bluuebunny/crossref_metadata_2025 huggingface dataset:
Background Enhancement Patterns on Screening Breast MRI – Impact on Biopsy Rate and Short Interval Follow Up.
Authors: Hambly N. , Morris E.
Published: December 2010

Introduction:Marked background enhancement is one of the perceived limiting factors on breast MRI possibly obscuring suspicious lesions and reducing both the sensitivity and specificity of detection of small cancers. Background enhancement on MRI refers to the normal physiological enhancement of the woman's fibroglandular tissue and varies with age, hormonal status, menstrual cycle and breast glandularity. Patterns of enhancement vary greatly and marked enhancement can occur even when the MRI is performed in week 2 of the menstrual cycle or in postmenopausal women.Aim:To determine what impact background enhancement has on biopsy rate and short interval follow up rate.Method:231 consecutive baseline screening MRIs, in women aged between 16 -79 (average age 48) performed between Jan 2003 and Oct 2004 were selected. Indications for screening included significant family history (46.3%), personal history of contralateral mastectomy (25.1%), personal history of atypia (19.5%), other (3.0%), personal history of mantle radiation (2.2%), personal history of breast cancer > 5 years previously (2.2%), personal history of ovarian cancer (1.7%).The background enhancement in each case was described as minimal (< 25% glandular tissue showing enhancement), mild (25-50% enhancement), moderate (50-75% enhancement) and marked (> 75% enhancement).The BIRADS category, positive predictive value of biopsy and false positive rate was determined and compared for enhancement patterns.Results:Of 231 screening studies 26.6% demonstrated minimal enhancement, 32.8% mild enhancement, 22.7% moderate enhancement and 17.9% marked enhancement. Patients with minimal/mild enhancement had significantly higher rate of normal (BIRADS 1/2) examinations than those with moderate/marked enhancement (47.8% vs. 30.5%, p = 0.008). Patients with minimal/mild enhancement had significantly lower rate of six month followup (BIRADS 3) (38.97% vs. 53.68%, p = 0.02). No significant difference in biopsy rate was noted (11.7% minimal/mild enhancement vs. 14.7% moderate/marked enhancement, p = 0.5). No significant difference in positive predictive value of biopsy between the groups was noted (35.7% for moderate/marked enhancement vs. 25% for minimal/mild enhancement, p = 0.52).Conclusion:No significant association was found between background enhancement pattern and biopsy rate or false positive rate. However, patients with moderate/marked enhancement were significantly more likely to be given a category of BIRADS 3 and assigned to short interval follow up. The increased rate of BIRADS 3 in patients with moderate/marked background enhancement may be related to reduced reader confidence in these patients. Recognition of normal background enhancement patterns is important to limit the number of patients undergoing short interval followup.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 4020.
Dynamic Breast Magnetic Resonance Imaging in the Assessment of Breast Glandular Tissue in High Risk Patients after Prophylactic Ovariectomy.
Authors: Vilar V. , Dromain C. , Caramella C. , Delaloge S. , Caron O. , Uzan C. , Balleyguier C.
Published: December 2010

Purpose: To evaluate the enhancement of breast glandular tissue by dynamic magnetic resonance imaging (MRI) in high risk patients, before and after prophylactic ovariectomy.Materials and Methods: A retrospective study was performed on 28 patients with high risk for breast and ovarian cancer who underwent preventive bilateral ovariectomy. Seven patients who were at hormone replacement therapy and chemotherapy were excluded. From the 21 patients included, 12 were BRCA 1; 6 were BRCA 2; 1 was BRCA 1 – 2; and 2 patients had strong familiar history. All patients underwent contrast breast MRI before and after surgery, for breast cancer screening. MRI was performed at GE Healthcare 1.5 Tesla equipement before (mask) and after 5 phases of contrast injection. All pre contrast images were subtracted from the post contrast images. The second subtracted sequences of MRI images were analyzed qualitatively according to the type of glandular enhancement (1. no enhancement, 2. mild homogeneous, 3. strong heterogeneous, 4. strong homogeneous, 5. mild heterogeneous), and quantitatively by measuring signal intensity of glandular tissue in the upper outer quadrant of the breast in the second sequence of post-contrast images. Patients were divided into 2 groups: with and without the appearance of a lesion between the first and second MRI, for comparative analysis with the fibroglandular tissue.Results: Mean age was 44 years (39-51). In qualitative analysis at MRI before ovariectomy (MRI 1), 33% of the patients showed type 3 enhancement; 33% type 5; 29% type 4 and 5% type 1. At MRI post-ovariectomy (MRI 2), 67% of the patients presented enhancement type 1; 19% type 4 and 14% type 3. Twenty patients showed a higher enhancement at MRI 1 and for 1 patient the enhancement was the same at MRI 1 and 2. In quantitative analysis, the average of enhancement of the glandular tissue at MRI 1 was 356.1 and for MRI 2 was 65.2. Three patients presented an enhanced nodule on the first and second MRI. The average of enhancement of the lesions at MRI 1 was 890.9 and at MRI 2 was 452.9.Conclusion: Ovariectomy substantially reduces the physiological fibroglandular breast enhancement. The significant decrease of the glandular breast enhancement in MRI can help to detect organic focal breast lesion, such as atypical ductal hyperplasia and to differentiate it from functional foci. The comprehension of this changes can lead to decrease the number of false positives in breast MRI interpretation.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 4024.
Adjuvant UK: A UK Prognostic Model for Early Breast Cancer That Includes Mode of Detection.
Authors: Wishart G. , Azzato E. , Pharoah P. , Greenberg D. , Rashbass J. , Kearins O. , Lawrence G. , Caldas C. , Ravdin P.
Published: December 2010

AIM: To develop and validate a prognostication model to predict overall survival for women treated for early breast cancer in the UK based on cancer registry data. Unlike SEER, this dataset includes accurate information on mode of detection as well as systemic therapy.METHOD: Using the Eastern Cancer Registration & Information Centre (ECRIC) dataset, information was collated for 5818 women diagnosed with invasive breast cancer in East Anglia from 1999-2003. All patients underwent surgery, had records of pathological staging (tumour size, grade, lymph node status, ER status), systemic treatment (chemotherapy, hormone therapy, both), mode of detection (screen-detected, symptomatic) and follow up, censored on 31 December 2007. A model was derived from these data using Cox proportional hazards, with ER positive and ER negative tumours modelled separately, and this was subsequently validated in an external dataset of 5468 patients from the West Midlands Cancer Intelligence Unit (WMCIU). Validation was performed by comparing (a) observed and expected mortality (overall & breast cancer specific) at 5 & 8 years and (b) receiver operating characteristic (ROC) curves in both ECRIC & WMCIU datasets.RESULTS:ECRIC datasetDifference in overall observed/expected mortality <1% at 5 years (14.8% vs 14.9%) and 8 years (18.9% vs 18.9%). Area under ROC curve (AUC) was 0.81.Difference in breast cancer specific observed/expected mortality <1% at 5 years (10.6% vs 11.0%) and 8 years (12.9% vs 13.5%). AUC was 0.84.WMCIU dataset:Difference in overall observed/expected mortality < 1% at 5 years (15.8% vs 16.5%) and 8 years (17.5% vs 17.8%). AUC was 0.79.Difference in breast cancer specific observed/expected mortality <2% at 5 years (11.0% vs 12.6%) and 8 years (12.2% vs 13.6%). AUC was 0.82.Overall model fit was good across all subgroups although the ER positive model provided better discrimination (AUC 0.82) than ER negative (AUC 0.75). There was no significant difference between the ROC curves generated with ECRIC and WMCIU data (ER positive X2 = 0.17, p=0.68; ER negative X2 =0.00, P=0.95).CONCLUSION: We have developed a prognostication model for early breast cancer based on data from a UK cancer registry that has included mode of detection for the first time. The model is well calibrated, provides a high degree of discrimination and has been validated in a second patient cohort. This model will underpin a new web-based prognostication and treatment benefit tool for early breast cancer based on UK data (Adjuvant UK).

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 4033.
Identification of ER Negative Tumors in Which MAPK Inhibition Restores Both ER Expression and Anti-Estrogen Responses.
Authors: El-Ashry D. , Brinkman J. , Miller P.
Published: December 2010

Background: Approximately 35% of breast cancer lacks expression of the estrogen receptor α (ERα). ERα negative breast cancer carries a poorer prognosis compared to ERα+ breast cancer, but more importantly, it precludes the use of less toxic anti-estrogen therapies. Our previous data suggest that many ERα- breast tumors have the potential to express ERα, but ERα expression is repressed by hyperactive MAPK as a result of altered growth factor signalling pathways. We have shown that hyperactive MAPK results in down-regulation of ERα and treatment that inhibits MAPK activity results in re-expression of ERα protein and restoration of anti-estrogen sensitivity in both in vitro cell line and ex vivo primary culture model systems.Results: Using a panel of 9 primary cultures established from ERα- breast tumors, we have examined the incidence of MAPK inhibition mediated restoration of ERα expression. Of the 9 primary cultures examined to date, 5 are from triple negative tumors and 4 are from ERα-/PR-/H2N+ tumors. Of the 5 triple negative cultures analyzed, 3 re-express ERα upon MAPK inhibition, while 2 do not, and all 4 ERα-/H2N+ cultures re-express ERα. Although each of these primary cultures exhibited wide variation in the level of hyperactive MAPK, in those that re-expressed ERα, it was successful inhibition of MAPK activity that led to the re-expression of ERα. To establish determinants of the subset of ERα- breast tumors in which MAPK inhibition will restore ERα expression and more importantly, those in which MAPK inhibition restores both ERα expression and anti-estrogen responses, we have performed both proteomic and genomic profiling of these 9 primary cultures in the absence and presence of MAPK inhibition. Key differences between those primary cultures that re-express ERα vs those that do not, and interestingly, between triple negative primary cultures that re-express ERα vs those that express Her2 and re-express ERα upon MAPK inhibition have been identified. The role of these in determining restoration of anti-estrogen responses as well are being established.Discussion: Our overall goal is to understand the mechanisms underlying ERα- breast cancer and to develop novel therapies for ERα- breast cancer based on the re-expression of ERα and restoration of anti-estrogen responses. Having shown that hyperactive MAPK can repress ERα expression, and more importantly that inhibition of MAPK can restore functional ERα expression, we now need to establish the subsets of ERα- breast cancer in which each of this mechanism is responsible for the lack of ERα expression. This knowledge will allow us to determine which ERα- breast cancer patients may benefit from the combined inhibition of MAPK and ERα. The ability to use drugs with low toxicity to re-express ERα in ERα- tumors and thus render them sensitive to anti-estrogens would be a major leap forward in the treatment of ERα- breast cancer.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 404.
Secreted Frizzled Receptor Protein 1 (sFRP-1) as Both a Potential Novel Biomarker of Triple Negative Breast Cancer (TNBC), and Its Sensitivity Against Taxane/Anthracycline Containing Neoadjuvant Chemotherapy.
Authors: Liedtke C. , Ruckert C. , Goette M. , von Wahlde M. , Kiesel L. , Symmans W. , Pusztai L.
Published: December 2010

Background: Patients with triple negative breast cancer (TNBC) carry an unfavorable prognosis despite an increased response to chemotherapy compared to other breast cancer subtypes. While tumor grade/proliferation is one major determinant of chemosensitivity in TNBC, we have recently suggested additional (molecular) features to have a significant impact as well.Material and Methods: Differential gene expression analysis was performed between TNBC (n=27) versus non-TNBC (n=106) using gene expression profiles (Affymetrix U133A) from breast cancers of patients treated with neoadjuvant taxane/anthracycline chemotherapy at MDACC (Hess et al. 2006). Differentially expressed genes were validated using profiles of 286 patients with breast cancer who did not receive systemic therapy (Wang et al. 2006). Genes were tested for (a) an association with relapse free survival, (b) an association with response to neoadjuvant chemotherapy and (c) correlation with Ki67 expression. siRNA knockdown experiments using triple negative and non-triple negative cell lines were carried out using both (a) a proliferation assay and (b) a chemosensitivity assay.Results: The secreted frizzled receptor protein 1 (sFRP-1), a member of the wnt-pathway, was identified as the top differentially overexpressed gene in TNBC (i.e. false discovery rate [FDR] < 0,0001) and successfully validated in the independent dataset. In TNBC, sFRP-1 expression showed no association with relapse-free survival, but significantly correlated with an increased sensitivity to neoadjuvant chemotherapy. Importantly, it did not correlate with expression of Ki67 in TNBC. siRNA-mediated knockdown of sFRP-1 expression intriple negative MDA-MB 468 breast cancer cell lines was associated with increased proliferation and most importantly decreased sensitivity to paclitaxel in vitro.Conclusion: We suggest sFRP-1 as a novel marker of (a) the triple negative breast cancer phenotype in general and (b) its chemosensitivity to taxane-containing chemotherapy independent of Ki67 expression. Preliminary siRNA-based cell culture experiments suggest sFRP-1 to play a causal role with regard to chemosensitivity of TNBC rather than solely constituting a novel marker thereof.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 4047.
The Impact of Estrogen Receptor-beta on the Actions of Endoxifen in Breast Cancer Cells.
Authors: Hawse J. , Wu X. , Subramaniam M. , Negron V. , Lingle W. , Goetz M. , Spelsberg T. , Ingle J.
Published: December 2010

Background: We have previously demonstrated that endoxifen is the most potent tamoxifen metabolite responsible for eliciting the anti-estrogenic effects of this drug in breast cancer cells expressing ERα (Wu, Hawse et al. Cancer Res 2009;69:1722). These studies support the clinical findings which demonstrate that women with genetically impaired CYP2D6 metabolism have significantly reduced endoxifen levels and a higher risk of breast cancer recurrence. More recently, data has emerged from several laboratories suggesting that expression of ERβ in breast tumors leads to increased benefits from tamoxifen therapy. However, the role of ERβ in mediating endoxifen action has yet to be explored.Methods: Using U2OS osteosarcoma cells, as well as Hs578T and MCF7 breast cancer cells, stably expressing ERβ, we examined the effects of endoxifen on ERβ protein levels by western blotting, ERβ transcriptional activity by real-time RT-PCR, and on ERβ inhibition of breast cancer cell growth through the use of proliferation assays.Results: We have discovered that endoxifen stabilizes ERβ protein, unlike its targeted degradation of ERα, and induces ERα/ERβ heterodimerization in a concentration dependent manner. We have also found that endoxifen is a more potent inhibitor of estrogen induced gene expression in MCF7 cells stably expressing ERβ. Concentrations of endoxifen that mimic patients with impaired CYP2D6 activity (20-40 nM) do not significantly alter E2-induced cell proliferation in MCF7 cells expressing only ERα. However, these concentrations of endoxifen significantly reduce the estrogen induced proliferation rate of MCF7 cells expressing ERβ.Conclusions: Our data demonstrate that endoxifen stabilizes ERβ protein, induces ER heterodimerization and more potently inhibits the actions of estrogen in breast cancer cells expressing ERβ. The presence of ERβ sensitizes cells to the anti-estrogenic effects of endoxifen and growth inhibition occurs at substantially lower endoxifen concentrations (20-40 nM) than in cells that express only ERα (100 nm). Our data suggest that tamoxifen therapy may be beneficial even in patients with reduced CYP2D6 metabolism when ERβ is expressed. These studies also suggest that the increased effectiveness of tamoxifen therapy in patients whose tumors express ERβ may be due to the effects of endoxifen. Finally, these findings underscore the need to further elucidate the role of ERβ in the biology and treatment of breast cancer.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 405.
The Role of the Systemic Inflammatory Response in Long Term Outcome from Operable Breast Cancer.
Authors: Henley N. , Tovey S. , Doran C. , McMillan D.
Published: December 2010

Aims: It is recognised that the presence of a systemic inflammatory response has independent prognostic value in a number of advanced and operable tumours. There is also evidence in advanced breast cancer (Al Murri et al., 2006). However, results in operable breast cancer have been conflicting. Therefore, we prospectively examined the relationship between the systemic inflammatory response and long term survival in primary operable breast cancer.Methods: Patients with invasive primary operable breast cancer (n=154) treated at Royal and Western Infirmaries, Glasgow between October 2000- January 2002 were studied. The systemic inflammatory response, as evidenced by elevated CRP and Il-6, was measured pre-operatively and approximately 12 months later. An elevated CRP was defined as >10mg/l, and IL-6 as >5ng/l. Demographic and clinical data including age, tumour size, histological grade, lymph node status oestrogen receptor (ER) status and HER2 status were recorded. The Nottingham prognostic index (NPI) was calculated and subdivided into good (0-3.4), intermediate (3.41-5.4) and poor (>5.41) prognosis. Patients were followed-up until 1st March 2009.Results: Mean age was 59 (range 34-87). HER-2 was positive in 18 patients (12%), ER status was positive in 121 patients (79%). NPI was good/ intermediate/poor in 55 (36%)/ 73 (47%)/ 26 (17%). Circulating concentrations of interleukin-6 were significantly correlated with those of C-reactive protein (rs=0.449, p<0.001). Median follow-up was 8 years (range 7 – 8.5 years). During this period, there were 20 breast cancer deaths and 16 of other causes. On univariate analysis, NPI (p<0.001), pre-operative CRP (p=0.03), pre-operative IL-6 (p=0.05) and post-operative CRP (p=0.05) predicted breast cancer specific survival. In contrast, post-operative IL-6, ER and HER2 status did not. On multivariate analysis, NPI (HR 7.78 95% CI: 3.44-17.61 p<0.001) and pre-operative CRP (HR 6.36 95% CI: 2.11-19.18, p=0.001) predicted breast cancer specific survival.Conclusions: The results of the present study suggest that the presence of a systemic inflammatory response prior to surgery, in particular CRP, predicts breast cancer specific survival independent of tumour based factors. Therefore measurement of the systemic inflammatory response may be useful in the pre-operative assessment of patients with primary operable breast cancer.We acknowledge the advice and support of Timothy G Cooke who died 20th July 2008.ReferenceAl Murri AM, Bartlett JM, Canney PA, Doughty JC, Wilson C, McMillan DC. Evaluation of an inflammation-based prognostic score (GPS) in patients with metastatic breast cancer. Br J Cancer. 2006;94:227-30.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 4050.
A Genomic Biomarker for Breast Cancer Development in High-Risk Women.
Authors: Bild A. , Sun Y. , Soldi R. , Conner T. , Walker D. , Werner T. , Spira A. , Andrulis I. , Buys S. , Johnson E.
Published: December 2010

BackgroundFamily history is an important factor contributing to a woman's risk of breast cancer development. This increased risk reflects the participation of inherited genetic components such as breast cancer susceptibility genes. However, many of the genetic components contributing to breast cancer remain unknown, and a number of women with a family history of breast cancer never develop breast cancer despite their high-risk status, while other high-risk woman go on to develop breast cancer. Thus, it is clear that we lack crucial pieces of information to help define a person's true risk of developing breast cancer.Methods and FindingsWe hypothesize that there are many undiscovered germline variations in genes that lead to altered gene expression patterns predictive of breast cancer development in high-risk women. We have developed a genomic model capable of predicting which high-risk women, both BRCA1/2 mutation carriers and BRCAX women, will actually develop breast cancer. Specifically, we use exon-level genome-wide expression profiling of peripheral blood mononuclear cells (PBMCs) to develop a model of disease risk (n=118 samples of either control or high-risk women). From this data, we generate a biomarker consisting of genes that most correlate to cancer development in women with strong family histories of breast cancer. Using an internal independent dataset, our biomarker can predict the high-risk women who will or will not develop cancer with over 85% accuracy. Further, we have validated this model on an independent external cohort (n=36) which was obtained and processed at sites unique to our training dataset. Our genomic biomarker can accurately predict breast cancer development in high-risk women with over 78% accuracy using this external dataset. Therefore, from our analyses, we can calculate with high accuracy an individual woman's true risk of developing breast cancer. This method provides a personalized approach to hereditary breast cancer risk assessment that is not currently available. This personalized risk assessment will aide clinicians in counseling their patients regarding specific management options based on a patient's individual risk of breast cancer. Lastly, these studies have also identified novel genes associated with breast cancer risk, which may provide a basis for targeted therapies that may help modify the risk of breast cancer development in high risk patients.ConclusionsTogether, these studies deliver both a non-invasive biomarker for hereditary breast cancer risk and a characterization of genes that contribute to breast cancer development. Overall, we expect these experiments to identify the genetic changes that underlie breast cancer predisposition, and assist clinicians and patients in determining the appropriate preventative measures based on their personal risk of developing breast cancer.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 4059.
Ki-67 Expression after Tamoxifen Given Preoperatively Predicts Disease Free Survival and Overall Survival in Women with Operable ER-Positive Breast Cancer.
Authors: Decensi A. , Decensi A. , Guerrieri-Gonzaga A. , Gandini S. , Serrano D. , Cazzaniga M. , Mora S. , Johansson H. , Lien E. , Lien E. , Luini A. , Pelosi G. , Pelosi G. , Bonanni B.
Published: December 2010

We have previously shown in a window of opportunity, 3-arm, double-blind phase II randomized trial conducted in a single Institution that tamoxifen given for 4 weeks between tru-cut biopsy and surgery at a daily dose of 5 or 1 mg day was equivalent to 20 mg in decreasing ki-67 expression. A total of 31 premenopausal and 89 postmenopausal women with operable stage I-II ER-positive breast cancer were recruited (Decensi A et al, JNCI 2003;95:779-90). Here we analyzed the prognostic and predictive significance of ki-67% after a median follow-up of 86 months (11-111) in the same cohort of 120 women who underwent adjuvant treatment according to St. Gallen guidelines. At the current follow-up time DFS was 76% and OS was 92%. The median (IQ range) post-treatment ki-67% value was 12% (7-17) in women free of recurrence versus 20% (11-29) in women who relapsed (p=.0014). Post-treatment (surgery) ki-67% was positively associated with worse prognosis in univariate analysis (log-rank test =.0009, with ki-67% categorized in three groups: <7 (n=24), 7-19 (n=63), ≥20 (n=29); Figure).Post-treatment ki-67% was significantly associated with breast cancer death as well (p=.005, ki-67% <20 versus ≥20). In multivariate Cox analysis of DFS, both pre- and post-treatment ki-67 used as continuous variables were significantly associated with a higher risk of recurrence independent of adjuvant treatment, estrogen receptor level and pre-surgical tamoxifen dose (p=.002 and .04, respectively). Women with post-treatment ki-67% in the middle tertile (7-19%) and top tertile (≥20%) had a HR=2.1 (0.5-9.7) and HR=6.7 (1.5-30.2) of recurrence compared with women in the bottom tertile, respectively. There was no significant difference among tamoxifen dose groups on recurrence or death. In multivariate Cox analysis of OS, both pre- and post-treatment ki-67 used as continuous variables were significantly associated with a higher risk of breast cancer death (p=.0453 and .003, respectively). The risk of breast cancer death was 5 times higher in women with post-treatment ki-67 >20% than in women with ki-67<20% (HR=5.26, 1.22-23, p=.03). Our findings indicate that ki-67 is a prognostic and predictive marker in women with operable ER-positive breast cancer. The modulation of Ki-67% after a few weeks of hormonal intervention before surgery is a promising surrogate biomarker for outcome prediction and provides the opportunity to screen active agents in a cost-effective manner.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 406.
Screening for BRCA1 Mutations in Patients with Triple Negative Breast Cancer and Basal Phenotype.
Authors: Somer R. , Farengo-Clark D. , Patel A. , Schwarting R. , Grana G. , Rodler E.
Published: December 2010

Introduction: Most models estimating risk of carrying germline BRCA mutations are based on family history and age of cancer onset. Additional histologic criteria may be useful to identify patients at risk for harboring BRCA1 mutations. BRCA1 positive breast tumors are significantly more often estrogen receptor, progesterone receptor, and erbB-2 (HER2-neu) negative (“triple negative” (TN)) and are strongly associated with a basal phenotype confirmed by DNA microarray. Basal-like tumors express cytokeratins 5,6, and 14 or 17 by immunohistochemistry (IHC). It is unknown whether testing for BRCA1 mutations in TN basal-like breast cancer patients will identify additional carriers of mutations in this gene.Methods: A retrospective evaluation of patients with triple negative breast cancer seen at Cooper Cancer Center between 2004 and 2006 was performed after IRB approval. IHC was performed to identify the subset of basal-like breast cancers.Results: A total of 89 women were identified as diagnosed with TN breast cancer, while 53 had pathology available for review. 21 patients were identified as basal phenotype, whereas 32 were non-basal. Of the 21 patients who had basal-phenotype tumors, six underwent genetic testing and 3 were found to have a deleterious BRCA1 mutation. Of 32 non-basal patients, 9 patients underwent genetic testing and 2 were found to have a deleterious BRCA1 mutation. There was no difference between basal subtype and BRCA1 postivity [Fisher's Exact Test p=0.32]. Of note, 1of 5 (20%) patients with basal phenotype had no family history but tested positive for BRCA1 mutation and would have been missed by relying on conventional methods.Conclusion: Although there is a trend towards a higher percentage of TN breast cancer patients with basal subtype having BRCA 1 mutations (50 vs 22%), this was not found to be statistically significant (Fisher's exact p=0.32). However, this dataset is limited by a small sample size. Only 40% of TN breast cancer patients had a basal phenotype by IHC in this study, while other studies have shown 70- to 85% rate. Ongoing evaluation of patients with basal-like subtype and triple negative tumors should continue to investigate this relationship, especially in light of newer therapeutics that may impact this population. This data also suggests that patients with basal phenotype and triple negative disease should be considered for referral for genetic testing even in the absence of family history.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 4075.
A Phase II Study of the Combination of Endocrine Treatment and Bortezomib in Patients with Endocrine Resistant Metastatic Breast Cancer.
Authors: Trinh X. , Van Laere S. , Deleu I. , van de Velde H. , Van der Auwera I. , Limame R. , Wojtasik A. , Maes H. , De Mesmaeker P. , van Dam P. , Prové A. , Vandebroek J. , Rasschaert M. , Vermeulen P. , Dirix L.
Published: December 2010

Background: The introduction of anti-hormonal treatment has been a major revolution in breast cancer management. The majority of patients with metastatic hormone receptor-positive breast cancer eventually suffer from progression of their disease despite different types of anti-hormonal treatment. Preclinical studies have indicated bidirectional inhibitory interactions between ER and NF-kappaB, governing in part endocrine-resistance and enhanced growth. The hypothesis that inhibition of NF-kappaB activation by a proteasome inhibitor might reverse the sensitivity to endocrine therapy was the rationale to initiate a phase II trial.Aim, Material and Methods: The aim is to investigate whether the addition of open label bortezomib to either a SERM or an AI will result in documented activity in patients with progressive and measurable disease on the identical endocrine agent. This endpoint is evaluated according to RECIST criteria every 8 weeks. The patients are stratified into 2 treatment groups, according to their current endocrine treatment either tamoxifen or an AI. Bortezomib is administered on days 1, 8, 15, 21 of a 6 week regimen at a dose of 1.6 mg/sqm on each treatment day. Consecutive tumour biopsies (if accessible) and blood samples are collected for translational purposes. Clinical benefit was predefined by the patients obtaining at least either stable disease, or a partial or a complete response according to RECIST criteria, lasting for at least four weeks. A two-stage statistical approach showed that this should at least occur in 1 patient in the first cohort of 14 patients, prior to further accrual.Results: At present N=9 patients have been enrolled. Eight patients received AI+bortezomib, while one received TAM+bortezomib. N=5/9 patients had progressive disease, N=4/9 patients had stable disease of which two patients (N=2/9) had stable disease continuing for more than 4 weeks. There were no objective antitumour responses observed. N=4/9 had grade III diarrhoea. Median time to treatment failure was 69 days [35-140], either due to toxicity (N=3/9) or (eventual) progression (N=6/9).Conclusions: Preliminary results shows that the clinical benefit rate is 22% (N=2/9). Although preliminary, these results warrant further accrual and suggest that the biological hypothesis seems clinically valid. Further recruitment is ongoing.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 4099.
Final Overall Survival (OS) Results from the Randomised, Double-Blind, Placebo-Controlled, Phase III AVADO Study of Bevacizumab (BV) Plus Docetaxel (D) Compared with Placebo (PL) Plus D for the First-Line Treatment of Locally Recurrent (LR) or Metastatic Breast Cancer (mBC).
Authors: Miles D. , Chan A. , Romieu G. , Dirix L. , Cortés J. , Pivot X. , Tomczak P. , Juozaityte E. , Harbeck N. , Steger G.
Published: December 2010

Background: Anti-VEGF monoclonal antibody BV significantly improves efficacy in combination with standard therapies in multiple tumour types, with limited impact on toxicity. Three randomised phase III trials (E2100, AVADO and RIBBON-1) in mBC have demonstrated that BV + 1st-line chemotherapy (CTx) significantly improves progression-free survival (PFS) and overall response rates (ORR). We present mature OS data from AVADO.Methods: Patients (pts) with HER2-negative LR/mBC were randomised to D 100mg/m2 + PL, D + BV 7.5mg/kg or D + BV 15mg/kg. D was given q3w for ≤9 cycles. BV or PL was given q3w until disease progression/unacceptable toxicity. After progression, pts were offered BV with 2nd-line anticancer therapy in a post-study treatment phase. The primary endpoint was PFS; secondary endpoints included OS, time to treatment failure, ORR, duration of response and safety. An exploratory analysis conducted in pts receiving post-progression CTx compared OS in pts receiving 2nd-line BV with those who did not.Results: 736 pts were enrolled March 2006–April 2007. The primary analysis (data cut-off October 2007; median follow-up 10.2 months) showed significant improvements in PFS and ORR for both BV-containing arms compared with PL + D. Mature OS data (data cut-off April 2009; median follow-up 25 months) are shown (Table). There was no difference in median OS between the study arms (range 30–32 months). Recognising the limitations of the non-randomised comparison in exploratory analyses, results suggest that use of BV with 2nd-line therapy is a possible reason for lack of OS difference. Updated PFS and ORR were superior for the 15mg/kg BV arm compared with PL + D. Results for the 7.5mg/kg BV arm also indicated a, less pronounced, treatment benefit. BV had limited impact on the safety profile of docetaxel. Increased SAEs in the 15mg/kg BV arm may be due to more pts receiving 9 cycles of D than in the PL arm (51% vs 42%).Conclusions: There was no difference in OS between study arms. Exploratory analyses suggest that use of 2nd-line BV with CTx may influence OS. These mature data confirm the improvement in PFS and ORR with BV 15mg/kg combined with D compared with PL + D.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 41.
Does 3D-Ultrasound Imaging Improve Target Delineation for Breast Boost Radiation Therapy Planning.
Authors: Young A. , Chadha M. , Geraghty C. , Harrison L.
Published: December 2010

Background: Conformal volume-based breast dosimetry using CT-simulation is the most widely used technique for treatment planning. As part of a prospective protocol we studied the role of 3D-Ultrasound (US) technique for Image-Guided Radiation Therapy in breast cancer. In this paper we explore if there is additional value to utilizing seroma images on 3D-US for defining the target volume for breast boost.Materials and Methods: The IRB-approved prospective study used the ClarityTM 3D-US breast system (Resonant Medical). At simulation, concurrent CT and 3D-US images were obtained then fused. The boost lumpectomy volume (GTV) was contoured on CT by a single physician blinded to the results of the US (MC). To create the planning target volume (PTV), the GTV was expanded circumferentially by 1.5 to 2.0 cm, with the exception of its relation to skin and chest wall which were excluded from the PTV. For this analysis, the fused images of the concurrent CT simulation and 3D-US obtained on 25 patients were evaluated for concordance in the coverage of the CT contoured GTV and PTV to that of the 3D-US seroma image.Results: In 1 patient the CT images were limited in identifying the lumpectomy cavity because of the dense nature of the breast. The cavity not appreciated on CT image was visible on US. In this case the application of 3D-US would have provided superior target definition. In 22 of the remaining 24 patients the seroma cavity seen on the US was also seen on CT and was included in the lumpectomy GTV contoured on the images from CT simulation. In 2 patients, approximately 5% of the US seroma was not part of the GTV but was encompassed in the expanded PTV. In these 2 patients the US seroma was in proximity to the edge. This eccentric location of the US seroma cavity in 1 out of the 2 patients would have resulted in a change in the PTV. While in the other patient even though the seroma cavity was close to the edge of the PTV, the proximity to the chest wall would have precluded any major modification of the PTV. Overall, in all cases the 3D-US seroma cavity was encompassed in the expanded PTV. In 2/25 patients the target volume would have been revised based on the additional data provided by the 3D-US images.Conclusions: In summary, from this experience we observed that the use of 3D-US images in addition to CT simulation can alter the target delineation in 8% of cases. The 3D-US is a non-ionizing imaging modality that may add benefit for improving target delineation when used concurrently with CT simulation.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 4107.
Examination of Immune Competence in Breast Cancer Patients at Six Months Post Surgery and Adjuvant Therapy through Assessment of T Cell and Dendritic Cell Functionality.
Authors: Gendler S. , Pathangey L. , Lakshminarayanan V. , Mukherjee P. , Pockaj B. , Suman V. , Loserth L. , Markovic S. , Ingle J.
Published: December 2010

It has long been known that immune suppression is induced by the presence of a tumor, resulting in compromised T cell and dendritic cell (DC) functionality. What is not well understood is how soon the immune system recovers normal functionality following surgical removal of tumors and adjuvant therapy. This timing is of critical importance, as the effectiveness of therapeutic immune strategies relies on optimal presentation of antigens and activation of T cells. Blood from breast cancer patients was collected prior to surgery and at 3 and 6 months post adjuvant therapy. Patients were treated with surgery (breast conservation or mastectomy) and chemotherapy (CT) (n=22) or surgery without chemotherapy (n=38). Most patients had radiation therapy (RT). Blood from 22 healthy woman of similar age served as controls. T cell functionality (TCF) was determined following stimulation with plate-bound anti-CD3 (1 µg/ml) and anti-CD28 (0.5 µg/ml) and proliferation was measured by 3H-thymidine uptake. DC functionality (DCF) was determined by ability to present allo-antigens in a mixed lymphocyte reaction. Values for the normal samples determined the normal ranges (NR). Responses were grouped into 4 categories based on their status at 6 months: Group 1 - remained in NR; Group 2 - climbed into NR; Group 3 - remained below NR; Group 4 - fell below NR. Analysis of immunosuppression will be based on lymphocyte subsets, cytokine, COX-2, and PGE2 metabolite levels.T Cell Proliferation AllWith CTNo CT N=60N=22N=38Group 120%14%24%Group 223%23%24%Group 338%41%37%Group 418%23%16% DC Function AllWith CTNo CT N=58N=20N=38Group 143%35%47%Group 219%25%16%Group 317%15%18%Group 421%25%18% Forty-three percent of patients had normal levels (Groups 1 and 2) for TCF and 62% for DCF by 6 months post adjuvant therapy, suggesting that breast cancer patients are potentially amenable to vaccine therapy. 28% of patients had both TCF and DCF in NR; 50% had either TCF or DCF in NR; and 22% had neither in NR. The percentage of patients who had TCF in NR by adjuvant therapy was: 35% CT+RT (6/17); 33% CT (1/3); 52% (17/33) RT; and 20% (1/5) neither. The percentage of patients who had DCF in NR by adjuvant therapy was: 67% (10/15) CT+RT; 33% (1/3) CT; 67% (22/33) RT; and 40% (2/5) neither. Proper understanding of the effects of tumor, chemotherapy and radiation therapy on immune function, especially the effects on T cells and DCs, may enable us to identify the appropriate patients in whom to study immunotherapy approaches in women with early breast cancer and to examine strategies to counteract cancer-related defects in immune function. (supported by P50CA116201)

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 4131.
Peri-Operative Suppression of Immune-Regulatory Blood Cells Predicts Wound Complications in Breast Cancer Patients.
Authors: Hogan B. , Peter M. , Thorpe R. , Achuthan R. , Carter C. , Horgan K. , Hughes T.
Published: December 2010

IntroductionWe have previously demonstrated an association between post-operative wound complications and systemic breast cancer recurrence (p<0.0001), Murthy et al (2007) Br J Cancer 97, 1211-7. The aim of this study was to examine the potential role of the immune system in establishing this association, and therefore whether immune factors might be used to predict either wound complications or cancer recurrences.MethodsPatients with primary operable breast cancer were prospectively recruited to the study. Serial blood investigations were performed pre-operatively, peri-operatively and post-operatively. Absolute numbers of various lymphocyte cell populations were measured using multi-colour flow-cytometry including CD45+lymphocytes, CD19+ B lymphocytes, CD3+ T lymphocytes, CD4+ helper T cells, CD8+ cytotoxic T cells and CD56+ NK cells. We also measured the levels of the NK cytotoxicity receptors NKp30 and NKp46 on the NK cell population.ResultsOne hundred and nine patients were recruited to the study and there was a wound complication rate of 13.4%. Absolute numbers of CD3+ T lymphocytes, CD4+ helper T cells, CD8+ cytotoxic T cells and CD56+ NK cells were significantly lower 4 hours post-operatively compared to pre-operative levels (p<0.05), although levels had typically recovered after 24 hours. However, NKp30 expression remained significantly reduced at 24 hours (p<0.05). Mastectomy patients had a significantly greater fall in T lymphocyte numbers than those having breast conserving surgery (p<0.05). Patients who went on to develop wound complications post-operatively had a significantly greater fall in their CD4+ helper T cells at 4 hours post-operatively, than those patients who did not go on to develop wound complications (p<0.05).ConclusionsBreast cancer surgery results in severe disruption to the immune system, with dramatic changes in levels of immune regulatory blood cells populations. Changes are predominantly immuno-suppressive. The greater the immune disruption as a result of surgery, the more likely the patient is to develop a wound complication. We believe that this peri-operative immuno-suppression may also provide a window of opportunity for the successful dissemination of tumour cells post-operatively thereby increasing the risk of future metastases; we are maintaining follow up on this patient cohort in order to test correlations between peri-operative immuno-suppression and systemic recurrences.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 4132.
Estrogen Receptor beta Exerts Growth-Inhibitory Effects on Human Mammary Epithelial Cells.
Authors: Springwald A. , Lattrich C. , Ortmann O. , Treeck O.
Published: December 2010

In mammary epithelium estrogen receptor beta (ERbeta) is widely expressed. Its expression is reported to decline during carcinogenesis of the breast and other tissues. In this study, we examined the consequences of a loss of ERbeta expression in mammary epithelial cells. We knocked down ERbeta transcript levels in human mammary epithelial MCF-10A cells and in MCF-7 breast cancer cells by means of stable transfection with a specific shRNA plasmid. ERbeta knockdown resulted in a significant growth increase of both cell types in a ligand-independent manner. This effect was accompanied by elevated cyclin A2 expression in MCF-10A cells and by decreased expression of growth-inhibitory p21/WAF and epithelial cell marker cytokeratine 8 in both cell lines. Transfection of ERbeta shRNA did not alter the absent proliferative estrogen response of MCF-10A cells, but conferred sensitivity to selective estrogen receptor modulator tamoxifen to this cell line. In contrast, ERbeta knockdown diminished estrogen responsiveness of MCF-7 breast cancer cells and also weakened the effect of tamoxifen on this cell line. These ligand-dependent effects only observed in MCF-7 cells exhibiting a high ERalpha/beta ratio were accompanied by smaller estrogenic repression of p21/WAF expression, an impaired tamoxifen-triggered induction of this gene and by relative downregulation of ERalpha and cyclin A2 transcript levels. Our data suggest that ERbeta exerts antiproliferative effects both on MCF-10A and MCF-7 cells in a ligand- and ERalpha-independent manner by regulation of p21/WAF or cyclin A2 gene expression. Knockdown of ERbeta in both cell types was sufficient to significantly decrease transcript levels of epithelial cell marker cytokeratin 8. The results of this study support the hypothesis that ERbeta acts as a tumor suppressor in mammary epithelium.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 4152.
GPR30 and HER-2 Expression in Invasive and Metastatic Breast Cancer.
Authors: Arias-Pulido H. , Joste N. , Lomo L. , Chaher N. , Lee S. , Verschraegen C. , Meisner A. , Martinez C. , Prossnitz E. , Royce M.
Published: December 2010

Background: GPR30 expression, a new estrogen receptor, has been previously associated with HER-2, tumor size and metastasis in invasive breast cancer (BC) but its role in paired normal (N), invasive (I), and metastatic (M), samples is unknown. We described GPR30 and HER-2 expression in a collection of paired N/I/M samples, derived from the same individual.Materials and Methods: GPR30 and HER-2 expression was assessed by immunohistochemistry (IHC) in tissue microarrays, containing paraffin-embedded cores from 100 patients diagnosed with invasive BC. N and M samples were also available from the same patient. GPR30 expression was evaluated by an H-score (Intensity (0, negative; 1+, weak; 2+, moderate; 3+, strong) x Percentage of stained epithelial cells). HER-2 expression was evaluated per standard criteria. Log rank tests and Wald tests were employed to assess the clinical impact of these molecular targets on patient outcome based on Kaplan-Meier Product estimator and Cox Proportional Hazard Regression.Results: GPR30 was expressed in 50%, 76%, and 72% of N, I and M, respectively, samples. HER-2 (3+) was found in 14% and 18% of I and M samples, respectively. GPR30 expression in I cases correlated with expression in M cases, and HER-2 expression in I but not M cases. GPR30 expression in M cases correlated with expression in HER-2 expression in M cases. HER-2 expression in I cases correlated with expression in M samples (P<0.05 for all comparisons). GPR30 and HER-2 expression were not associated with grade or stage (P>0.05). GPR30 expression in I or M samples was not associated with either overall survival (OS) or BC-specific survival (BCSS)(P>0.5). HER-2 expression was marginally associated with OS in I (P=0.06; Hazard Ratios (HR): 1.91; 95%CI: 0.956, 3.83) but not in M (P=0.23) cases. HER-2 was significantly associated with BCSS in I cases (P=0.03; HR: 2.39; 95%CI: 1.07, 5.32) and marginally in M (P=0.08) cases.Discussion: A previous study suggested that GPR30 expression predicted the development of metastasis. We found high GPR30 expression in both the primary and metastatic sample but the difference was not significant. While GPR30 expression was not associated with OS or BCSS, HER-2 expression was marginally associated with OS and significantly associated with BCSS in invasive cases.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 4158.