the end of 2015. Citation Format: Witzel I, Loibl S, Laakmann E, Augustin D, Flock F, Dohmen H-H, Durmus G, Frank M, Hesse T, Ignatov A, Kühn T, Neunhöffer T, Park-Simon T-W, Schmidt M, Stefek A, Weide R, Würschmidt F, Fehm T, Moebus V, von Minckwitz G, Burchardi N, Mueller V. Brain metastases in breast cancer network Germany (BMBC, GBG 79): First analysis of 548 patients from the multicenter registry. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P6-17-08.

Background: Immune checkpoint blockade is increasingly discussed in breast cancer. We examined the prognostic significance of the immune checkpoint receptors cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) and programmed cell death protein 1 (PD-1) in node-negative breast cancer. Methods: Microarray based gene-expression data for CTLA-4 (221331_x_at) and PD-1 (207634_at) were analysed in four previously published cohorts (Mainz, Rotterdam, Transbig, Yu) of node-negative breast cancer patients not treated with adjuvant therapy (n=824). A meta-analysis of previously published cohorts was performed using a random effects model. Prognostic significance of CTLA-4 as well as PD-1 for metastasis-free survival (MFS) was examined in the whole cohort and in different molecular subtypes: luminal (ER+/HER2-), basal-like (ER-/HER2-) and HER2+. Independent prognostic relevance was analysed using multivariate Cox regression. Results: Higher RNA expression of CTLA-4 was related to better MFS in a meta-analysis of the whole cohort (HR 0.58, 95% CI 0.38-0.88, P=0.0102). Prognostic significance was most pronounced in the HER2+ positive molecular subtype (HR 0.23, 95% CI 0.08-0.65, P=0.0062) as compared to luminal (HR 0.68, 95% CI 0.39-1.18, P=0.1744) and basal-like (HR 0.53, 95% CI 0.25-1.15), P=0.1087) carcinomas of the breast. PD-1 RNA expression, however, was not associated with outcome in the whole cohort of patients (HR 0.88, 95% CI 0.32-2.43, P=0.1853). A trend for improved survival was noticed in basal-like breast cancer (HR 0.40, 95% CI 0.15-1.08, P=0.0701). Neither luminal (HR 0.81, 95% CI 0.28-2.36, P=0.2122) nor HER2+ (HR 0.85, 95% CI 0.27-2.68, P=0.7759) patients showed an association of PD-1 with MFS. CTLA-4 showed independent prognostic significance (HR 0.393, 95% CI 0.224-0.688, P=0.001) in multivariate analysis. In addition to CTLA-4, only histological grade of differentiation (HR 2.335, 95% CI 1.490-3.660, P Conclusions: The immune checkpoint receptor CTLA-4 has independent prognostic significance in node-negative breast cancer. Higher expression of CTLA-4 is associated with improved outcome. The prognostic impact of CTLA-4 differs between molecular subtypes and is most pronounced in HER2+ breast cancer. Citation Format: Schmidt M, van de Sandt L, Heimes A-S, Battista M, Lebrecht A, Almstedt K, Hoffmann G, Rahnenführer J, Hengstler JG. Prognostic significance of immune checkpoint receptors in node-negative breast cancer. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P2-08-07.

Background: The BOLERO-2 study demonstrated a relevant benefit for patients who received everolimus in addition to exemestane in patients who progressed during/after a non steroidal aromatase inhibitor (NSAI), which led to approval of everolimus in this indication. However, in routine use a high rate of intolerability due to side effects is reported. The most common side effect of everolimus is mucositis with a reported high rate of intolerability especially during the first 12 weeks of treatment. Mucositis is also considered to be the leading cause for treatment discontinuation not related to tumor progression. In the neoadjuvant GeparQuinto study, a dose-escalation schema was successfully used to improve tolerability of everolimus together with cytotoxic agents. Methods: DESIREE (NCT02387099) is a randomized, double-blind, phase II study of everolimus in addition to exemestane in patients who progressed during or after NSAI. Patients will be randomized in a 1:1 ratio to receive either everolimus 10 mg/day (week 1-3: 4x2.5 mg/day, blinded; week 4-24: 10mg/day, open according to label) or an escalating dose of everolimus as follows: week 1: 1x2.5 mg verum + 3x placebo/day; week 2: 2x2.5 mg verum + 2x placebo/day; week 3: 3x2.5 mg verum + 1x placebo/day; week 4-24: 10 mg/day (open according to label). The primary aim of the study is to evaluate the incidence of the first episode of mucositis grade 2-4 (WHO's oral toxicity scale) any time during a 12 week period after start of everolimus treatment. Secondary objectives are to compare the cumulative rate of mucositis grade 2-4 at 24 weeks after start of treatment, the cumulative rate of mucositis grade 1 and any grade at 12 and 24 weeks after start of treatment, the rate of patients on 10 mg daily at 12 weeks and 24 weeks after start of treatment, the clinical benefit rate after 24 weeks, safety, time to onset of grade ≥2 mucositis, the cumulative everolimus dose at 4 weeks, the relative dose intensity for everolimus, and quality of life using the FACT-B and the QSDQ questionnaire. Biomaterial (whole blood, serum, plasma and optional primary tumor/metastasis tissue) will be collected to evaluate potential biomarkers predicting safety and compliance. Overall, 156 evaluable patients (78 in each arm) are required to detect a clinically relevant difference of 20% in the mucositis rate between treatment arms using a continuity-corrected χ2-test on a significance two sided level alpha of 0.2 and a power of 90%. The rate was estimated to be 40% and 20% in the control arm and the treatment arm, respectively. Results: The study will be conducted in up to 60 German centers. Recruitment will start in June 2015. Enrollment is planned to be completed within 24 months. Conclusion: The combination of everolimus and exemestane has shown to improve the outcome of patients with metastatic breast cancer. In the DESIREE trial a dose-escalating schema will be employed to enhance patient compliance and tolerability necessary to achieve an adequate dose-intensity. Citation Format: Loibl S, Furlanetto J, Barinoff J, Bauerschlag D, Herr D, Lübbe K, Maass N, Müller V, Mundhenke C, Schmidt M, Schwedler K, Thill M, Gkantiragas I, Burchardi N, von Minckwitz G. DESIREE - A multicenter, randomized, double-blind, phase II study to evaluate the tolerability of an induction dose escalation of everolimus in patients with metastatic breast cancer. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr OT1-03-08.

Background Retrospective analyses have shown survival differences in patients with primary metastatic breast cancer (PMBC) in comparison to patients with secondary metastatic breast cancer (SMBC). A subgroup analysis of 2,708 patients with metastatic breast cancer from two prospective non-interventional studies (NIS) allows exploring this hypothesis. The Her2-positive-NIS investigated trastuzumab therapy for Her2 positive metastatic disease additional to predominantly first line chemotherapy. The second NIS observed bevacizumab therapy for predominantly Her2 negative disease additional to chemotherapy as first- line option. Methods Information on PMBC or SMBC was available in 2,401 patients diagnosed with metastatic breast cancer and treated with targeted therapy between 2000 and 2011. Previously endocrine therapy in palliative status was allowed. Progression free survival (PFS) was defined as the time from the start of antibody therapy to disease progression or death; overall survival (OS) was defined as the time from the start of targeted therapy with antibody to death of any cause; PFS and OS were estimated by Kaplan-Meier product limit method. Results 570 (24%) women had PMBC and 1831 (76%) had SMBC. Median follow-up was 23 months (range: 0.2 - 137). Median PFS in patients with PMBC and SMBC was 13.0 and 10.4 months, respectively (p = 0.0043); median OS for women with PMBC and SMBC was 33.5 and 28.5 months, respectively (p = 0.0023). 5 years after start of the study treatment 28% patients with primary and 23% with secondary metastatic disease were alive. Conclusion In a population of breast cancer patients treated with modern antibody therapies, woman with PMBC live significantly longer than those with SMBC. A possible explanation for this observation might be a negative selection process of patients suffering from secondary disease in spite of adequate primary treatment. Every fourth patients with metastatic breast cancer is a long survivor. The clinical-pathological characteristics and risk factors for this group are unclear. Prospective trials including translational investigations for these patient populations are needed. Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P3-06-04.

Background: Bevacizumab (BEV) combined with chemotherapy (CT) significantly improves progression-free survival (PFS) and response rate (RR) vs CT alone in the 1st-line treatment of HER2-negative metastatic breast cancer (mBC). BEV was continued for ≥1 year in 20% of patients (pts) in the global ATHENA safety study. We examined a similar subgroup of pts with long-term antibody treatment in a large German observation study. Meanwhile, long-term follow-up of up to 4.5 years allows to present the final results, including mature data on overall survival. Methods: Pts who had received no prior CT for their mBC received BEV-PAC per the European label. Efficacy and safety were documented in detail for up to 1 year (or until progression, death, or BEV discontinuation if earlier), with subsequent retrieval of long-term follow-up. Results: By April 2013, final data were available for 865 pts, of whom 167 (20%) were recorded to have received BEV for ≥1 year. Baseline characteristics of this subset relative to the overall population are summarized in the table. CharacteristicOverall population (n = 865)Subgroup treated with BEV for ≥1 year (n = 167)Median age, years (range)58(26-87)57(28-79)Age ≥65 years,%3230Metastatic at diagnosis,%2018Disease-free inreval 1,%98Prior (neo)adjuvant chemotherapy,%6661*n = 591, initially M0. #n = 113 Dose reductions of long-term BEV occurred in only 3% of cycles and (at least once) in 17% of pts. In the majority of pts treated for ≥1 year, BEV was continued as a single agent after discontinuation of CT. The overall best RR in pts treated for ≥1 year was 80% (complete response in 19%). Median PFS, based on observed events in 83% of all pts, was 18.4 months compared to 9.6 months in the overall population. Overall survival data, based on 524 deaths (61%), show a median of 21.6 months. In the subgroup with prolonged therapy, median OS amounts to 35.7 months with 40% of pts still alive after 4 years. Safety of long-term BEV is as previously reported with only a slightly increased hypertension rate (35%, grade ≥3: 11%), and no cases of gastrointestinal perforation, arterial thromboembolic event, or reversible posterior leukoencephalopathy syndrome. Conclusions: According to the mature data of this study, including those on OS, a notable proportion of pts appear to derive benefit from prolonged exposure to first-line BEV-containing therapy. Although baseline characteristics exhibited only moderate indicators for an a priori more favorable prognostic profile in the subset of pts treated for ≥1 year, of course one has to keep in mind, that time-related efficacy data are biased towards improved outcome in those able to continue BEV for ≥1 year. However, the median survival of almost 3 years suggests that in a considerable number of pts continued first-line BEV-PAC correlates to favorable outcomes with limited adverse effects. Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P2-16-03.

Background: Risk assessment in node-negative (N0) breast cancer patients (pts) is routinely performed by established clinico-pathological algorithms (CP) (Schmidt et al. Ann Oncol 2009). ASCO guidelines also recommend invasion markers uPA/PAI-1 (UP; Harris et al. JCO 2007). The prospective randomized multicenter NNBC 3-Europe trial had two questions: 1) Does UP compared to the CP algorithm improve identification of low-risk node-negative breast cancer patients? 2) Does addition of taxanes to adjuvant chemotherapy improve DFS probability in high-risk node-negative breast cancer? Data are mature to report on the second question only. Methods: Between 2002 and 2009, 4,147 node-negative breast cancer pts were recruited. Risk assessment was performed by CP (43 %) or by UP (57 %). High-risk pts were randomized to receive FE100C*3-Doc*3 (FEC-D) versus FE100C*6 (FEC) as adjuvant chemotherapy. Primary endpoint was disease-free survival probability (DFS). It was planned to accrue 2,572 pts allowing detection of 4% reduction in recurrence probability at 5 years with a power of 80 % at a level of α=0.05 (LogRank) after a minimum observation time of 2.5 years for every recruited patient. Results: In the intent-to-treat population (ITT, n=2,541), 1,286 high-risk pts received FEC-D, and 1,255 received FEC. In July 2012, 90.8 % of patients had reached the minimum observation time – median follow-up was 43.8, and 44.4 months. Main patients' characteristics were distributed similarly between the two chemotherapy arms: median age was 53 yrs, 37.4 % of pts were premenopausal, median tumor size was 1.90 cm, grade 3 was seen in 53.8 %, hormone receptor status was negative in 30.1 % and HER2 was overexpressed in 20.0 %. Mastectomy was performed in 11 %. All planned courses of FEC-D were given in 84.4 %, of FEC in 91.4 % (p < 0.0001). DFS after 2.5 yrs was for FEC-D 97.5 % (+/− 0.45 %) and for FEC 97.3 % (+/− 0.47 %). Overall-survival probability (OS) after 2.5 yrs was 99.0 % (+/− 0.29 %) for FEC-D and 99.5 % (+/− 0.20 %) for FEC. The survival differences between the two arms were not statistically significant. Per-protocol analysis did not reveal any additional results. Conclusions: Use of taxanes does not further reduce the rate of early recurrences and led to significantly more treatment discontinuation. Today, with adequate adjuvant chemotherapy even high-risk node-negative breast cancer patients have an excellent prognosis. Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P1-13-13.

Background: A genomic index for sensitivity to endocrine therapy (SET) was previously derived from genes strongly positively and negatively coexpressed with estrogen receptor (Symmans et al. 2010, JCO 28:4111). This SET index has been reported to predict survival benefit from adjuvant endocrine therapy independently of prognosis. Materials and Methods: Affymetrix gene expression profiling was performed on 307 ER positive primary breast cancers from a retrospective cohort treated solely with endocrine therapy. In a blinded study the SET index' ability to predict survival was analyzed in this previously unpublished dataset. Affymetrix profiles (CEL files) were anonymized and provided to the developers of SET index (Nuvera Biosciences) without any clinical information. The 261 profiles that passed QC were categorized into SET classes based on the published prespecified cutoffs. Samples categorized as ER negative based on gene expression were excluded. Subsequently classifications were unblinded and clinical associations analyzed according to a predefined protocol. Exploratory analyses were performed on the relationship of SET index to other genomic signatures. We also assessed a potential pure prognostic value of SET index in an additional dataset of 164 node negative ER positive patients that did not receive any adjuvant treatment. Results: A lower SET index significantly correlated with higher grade (p = 0.009) and negative PgR status (p = 0.016). We detected no significant differences for age, tumor size, lymph node status, and HER2 status between patients with high, intermediate, and low SET index. In the lymph node negative (LNN) cohort (n = 120) we observed a significant difference in DFS (5yr DFS 77.1±10.2% vs 94.4±2.2%; HR 4.20, 95% CI 1.72–10.2; p = 0.002) and DMFS (HR 3.18, 95% CI 1.20–8.47; p = 0.020) for patients with low SET index. In contrast, we found no prognostic value of SET index in lymph node positive patients (n = 95). In multivariate analyses of LNN patients including SET, age, tumor size, histological grade, and PgR status, only SET was significantly associated with DFS (HR 3.37, 95% CI 1.25–9.01; p = 0.016) and DMFS (HR 3.03, 95% CI 1.08–8.55; p = 0.036). In exploratory analyses SET index was not correlated to other genomic signatures related to proliferation (as Recurrence Score, GGI, and NKI70) or immune response (e.g. 7IGS, SDPP). When we included all these genomic signatures as continous scores in a multivariate stepwise Cox regression model in the LNN endocrine treated cohort, only SET remained as significant (p = 0.025) while Recurrence Score displayed a strong trend (p = 0.054). We also verified that SET had no pure prognostic value in an additional dataset of 164 ER positive patients that did not receive any adjuvant treatment. Conclusions: In a blinded analysis the predictive ability of SET index was prospectively validated in an independent cohort of node-negative patients. Our exploratory analysis demonstrates that SET index is unrelated to other genomic signatures and delivers independent information on the response of patients to endocrine therapy rather than prognosis. Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P2-10-17.

Background: FN is a dose-limiting toxicity of chemotherapy which can affect patient (pt) outcomes. EORTC guidelines recommend G-CSF primary prophylaxis when the overall FN risk assessment is ≥20%. We evaluated risk assessment and guideline adherence (prophylaxis in high-risk pts) in clinical practice among breast cancer patients. Methods: Eligible pts were enrolled consecutively at centres selected based on experience and geographical spread. Key inclusion criteria were diagnosis of solid tumour or lymphoma, physician-assessed overall FN risk ≥10% (chemotherapy risk plus individual risk factors per EORTC guidelines), and planned primary (PPP) or secondary (PSP) prophylaxis with pegfilgrastim. Pegfilgrastim administered >3 days after chemotherapy completion was classified as therapeutic use. The primary objective was to describe the proportion of pts with an investigator-assessed overall FN risk of >20% or 10–20% receiving PPP or PSP. Secondary objectives included evaluating FN incidence and chemotherapy dose reductions/delays. Data from breast cancer patients only are reported, and prophylaxis in high-risk patients was determined to evaluate guideline adherence. Results: 1003 pts were enrolled from Nov 2007 to Sept 2010. Mean (±SD) age was 54.4 (±11.2) years, almost all pts (99%) were female. Treatment intent was deemed curative by the investigator in 91% of pts. The investigator-assessed FN risk was >20% in 505 (50%) pts and 10–20% in 414 (41%) pts. Despite eligibility criteria, 84 (8%) pts were assessed by investigators as Conclusion: Adherence to G-CSF guidelines, and reasons for differences between planned and administered prophylaxis warrants further investigation. Comparison of outcomes between prophylaxis groups should be interpreted with caution; however, FN incidence remained low with pegfilgrastim PP among breast cancer pts in German clinical practice. Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P1-15-02.

Background: The prognostic significance of CD8 positive cytotoxic lymphocytes in breast cancer is well accepted. However, the role of CD4 positive lymphocytes is ambiguous. We examined the prognostic relevance of CD4 positive tumor-infiltrating lymphocytes in previously published node-negative breast cancer cohorts using RNA expression. Methods: Microarray based gene-expression data for CD4 (203547_at) were analyzed in four previously published cohorts (Mainz, Rotterdam, Transbig, Yu) of node-negative breast cancer patients not treated with adjuvant therapy (n = 824). A meta-analysis of previously published cohorts was performed using a random effects model. Prognostic significance of CD4 on metastasis-free survival (MFS) was examined in the whole cohort and in different molecular subtypes (ER+/HER2−, ER−/HER2−, HER2+). Independent prognostic relevance was analyzed using multivariate Cox regression. Results: Higher RNA expression of CD4 was related to better MFS in a meta-analysis of the whole cohort (HR 0.66, 95% CI 0.49–0.90, p = 0.0074). Prognostic significance was most pronounced in the HER2+ positive molecular subtype (HR 0.32, 95% CI 0.14–0.75, p = 0.0091) as compared to ER+/HER2− (HR 0.62, 95% CI 0.29–1.32, P>0.05) and ER−/HER2− (HR 0.61, 95% CI 0.28–1.35, P>0.05) carcinomas of the breast. CD4 showed independent prognostic significance (HR 0.60, 95% CI 0.37–0.96, p = 0.032) in multivariate analysis. In addition to CD4, only histological grade of differentiation (HR 2.43, 95% CI 1.50–3.94, P < 0.001) and tumor size (HR 1.87, 95% CI 1.19–2.94, p = 0.007), but not age, HER2 status or hormone receptor status retained an independent prognostic association with MFS. Conclusion: CD4 positive tumor-infiltrating lymphocytes have independent prognostic significance in node-negative breast cancer. A higher expression is associated with improved outcome. Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P2-10-13.

Background: In recent years, surgeons have utilized Harmonic instruments to perform breast cancer resection. Retrospective and prospective studies have demonstrated that the use of this surgical device for mastectomy and axillary dissection can reduce perioperative blood loss, seroma formation, and duration and total amount of drainage. No study has analyzed the feasibility of Harmonic instruments in breast-conserving surgery. We conducted a prospective, randomized clinical trial comparing Harmonic instrument and conventional surgery in the performance of breast conserving surgery and axillary procedures to determine differences in surgical procedures, postoperative outcome, and complications. Methods: One hundred and forty seven patients with operable breast cancer who underwent breast-conserving surgery at a single institution between December 2009 and March 2011 were included in the analysis. Surgery was performed in 73 patients with the Harmonic Focus® device and in 74 with scissors and electrocautery. Charts were reviewed for patient demographics, histopathologic reports, surgical procedures, length of stay, volume and duration of postoperative drainage, complications such as seroma and hematoma formation, and postoperative pain. Results: We found a statistically significant difference in size of resected breast tissue (p Discussion: This is the only study comparing the Harmonic instrument with traditional surgery in both breast conserving surgery and axillary procedures. From this study, we conclude that despite higher costs, the Harmonic device is safe to use and provides key benefits in intraoperative technique, postoperative outcome, and rate of complications in breast cancer surgery. Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-15-09.

Background: The EndoPredict (EP) score is a multigene classifier to predict the likelihood of distant recurrence in ER-positive, HER2−negative breast cancer patients treated with adjuvant endocrine therapy. Two large randomized phase III trials involving endocrine therapy only (n > 1700) demonstrated additional prognostic information of the EP score independent from clinicopathological parameters by classifying 49% as low risk. However, the predictive role of the EP is not clear. Therefore, we examined whether the EP Score also predicts sensitivity towards neoadjuvant chemotherapy in ER-positive, HER2−negative breast cancer patients. Methods: Four publicly available gene expression data sets (Affymetrix HG-U133A) were retrieved from the gene expression omnibus (GEO) data repository. All analyzed breast cancer patients were treated with anthracycline or taxane/anthracycline-based neoadjuvant chemotherapy. Microarray cel files were MAS5 normalized with a global scaling procedure and a target intensity of 500. The analysis was restricted to ER-positive, HER2−negative breast cancer patients according to pre-specified cut-off levels for the respective ESR1/ERBB2 Affymetrix probesets. The EP score was calculated and patients were classified as having low or high risk according to the pre-specified validated cut-off value. Pathological complete response (pCR) — defined as no residual invasive cancer in the breast or lymph nodes — was used as the primary endpoint for the assessment of treatment response. Results: The EP Score was examined in 221 ER-positive, HER2−negative breast cancer patients treated with neoadjuvant therapy. Among the 221 patients, 61 tumors (27.6%) were classified as EP-low-risk, whereas 160 tumors (72.4%) were EP-high-risk. Only one of the EP-low-risk tumors achieved a pCR after neoadjuvant therapy, whereas 24 of the 25 pCR events were classified as EP high risk. The sensitivity of the EP score was 96% and the negative predictive value 98% with an area under the receiver operating characteristic curve of 0.73. Conclusions: The EP Score is a predictor of chemosensitivity in the neoadjuvant setting. The test correctly identified all but one of the patients achieving a pCR suggesting that the benefit of cytotoxic chemotherapy is limited to the EP high risk group. Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-06-26.

Background: First-line bevacizumab (BEV) combined with weekly paclitaxel (PAC) significantly improves progression-free survival (PFS) and response rate (RR) vs PAC alone in HER2−negative metastatic breast cancer (mBC), as shown in E2100. The benefit of BEV combined with other chemotherapy (CT) agents was demonstrated in AVADO and RIBBON-1. BEV was continued for ≥1 year in 21% of patients in the global ATHENA safety study and in 42% of patients in the JO19901 single-arm Japanese study of BEV-PAC. We analyzed data from the subgroup of patients treated for ≥1 year in a German routine oncology practice study to provide insight into the safety and efficacy of prolonged first-line BEV-PAC. Methods: Patients who had received no prior CT for their mBC received BEV-PAC per the European label. Efficacy and safety were documented for up to 1 year (or until progression, death, or BEV discontinuation if earlier) with additional long-term follow-up. Data from patients treated with BEV for ≥1 year were extracted for this analysis. Results: By Jan 2011, data were available for 818 patients, of whom 157 (19%) had already received BEV for ≥1 year. Baseline characteristics of this subset relative to the overall population are summarized in the table. In 79% of those treated for ≥1 year, BEV was continued as a single agent after discontinuation of CT. The overall RR in patients treated for ≥1 year was 81% (complete response in 20%). Median PFS was 17.9 months (events in 44% of patients) vs 9.4 months in the overall population (events in 68%). Overall survival data are immature, as 81% of those treated for ≥1 year are still alive. The most common grade 3/4 adverse events in patients treated with BEV for ≥1 year were hypertension (11% of patients), pain (10%), and leukopenia (7%). There were no cases of gastrointestinal perforation, arterial thromboembolic event, or reversible posterior leukoencephalopathy syndrome in those treated for≥1 year. Analysis of adverse events according to time of onset is ongoing. Conclusions: A notable proportion of patients appear to derive benefit from prolonged exposure to first-line BEV-containing therapy. In the present analysis, baseline characteristics appeared more favorable in the subset of patients treated for ≥1 year than in the overall population. Efficacy data clearly have a bias towards improved outcome in those able to continue BEV for ≥1 year, as these patients had sustained disease control for ≥1 year. However, the efficacy of prolonged BEV-containing therapy is of interest and suggests that some patients achieve sustained disease control with continued first-line BEV-PAC with limited side effects. Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-14-04.

Background: Transient induction of HER2 expression upon inhibition of the estrogen-receptor (ERα) might be an underestimated rescue mechanism that enables ER+/HER2- breast cancer cells to survive and to grow in the presence of endocrine therapies. Besides protecting ER+/HER2- BC cells from endocrine treatment, transient HER2 expression is also considered to save circulating, disseminated and dormant ER+/HER2- BC cells from apoptotic cell death in hostile environments and might therefore be of importance for ER+ BC progression, metastasis formation and recurrence. Thus therapeutic strategies that efficiently target both ERα and HER2 simultaneously might improve treatment of patient with ER+/HER2- BCs. Methods: Circulating tumor cells (CTCs) were originally isolated from a patient with bilateral ER+/HER2- metastatic BC. These cells have been treated with either the proteasome inhibitor (PI) carfilzomib or fulvestrant alone or with both drugs in combination. The consequences of these drugs upon ERα and HER2 expression were monitored by western blotting or qPCR. Induction of cell death upon treatment was measured by PARP and caspase cleavage or by quantification of SubG1 cells using propidium iodide staining or by the use of colony forming assays. Results: Fulvestrant treatment of CTCs decreases the amount of ERα but immediately increases HER2 expression whereas carfilzomib markedly inhibits the expression of both ERα and HER2 simultaneously. Combined treatment of CTCs with carfilzomib + fulvestrant cause reduced expression of HER2 and lead to a much stronger decrease of ERα than carfilzomib or fulvestrant alone. Fulvestrant causes no significant reduction of proliferation and no induction of cell death. Conversely the combination of carfilzomib and fulvestrant causes a significant induction of apoptotic cell death and a massive reduction of colonies in colony forming assays. Conclusion: These findings suggest that rapid and transient up-regulation of HER2 expression following endocrine treatment might be an important so far underestimated adaptive mechanism which enables ER+/HER2- BC cells to sustain proliferation in the presence of ERα-inhibitory drugs and to stay alive during the metastatic process. Furthermore these data also lead to the assumption that PIs such as carfilzomib in combination with ERα degraders could be a potential therapeutic strategy for efficient targeting of metastatic ER+/HER2- BC cells. Citation Format: Thaler S, Roßwag S, Pantel K, Sleeman JP, Schmidt M, Cotarelo CL. Targeting of endocrine therapy-induced estrogen-receptor/HER2-cross-talk in circulating tumor cells from metastatic ER+/HER2- breast cancer: Implications for treatment of ER+/HER2- breast cancer [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P3-01-23.

Background: PELICAN is a phase III trial of pegylated liposomal doxorubicin (PLD) vs capecitabine (CAP) as first-line therapy of patients (pts) with MBC. In the elderly pts, a CGA and analyses on biologic parameters on inflammation and nutrition, incorporated in the Prognostic Inflammatory and Nutritional Index (PINI) and Glasgow Prognostic Scale (GPS), were performed and results correlated to time to progression (TTP), overall survival (OS), time to treatment failure (TTF), and toxicity. Methods: CGA comprised data on activities of daily living (ADL), instrumental activities of daily living (IADL), ECOG and Karnofsky performance score (KPS), comorbidity (CIRS-G), and comedication. According to CGA, pts were classified into groups 1-3, ranging from fit to frail (Balducci 2000). For biologic assessment, blood samples of 86 pts were collected and centrally analysed. PINI was defined as [(CRP (mg/L) x A1GP (mg/L)]/[albumin (g/L) x prealbumin (mg/L)]. For GPS: pts were allocated a score of 2 if CRP >10mg/L and albumin Results: 210 pts (PLD:105; CAP:105) were randomized, stratified by age & prior anthracycline. Median age was 62y (22-85). Pts received a median of 5 cycles each of PLD and CAP. Median TTP was comparable in both arms (6.7mo PLD vs 7.1mo CAP, p=0.346). There was an insignificant trend for higher GPS and PINI in pts ≥65y. According to CGA, 102 pts were considered fit (group 1, 72%), 21 compromised (group 2, 15%), and 19 frail (group 3, 13%). CGA did not correlate with therapy efficacy. Pts with good KPS of ≥90% (n=46) had a significantly longer TTP vs pts with a worse KPS (n=44) (median KPS 100%: 8.7mo; 90%: 8.6mo; ≥90%: 3.8mo; p=0.0027). Pts with GPS of 0 or 1 had a significantly longer TTP and TTF vs pts with a GPS of 2 (median TTP score 0: 8.3mo, score 1: 9.2mo, score 2: 2.8mo, p=0.0002). Grouping of pts by PINI values (10) showed no correlation with TTP or TTF, and a nonsignificant trend for shorter OS for pts with PINI >10 (p=0.056). Among the analyzed laboratory parameters as single values, CRP showed a significant correlation with TTP (median TTP ≥5mg/L: 10.4mo, > 5 mg/l: 6.2mo, p=0.031) as did serum amyloid A (SAA, median TTP ≥6.8mg/L: 10.4mo, >6.8mg/L: 6.0mo, p=0.026) and serum albumin (median TTP ≥35g/L: 8.3mo, Conclusions: First-line chemotherapy of MBC pts using PLD or CAP showed similar efficacy with respect to TTP. Incorporation of CGA into a phase III trial is feasible and yields information that would otherwise have been missed. Prospective analyses of parameters of systemic inflammation and nutrition may have prognostic significance regarding TTP, TTF, and potentially OS in pts with breast cancer. Results of CGA, PINI and GPS need to be validated prospectively in treatment algorithms for this population. Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P6-11-06.

Background: Both progression-free survival (PFS) and response rate (RR) are significantly improved when bevacizumab (Bev) is combined with first-line chemotherapy for locally recurrent/metastatic breast cancer (LR/mBC), as demonstrated in three randomized, phase III trials (E2100, AVADO, RIBBON-1). The benefit derived from Bev appears to be similar in triple-negative (TN) and non-TN LR/mBC according to subanalyses of the randomized trials. We conducted a subpopulation analysis of TN patients treated with first-line Bev-paclitaxel (Pac) in a large German observational study in the context of routine oncology practice. Methods: Patients with HER2-negative LR/mBC received first-line Bev-Pac according to the European label at the time of study design. Safety and efficacy data were collected for up to 1 year (or until progression, death, or Bev discontinuation if earlier). The endpoints were safety and efficacy (PFS and RR). We conducted an exploratory analysis of the subset of patients with TN disease. Results: Of the 567 patients for whom data are currently available, 115 (20%) had TN disease. Baseline characteristics and efficacy are summarized below. Overall survival data are immature and follow-up is continuing. Conclusions: In this ongoing study, first-line Bev-Pac demonstrated a 51% RR and median PFS of 7.7 months in patients with TN LR/mBC. These data are consistent with findings from retrospective analyses of randomized phase III trials and a subpopulation analysis of the ATHENA study. Prospective trials evaluating Bev combination regimens in TN breast cancer are ongoing. Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-16-12.

Background: In patients (pts) with metastatic bone disease (MBD) due to breast cancer (BC) the incidence of skeletal related events (SRE) and the severity of bone pain are effectively reduced by bisphosphonates (BP), the current standard of care in this setting. As treatment with some BP may adversely affect kidney function, renal safety is a key determinant of BP risk/benefit assessment. Ibandronate (IBA), a BP available in IV and oral formulations, demonstrated sustained effects on skeletal related events (SRE) and bone pain with a low incidence of renal adverse events in randomized clinical trials (RCT). To verify those results under real life conditions, a large-scale non interventional study (NIS) was conducted in Germany. Patients and methods: In accordance with the terms of the marketing authorization, BC pts with MBD received 6 mg IBA IV every 3-4 weeks or 50 mg of oral IBA once daily (at the physician's discretion) for up to 24 weeks. Pain severity (VAS), use of analgesics (classified according to the WHO analgesic ladder), SRE, and renal function (estimated creatinine clearance, eCrCl; Cockcroft-Gault formula) were recorded in 4-week intervals. Results: 3515 pts, mean age 63.6 +11.7 years, were evaluated. 2320 (66%) were BP-naïve; 418 (12%) had received prior treatment with IBA, 777 (22%) with other BPs, including zoledronic acid (ZOL) (503; 14%) and pamidronate (256; 7%). At baseline, mean pain severity (VAS) was lower for pts pretreated with IBA (2.5 ± 2.1) compared to BP-naïve pts (3.2 ± 2.4; p < 0.0001) or pts pretreated with other BPs (3.1 ± 2.4; p < 0.0001) and analgesic use was less frequent (38% vs 47% and 44%, respectively). For the 2678 pts evaluable for renal function throughout the observation period, mean eCrCl at baseline was significantly lower in pts pretreated with ZOL (72.4 ± 31.7 ml/min) than in BP-naïve (79.5 ± 30.7 ml/min; P Conclusion: In this NIS in BC pts with MBD, the largest-to-date study with IBA, IV and oral IBA showed marked and sustained pain relief with an overall reduction of analgesic use and a renal safety profile comparable to results of RCT. Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P1-13-02.

Background: Aurora kinase A (AURKA) is important for cell cycle progression. Inhibitors that target AURKA are currently in clinical development. We examined the prognostic impact of AURKA in three cohorts of node-negative breast cancer patients without adjuvant systemic therapy (n=766). Methods: AURKA (probe set ID 204092_s_at) was analysed utilizing microarray based gene-expression data of three independent and previously published cohorts of node-negative breast cancer patients (Mainz, Rotterdam, TRANSBIG). In addition to AURKA, we examined the prognostic impact of age, histological grade, tumor size, estrogen receptor (ER) and HER2. Metastasis-free survival (MFS) was analyzed with univariate and multivariate Cox regression. Results: AURKA displayed a strong positive correlation with grade (P Conclusion: AURKA has prognostic relevance in three independent cohorts of node-negative breast cancer patients making it a suitable target for therapy with Aurora kinase inhibitors. Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P3-10-22.

Background: In women with HER2-positive breast cancer progressing during trastuzumab treatment, continuation of trastuzumab plus capecitabine (XH) showed a significantly improved overall response rate and time to progression compared with capecitabine (X) alone (von Minckwitz et al, J Clin Oncol 27:1999-2006, 2009). Here we report the final analysis of overall survival (OS). Methods: Patients (pts) with HER2-positive, locally advanced or metastatic breast cancer who progressed during treatment with trastuzumab with or without adjuvant and/or 1st-line metastatic chemotherapy were prospectively randomized to X (2500 mg/m2 on days 1-14, q3w) or XH (6 mg/kg, q3w). OS was a pre-specified secondary endpoint of the study. Results: Median follow up at June 2010 was 20.7 months. 59 of 74 and 60 of 77 pts died in the X alone and XH arm, respectively. Median OS was 20.6 and 24.9 months with X alone and XH, respectively. This difference was not statistically significant (HR=0.94 [0.65-1.35]; p=0.73). Cox proportional hazard model showed performance status, hormone receptor status and metastatic site as independent prognosticators for survival. No difference between treatment arms was observed for pts with a clinical response or clinical benefit, respectively. Pts who continued/restarted anti-HER2 treatment (trastuzumab or lapatinib) after 2nd progression (N=88) had an OS of 18.8 compared with 13.3 months for those who did not receive 3rd line treatment with anti-HER2 agents (N=52) (HR 0.63; p=0.02). Pts who continued treatment after progression as initially randomized (anti-HER2 treatment after XH (N=31) or no anti-HER2 treatment after X alone (N=53)) had an OS of 26.7 months and 20.4 months (HR 0.71; p=0.2). Conclusions: Final OS analysis of the GBG-26 study could not demonstrate a survival benefit for treatment beyond progression with trastuzumab. However in a post-hoc analysis, pts receiving anti-HER2 treatment as 3rd line therapy showed a better OS than those not receiving this targeted treatment. Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P6-14-05.

Background: Molecular tests predicting outcome of breast cancer patients may be useful for treatment decisions in addition to standard clinicopathologic features. Methods: Using human genome HG-U133A array and qRT-PCR datasets, we developed and validated a gene-expression signature predicting the likelihood of distant recurrence in postmenopausal, early-stage breast cancer patients with estrogen receptor-positive, HER2-negative tumors treated with adjuvant endocrine therapy. RNA levels assessed by qRT-PCR in formalin-fixed paraffin-embedded tumor specimens were used to calculate a risk score (T5) and to determine a risk group (low or high) for each patient. The prospectively defined T5 risk score was then validated independently in patients from two large randomized phase III trials. Distant recurrence-free survival and overall survival were analyzed with Cox models adjusted for clinicopathological factors. The primary endpoint was time to distant recurrence. Results: In a training set of 964 tumors, we identified a gene-expression signature consisting of three proliferation-related genes (BIRC5, UBE2C, DHCR7), five estrogen-regulated genes (RBBP8, IL6ST, AZGP1, MGP, STC2), and three reference genes (CALM2, OAZ1, RPL37A). For the validation, RNA analysis was possible in 1702 of 1725 (99%) tumors of both validation sets. Women were classified as having low risk (n=832; 49%) or high risk (n=870; 51%) by the T5 risk score. The T5 risk score provided prognostic information independent from clinicopathologic risk as estimated by Adjuvant!Online or Ki67 labeling index. Patients with a higher T5 risk score had a significantly shorter time to distant recurrence (adjusted hazard ratio, 1.24; 95% confidence interval [CI], 1.15 to 1.33; P Conclusions: Using formalin-fixed paraffin-embedded tumor specimens, the multigene T5 risk score provides prognostic information independent of Adjuvant!Online or Ki67 labeling index. By combining the T5 risk score with clinicopathological risk, we were able to accurately identify breast cancer patients with low risk or high risk for distant recurrence. Using this new easy-to-use multigene tool in clinical practice will assist in optimizing adjuvant therapy by reducing both undertreatment and overtreatment and thus improves outcome and quality of life of patients with early-stage breast cancer. Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P3-10-07.

Background:The use of herbal preparations (HEP) to alleviate climacteric disorders is expected to increase as women seek alternatives to menopausal hormone therapy to avoid the associated breast cancer risk. Data are sparse on the long-term effects of HEP containing phytoestrogens and black cohosh on breast cancer risk. Methods: Within a German case-control study, associations between patterns of HEP use and incident breast cancer were investigated in 10,121 postmenopausal women (3,464 cases, 6,657 controls). Information on HEP use was collected in face-to-face interviews supported by a list of brand names. Multivariate logistic and polytomous regression analyses were done. Findings: Ever use of HEP (9.9%) was inversely associated with invasive breast cancer [odds ratio (OR), 0.74; 95% confidence interval (CI), 0.63-0.87] in a dose-dependent manner (OR, 0.96 per year of use; P = 0.03). Classes of HEP did not differ significantly (Pheterogeneity = 0.81). Risks for invasive ductal (OR, 0.72; 95% CI, 0.60-0.87) and combined lobular/mixed/tubular tumors (OR, 0.76; 95% CI, 0.58-1.01) were similarly reduced by any HEP use but not for in situ carcinomas (1.34; 95% CI, 0.86-2.09). There were no substantial differences in associations of HEP use by estrogen receptor status (ER+ OR, 0.74; 95% CI, 0.62-0.89; ER− OR, 0.68, 95% CI, 0.50-0.93) and progesterone receptor status of the tumor. Interpretation: Our findings support the hypothesis that HEP use protects from invasive breast cancer in postmenopausal women. Among conceivable modes of action, those independent of estrogen receptor–mediated pathways seem to be involved (i.e., cytotoxicity, apoptosis). (Cancer Epidemiol Biomarkers Prev 2009;18(8):2207–13)

Background Inhibition of COX-2 has been shown to attenuate the metastatic process in pre-clinical models of human breast cancer (BC). The primary aim of this study was to assess the effect of 2 years adjuvant therapy with the COX-2 inhibitor celecoxib compared with placebo in HER2-ve primary BC patients. Patients & Methods Patients were randomised in a 2:1 ratio to receive celecoxib 400mg once daily or placebo for 2 years. Patients had to have completely resected BC with prior local and systemic adjuvant treatment according to local practice. Concurrent radiotherapy was permitted and hormone receptor +ve patients received endocrine therapy according to local practice. Patients with HER2+ or node negative, T1 and grade 1 disease were excluded. Median age of patients was 55 years (IQR: 49-63). 50% of patients had tumours >2cm; 42% were grade 3; 48% had node +ve disease. According to local assessment 73% were ER/PgR +ve. Primary endpoint was Disease Free Survival (DFS); defined as time from randomisation to date of first event, with events contributing to analysis defined as recurrence (distant/local), new primary BC (ipsilateral/contralateral) and death. Secondary endpoints included Overall Survival (OS), toxicity, cardiovascular mortality and incidence of second primaries. Subgroup analysis by hormone receptor status was pre-planned. Survival endpoints are analysed using Cox-proportional hazards and log-rank tests; restricted mean survival is used where proportional hazards do not hold. Results Between January 2007 and November 2012, 2639 patients were randomised (1763 celecoxib; 876 placebo) from 181 centres across the UK and Germany. At 13th April 2017, median follow up was 60 months (IQR: 48-72) with 428 DFS events reported. Unadjusted survival analysis results are presented below, with hazard ratio 5 year survival estimate (95% CI)Hazard ratio (95% CI)p-valueDFS (all patients) Celecoxib83% (81, 85)1.02 (0.83 – 1.24)0.88Placebo83% (80, 86)1- DFS within ER+ Celecoxib87% (85, 89)0.89 (0.69 – 1.16)0.40Placebo86% (83, 89)1- DFS within ER- Celecoxib72% (68, 76)1.17 (0.85 – 1.61)0.33Placebo75% (69, 80)1- OS (all patients) Celecoxib90% (88, 91)0.97 (0.75 – 1.25)0.81Placebo90% (88, 92)1- The interaction between ER status and treatment was not significant; p=0.36. In the celecoxib and placebo groups there were 17 and 8 deaths respectively in patients who had not relapsed. These were due to cardiac (n=3; 2) and other (n=14; 6) in the celecoxib and placebo groups respectively; none were GI related. In total 304 serious adverse events were observed in 265 patients (186/1763 celecoxib; 79/876 placebo). In the celecoxib and placebo groups respectively these were related to cardiac (n=12; 7), GI (n=9; 2) and other (n=193; 81). Work is ongoing to determine whether a subset of ER+ patients whose primary tumours show the characteristics of a COX-2 signature receive greater benefit from celecoxib. Conclusions There is no benefit of celecoxib in the ITT population. Further exploratory studies focussing on the ER+ subpopulation are ongoing. Celecoxib treatment is not associated with significant toxicity when compared to placebo in this population of BC patients. Citation Format: Coombes RC, Tovey H, Kilburn L, Mansi J, Palmieri C, Bartlett J, Hicks J, Makris A, Evans A, Loibl S, Denkert C, Murray E, Grieve R, Coleman R, Schmidt M, Klare P, Rezai M, Rautenberg B, Klutinus N, Rhein U, Mousa K, Ricardo-Vitorino S, von Minckwitz G, Bliss J. A phase III multicentre double blind randomised trial of celecoxib versus placebo in primary breast cancer patients (REACT – Randomised EuropeAn celecoxib trial) [abstract]. In: Proceedings of the 2017 San Antonio Breast Cancer Symposium; 2017 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2018;78(4 Suppl):Abstract nr GS3-03.

Background: Urokinase plasminogen activator (uPA) and its inhibitor (PAI-1) determined by ELISA from fresh-frozen tumor tissue have been evaluated as prognostic factors in prospectively randomized trials in node-negative breast cancer. However, uPA and PAI-1 mRNA expression might reveal new clinically relevant information using paraffin-embedded tumor tissue. Methods: We evaluated uPA and PAI-1 mRNA expression with the Affymetrix HG-U 133A array also within molecular subgroups of breast cancer in a finding cohort of breast cancer patients (cohort A; n = 548). We validated mRNA expression in a cohort of HER2 positive breast cancer patients (cohort B, n = 347). For uPA and PAI-1, mRNA values below and above the median were compared in two different probesets. “Luminal”, “triple-negative” and “HER2 positive” subcohorts were defined by ESR1 and ERBB2 mRNA expression using pre-defined cut-offs. Results: In cohort A, elevated PAI-1 mRNA expression was associated with shorter disease-free survival (DFS) and overall survival (OS), elevated uPA mRNA expression was associated with shorter OS. Regarding different molecular subgroups, 71% (n = 388) of tumors had a luminal, 13% (n = 69) a HER2 positive and 17% (n = 91) a triple-negative subtype. Only in the HER2 positive subgroup, elevated PAI-1 mRNA expression was associated with shorter disease-free survival and overall survival (p = 0.003 for DFS and p = 0.012 for OS). Same results were found for uPA in HER2 positive patients (DFS, p = 0.027 and OS, p = 0.011). In contrast, no association between both markers and DFS and OS could be seen in the luminal or triple-negative subgroups. In the HER2 positive validation cohort B elevated uPA and PAI-1 mRNA expression showed a strong association with shorter DFS (p = 0.013 for PAI-1, p = 0.001 for uPA). Conclusion: Results of this study demonstrate that the prognostic impact of uPA and PAI-1 mRNA expression was observed mainly in patients with HER2 positive tumors. Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P6-06-44.

Background The cell surface receptor RANK (receptor activator of NFκB), its ligand (RANKL) and the decoy receptor of RANKL osteoprotegerin (OPG) play an important functional role in bone physiology and in bone metastasis by regulating osteoclasts. Just recently it was shown that tumor-infiltrating lymphocytes can stimulate breast cancer metastases through RANK-RANKL signalling. Material and methods: We analyzed gene expression of RANK, RANKL and OPG in a combined Affymetrix dataset of 307 ER positive breast cancers from our institutions which were either untreated of treated with chemotherapy. Kaplan Meier analysis of disease free survival and Cox regression analysis was applied to examine the prognostic value of the different markers. Results: We observed no significant difference in survival when samples were analyzed according to either RANK or RANKL mRNA expression. In contrast when samples were stratified in quartiles of OPG expression a positive linear relationship of survival with the expression of OPG was observed. Moreover since OPG demonstrated a bimodal type of expression a cutoff value can be derived from the expression data. Using this cutoff value a hazard ration of 2.14 (95% CI 1.27−3.61; P=0.004 for low OPG expression was detected. OPG expression correlated with lower proportion of grade 3 tumors (15.7% vs 27%; P=0.022) and a higher proportion of PgR positive samples (86.2% vs 71.4%; P=0.002). No significant differences were observed for lymph node status, age, tumor size and HER2 status. In multivariate analysis only lymph node status remained significant while OPG, Ki67, age, grade, and PgR only displayed a trend towards significance. Conclusion: Expression of osteoprotegerin seems to correlate with good prognosis in ER postive breast cancer. These data are in line with in vitro studies demonstration that OPG inhibits RANKL induced migration of tumor cells. Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-01-12.

Background: Infiltration of cytotoxic T lymphocytes (CTL) is a common feature in breast cancer. We examined the prognostic impact of CD8 positive CTL in formalin-fixed-paraffin-embedded (FFPE) tissue using immunohistochemistry (IHC) in node-negative breast cancer and validated our findings in previously published cohorts using RNA expression. Methods: CD8 was evaluated in FFPE tissue of 339 medically untreated node-negative breast cancer patients utilizing IHC. Results were validated utilizing microarray based gene-expression data of four cohorts of medically untreated node-negative breast cancer patients (n=824). Impact of CD8 on metastasis-free survival (MFS) was analyzed with univariate and multivariate Cox regression. Meta-analysis of previously published cohorts was performed using a random effects model. Prognostic significance was examined in the whole cohort and in different molecular subtypes (ER+/HER2−, ER−/HER2−, HER2). Correlation between RNA expression and IHC was analyzed according to Spearman. Results: Immunohistochemical detection of CD8+ CTL was associated with MFS in univariate (hazard ratio [HR] 0.76, 95% confidence interval [95% CI] 0.59−0.98, P=0.037) as well as in multivatiate analysis (HR 0.61, 95% CI 0.46−0.81, P=0.001). Protein levels of CD8 had a good correlation with RNA expression (r=0.488). Higher RNA expression of CD8 was related to better MFS in a meta-analysis of the whole cohort (HR 0.74, 95% CI 0.60−0.90, P=0.0023). However, prognostic significance was confined to ER+/HER2− (HR 0.71, 95% CI 0.51−0.99, P=0.0201) and HER2+ (HR 0.60, 95% CI 0.38−0.97, P=0.037) but not to ER−/HER2− molecular subtypes (HR 0.77, 95% CI 0.52−1.14, P=0.1965). Conclusion: CD8 positive CTL have independent prognostic significance in node-negative breast cancer. Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-01-13.

Background: Both the prognosis and the therapeutic options in triple negative breast cancer (TNBC) are rather limited. Current prognostic gene expression profiles for breast cancer mainly reflect proliferation status and are most useful in ER-positive cancers. The identification of prognostic gene signatures from TNBC cohorts in previous studies was hindered due to relatively small sample sizes. Materials and Methods: All currently available TNBC gene expression datasets generated on Affymetrix U133 gene chips were assembled. To minimize inter-laboratory variation we analyzed only highly comparable arrays and data set-biased genes were filtered. Supervised analysis was applied to identify a prognostic signature from a finding cohort of 394 TNBC and validation was performed in an independent cohort of 261 TNBC. The genes from the prognostic predictor were analyzed for their correlation to known molecular phenotypes among TNBC. Results: Two supervised prognostic signatures consisting of 264 and 26 probesets, respectively, were obtained when applying different cutoffs for false discovery rates of 25% and < 3.5% in the finding cohort. In multivariate analysis in the independent validation cohort hazard ratios of 4.03 (95% CI 1.71−9.48; P=0.001) and 4.08 (95% CI 1.79−9.28; P=0.001), respectively, were obtained for the two signatures. When compared to 16 metagenes for previously described molecular phenotypes in TNBC the prognostic signatures displayed highest correlation to metagenes for IL-8/inflammation, VEGF/angiogenesis, and Histones. A subset of genes in the 264-probeset signature was inversely associated with a poor prognosis (29/264=11.0%). Most of these “good prognosis” genes are correlated with immune cell metagenes (21/29=72.4%). In contrast both identified supervised prognostic signatures did not correlate to previously published prognostic signatures (recurrence score, genomic grade index, Amsterdam signature, wound response signature, 7-gene immune response module, stroma derived prognostic predictor, and a medullary like signature). Regarding the response of TNBC to neoadjuvant chemotherapy the predictive value of the B-cell metagene was superior to the 264- and 26-probeset signatures. However combination of the B-cell metagene and the signatures increased the AUC in ROC-analysis from 0.606 to 0.656. Conclusions: The use datasets consisting only of TNBC allows identification supervised prognostic signatures for TNBC which are unrelated to previously known prognostic signatures. Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr PD03-02.

Background: Gene expression profiles in breast cancer mainly reflect ER, PgR, HER2 status, and proliferation. Triple negative breast cancers (TNBC) seem to constitute a heterogenous entity and their relationship to basal-like breast cancers (BLBC) is not fully clear. Further understanding of TNBC is important since new treatments are needed. Materials and Methods: We assembled an Affymetrix dataset of n=579 TNBC (finding cohort n=394, validation cohort n=185). Unsupervised methods identified principal metagenes differing among TNBC in this clean cohort. Clinical data were correlated to these metagenes for the basal-like and apocrine phenotypes, different types of immune cells, the claudin-CD24 signature, markers of blood and adipocytes, as well as signatures for stroma, angiogenesis, and inflammation. Results: 73% of TNBC displayed a basal-like phenotype correlating with poor histological grading and younger age. However, no effect of basal-like phenotype on survival was observed. A good prognosis was obtained for expression of immune cell metagenes and a poor prognosis was associated with metagenes for inflammation, angiogenesis and stromal cells. A combination of high B-cell metagene and low IL-8 metagene expression identified 27% of TNBC with a favourable prognosis (HR 0.25, 95% CI 0.12-0.48; P Conclusion: All published prognostic signatures assign TNBC to a poor prognosis group. Use of a subtype-specific dataset not confounded by the main variables of breast cancer allowed the identification of a novel prognostic signature for TNBC. Targeting the IL-8 pathway could be a valuable therapeutic opportunity in TNBC. Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr S5-5.

Background: Bisphosphonates (BP), the current standard of care for patients with metastatic bone disease (MBD) due to breast cancer (BC), effectively reduce the incidence of skeletal related events (SRE) and the severity of bone pain. However, BP treatment may adversely affect kidney function and thus renal safety is a key determinant of BP risk/benefit assessment. In randomized clinical trials (RCT), Ibandronate (IBA), a third generation amino-BP available in IV and oral formulations, has demonstrated sustained effects on SRE and bone pain with a low incidence of renal adverse events. To verify those results under real life conditions, a large-scale non interventional study (NIS) is currently being conducted in Germany.Patients and methods: In accordance with the terms of the marketing authorization, BC patients with MBD receive 6 mg IBA IV every 3–4 weeks or 50 mg of oral IBA once daily (at the physician's discretion) for up to 24 weeks. Pain severity (VAS), analgesic use (WHO analgesic ladder), SRE, and renal function (serum creatinine levels and estimated creatinine clearance, eCrCl; Cockcroft-Gault formula) are recorded in 4-week-intervals.Results: For this interim analysis, data for 2676 patients, mean age 63.6 ±11.8 years, were evaluated. 1735 (65%) were BP-naïve; 317 (12%) had received prior treatment with IBA, 614 (23%) with other BPs, including zoledronic acid (ZOL) (393; 15%) and pamidronate (PAM) (210; 8%). At baseline, mean pain severity (VAS) was lower for patients pretreated with IBA (2.7 ± 2.2) compared to BP-naïve patients (3.3 ± 2.4; p < 0.0001) or patients pretreated with other BPs (3.2 ± 2.4; p= 0.0041) and analgesic use was less frequent (38% vs 49% and 47%, respectively).In patients reporting pain at baseline, mean pain severity decreased on IBA treatment throughout the observation period (VAS: 3.2 ± 2.4 to 2.4 ± 2; p < 0.0001). In parallel, there was an overall reduction of analgesic use with 57% of patients not requiring analgesics at the end of the observation period. SRE were rare (7%), irrespective of prior treatment. For patients evaluable for renal function throughout the observation period, mean eCrCl at baseline was significantly lower in patients pretreated with ZOL (70.7 ml/min) than in BP-naïve (78.9 ± 30.9 ml/min; p < 0.0001) or IBA-pretreated patients (78 ± 29.1 ml/min; p=0.00386). Changes in renal function during IBA treatment were small and balanced across all subgroups. Overall tolerability of IBA treatment was rated as very good or good by 96% of physicians and patients alike.Conclusion: In this interim analysis of a large scale NIS in BC patients with MBD, IV and oral IBA showed marked and sustained pain relief with an overall reduction of analgesic use and a renal safety profile comparable to results of RCTs. Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 1108.

Background: Based on interim analysis of Chemo N0, the first prospective multicenter randomized clinical trial using biomarkers uPA/PAI-1 for risk stratification and chemotherapy selection in N0 breast cancer, uPA/PAI-1 are now recommended for routine use by ASCO and AGO guidelines. Final 10-year follow-up of Chemo N0 has validated the clinically relevant long-term prognostic and predictive impact of uPA/PAI-1 in N0 breast cancer. To quantify the clinical utility of this impact, outcomes in subgroups defined by these biomarkers were compared to predictions of Adjuvant Online™, the leading validated tool based on established clinical factors. Methods: In Chemo N0, patients (n=647, 1993-98) were prospectively stratified according to uPA/PAI-1: High-risk patients were randomized (6x CMF vs. observation), low-risk patients observed. Retrospectively, we divided recruited patients into two groups (chemotherapy vs. no adjuvant therapy) and stratified using uPA/PAI-1 (low vs. high). Individual 10-year OS was calculated by www.adjuvantonline.org (Version 7.0); these estimated values were compared to the observed Chemo N0 10-year OS. Results: 383/647 patients have complete 10-year follow-up data. Median 10-year OS estimated by Adjuvant Online™ was 80.5% taking into account administered adjuvant therapy; 10-year Chemo N0 follow-up revealed observed 10-year OS of 74.7% (97/383 died). 77.8% (298/383) did not receive chemotherapy: In high-risk, untreated patients (n=137), 10-year OS was 72.3% vs. estimated 81.1%. In CMF-treated patients (all high-risk, n=85), observed OS was 61.2% vs. estimated 76.4%. Conclusions: For the first time, risk assessment by novel biomarkers is compared to that by Adjuvant Online™ in final data from a randomized prospective clinical trial. In patients with high uPA/PAI-1, the individual 10-year risk calculated by Adjuvant Online™ seems to be underestimated compared to observed patient outcome. Tumor-biological information provided by validated biomarkers thus has the potential of enhancing epidemiology-based risk estimation using only established factors. Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 4038.

Background: A number of molecular signatures have been published to aid breast cancer prognosis and therapy response prediction. A 76-gene signature has been developed in a node-negative patient cohort that did not receive systemic therapy.1 Another prognostic 97-gene signature measures predominantly proliferation-associated genes.2 Finally, a 21-gene signature has been developed for node-negative and estrogen receptor–positive breast cancer patients treated with tamoxifen in the adjuvant setting.3 Here we compare the prognostic performance of all three published algorithms in a cohort of 189 node-negative breast cancer patients treated with tamoxifen.Materials and Methods: Fresh-frozen tumors from node-negative patients were profiled on HG-U133a arrays. In addition, HG-U133a datasets with clinical annotation were downloaded from GEO (http://www.ncbi.nlm.nih.gov/geo/). All patients received tamoxifen only as adjuvant treatment after surgery. We determined the molecular subclass on the basis of ESR1 and ERBB2 mRNA expression; only ESR1-positive and ERBB2-negative tumors were considered for further analysis, leaving 141 in-house and 48 public datasets. After mapping of the gene signatures to the HG-U133a platform, we performed Cox regression and ROC curve analysis with distant metastasis as endpoint.Results: Cox regression analysis yielded a significant outcome association for all three gene signatures (76-gene signature: P = 0.0018; 97-gene signature: P = 0.0294; 21-gene signature: P = 0.0025) in the whole ESR1+/ERBB2– cohort. However, the 97-gene signature did not yield a significant result by ROC analysis for distant metastasis at 5 years (76-gene signature: AUC = 0.645, CI = 0.505–0.785; 97-gene signature: AUC = 0.608, CI = 0.462–0.754; 21-gene signature: AUC = 0.744, CI = 0.633–0.854). In addition, when the analysis was restricted to grade 2 tumors (n = 114), only the 21-gene signature remained prognostic by Cox regression (76-gene signature: P = 0.2405; 97-gene signature: P = 0.9001; 21-gene signature: P = 0.047) as well as by ROC curve analysis at 5 years (76-gene signature: AUC = 0.575, CI = 0.398–0.751; 97-gene signature: AUC = 0.532, CI = 0.347–0.717; 21-gene signature: AUC = 0.699, CI = 0.549–0.849).Discussion: While all three gene signatures reveal a significant outcome association in our whole patient cohort by Cox regression analysis, only the 21-gene signature remains significant in a subset analysis of grade 2 tumors. Since this group is clinically particularly challenging, further comparison between prognostic gene signatures in homogeneously treated patient cohorts is needed. Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 104.

Background:Tumors of the luminal B subtype of ER postive breast cancer are characterized by high proliferation as compared to the luminal A subtype. The luminal B have a worse prognosis. We aimed to identify genes specifically expressed in the luminal B subtype of breast cancers and analyze the prognostic impact of these genes and their relationship to endocrine therapy.Methods:121 genes overexpressed in LumB tumors were identified in a test set of 171 Breast cancer samples and reproducibly obtained in four independent validation datasets. The scaffold protein NHERF1 was analyzed in a large scale meta-analysis of microarray datasets encompassing n=3030 breast cancer samples.Results:NHERF1 is an ER regulated gene located on chromosome 17 coding for a scaffold protein involved in growth factor signal transduction. NHERF1 expression among ER positive tumors is associated with larger tumor size, higher histolocigal grading, and HER2 expression. A prognostic value of NHERF1 was observed among ER positive tumors (univariate HR 1.49, 95% CI 1.23-1.80, P Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3164.

Background: Utilizing microarray based gene-expression analysis of fresh-frozen tissue we could recently demonstrate the prognostic impact of a B cell metagene in node-negative breast cancer (Cancer Res 68: 5405-5413, 2008). In the present study we investigated the prognostic significance of immunoglobulin kappa c (IGKC) in formalin-fixed paraffin embedded (FFPE) breast cancer specimens of 363 node-negative breast cancer patients which were not treated in the adjuvant setting.Methods: RT-PCR was used to analyze mRNA expression of IGKC in FFPE tissue of 363 patients with node-negative breast cancer. Additionally to IGKC we examined the prognostic impact of age, histological grade, tumor size, estrogen receptor (ER) and progesterone receptor (PR). Metastasis-free survival (MFS) was analyzed with uni- and multivariate Cox regression.Results: Median follow-up was 10 years. 69 patients (19%) developed distant metastasis. Univariate analysis showed significant results for histological grade (HR 2.2, 95% CI 1.54-3.19, p Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 4067.

Abstract #3114 Background: At present, academic medical centers conducting investigator initiated trials (IIT) have the choice of relying on paper or going electronic regarding electronic data capture and data management solutions. Many of the electronic systems have originally been developed for requirements of the pharmaceutical industry and all of the currently available systems would thus be isolated in the existing IT setting of the medical centers. We here suggest an approach to integrate a data management system into existing hospital information systems (HIS) and to provide the end users (e.g. physicians, investigators, study nurses) with consolidated user interfaces and integrated systems. Methods: In 2005, a collaboration between the Department of Obstetrics and Gynecology, Klinikum rechts der Isar, Technical University of Munich and Siemens Medical Solutions was initiated as a pilot project. An integrated platform for clinical trials was developed and implemented to support the HEDON trial (Herceptin-Docetaxel-Neoadjuvant), an investigator initiated clinical phase II trial concerning primary systemic therapy of breast cancer. The HEDON trial started in February 2006 and will have recruited all of a total of 90 planned patients in the end of this year. In addition, a translational research trial (TransHEDON) evaluating therapy response markers is an essential component of the clinical trial. Using the integrated platform, clinical data as well as laboratory data are automatically populated from the HIS into the electronical study CRFs, after data confirmation by investigators or study nurses (depending on their defined role) using the validation buffer of the system. The existing PACS and HIS are made accessible from the platform which has special capabilities to greatly improve user interactions with all relevant systems. To quantify the benefit of such an integrated system, an integrated evaluation project was set up including the quantification time savings for all involved personnel. For a pre-defined time period, data capture and data management processes are performed paper based and electronically. This allows thorough and critical analysis of all involved processes Resukts; Errors that occur specially due to manually entering laboratory data can be dramatically minimized using such an electronical data capture system. Another important benefit is time savings. The data will be presented. Conclusion:This single-sign-on electronic platform for clinical trials has been validated in its whole and fulfils the regulatory requirements as specified by GCP or 21 CFR part 11 compliance for trials of the FDA. We consider this integrated system to be very helpful for implementation of clinical trials into a routine hospital setting. Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 3114.

Abstract #1092 The American Society of Clinical Oncology 2007 Update of Recommendations for the Use of Tumor Markers in Breast Cancer suggests the use of the invasion markers urokinase-type plasminogen activator (uPA) and its inhibitor PAI-1 for risk assessment (Harris et al. JCO 25:5287). In 2003, we launched the NNBC 3-Europe trial for node negative breast cancer patients with the following questions: 1) Comparison of risk-assessment: invasion markers uPA/PAI-1 vs. clinico-pathological factors. 2) Comparison of adjuvant FEC-Docetaxel (anthracycline-taxane sequence) vs. standard FEC in high-risk patients? 3) Use of fresh tissue sampling was necessary within this trial. Quality assurance for the testing procedure was prospectively performed. Study Design: Risk assessment is performed either by clinico-pathological factors or by uPA/PAI-1. High-risk patients receive adjuvant chemotherapy according to randomisation. Adjuvant endocrine therapy is given according to guidelines. All laboratories take part on the central quality assurance program. In addition, we also analyzed feasibility of uPA/PAI-1 determination from core needle biopsies. Results:To date, from 131 participating centres, 3060 patients have been registered for the trial. Biological risk assessment was performed in 2000 patients. Using both tumor grade and uPA/PAI-1 results, the low-risk group comprises 38% of the patients, whereas 62% were considered high-risk and randomized for the adjuvant chemotherapy question. All centres show a well established collaboration with the pathologists who were responsible for histological confirmation of the frozen tumor samples and for the weekly shipment to the central labs. Mean coefficients of variation (CV) for between-lab and within-lab are 12% for uPA and PAI-1. Using core needle biopsies (n=36) in comparison to larger tumor pieces, we found a rather good correlation (r=0.89; p Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 1092.