bluuebunny/crossref_metadata_2025 huggingface dataset:
P4-09-05: CA15-3 Adds Prognostic Information in “Luminal” Type Breast Cancer – A Single Center Experience Evaluating 700 Patients with a Median Follow-Up of 5 Years.
Authors: Kantelhardt EJ , Vetter M , Steer S , Ruider T , Holzhausen H-J , Strauβ H-G , Groβe R , Thomssen C
Published: June 2012

Background: Prognostic markers are essential for the decision about individual therapy for patients with newly diagnosed breast cancer. Biological meaningful cancer types are revealed by gene expression analysis. Steroid hormone receptor (HR) and HER2 status of the tumor by immunohistochemistry (IHC) are more easily available and predominantly resemble these cancer types. The “luminal” type as HR pos. and HER2 neg., the “luminalHER2” type as HR pos. and HER2 pos., the “HER2” type as HR neg. and HER2 pos. and the “triple neg.” type as HER neg. and HER2 ***neg.. We evaluated preoperative serum CA 15–3 and CEA within IHC-cancer types and outcome using our own cohort from 1999–2010.

Material and Methods: Since 1999 all patients with breast cancer were entered in our tumor registry. Patients were treated by surgery and adjuvant therapy according to national guidelines (www.ago-online.de). Data was entered into SPSS by a specially trained study nurse. Follow-up was obtained yearly using our own out-patient clinic, information from general practitioners and the general cancer registry. Informed consent was taken from the patients at time of diagnosis. CA 15–3 and CEA were defined elevated if above 25 U/ml or 4,6 μg/l respectively.

Results: Preoperative serum CA15-3 available for 1149 patients. Patients with elevated results showed a reduced 5year disease-free survival (DFS) of 74.4% as compared to 84.7% (p<0.001). CA15-3 remained an independent prognostic factor in multivariate analysis (nodal status, grading, HR). Within HR positive disease, normal CA15-3 values predict a significantly better 5years DFS of 89.9% compared to 79.7% (p<0.001). No significant difference was seen in HR negative patients (5year DFS 57.5% vs 68.4%; p=n.s.). Patients with HER2 status available (n=700) were classified into biological tumor types by IHC. Significant differences in DFS were seen for “luminal” (n=435) tumors only (5year DFS 81.0% vs 91.8% (p<0.001). Only trends of differences in DFS were seen in the other less frequent tumor types “luminalHER2” (n=90) or “triple neg.” (n=112) types. Within the “HER2” (n=63) group no discrimination by serum levels of CA15-3 was seen. CEA did not add information on prognosis.

Discussion: Decision on adjuvant therapy is increasingly based on tumor biology. Particularly, in “luminal” tumors additional prognostic information is needed to decide on adjuvant chemotherapy. CA15-3 may be an easily available, independent marker to identify patients with a higher risk of recurrence within this group. Prospective validation and comparison to molecular typing is needed.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-09-05.
P4-09-07: Breast Cancer Outcome by Combined Immunohistochemical ER/PR/HER2 Receptor Phenotype.
Authors: Brouckaert O , Laenen A , Reynders A , Vanderhaegen J , Amant F , Leunen K , Smeets A , Berteloot P , Van Limbergen E , Weltens C , Moerman P , Paridaens R , Wildiers H , Vergote I , Christiaens M-R , Neven P
Published: June 2012

Introduction: We previously reported that molecular prognostic subgroup classification of operable breast cancers resembles the classification based on combined immunohistochemical expression of ER, PR and HER-2 receptors. This update represents a larger cohort with more detailed follow-up (median 5.5y).

Patients and methods: A prospective cohort of 4334 breast-cancer patients primary operated at our institution between 2000 and 2009. Steroid receptors were considered positive for any nuclear staining, HER-2 for strong (3+) membrane staining or positive fluorescence in situ hybridization. Survival curves were calculated for six predefined breast cancer subgroups: ER + PR + HER-2 - (PPN), ER + PR - HER-2 - (PNN), ER + PR + HER-2 + (PPP), ER - PR - HER-2 - (NNN), ER - PR - HER-2 + (NNP), and ER + PR - HER-2 + (PNP) (figures not allowed in abstract). Disease free interval (DFI), distant metastatis free interval (DMFI), overall survival (OS) and breast cancer specific survival (BCSS) were calculated. A long-rank test was used for testing the null hypothesis of no difference between the groups.

Results:

Conclusion: Combined receptor expression for ER/PR/HER2 determines specific breast cancer subtypes with significant different clinical behavior. Both predictive and prognostic factors need to be considered when interpreting these results.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-09-07.
P4-09-13: External Validation of Adjuvant! Online Breast Cancer Prognosis Tool. Improvement Is Still Needed.
Authors: Hajage D , De Rycke Y , Bollet M , Savignoni A , Caly M , Pierga J-Y , Horlings HM , Van de VjverMJ , Vincent-Salomon A , Sigal B , Senechal C , Asselain B , Sastre X , Reyal F
Published: June 2012

Introduction: AdjuvantOnline is a web-based application designed to provide 10 years survival probability patients with breast cancer. Few validation studies have underlined some limitations, particularly an overestimation of the prognosis among certain subgroups of patients. Moreover, several predictors such as HER2 over expression status and proliferation markers have not been assessed in Adjuvant! original study. We provide the validation of AdjuvantOnline algorithm on two breast cancer datasets collected from two large European cancer centres, and we determined whether the accuracy of AdjuvantOnline is improved by others well known prognostic factors.

Material and Methods: The French data set is composed of 456 women with early breast cancer, treated at the Institut Curie between 1995 and 1996. The dutch data set is composed of 295 women less that 52 years treated at the Netherlands Cancer Institute between 1984 and 1995. Agreement between observation and Adjuvant! prediction was checked by testing that the calibration slope was equal to 1. Logistic models were performed to evaluate whether risk factors adds significant prognostic information, including AdjuvantOnline a priori information as an offset.

Results: Ten years survival status was known for 383 patients in the French data set and 247 patients in the Dutch data set. Adjuvant! prediction was globally well calibrated in the French data set (observed survival 86%, predicted survival 85%), but was overestimated in high grade, HER2 positive and Ki67 > 20% subgroups. HER2 status, Mitotic Index, Ki67 and treatment type were strongly associated with 10-year survival, even considering AdjuvantOnline a priori information. In the Dutch data set, the overall 10-year survival was overestimated by AdjuvantOnline (observed 66%, predicted 79%), particularly in patients less than 40 years old.

Conclusion: AdjuvantOnline needs to be updated to adjust overoptimistic results in young and high grade patients, and should consider candidates, such as Ki67, HER2 and Mitotic Index.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-09-13.
P4-09-15: Clinical and Pathological Predictors of Outcome among Patients with Triple Negative Breast Cancer.
Authors: Pearlstone DB , Gray M , Garofalo RM , Trapani M , Nyirenda T , Hazelwood VA
Published: June 2012

Background

Triple negative (TN) breast cancer portends a poor prognosis, yet outcomes remain heterogeneous. Specific characteristics of individual TN patients that can predict outcome compared to non-triple negative (NTN) patients have yet to be well defined.

Methods: A prospective oncology database at Hackensack University Medical Center was queried for all patients with invasive breast cancer between 2000 and 2005. Patients with triple negative breast cancer were identified and compared to non-triple negative patients. Numerical values were compared by Mann-Whitney test; categorical values by Chi square/Fisher exact test. Log-rank test was used to compare outcome statistics, univariate and multivariate analyses were used to identify factors affecting relapse-free survival and overall survival.

Results: A total of 2,216 patients were identified (TN: n=260; NTN: n=1956). There was no difference in race (white vs non-white; TN: 82.3% vs NTN: 87.3%) or family history of breast cancer (TN: 19.2% vs NTN: 16.8%). TN patients were significantly younger (median age; TN: 55.5 vs NTN 57.7; p< .05), had larger tumors (mean cm; TN: 3.1 vs NTN: 2.4; p<.05), had higher grade tumors (% grade 3; TN:74.6 vs NTN: 26.1; p<.0001), had higher proliferation index (>20% nuclei stained for Ki-67; TN: 30.8% vs NTN: 6.7%; p<.0001), but were less likely to have lymph node metastases at presentation (TN: 35.7% vs NTN: 48.0%; p < .0001). Among node negative patients, overall survival (OS) and disease free survival (DFS) were not significantly different between TN and NTN. Among node positive patients, both OS and DFS were worse for TN patients compared to NTN patients (p < .0008). By univariate analysis, age, nodal status, tumor size and tumor grade were all independent predictors of DFS, but proliferation rate and TN status were not. In a multivariate analysis, however, age, TN status, nodal status and tumor size were all significantly associated with DFS.

Conclusion: Although TN patients have poor outcomes overall, among node negative patients, TN status does not affect overall nor disease free survival. Among node positive patients, however, TN status has a significant impact on outcome. In this analysis, well established predictors of outcome, including age, tumor size, tumor stage and nodal status were all independent predictors of outcome, but TN status and Ki-67 level were not.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-09-15.
P4-09-22: Multicenter Quality Assurance Profile Review of Lobular Breast Carcinomas Versus the 21 Gene Recurrence Score: Assessment of Clinical Relevance.
Authors: Dabbs DJ , Hicks D , Tubbs R , Bhargava R , Brufsky A
Published: June 2012

Introduction

The oncotype dx (ODX) 21 gene recurrence score (RS), was intended to predict the 10 year risk of breast cancer recurrence for estrogen receptor positive, lymph node negative disease (NEJM 2004;351:2817–26). The proportions of patients that were reported as low, intermediate and high risk were 51%/22%, 27%. This population of patients was dominated by ductal carcinomas (DCA), with a paucity of classical lobular carcinomas (CLCA), which are ER-rich and low grade histology. To date, there is dearth of data regarding the relationship of the RS and CLCA, and the clinical relevance regarding the ODX RS in CLCA has not been studied.

Methods

As part of quality assurance review at three institutions, the ODX RS for all CLCA were reviewed in order to determine the perceived clinical relevance of the ODX RS compared to the proportions of RS reported (NEJM 2004;351:2817–26).

Results

A total of 219 cases of ODX were performed on CLCA. The low/intermediate/high risk profile was 58%/40%/2%.

Conclusions (1) The risk profile of ODX for CLCA is very different from the original DCA dominant population previously reported. (2) If the goal of ODX is to identify candidates for chemotherapy, then a high RS is a rare event for CLCA, and as such, the judgment to administer chemotherapy to an individual patient would most likely depend upon factors other than the ODX, because 98 percent of patients are combined low/intermediate risk RS.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-09-22.
P4-10-09: Hormone Replacement Therapy: A Benign Breast Disease's Risk?
Authors: Hamy AS , Gronier H , Porcher R , Giacchetti S , Cuvier C , De Roquancourt A , Espie M
Published: June 2012

Background Use of hormone replacement therapy (HRT) has been associated with increased risk of breast cancer. However relations between use of HRT and benign breast lesions remain contradictory (1-3). Our goal was to investigate whether HRT and its duration use are associated with benign breast lesions.

Materials & Methods From 2001 to 2007, 2708 patients with a non palpable breast lesion were referred to our breast care center. Radiological abnormalities were screened either by breast ultrasound mammography, or MRI. Of those, 1 329 patients had a biopsy in our Radiology Unit. We focus on the postmenopausal women (n=739). Clinical data including HRT use extent were reported. Patients with previous history of benign breast disease or malignancy were excluded.

Results The analysis was performed on 739 patients. Biopsies yielded a benign result in 414 (56%) of them (fibroadenoma n=71, blunt duct adenosis n=94, fibrocystic disease n=92, epithelial hyperplasia n=56, others n=101), high risk lesions in 57 (8%) (atypical ductal hyperplasia n=28, atypical lobular hyperplasia n=19, lobular carcinoma in situ n=10) and malignancy in 268 (36%) (not studied here). For the 471 patients with benign or high risk breast lesion, the median age was 59 years, 80% had at least one pregnancy and 27% had a family history of breast cancer. Among these 471 patients, 47% of them had used an oral contraceptive and 53% of them a HRT, with a mean duration of use of 2 years.

The HRT use was not significantly associated with the type of benign or high risk breast disease. No association was observed either between breast disease and duration of HRT (less or more than 7 years respectively).Older age was significantly correlated to atypia or carcinoma (median age at diagnosis 59 y.o in benign lesions, 60 y.o in high risk lesions and 62 y.o in malign lesions, with significant difference) (p<0,0001).

Conclusion In this study of postmenopausal women, HRT and its duration use were not associated with the type of benign or high risk breast lesion. Although a lack of statistical power may be invoked to explain the results, we are called to believe that the magnitude of an effect of HRT use is small, if ever it exists.

1. Finkelde DT, Kitchen PR, Hayes PR, McKinlay MR, Henderson MA 2001 Symptomatic benign breast disease and hormone replacement therapy. Breast 10:127–130

2. Gayet A, Esteve J, Seradour B, Piana L, Jacquemier J 2003 Does hormone replacement therapy increase the frequency of breast atypical hyperplasia in postmenopausal women? Results from the Bouches du Rhone district screening campaign. Eur J Cancer 39:1738–1745

3. Rohan TE, Miller AB 1999 Hormone replacement therapy and risk of benign proliferative epithelial disorders of the breast. Eur J Cancer Prev 8:123–130

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-10-09.
P4-11-05: Association between Bisphosphonate Use in Metastatic Breast Cancer (MBC) and Overall Survival.
Authors: Mathew A , Mathew IE , Rosenzweig MQ , Brufsky AM
Published: June 2012

Background

Pre-clinical studies on bisphosphonates in breast cancer have suggested an anti-tumor effect in addition to its bone protective role. However, randomized controlled trials of bisphosphonates versus placebo have found little evidence of increased overall survival (OS) in MBC. We conducted a retrospective single institution cohort study of MBC patients to evaluate the association between bisphosphonate use and overall survival.

Methods: Baseline demographic and tumor specific data were collected on newly diagnosed MBC patients between January 1998 and December 2009. Other variables included the number and sites of each metastasis, use of baseline neoadjuvant and adjuvant chemotherapy, and use of hormonal therapy. Bisphosphonate use was defined as present if it was administered for a period of at least 3 months in the metastatic setting. Overall survival was determined from the date of diagnosis of first metastatic disease. Survival analysis was performed using the Kaplan-Meier method and Cox proportional-hazards model.

Results: Data were available on 737 patients with MBC, of whom 434 died during a median follow-up of 2 years; median age was 50.3 years. 92% of patients were Caucasian; 32% were both ER and PR-negative; and 31% were HER2−positive. Over 67% of MBC patients had bone metastasis and nearly 80% received bisphosphonates. Multivariate analysis found an overall survival benefit for bisphosphonate use, with a hazard ratio of 0.63 (95% confidence interval: 0.48−0.84; p<0.002), when adjusted for variables with significant effect on survival on univariate analysis and other known prognostic variables. These variables include age, stage at diagnosis, race, hormone receptor status, HER2 status, and number of metastatic sites, presence of bone metastasis and the use of adjuvant therapy. The administration of adjuvant therapy did not yield a significant survival advantage in the analyses.

Conclusion: This retrospective cohort study provides evidence for an OS benefit with the use of bisphosphonates in MBC even after controlling for other significant prognostic factors.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-11-05.
P4-11-07: Feasibility and Acceptability of Offering Breast Cancer Risk Estimation in the Context of the UK National Health Service Breast Cancer Screening Programme: A New Paradigm for Cancer Prevention.
Authors: Evans DGR , Astley S , Stavrinos P , Sahin S , Ingham S , McBurney H , Eckersley B , Wilson M , Beetles U , Harvie M , Warren R , Sergeant J , Hufton A , Warwick J , Newman W , Buchan I , Cuzick J , Howell A
Published: June 2012

Background: Currently there are no real attempts internationally to tailor breast screening programmes to individual risk Methods: We have assessed the feasibility of collecting breast cancer risk information during routine mammographic screening in the National Health Service Breast Screening Programme (NHSBSP) in England, in order to consider, ultimately, adapting the screening interval to risk of breast cancer and introducing preventive strategies in women at high risk. The study Predicting Risk Of Cancer At Screening (PROCAS) aims to recruit 60,000 women over 3 years. Results: 26,000 women (June 8th 2011) have so far given consent to join the study. Thirty six percent of the first 20,000 women in nineteen screening sites in Manchester consented to enter the study and completed a risk factor questionnaire. The median 10 year breast cancer risk was 2.65%, with 926 (9.26%) of the first 10,000 women having a 10 year risk of ≥5% and 92 (0.92%) having a 10 year risk of ≥8% (Tyrer-Cuzick), IQR:1.35. 832 (8.32%) women had a mammographic density of 60% or greater (Visual Analogue Scale). We collected saliva samples from 1019 women for genetic analysis and will extend this to 18% of participants. Of those who agreed to participate in the study, 94% indicated that they wished to know their breast cancer risk. Women with a 10-year risk of ≥8%, and women with a 10-year risk of ≥5% and mammographic density ≥60% were invited to attend or be telephoned to be counselled. To date 138 have accepted with 135, so far, having received risk counselling. Nineteen percent of the high-risk women identified subsequently decided to enter a randomised breast cancer prevention study with either a dietary or drug intervention (IBIS2, anastrazole vs placebo). Results from the first 1,000 women who provided DNA samples suggest that the risk information from the 18 validated SNPS may enhance existing risk models. Conclusion: This study demonstrates that it is feasible to determine individual breast cancer risk and offer women appropriate risk-reducing interventions within the context of a population-based mammographic screening programme.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-11-07.
P4-11-08: Changes in the Distribution of Loco-Regional and Distant Breast Cancer Recurrences over the Last 20 Years: Implications for Patient Care and Future Research.
Authors: Bouganim N , Clemons M , Amir E
Published: June 2012

Introduction: Improvements in adjuvant therapy have led to a sustained fall in recurrences after early breast cancer. The differential reduction of both local-regional and systemic recurrences is poorly understood. This study aimed to explore changes in the distribution of loco-regional and distant recurrences in clinical trials reported over the last 20 years. We also aimed to determine the relative impact of adjuvant chemotherapy and endocrine therapy.

Methods: A MEDLINE search for adjuvant, Phase III randomized breast cancer clinical studies between January 1990 and March 2011 was performed. Neo-adjuvant, single agent biologics and studies that did not report the proportion of loco-regional and distant recurrences were excluded. Change in the frequency of recurrences was assessed as the non-parametric correlation between the number of loco-regional recurrences (as a proportion of all recurrences) and time. Studies were weighted by sample size. Pre-specified subgroup analyses were assessed using the interaction test and included type of surgery performed, radiotherapy use, menopausal status and type of systemic therapy delivered. Definition of local and distant recurrences differed between studies. For consistency, loco-regional recurrences were classified as recurrences limited to the ipsilateral breast, chest wall, axillary, supraclavicular and internal mammary lymph nodes. Any other recurrence was defined as distant, with the exception of contralateral breast cancer; that was excluded from this analysis.

Results: Fifty-three randomized clinical trials with a total of 86,598 patients were included in the analysis. Between 1990 and 2011, the proportion of loco-regional recurrences has decreased from approximately 50% to 10% (Spearman's rho = −0.40, p<.001). There was no interaction between type of surgery (mastectomy vs. lumpectomy, p=0.40), adjuvant radiotherapy use (p=0.63) and menopausal status (p=0.95) and the correlation of loco-regional recurrences and time. Chemotherapy use showed a larger negative correlation compared with endocrine therapy (rho = 0.49 vs rho = 0.24, p=0.008).

Conclusion: Advances in treatment of early breast cancer have differentially reduced the proportion of loco-regional recurrences compared with distant recurrences. In recent trials, loco-regional recurrences account for less than 10–15% of all recurrences. These falling event rates may affect patient care, especially when deciding on treatments influencing loco-regional control. This change may also impact on the design of clinical trials assessing loco-regional therapy such as surgery and/or local radiation therapy.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-11-08.
P4-11-12: Molecular Phenotype of Breast Cancers in a Large Cohort of Young Women According to Time Interval Since Pregnancy.
Authors: Collins LC , Gelber S , Marotti JD , Cole K , Kereakoglow S , Ruddy KJ , Brachtel EF , Schapira L , Come SE , Borges VF , Schedin PJ , Warner E , Winer E , Partridge A
Published: June 2012

Background: The increase in breast cancer risk during pregnancy and post partum is well recognized. The cross-over to protective effect does not occur until many years later and varies with age at first birth. Recently, a genomic signature specific to the pregnant compared with the non-pregnant breast has been identified; this signature remains present in the postmenopausal parous breast. Given this, we investigated whether time interval since pregnancy affects the phenotype of breast cancers arising in young women compared with nulliparous women. Methods: We examined molecular phenotype, according to histologic grade and biomarker status, in relation to time since pregnancy in an ongoing prospective cohort study (n=355) of young women (≤40yrs) with breast cancer. Medical records were reviewed for tumor stage and receptor status. Parity was ascertained from questionnaires completed within 9 months of diagnosis. Tumor grade was determined by central pathology review. Using tumor grade and biomarker expression, cancers were categorized as luminal A (ER+ and/or PR+, HER2−, histologic grade 1 or 2); luminal B ( ER+ and/or PR+, HER2+, or ER and/or PR+, HER2− and grade 3); HER2 type (ER-, PR-, HER2+); and triple negative (ER-, PR-, HER2−).

Results: The median age of the study population is 37 years (range 17–40). Overall, 80% of women had stage 1 or 2 disease; 67% of cancers were ER positive and 32% showed HER2 overexpression. The distribution of breast cancer molecular phenotypes by time interval since last pregnancy is shown in the table.

Distribution of molecular phenotype by interval between last pregnancy and diagnosis

In our large cohort of parous young women, we found no differences in the distribution of molecular phenotype according to time interval since pregnancy. However, nulliparous young women were more likely to develop luminal A cancers compared to parous women (40% vs. 29%; unadjusted chi square p-value=0.03) and appeared less likely to develop HER2−type and triple negative cancers (7% vs. 13%, p-value=0.09 and 17% vs. 23%, p-value=0.22 respectively). There were no differences in the distribution of luminal B cancers. Conclusions: The distribution of molecular phenotypes is similar among parous young women regardless of the time interval since parturition. Nulliparous young women appear more likely to develop luminal A cancers compared to parous women. Whether the difference in molecular phenotypes of pregnancy-associated breast cancers vs. cancers arising in nulliparous women is due to the effects of genomic alteration remains to be investigated. Effects of a prior pregnancy appear consistent across a 5-year period, in keeping with the concept of genomic alterations identified in the normal pregnant breast and thereafter.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-11-12.
P4-12-15: Cancer Survivor Care: An Evaluation of a Group Visit Model of Care for Breast Cancer Survivors at Duke Cancer Center.
Authors: Trotter KJ , Schneider S
Published: June 2012

Background: The number of cancer survivors is growing, currently with about 2.5 million breast cancer survivors. (ACS, 2010). The Institute of Medicine report on survivor care stressed care coordination along with attention to survivor concerns as key issues in improving follow-up care. Group visits have been shown to improve access, health outcomes and health care utilization rates, as well as self-management skills.

Purpose: This prospective program evaluation measured patients opinions about participation in a group visit clinic.

Methods: A new model of breast cancer survivor care was designed and piloted at Duke Cancer Center. Breast cancer survivors who were three years post diagnosis attended the clinic together in groups of six. An interdisciplinary group visit format in the initial part of the appointment provided surveillance, education and support, as well as formation of an individualized survivorship care plan to be shared with the primary care provider. The first hour included review of their personal care plan and a 45-minute facilitated discussion. Afterwards, individual visits with the nurse practitioner, and dietitian, physical therapist or social worker occurred.

A 22-item questionnaire designed to ascertain opinions regarding logistics and the style and function of care delivered is being administered to participants. The evaluation tool was approved by a panel of experts; pilot tested and has a reading level of 7.8. Data collection will be complete July 1, 2011. 103 surveys have been collected with a 91% response rate. Preliminary analysis shows high satisfaction with the group visit format. What participants liked most about the program was sharing with other survivors. 86% of participants indicate that they will share the survivorship care plan with their PCP. Only 10% responded that they were not sure they needed this type of follow up now and expressed that it would be more helpful immediately after finishing treatment. Final data will be presented. Barriers and facilitators to implementing this type of care model will be presented.

Conclusions: The group visit model applied to a breast cancer survivor clinic appears feasible and highly satisfactory to participants. Implications for practice: Group visits can be considered one option for survivor care.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-12-15.
P4-16-03: Proof of the Anti-Tumour Effect of Zoledronic Acid (ZA) in Naive Bone-Only Metastatic and Locally Advanced Breast Cancer: Results from the Biological Window Therapy.
Authors: Foroni C , Andreis D , Maldotti M , Cappelletti MR , Generali DG
Published: June 2012

Background: Pre-clinical studies have demonstrated an anti-tumour activity of ZA in several cancers. However, the clinical evidence of the ZA anti-tumour effect is still uncertain. Our study aimed to prove that ZA has anti-tumour activity administered alone as a biological window of 14 days therapy on naïve bone-only metastatic and locally advanced breast cancer.

Material and Methods: 27 patients with locally advanced breast cancer (Group 1) and 12 patients at their first relapse with bone metastasis only (Group 2) received 4 mg single dose of ZA before starting with any treatment. In Group 1, Ki67 expression was evaluated in tumour specimens obtained before and after ZA administration (basal, day 14). In Group 2, circulating tumour cells (CTCs) were evaluated at baseline, 48 hrs and day 14. In Group 1 and 2, the apoptosis and necrosis of tumour cells were tested on blood samples by M30 and M65 ELISA assay respectively. Parallel, the anti-antitumor activity of ZA, given at the dose of 250 mg/mouse every 4 days for 6 cycles, in MDA-Mb-231 bearing mice was evaluated.

Results: A significant reduction of Ki67 expression after 14 days (mean, 25.33% to 21.78%; p=0.035) was observed in Group 1. In Group 2, we detected a significant reduction of CTC number after 48 hrs (median, 16 to 7; p=0.045), followed by a tendency to increase on day 14 (p=0.093). The M30/M65 analysis performed on paired blood samples in Group 2 showed after 14 days a significant increase in M65 (median, 97 to 112 U/L; p=0.018), more accentuated than the concomitant increase in M30 (p=0.308). In MDA-MB-231 bearing mice, ZA treatment caused a significant inhibition of tumor growth.

Conclusions: These results are the first prospective clinical data showing a direct anti-tumour effect (either on the tumour or on CTCs) of ZA, confirming the in vitro data. By directly affecting the proliferation, ZA could have an anti-tumour effect via induction of necrosis, suggesting its possible use in combination with conventional oncologic treatments of breast cancer.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-16-03.
P4-17-03: Incidence Rate of Asymptomatic Brain Metastases in Patients with HER2+ Metastatic Breast Cancer Screened for EGF111438/CEREBEL Study.
Authors: Pivot X , Hackmann J , Manikhas A , Moore Y , Parikh R , Kothari D , Joshi A , Aktan G , Coleman R
Published: June 2012

BACKGROUND Brain metastases constitute a major clinical concern occurring in 28–43% of women treated for HER2+ metastatic breast cancer (MBC). The initial diagnosis of Central Nervous System (CNS) lesion is commonly based upon development of neurological symptoms. Data to address incidence of CNS lesions in asymptomatic patients are rare and not generally reported. EGF111438/CEREBEL study requires systematic screening for brain metastases by brain MRI and the ineligible patients due to detection of asymptomatic brain metastases are reported here.

METHODS The patients screened for CEREBEL have consented to a multi-centre, open-label Phase III study in subjects with HER2+ metastatic breast cancer who received prior treatment with anthracyclines or taxanes and trastuzumab in the adjuvant or metastatic setting. Eligible subjects must not have any history of CNS metastases (including leptomeningeal involvement) or have evidence of benign or malignant brain tumours and/or clinical evidence of spinal cord metastases at baseline. In order to confirm this, subjects must have negative brain MRI at baseline as confirmed by independent review (IR). All MRI scans of the brain had to be acquired with gadolinium contrast agent (T1 only) and 3mm slice thickness without gaps. The primary endpoint of CEREBEL is the incidence of CNS lesions as first site of relapse. A total of 650 subjects (325 per group) are expected to be randomized between capecitabine + trastuzumab versus capecitabine + lapatinib.

RESULTS The study is currently recruiting with 322 patients enrolled as of 15.06.2011. At this date, the total number of patients screened were 492 and 170 of these patients were screen failures. There are 15 patients in screening. The percentage of screen failures due to asymptomatic brain metastases assessed by investigators was 13.5% (23/170). The overall incidence of clinically asymptomatic brain metastases assessed by investigators for this study is 4.7% (23/492).

CONCLUSIONS Around 5% of all screened patients (14% of screen failures) in this study who were clinically thought to be free of brain lesions actually had brain metastases verified by brain MRI. We therefore conclude that screening for occult brain metastases by MRI is important for trials with defined CNS endpoints.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-17-03.
P4-18-01: Prognostic Role of Tumor and Stromal p16/INK4A in Ductal Carcinoma In Situ (DCIS).
Authors: Linke SP , Bremer TM , Wärnberg F , Zhou W , Goldstein L , Jirström K , Amini R-M
Published: June 2012

Background

DCIS is a pre-invasive breast disease diagnosed in ∼50,000 women/year in the US. The standard treatment recommendation is for mastectomy or breast-conserving surgery plus radiotherapy (BCS+RT). However, only ∼10% of BCS patients avoid recurrence due to RT, and RT provides little or no survival benefit. Thus, it is important to stratify patients by risk to help tailor treatment to maximize benefit and minimize unnecessary adverse effects and costs.

Material and Methods: Tissue microarrays that included tumor cores from all women diagnosed with pure DCIS between 1986 and 2004 in Uppland and Västmanland, Sweden (n=458) were stained and scored for p16/INK4A (clone E6H4) using immunohistochemistry (IHC). Semi-quantitative manual scoring was done separately in the tumor epithelial cells and peri-tumoral carcinoma-associated fibroblasts (CAFs). Based on previous reports, thresholds to define low and high p16 levels were set at 25% and 5% positive staining, respectively, for the tumor and stromal components. HER2 IHC scoring was reported previously. Clinicopathologic characteristics of the set of patients for which complete marker data was obtained (n=176) were not statistically different than the full set of patients. For survival analysis, local ipsilateral DCIS and invasive recurrences, as well as lymph node and distant metastases, were included as events (patients were censored if they had contralateral breast or other cancers as the first event).

Results: Patients were stratified into low and high risk groups using a multivariate model incorporating tumor p16, stromal p16, HER2, and margin status. Patients treated with BCS only (n=67) had a baseline 10-year recurrence rate of 31%. The model identified high and low risk groups with 10-year recurrence rates of 38% and 7%, respectively (hazard ratio=7.1, p=0.009). Similar results were achieved when BCS only and BCS+RT patients were combined (n=130). Interestingly, all six recurrence events in the set of patients who received full mastectomies (n=43) were in the model high risk group.

Discussion: Previous studies have shown mixed results when p16 was used as a single marker. Our results suggest a prognostic interaction between tumor and stromal p16, as well as HER2, that will form the basis of a more comprehensive multi-marker IHC-based prognostic panel to assess risk of recurrence in DCIS patients.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-18-01.
P5-01-16: The Detection of Circulating CK19 mRNA-Positive Cells in the Blood and the Mitotic Index of the Primary Tumor Have Independent Prognostic Value in Early Breast Cancer.
Authors: Tryfonidis K , Perraki M , Apostolaki S , Kafousi M , Agelaki S , Georgoulias V , Stathopoulos EN , Mavroudis D
Published: June 2012

Background: Previous studies have shown that the molecular detection of CK-19 mRNA in peripheral blood and the mitotic index of primary tumor have prognostic value in early breast cancer. The aim of the present study was to assess the association between these variables. Patients and Methods: The primary tumors of 223 operable breast cancer patients (92 premenopausal and 131 postmenopausal) were evaluated for the mitotic activity index (MAI) classified either as <5/10, 6–10/10 and >10/10 or <10/10 and >10/10 mitoses/hpf using a standardized protocol as previously reported (Baak JP et al, Breast Cancer Res Treat 2009, 115:241–254). Peripheral blood was also collected before the start and after the end of adjuvant chemotherapy for detection of CK-19 mRNA-positive cells by RT-PCR as previously described (Stathopoulou et al, Clin Cancer Res 2003, 9:5145–5151). Results: After a median follow-up of 118 months, 75 (33.6%) patients have experienced disease relapse and 56 (25.1%) have died of breast cancer. MAI was strongly associated with disease-free survival (DFS) and overall survival (OS) (p<0.001 for both DFS and OS). Detection of CK19 mRNA-positive cells in the peripheral blood before but not after adjuvant chemotherapy was marginally associated with worse DFS (p=0.055) and OS (p=0.059). There was no correlation between MAI and CK19 mRNA detection before chemotherapy. Cox regression analysis revealed that both MAI and CK19 mRNA-positive cell detection before adjuvant chemotherapy were independent variables associated with decreased DFS (p<0.001 and p=0.038, respectively) and OS (p<0.001 and p=0.029, respectively). The interaction test showed no significant association between MAI and detection of CK19 mRNA-positive cells.

Conclusion: MAI of the primary tumor and detection CK-19 mRNA-positive cells in the blood before the start of adjuvant chemotherapy in women with early breast cancer are two independent prognostic factors associated with clinical outcome.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P5-01-16.
P5-03-04: Circadian Modulation of Estrogen Receptor alpha Promoter Activity.
Authors: Villa LM , Santos CS , Cousins S , Finkielstein CV
Published: June 2012

The circadian clock is a molecular mechanism that synchronizes environment variations with physiological changes. Disruption of the circadian clock has been linked to increased risk in diseases and a number of disorders (e.g. jet lag, insomnia, coronary heart disease and cancer). Period 2 (PER2), a circadian protein, is at the center of the clock's function. The complete loss of per2 has been shown to be common in several types of cancer including breast and ovarian. Epidemiological studies established a correlation between circadian disruption and the development of estrogen dependent tumors. The expression of estrogen receptor alpha (ERa) mRNA oscillates in a 24-hour period and, unlike PER2, ERa peaks during the light phase of the day. Because up regulation of ER relates to tumor development, defining the mechanisms of ERa expression will contribute to our comprehension of cellular proliferation and regulation of normal developmental processes. The overall goal of this project is to investigate the molecular basis for circadian control of estrogen receptor alpha (ERa) transcription.

Transcriptional activation of ERa was measured using a reporter system in Chinese Hamster Ovary and MCF-7 cell lines. Data show that PER2 influence ERa transcription through a non-canonical mechanism independent of its circadian counterparts. Promoter analysis using the Alibaba 2.0 suite revealed the presence of various putative novel partners. Among them, OCT-1 was found to directly interact with Per2, in vitro, and with corresponding response element within the ERa promoter. The predicted binding site of OCT-1 is 267 bps upstream of the promoter. Pull-down assays were used to map direct interaction of various PER2 recombinant proteins and the DNA-binding domain of OCT-1. Radiolabeled OCT-1 was found to bind to the C-terminal end of PER2. Electrophoretic mobility shift assays were used to determine direct binding of OCT-1 to the promoter. ChIP assay confirmed the recruitment of PER2 to the estrogen promoter by OCT-1. Fluorescence anisotropy and surface plasmon resonance were employed to establish the binding affinity of OCT-1 to the ERa promoter and PER2. OCT-1 was found to have a high affinity for the ERa promoter (∼10 nM). Furthermore, OCT-1 recruits PER2 to the estrogen promoter. Preliminary data suggests that the transcription factor PER2 might play a role in the regulation of the estrogen promoter.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P5-03-04.
P5-04-01: BP1, a Homeotic Transcription Factor, Indirectly Upregulates ER-alpha in ER Positive Cell Lines.
Authors: Berg PE , Awate S , Kirolikar SP
Published: June 2012

Background: BP1 is a member of the homeobox gene family of transcription factors. Our recent studies have shown that BP1 may play a role in breast cancer cell survival, aggressiveness and metastasis. BP1 protein (pBP1) is expressed in 80% of invasive ductal breast tumors, including in 57% of ER positive and 89% of ER negative tumors. BP1 overexpression results in a more aggressive phenotype in breast cancer cells and larger tumors in women with breast cancer. We also observed the presence of mammary tumors in mice without estrogen supplementation after injection of MCF7 cells overexpressing BP1 compared with unsupplemented mice receiving MCF7 cells containing an empty vector. Our goal was to determine the pathway by which BP1 might cause this decrease in estrogen dependence of MCF7 cells. ER-alpha is a nuclear hormone receptor activated by estradiol and is a major target of anti-estrogen therapy. It is tightly regulated at the genomic and the non-genomic levels. Increased stability of ER-alpha is one mechanism by which cells can become ER-alpha independent. One way of altering ER-alpha stability is via p300 and BRCA1 since p300 acetylates and stabilizes ER-alpha, while BRCA1 destabilizes ER-alpha. We have previously shown that BP1 transcriptionally down-regulates BRCA1, suggesting that pBP1 may participate in this pathway.

Materials and methods: Protein levels of BRCA1, p300, pS2, pBP1 and ER-alpha were determined by Western Blot analysis. RNA levels were measured by real-time PCR. A ChIP assay was performed to confirm binding of pBP1 to the EP300 gene and binding of ER-alpha to BP1. MCF7 parental cell lines were treated with 10nM E2 to induce the expression of pBP1. Experiments involving effects of siBP1 on p300 expression are underway.

Results: Our data demonstrate that MCF7 ER+ cells engineered to overexpress BP1 exhibit increased ER-alpha protein, increased p300 and decreased BRCA1 protein as compared with control cells containing an empty vector and grown under the same conditions. Increased ER-alpha levels correlated with increased pS2 expression, a known downstream target of ER, demonstrating that there is more functional ER-alpha protein in cells overexpressing BP1. ChIP assays showed that pBP1 binds in the first intron of EP300-Histone acetyltransferase (p300). ChIP assays also showed that ER-alpha binds to an estrogen response element (ERE) upstream of the BP1 gene. MCF7 parental cell lines, when serum starved and treated with E2, showed increased BP1 expression compared with cells grown in charcoal stripped serum. Thus, BP1 may increase the stability of ER-alpha by decreasing BRCA1 and increasing p300. Conclusions: pBP1 binds and transcriptionally activates p300, leading to increased ER-alpha stability, while also partially repressing BRCA1. Moreover, ER-alpha, via a feed-forward mechanism, binds to the BP1 gene and increases its expression.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P5-04-01.
P5-05-04: Bipshenol-A Modulates Estrogen Receptor Alpha and Compromises Tamoxifen-Induced Apoptosis.
Authors: Goodson III WH , Luciani MG , Sayeed A , Jaffee I , Moore II DH , Dairkee SH
Published: June 2012

BACKGROUND: Antiestrogens are important for treatment and prevention of breast cancer. Such drugs influence multiple pathways involved in regulation of cell survival — including apoptosis. In previous work, exposure of human, benign, high-risk donor breast epithelial cells (HRBECs) to the xenoestrogen (XE) bisphenol-A (BPA) induced gene expression changes predicting disruption of apoptotic signaling (Dairkee et al, Cancer Research 2008). Therefore, we hypothesized that antiestrogenic, pro-apoptotic effects of tamoxifen are likely modulated under the influence of XEs. Because the environmental XE, BPA is routinely detected in a large percentage of random human blood samples, we employed functional tests to determine whether BPA interferes with tamoxifen-induced apoptosis in XE-exposed HRBECs.

METHODS: With IRB approval and written informed consent, HRBECs were obtained by random periareolar fine needle aspiration (RPFNA) from women who were high risk on the basis of personal or family history of breast cancer, breast density, or high-risk histology. Primary HRBEC cultures were treated for one week with a low concentrations of BPA corresponding to levels reported in human body fluids. Estrogen Receptor (ER)-α and -β were measured by Western blot (n=6). Altered response to tamoxifen toxicity was measured by analyzing quantitative changes in the Annexin V-positive apoptotic cell population (n=8).

RESULTS: After one-week exposure to BPA, ER-α increased in six of six cases, ER-β decreased in 5 of 6 cases, and we observed a 25 percent reduction in tamoxifen-induced apoptosis. Prior exposure to BPA at the high concentration range detected in the human population reduced tamoxifen efficacy by 50 percent. There was a dose response of evasion of apoptosis to increasing concentrations of BPA (P<0.001, two-sided test for slope fixed effect in a linear mixed effects regression). Prior exposure to another XE, methylparaben (MP), caused a similar dose-dependent decline in tamoxifen-induced apoptosis (p<0.001).

CONCLUSION: BPA and MP, in a range of concentrations measured in humans, prevent tamoxifen-induced apoptotic cell death. For BPA this involves upregulation of ER-α and downregulation of ER-β. Because these XEs are widespread in the American population, they may play a role in limiting the prophylactic and therapeutic efficacy of tamoxifen and other hormone-based therapies.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P5-05-04.