bluuebunny/crossref_metadata_2025 huggingface dataset:
Multiple Biomarker Expression in Circulating Tumor Cells (CTCs) from Metastatic Breast Cancer (MBC) Patients (Pts).
Authors: Somlo G. , Hsieh H. , Curry D. , Frankel P. , Krivacic R. , Lau S. , Lazarus N. , Baker N. , Swain-Cabriales S. , Bruce R.
Published: December 2010

Background: Numeration of CTCs from MBC pts is predictive of outcome. Quantitative changes in CTC-s are currently tested for their potential to monitor therapy (Rx). Biomarker characterization of CTCs may be a useful adjunctive guide for Rx selection. Method: At the COHCC between 5/1/2008 and 4/31/09, consecutively treated pts with newly diagnosed/or progressing MBC were accrued. Blood samples (10-40 ml) were procured prior to or during systemic Rx, and were sent to PARC for analysis. A novel high-speed scanning instrument located CTCs from cytokeratin (CK) labeling enabling high resolution images to be selectively acquired using digital microscopy. From these images, CTCs were identified by CK, DAPI (nuclear marker) and CD45, and protein expression levels were determined for HER2, ER, ERCC1 and EGFR. Cell lines with expression of each marker were used for normalization of the cell intensities, and a scoring system was used to account for relative number and expression levels of markers on the CTCs. Results: Of 21pts tested 81% were found to have detectable CTCs. CTCs were further analyzed from 13 such pts, some of whom had multiple specimens. Expression of EGFR and ERCC1 were detected in 77% and 92% of specimens tested. Expression of HER2 was detected in 47% and ER in 91% in samples tested. Discordance rates for the expression of the above 4 markers on the primary tumors vs. CTC were measured either before, during systemic treatment, or at progression on therapy. We observed significant discordance rates for all markers tested:ER 36%; ERCC1:20%; EGFR:60%; and HER2: 50%, respectively. Conclusions: Multiplex tumor marker testing of CTCs from pts with MBC is feasible. Following additional validation of expression patterns and the high discordance rates observed between CTCs and primary or metastatic tumor sites, prospective trials incorporating CTC expression into personalized treatment strategies may be justified.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3007.
Predictive Factors for Positive Non-Sentinel Nodes Following a Positive Sentinel Node Biopsy: NSABP B-32.
Authors: Julian T. , Anderson S. , Golesorkhi N. , Fourchotte V. , Zheng P. , Mamounas E. , Brown A. , Boudros E. , Bear H. , Costantino J. , Wolmark N.
Published: December 2010

Background: Following a positive sentinel node biopsy (SNB), current guidelines recommend an axillary dissection (AD) regardless of SN metastatic tumor size. In the majority of clinically node negative patients the risk for positive non-sentinel axillary nodes (NSN) is low. Predictive factors for positive NSNs following a positive SNB are analyzed in NSABP B-32 with inclusion of SN metastatic tumor size.Materials and Methods: After stratification, women with operable invasive breast cancer and clinically negative nodes were randomized to Sentinel Node Resection (SNR) with immediate conventional Axillary Dissection (AD) [Group 1] or to SNR without AD [Group 2]. Group 2 patients with positive SNs underwent AD. A multivariate analysis of SN positive patients from both groups for whom both a SNR and an AD had been performed was used to assess the need for AD following SNB. Nodes were classified as either SNs or NSNs, defined as all axillary dissection nodes plus any intramammary or other nodes that were not resected as SNs.Results: Between May 1999 and February 2004, 5,611 patients were entered into NSABP Protocol B-32. There were a total of 1,361 SN positive patients with AD from both groups. Data from 1,166 patients were available for multivariate analysis which included SN metastatic tumor size in 735 patients: 424 patients with macrometastaes (>2 mm) and 311 with micrometastases (<2 mm). In 626 patients SN metastatic size was unknown.In patients with positive SNB, results from the final multivariate model based on 653 patients with known covariate values indicated clinical tumor size was a significant predictor for positive NSN (p=0.044, OR: 1.17). Lymphovascular invasion was a significant predictor for positive NSN (p=0.0004, OR: 1.85). SN metastatic tumor size (Macro vs Micro) was a highly significant predictor for positive NSN (p<0.0001, OR: 3.42). Age at study entry, treatment type, proposed type of surgery, HER-2 status, and location of tumor were not significant multivariate predictors for positive NSN. Predictive modeling for positive NSN probability will be presented.Conclusion: Completion AD following a positive SNB, although helpful in prognosis and treatment planning, may not be required in patients with small tumors, absence of lymphovascular invasion, and micrometastases.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 301.
Implementation of a Composite Score System for Evaluation of Circulating Tumor Cells in Blood of Breast Cancer Patients.
Authors: de Albuquerque A. , Kaul S. , Breier G. , Krabisch P. , Stryzhevska N. , Bastert G. , Fersis N.
Published: December 2010

Background: We present a highly specific immunomagnetic separation technique in combination with an improved multimarker gene panel for molecular identification and characterization of circulating breast tumor cells in blood. Here we describe the creation of a new composite score in order to manage results and avoid false positives. Methods: Blood from breast cancer patients with primary or metastatic disease was drawn into two 10mL EDTA-tubes. The high affinity antibodies BM7 (MUC1) and VU1D9 (EpCAM) were used for immunomagnetic tumor cell enrichment. Separated cells were lysed and used for mRNA isolation and c-DNA synthesis (Qiagen®). A real-time quantitative RT-PCR approach using MESA FAST SYBR Assay (Eurogentec®) and primers selected from the UniversalProbeLibrary system (Roche AG®) for the epithelial markers cytokeratin 19 (CK19), mammaglobin 1 (MG1), epithelial cell adhesion molecule (EpCAM), baculoviral IAP repeat-containing 5 (Sur), immunosuppressive CD276 (CD276), carcinoembryonic antigen-related cell adhesion molecule 5 (CEA), HER-2, aldehyde dehydrogenase 1 family, member A1 (ALDH1), hypoxia inducible factor (HIF-1alpha) and CD44 molecule (CD44) was used for tumor cell identification and characterization. The ß-actin transcript was used for internal control and matched calibrator probes containing 2 or 10 tumor cells were used for quantitative expression analysis of tumor associated genes present in blood.Results: Positivity rate was based on a score that consists of 2 different characteristics: Marker detection (ranking according to the number of positive markers, varies from 0 – negative - to 1 – positive - for each marker) in combination with the marker expression level (ranking according to the C(t) values and varies from 0 – less then 2 tumor cells – to 1 – more then 2 tumor cells). This composite score is based on the expression of CK19, MG1, EpCAM, Sur and CD276 and has a ranking from 0 to 10. Negative cases are classified with score 0 and 1, cases ranked with score 2 need retesting and finally scores above 2 indicate clearly positive patients. The additional surrogate markers CEA, HER-2, ALDH, HIF and CD44 are analysed in positive patients in order to obtain further information. In our group of metastatic breast cancer patients, 53% were classified as positive, 42% of the patients were negative and in 5% a retesting is needed. Conclusion: The results of the composite score clearly show an increase of sensitivity and specificity for this assay. The implementation of this test in the routine monitoring of patients should help us to evaluate treatment response and create individualized treatment schedules.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3015.
Examination of TIMP-1 Levels and Relapse-Free Survival for Patients in NCIC CTG MA.14 Who Received Adjuvant Tamoxifen +/- Octreotide LAR.
Authors: Ali S. , Chapman J. , Lipton A. , Leitzel K. , Pritchard K. , Pu X. , Wilson C. , Carney W. , Shepherd L. , Pollak M.
Published: December 2010

Background: Tissue inhibitor of metalloproteinase-1 (TIMP-1) has been shown to have diverse multifunctional roles in tumorigenesis. AL/KL/SA postulated from first line metastatic endocrine therapy that elevated pre-treatment serum TIMP-1 required additional therapy. NCIC CTG MA.14 adjuvant endocrine trial permitted examination of whether TIMP-1 serum levels post-chemotherapy was associated with RFS. Methods: NCIC CTG MA.14 is a trial where 667 postmenopausal patients were randomized to receive adjuvant tamoxifen +/- octreotide LAR with a stratification factor of no, concurrent or sequential chemotherapy. For the purposes of this work, this was simplified to whether a patient had or had not received adjuvant chemotherapy prior to the serum draw. Serum TIMP-1 was assessed on >90% of the trial patients. We examined the effect of baseline TIMP-1 levels on relapse-free survival (RFS) of 1) all types, 2) bone only, 3) all types of bone, and 4) non-bone, by timing of chemotherapy before the serum draw utilizing continuous TIMP (ng/ml) and categorical TIMP (<454, >454 ng/ml, based on 95% non-parametric cut-point for healthy post-menopausal females). Data were available on patient and tumour characteristics of age (years), pathologic tumour size (cm), pathologic lymph node status (# nodes), IGF-1, C-peptide, IGFBP-3. Step-wise forward Cox multivariate models were used where a factor was added if p<=0.05, and interaction terms were added for TIMP-1 level and timing of chemotherapy. Results: High (categorical) TIMP-1 was significantly associated with longer RFS (p=0.04) in the non-bone RFS multivariate model in the subgroup of patients who had prior chemotherapy, but not in those who did not. The interaction between TIMP-1 and administration of chemotherapy before serum draw was significant (p=0.02). High (continuous) TIMP-1 was significantly associated with longer bone only RFS (p=0.04) in subgroup who had no chemotherapy before the serum draw, but not in those who may or may not have had it after. There was no significant interaction effect. Categorical and continuous TIMP-1 were not associated with any other type of RFS. Conclusions: We found a predictive benefit with the assessment of serum TIMP-1 in hormone receptor positive patients, that high TIMP-1 levels after adjuvant chemotherapy were predictive of a reduction in non-bone RFS.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3022.
Human Circulating Endorepellin LG3 Fragment and Breast Density: A Potential Combination with Screening Mammography in Women with Dense Breast.
Authors: Lee J. , Lee J. , Yang E. , Han W. , Han W. , Lee C. , Noh D. , Noh D.
Published: December 2010

(Background) In vitro studies have reported that the endorepellin LG3 fragment (ERLG3F), which is matrix-derived endogenous anti-angiogenic factor, can be a circulating biomarker candidate for breast cancer. This study examined the plasma ERLG3F levels in breast cancer patients and normal healthy controls, and evaluated its potential roles as a screening biomarker for breast cancer (Methods) ERLG3F levels were determined by competitive ELISA in the plasmas from prospectively constructed independent cohorts (train and test set) from July 2006 to September 2007 for the Biomarker Discovery Project (funded by 21C Frontier Functional Proteomics Program of South Korea). The Breast Cancer Cohort (BC) (n=366) has plasmas drawn preoperatively; the Healthy Control Cohort (HC) (n=305), drawn when volunteers visited our Healthcare Center for medical check-up. Circulating ERLG3F, mammography, and the combining marker (CM) of both were evaluated for their performance of identifying breast cancer in the train set and verified again in the test set. (Results) Decreased ERLG3F levels were observed in the individual plasmas from BC compared to the HC. The decrease was significantly prominent in patients without axillary lymph node metastasis (negative vs. positive), young age (< 50 vs. ≥50 yr), and dense breast (mammographic density Gr 3 or 4 vs. 1 or 2). We used a cutoff level, which correspond to 2SD below the mean ERLG3F in total healthy controls. ERLG3F significantly improved the performance of mammography in distinguishing breast cancer cases from healthy controls, although it is less sensitive than mammography as a single marker. (Conclusion) We conclude that ERLG3F can be a useful diagnostic biomarker for breast cancer and its role can be even better when used for young women with dense breast for whom mammography is less helpful.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3026.
Inhibin A Is Downregulated during Chemotherapy in Patients with Early Breast Cancer.
Authors: Rack B. , Burkhardt N. , Jueckstock J. , Kuhn C. , Noeding S. , Hoenig A. , Friedrich M. , Schindlbeck C. , Sommer H. , Friese K. , Janni W. , Mylonas [.
Published: December 2010

Introduction: Inhibins are dimeric glycoproteins, composed of an alpha-subunit (INH-a) and one of two possible beta-subunits (ßA or ßB), with substantial roles in human reproduction and in endocrine-responsive tumours. In breast cancer tissue, the growth factor Inhibin A is involved in cell differentiation and proliferation, thus suggesting a possible role as tumor marker. Aims of this study were to determine the serological measurement of Inhibin A (a- bA) in breast cancer patients during chemotherapy.Material and Methods: A series of 28 high risk N0 and N+ breast cancer patients who underwent standardized chemotherapy (3 x FEC and 3 x Docetaxel) followed by two years of zoledronate in the German SUCCESS trial were prospectively evaluated before chemotherapeutic treatment as well as after chemotherapy and two years after chemotherapy for the serological expression of Inhibin A. For serological analysis the ultrasensitive Inhibin A ELISA (DSL – U.S.A.) was used according to manufactures instruction. For statistical analysis the Wilcoxon rang sum test was used for paired samples. Statistical significance was assumed at p< 0.05.Results: The concentration of Inhibin A showed a significant decrease between data obtained before chemotherapy and after chemotherapy (p<0.005) and two-year follow up (p<0.001). Interestingly, there were seen no differences between the time point four weeks after chemoherapy and at two years (p=0.744).Discussion: Therefore, chemotherapy decreases significantly the Inhibin A concentration during chemotherapy. This might reflect a suppression of ovarian function and might be a marker for chemotherapy-induced amenorrhoea. Moreover, it has been suggested that Inhibin A might be a tumour marker for breast cancer, and therefore a sudden increase of its concentration might be indicative of breast cancer recurrence.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3028.
Paradoxical Relationship of Chromosomal Instability with Breast Cancer Outcome: Identification of a Good Prognostic ER Negative/HER2 Negative Breast Cancer Cohort with Extreme Chromosomal Instability.
Authors: Swanton C. , Swanton C. , Juul N. , Li Q. , Eklund A. , Richardson A. , Szallasi Z. , Szallasi Z.
Published: December 2010

Chromosomal Instability (CIN) is widely held to be associated with poor prognosis in solid tumours, including breast cancer, that may be related to enhanced clonal evolutionary capacity and the rapid acquisition of multi-drug resistance. Paradoxically, CIN is also associated with tumour suppression in animal models that may result from excessive genomic instability leading to autonomous cell lethality, a phenomenon analogous to "mutational meltdown" in model systems.We addressed whether such a phenomenon could be identified in breast cancer through a meta-analysis of CIN frequency in 1850 breast cancer patients. We assessed the distribution of CIN across breast cancer subtypes and its relationship with clinical outcome and prognosis using a surrogate of CIN, the CIN70 expression signature that correlates robustly with CIN in breast cancers quantified by DNA image cytometry. Increasing degrees of CIN predicted for poor outcome in multivariate analysis of 1850 patients and occurred more frequently in younger patients. Furthermore, CIN was more common in higher risk disease subtypes, including basal-like breast cancer, consistent with this pattern of genomic instability being a major determinant of breast cancer outcome.However, in agreement with previous evidence documenting a role for CIN in mediating cell lethality and tumour suppression in animal models, we demonstrate that a subset of tumours with extremes of CIN, that occur predominantly within the ER negative and HER2 negative subgroup, are associated with unexpected good prognosis in untreated breast cancer patients.We define a threshold of CIN beyond which clinical outcome improves and the relationship between CIN and proliferation is uncoupled. This evidence defines a paradoxical role for CIN in limiting tumour cell viability in vivo and suggests therapeutic strategies to tilt the balance toward CIN-mediated lethality in high-risk breast cancer subtypes in order to improve patient outcome.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3030.
High Expression of Cyclin E Increases the Risk for Breast Cancer Death by 2-3 Fold in Node Negative Breast Cancer Patients.
Authors: Lundgren C. , Holmqvist M. , Holmberg L. , Holmberg L. , Blomqvist C. , Fjällskog M.
Published: December 2010

Background: A large proportion of women with lymph node negative breast cancer treated with chemotherapy do not benefit from such treatment. Several studies have suggested that proliferation markers can identify patients at increased risk for recurrence. However, current international guidelines do not recommend proliferation markers for clinical use due to lack of well-powered studies and standardization of methodology. We wanted to investigate the prognostic value of cyclin E1 in a large population-based study on node negative breast cancer patients.Study Design: In a population-based, case-control study, 190 women who died from breast cancer were defined as cases and 190 women alive at the time for the corresponding case's death as controls. Inclusion criteria were tumor size ≤ 50 mm, no lymph node metastases and no adjuvant chemotherapy. The study was designed to detect an odds ratio (OR) of 2.5 with a power of 90% at a significance level of 0.05. Tumor tissue was immunostained for cyclin E1 with a commercial antibody. Statistical analyses were performed for 200 (cyclin E 200) and 500 cells (cyclin E 500) counted, respectively.Results: Cyclin E1 was highly correlated to histologic grade, Ki-67 and cyclins A/B (rs= 0.5-0.6; p<0.001) and inversely correlated to hormone receptors (rs=-0.2--0.3; p<0.001). There was a statistically significant association between cyclin E1 and breast cancer death both in uni- and multivariate (adjusted for tumor size and age) analyses with OR 2.6 -2.7 for cyclin E 200 and OR 2.1-1.9 for cyclin E 500.Conclusions: We found that cyclin E1 is a prognostic factor for breast cancer death in a population-based node negative patient cohort. Patients with tumors expressing high levels of cyclin E have a 2-3 fold increased risk for breast cancer death.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3033.
Expression and Prognostic Value of L1-CAM in Breast Cancer.
Authors: Milde-Langosch K. , Schröder C. , Mueller V. , Schumacher U. , Wirtz R. , Altevogt P. , Krenkel S. , Jaenicke F.
Published: December 2010

Background: The L1 cell adhesion molecule (L1-CAM) has been shown to be involved in cell motility, invasion, chemoresistance and tumor growth in various experimental models, and is associated with impaired prognosis in melanoma, colon and ovarian cancer. However, limited information about its expression in breast cancer is available.Methods: To analyse its role and possible prognostic value in primary breast cancers, L1-CAM mRNA and protein expression using microarray analysis (n=167) and Western Blotting (n=106) was performed. Its cellular distribution was demonstrated by immunohistochemistry. The results were correlated with classical histological data, biologically relevant disease markers and clinical data including disease outcome.Results: L1-CAM mRNA overexpression and protein expression was observed in 14-15% of the carcinomas and its expression was confirmed by immunohistochemical staining. Generally tumour cells at the tumour stroma interface were L1-positive. High L1-CAM expression was associated with nodal involvement, high grading, human epidermal growth receptor 2 (Her-2), plasminogen activator inhibitor 1 (PAI-1) and vascular endothelial growth factor (VEGF) expression and a negative estrogen receptor (ER) status, but not with neuroendocrine markers. High L1-CAM mRNA expression in primary breast cancers was associated with a significantly shorter disease-free (p=0.011) and overall survival (p=0.005).Conclusions Our results indicate that L1-CAM is highly expressed in a relatively agressive subgroup of breast cancers. Given the emerging functional role of L1-CAM in promoting tumor growth and metastasis our results suggest that L1 may have this function in breast cancer as well. Thus, L1-CAM might be a suitable target for specific immunotherapies in selected high-risk breast cancer patients.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3044.
Preliminary Comparison between AQUA and Centralised ER/PgR Analysis within the TEAM Pathology Study.
Authors: Bartlett J. , Rimm D. , Brookes C. , Dolled-Filhart M. , Robson T. , van de Velde C. , Billingham L. , Campbell F. , Hasenburg A. , Hille E. , Kieback D. , Putter H. , Markopoulos C. , Christiansen J. , Gustavson M. , Mallon E. , Meershoek-Klein Kranenburg E. , Parideans R. , Seynaeve C. , Rea D.
Published: December 2010

Background: The Tamoxifen and Exemestane Adjuvant Multinational (TEAM) trial identified quantitative relationships between ER/PgR (Can Res 69:83S) and patient prognosis, and suggested that high ER expression might be predictive for benefit from aromatase inhibitors. Conventional immunohistochemical assays, even when performed centrally, appear to be more variable than fluorescent immunohistochemical assays analysed by automated image analysis such as the AQUA system. However, output from the two systems may provide subtly different results for biomarkers. We have performed analysis of ER/PgR using AQUA technology in the TEAM pathology cohort and will present data on the full anlaysis of ER/PgR by central IHC and AQUA in 4600 cases. Preliminary data on ER (using a single TMA core) are included in the current analysis.Patients & Methods: TMAs from 4598 TEAM patients were analysed by AQUA and image analysis using established algorithms performed. Comparisons between IHC and AQUA were performed and outcome data will be analysed and presented.Results: Interim AQUA results from a single core were available from 2319 TEAM cases. Analysis of results suggested incremental increases in AQUA scores across conventional Allred categories (2-8). Regression analysis suggests that multiple cores may be required adequate representation of ER expression. PgR data will be presented.Conclusion: Initial comparison between AQUA and ER expression by central IHC suggests these methods are comparable. Further data is required to assess the potential value of AQUA scores and will be presented.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3045.
Computerized Histologic Image-Based Risk Score (IbRiS) Classifier for ER+ Breast Cancer.
Authors: Ganesan S. , Madabhushi A. , Basavanhally A. , Xu J. , Bhanot G. , Barnard N. , Toppmeyer D.
Published: December 2010

Background: Measurement of estrogen receptor (ER) expression is a routine part of clinical evaluation of individual breast cancers and is used to guide treatment. However not all ER+ breast cancers show equal benefit from hormonal or other treatment. The RT-PCR based Oncotype Dx assay, has been recently shown to robustly stratify early stage ER+ breast cancer and identify those tumors that will have low recurrence rates when treated with adjuvant hormonal therapy alone. Interestingly, standard pathologic grading, based on visual analysis of tumor morphology by trained pathologists, has a strong correlation with Oncotype Dx recurrence scores; low recurrence tumors are mostly low grade, and high recurrence tumors are mostly high grade. However, a major problem with use of pathologist-assigned histologic grade as a prognostic tool is the lack of reproducibility of histological grading between different pathologists. Computer aided image analysis and machine learning techniques offer a way to obtain highly reproducible image-based classification of ER+ breast cancer. These analyses can be performed on digital images of routinely obtained breast cancer histology and be incorporated into a prognostic assay.Materials and Methods: High resolution digital images were obtained for a series of ER+ breast cancers for which associated Oncotype Dx Assay results were available. Regions of invasive breast cancer were identified and then processed by computer-assisted image analysis methods. An automated nuclear detection scheme based on Expectation Maximization algorithm was used to identify cancer cell nuclei, followed by graph-based feature extraction using the Voronoi graph, Delaunay Triangulation and Minimum Spanning Trees, and dimensionality reduction using Graph Embedding with Support Vector Machine based classification. The final manifold generated by GE was unwrapped into a linear space and a Euclidean distance metric was used to generate a single score (Image Based Risk Score- (IbRiS)) for each sample. Correlations between IbRS, clinical features such as grade, and Oncotype Dx Recurrance Score were determined.Results: Unsupervised analysis of image-based features of high resolution digital images of ER+ breast cancer histology leads to natural separation of tumors, with low grade tumors separating from high grade tumors. There is also a robust separation of tumors with high Oncotype Dx Recurrance Scores from tumors with low Recurrance Scores.Discussion: Unsupervised analysis of high resolution image-based features can stratify ER+ breast cancers in a fashion that correlates well with a gene-expression based prognostic assay, Oncotype Dx. These data suggest that tumors with distinct gene-expression profiles also have distinct image-based features that can be measured by computer-aided image analysis and used to build prognostic and predictive assays.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3046.
Tumor-Stroma Ratio within the Primary Tumor Is a Prognostic Factor in Early Breast Cancer Patients, Especially in Triple-Negative-Carcinoma Patients.
Authors: de Kruijf E. , van Nes J. , van de Velde C. , Putter H. , Smit V. , Liefers G. , Kuppen P. , Tollenaar R. , Mesker W.
Published: December 2010

Background: Breast cancer patients comprise a prognostic heterogeneous group. Stroma-tissue surrounding cancer cells plays an important role in tumor behavior. The prognostic value of the tumor-stroma ratio was evaluated.Material and Methods: A cohort of 574 patients with early breast cancer, primarily treated by surgery between 1985 and 1994 was analyzed. The percentage of intra-tumoral stroma was visually estimated on haematoxylin-eosin-stained histological sections from primary tumor tissue by two observers. Patients with more than 50% intra-tumor stroma were quantified as stroma-rich and patients with less than 50% intra-tumor stroma as stroma-poor.Results: Of all tumors, 68% were classified as stroma-rich and 32% as stroma-poor. Cohen's kappa coefficient revealed an almost perfect agreement in classification (kappa=0.85; 94% concordance in classification) between the two observers. For the total group of patients, stroma-rich tumors had a shorter relapse free period (RFP) (Hazard ratio (HR) 1.62; 95%CI 1.23-2.13; p=0.001) and overall survival (OS) (HR1.29; 95%CI 1.03-1.60; p=0.025) compared to stroma-poor tumors. Tumor-stroma ratio was an independent prognostic parameter for all patients (p<0.001) and in stratified analysis for patients that received systemic treatment. Importantly, within the triple-negative-cancer (TNC) subpopulation, patients with stroma-rich tumors had a 3.19 times higher risk of relapse (p=0.003) compared to patients with stroma-poor tumors, independently of grade and lymph node status. Five year RFP-rates for patients with stroma-rich compared to stroma-poor tumors were 56% and 81% respectively.Discussion: Tumor-stroma ratio was an independent prognostic factor for RFP in breast cancer patients, especially in TNC. The tumor-stroma ratio is easy to apply, of low cost and reproducible. It is a candidate parameter that can easily be implemented in routine daily diagnostics to optimize risk-stratification for breast cancer patients and should therefore be considered to be implemented in standard pathology reports.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3049.
Feasibility of Sentinel Lymph Node Biopsy in Multiple Unilateral Synchronous Breast Cancer: Results of a French Prospective Multi-Institutional Study (IGASSU 0502).
Authors: Giard S. , Penel N. , Chauvet M. , Mignotte H. , Martel P. , Tunon de Lara C. , Gimbergues P. , Dessogne P. , Classe J. , Fondrinier E. , Marmousez T. , Blanchot J.
Published: December 2010

Introduction: Controversy remains over whether to perform sentinel lymph node biopsy (SLNB) in multiple (multicentric or multifocal) unilateral synchronous breast cancer. Several small retro- or prospective studies, included pre- or post-operative (or both) multiple synchronous tumours (MST) have suggested that the test performance of SLNB is similar to that seen in unifocal disease. The purpose of this study was to evaluate the feasibility and accuracy of SLNB in preoperatively diagnosed invasive MST.Patients and Methods: The Interest of Axillary SLNB in Multiple Invasive Breast Cancer (IGASSU) study was a prospective multi-institutional study with initial breast surgery, SLNB, and systematic level I to II axillary lymph node dissection (ALND). Patients eligible for the IGASSU study had an operable invasive MST, defined as two or more physically separate invasive tumours in the same or different breast quadrant. The diagnosis of invasive MST was confirmed histologically in all patients by core needle biopsy before surgery. Detection of sentinel node was performed by using either blue patent or radiocolloide injection or both. Injection sites were subareolar.Statistical Analysis: Sensitivity (Se), negative predictive value (NPV), accuracy (A), false negative rate (FNR) and their 95%-confidence intervals (95%CI) were calculated with a classical 2x2 contingency table. A univariate analysis using odds ratio calculation was performed to identify the risk factors for false negative results.Results: Between March 1, 2006, and August 31, 2007, 216 patients were prospectively included from 16 institutions. Of these patients, 211 were evaluable. The SLNB identified rate were 93.4% (197/211). A mean number of 2.2 SLN (range, 1 to 8, ± 1.4) was successfully excised. The mean number of resected nodes in ALND was 12 (range, 1 to 39, ± 5.7). The FNR was 13.6% (14/103) [95%CI: 7- 20%], Se was 86 .4% (89/103) [95%CI: 79- 93%], NPV was 87% (94/108) [95%CI: 80-93%], A was 92.9% (183/197) [95%CI: 89- 96%]. For the 14 false-negative SN, all had ≤ 3 involved nodes in ALND. Table 1 shows patterns of management of the axilla.Table 1 ALND+ALND-Non-identified SLNB113Identified SLNB+4544Identified SLNB-1494 In a univariate analysis, tumour location (only external location vs other location) was the only clinico-pathological factor influencing the FNR (22% [11-33%] vs 7% [4-10%]), even then median aggregate histological tumour size was smaller in external tumours (17mm [range, 12-80] vs 34mm [range, 8- 90], p=0.016).Conclusion: With a FNR of 13.6% (95%CI: 7-20%), we do not recommend SLNB as a routine procedure for multiple unilateral synchronous breast cancer, even for small tumour foci.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 305.
The Influence of Mode of Presentation on the Pattern of Recurrence in Early Breast Cancer (EBC).
Authors: Mansell J. , Angerson W. , Wilson C. , Doughty J.
Published: December 2010

IntroductionAn early peak in recurrence exists in women with EBC. This may be explained underlying tumour biology or a reaction to surgery. Screen detection has been shown to have a prognostic effect independent of disease stage suggesting a connection with underlying tumour biology. We aim to determine if the pattern of recurrence differs between modes of presentation.MethodsData from 3 centres in Glasgow of consecutive women aged between 50-65yrs with early stage breast cancer (EBC) diagnosed between 1995 and 2001 were examined. Recurrence was defined as invasive disease at any site. Patients were grouped by mode of presentation into screen detection and symptomatic. Kaplan Meier and time dependent Cox analysis were performed.Results1534 women were included with a median follow up of 5.5yrs. Mode of presentation was screening in 1007 (65.6%) women. At 2.5yrs cumulative recurrence was 2.5% (95% CI 1.5-3.5) in the screen detected group and 9.7% (7.2-12.2) in the symptomatic group. At five years the corresponding rates were 7.8% (6.0-9.6) vs. 19.1% (15.6-22.6). Time dependent multivariate analysis showed that the difference in the risk of recurrence between the modes of presentation was significantly different within 2.5yrs from diagnosis compared with later than 2.5yrs (p = 0.019). The hazard ratio for recurrence within 2.5 yrs was 0.59 (95% CI 0.34-1.00, p=0.05) and after 2.5yrs was 1.23 (0.84-1.80, p =0.285).ConclusionsThese results suggest that women with screen detected cancers have a better prognosis initially following diagnosis independent of disease stage. However this additional benefit isn't maintained with longer follow up.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3056.
Incidence and Demographic and Tumor Characteristics of HER2-Positive Invasive Breast Cancer in a Large, Unselected Population, 2000-2006.
Authors: Fehrenbacher L. , Habel L. , Capra A. , Anthony A. , Li X. , Quesenberry C. , Fulton R.
Published: December 2010

BACKGROUND: Most HER2 data have been generated from populations of invasive breast cancer (IBC) patients with selected patient and tumor characteristics (clinical trials, referral centers). The average age of women in US adjuvant trastuzumab trials was 49. Pre 2000, clinical trials of adjuvant chemotherapy included 25-40% HER2+ cases. We used an unselected population of women with IBC to estimate the age-specific incidence rates of HER2+ IBC and compare patient and tumor characteristics of HER2+ and HER2- disease.METHODS: Among female members of Kaiser Permanente Northern California (KPNC) 20 years or older (n=1.22 million), we identified all those diagnosed with a new primary IBC in 2000-2006 (n=16,975). During this period, all IBCs were routinely tested for HER2 by KPNC's regional IHC laboratory (center of excellence for HercepTest, CLIA licensed and CAP certified) and those that were IHC 2+ were sent for FISH testing; IHC 3+ or IHC 2+/FISH+ (ratio ≥ 2.0) were considered HER2+. Demographic and tumor characteristics were derived from the KPNC tumor registry. *Complete HER2 testing available for 94% of IBCs.RESULTS: Incidence of HER2+ IBC increased less dramatically with age and peaked 20 years earlier than incidence of all IBC (Table 1). Among those with IBC, the percent that were HER2+ decreased with age. Of all HER2+ IBCs, 69% were ≥50 years old and 42% were ≥60 years old.Incidence of all IBC and incidence of HER2+ diseaseAge GroupPerson-YrsNo. of IBCsIBC rate/100KHER2+ IBCs*%HER2+HER2+IBC rate/100K20-39288153371024.6416424.75.6940-4917402132680154.0049819.728.6250-5915535284354280.2759914.538.5660-699677624280442.2649412.351.0570-796676013318497.002718.740.5980+3666531633445.381429.638.73ALL817729016975207.59216813.626.51 The percent of tumors that were HER2+ increased with stage (local=11%, regional=18%, distant =25%), tumor differentiation (well =2%, moderate =12%, poor=26%, undifferentiated=30%), and generally with size (<0.5cm=17%, 0.5-0.9cm=7%, 1-1.9 cm=10%, 2-4.9 cm=17%, >5 cm=20%). ER- and PR- tumors were more likely to be HER2+ (of all ER- tumors, 29% were HER2+; of all PR- tumors, 25% were HER2+; of all ER+ tumors, 10% were HER2+; of all PR+ tumors, 9% were HER2+). Among all IBC patients, percent of disease that was HER2+ was highest for Asians (20%) and lowest for Caucasians (12%).CONCLUSION: In a large, unselected, population of invasive breast cancer patients, the incidence of HER2+ disease was highest among women aged 60-69 years. Among breast cancer patients, HER2+ disease was highest in Asians. The percent HER2+ was 13.6% for all breast cancer patients in this population based cohort and highest (24.7%) in the 20-39 year age group.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3058.
The TGFBR1*6A/9A Polymorphism Is Not Associated with Differential Risk of Breast Cancer.
Authors: McInerney N. , McInerney N. , Colleran G. , Rowan A. , Barclay E. , Jones A. , Curran C. , Miller N. , Tomlinson I. , Sawyer E. , Sawyer E. , Kerin M.
Published: December 2010

Background: A polymorphic 9-bp deletion in exon 1 of TGFBR1 (TGFBR1*6A) has been identified as a low-penetrance cancer susceptibility allele. The strongest association in the initial studies was with breast cancer; however these studies included patients with different types of cancer, including colon, cervical and breast carcinomas, with only a small proportion being breast cancer patients. In subsequent case-control studies focussing on breast cancer alone, the results have been equivocal. Methods: In order to clarify whether TGFBR1*6A is associated with breast cancer risk, we have genotyped this polymorphism in 988 breast cancer cases and 1016 controls form the West of Ireland and also performed a meta-analysis of previously published data (5150 cases and 6344 controls). All statistical analyses were undertaken using STATA 9.2.Results: In our series from the West of Ireland, we found no association (genotypic odds ratio (OR) under a dominant model= 0.93, 95% confidence interval (CI) 0.73-1.19, p=0.57; allelic OR=0.93, 95% CI 0.74-1.15, p=0.49). Meta-analysis showed evidence of heterogeneity among studies. Using the random effects model, there was no evidence of an association of the *6A allele with breast cancer (genotypic OR under a dominant model= 1.10, 95% CI = 0.94 to 1.28, p=0.24, allelic OR=1.12, 95%CI 0.97-1.31, p=0.13) with breast cancer. Conclusions: This study confirms that TGFBR1*6A is not associated with breast cancer and highlights the problems associated with small case-control studies, especially where minor allele frequencies are low. It remains to be seen whether the association with other cancer types, including colorectal cancer, can be validated in large case-control series.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3062.
In Asian-American Women, Westernization Influences Estrogen Metabolism, but Not Total Endogenous Estrogen Production.
Authors: Ziegler R. , Fuhrman B. , Xu X. , Gail M. , Keefer L. , Veenstra T. , Hoover R.
Published: December 2010

Background: Historically, breast cancer incidence rates were 4-7 times higher in the United States than in China and Japan. When Asian women migrate to the U.S., their risk rises over several generations and approaches that in U.S. Whites. Endogenous estrogen has been postulated to explain these international differences. Methods: In a population-based case-control study in Chinese, Japanese, and Filipino women living in San Francisco-Oakland, CA; Los Angeles, CA; or Oahu, HI, we documented a six-fold gradient in breast cancer risk with Westernization. Using 12-h, overnight urines from 263 premenopausal mid-luteal and 168 postmenopausal control subjects, we investigated associations of estrogens and estrogen metabolites (EM) with Westernization, particularly birthplace in the East or West. Fifteen EM were measured concurrently by liquid chromatography-tandem mass spectrometry with high accuracy, precision, and sensitivity. Log-transformed EM measures, adjusted for age and ethnicity, were compared by robust linear regression. Results: Total urinary EM (pmol/mg creatinine) was not associated with birthplace in premenopausal or postmenopausal women. When the three estrogen hydroxylation pathways were expressed as percent of total EM, the relative difference by birthplace, West compared to East, was 15% lower for 2-pathway EM (p=0.009), 7% lower for 4-pathway EM (p=0.37), and 10% higher for 16-pathway EM (p=0.008). Similar results were observed for individual EM in each pathway, premenopausal and postmenopausal women, and the three ethnicities. In both menopausal groups, 2-pathway EM concentration decreased steadily with increasing risk across six categories of Westernization. Conclusions: Our results do not indicate that total estrogen production explains the increased breast cancer risk in Western societies. However, estrogen metabolism patterns, especially reduced 2-hydroxylation, may contribute to the elevated risk; and underlying mechanisms merit further exploration.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3063.
Prevalence of Methylenetetrahydrofolate Reductase C677T and A1298C Polymorphisms and Risk of Breast Cancer in Mexican Women: An Interim Report of a Case-Control Study.
Authors: Scott S. , Loyo T. , Borbolla J. , Villela L. , Perfecto Y. , Cardona S.
Published: December 2010

Background: Among other causes, Breast Cancer (BC) appears as a result of genetic and epigenetic changes. Modifications in important pathways, such as the reactions that provide essential cofactors for synthesis of S-adenosylmethionine (one-carbon metabolism), and those involved in DNA synthesis and methylation could be linked to the development of this disease. Research has been conducted about whether MTHFR polymorphisms are related to the development of breast cancer in at least 10 different races, with different outcomes. Two of the most frequently studied MTHFR polymorphisms are C677T and A1298C, and their frequency is unknown among Mexican women. Our objective was to determine the frequency of these two polymorphisms and to ascertain weather their presence increases the risk of developing BC in our population.Materials and methods: 37 blood samples from histopatologically confirmed breast cancer patients were collected from hospitals in Monterrey, Uruapan and Villahermosa, all in Mexico. 115 healthy control samples were obtained from the same cities, and were matched for age in 5-year interval-groups. Cases and controls were third generation Mexican and older than 18 years. Genotyping of every sample was performed by PCR and restriction with enzymes.Results: The C677T Polymorphism was found in 28 cases (75.67%), whereas 76/115 controls (66.08%) showed the polymorphic allele. Case-controls analysis was adjusted to the confounding variable age group, an odds ratio of 1.58 (IC 95% de 0.66 – 3.82) was obtained. We did not find a significant relationship between the presence of the C677T polymorphism and the presence of breast cancer. The A1298C Polymorphism was found in 19/37 cases (51.35%), while only 32/115 (27.82%) of the control samples contained the allele. The odds ratio adjusted for the same confounding variable was 2.72 (IC 95%: 1.26 – 5.87). These results suggest a significant association between the presence of the polymorphism and BC.Discussion: Despite the exhaustive research that has been conducted regarding MTHFR polymorphisms C677T and A1298C, their association to the risk of breast cancer remains unclear. Some research groups even suggest that the presence of the polymorphisms, either individually or combined, confers certain protection against the disease. Up until now there were no data in Mexican population. Differences in the results of different works probably lie in the nature of the populations analyzed and the experimental design. We believe that a bigger sample size will allow us to determine if A1298C polymorphism is indeed, a risk factor for BC in Mexican women.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3064.
Complementary and Alternative Medicine Use and All-Cause Mortality among U.S.-Born and Foreign-Born Breast Cancer Cases: Results from the Hereditary Breast and Ovarian Cancer (HBOC) Study.
Authors: Bessonova L. , Largent J. , Anton-Culver H.
Published: December 2010

Background: Complementary and alternative medicine (CAM) therapies are used by people from all racial/ethnic backgrounds, and CAM use among cancer patients has recently been a focus of much research. Breast cancer patients, specifically, have embraced many CAM therapies. Use of these therapies is estimated to be between 48 and 70 percent among women diagnosed with breast cancer in the United States (Nahlez and Tabbara, 2003). Mortality patterns in foreign-born women diagnosed with breast and other cancers who immigrate to the United States are complex (Singh and Hiatt, 2006). However, it is unclear how country of birth relates to CAM use and overall survival among women with breast cancer. Therefore, we investigated the associations between country of birth, CAM use, and overall survival in the Hereditary Breast and Ovarian Cancer (HBOC) study, a population-based sample of breast cancer cases in California, comparing U.S.-born versus foreign-born women.Methods: Five hundred and fifty incident breast cancer cases (average age=54.68; SD=11.12) were sampled from the HBOC Study, which were identified through the Orange County regional registry for the state California Cancer Registry. Cases were contacted via telephone, and information on health behaviors, disease history, and socio-demographic characteristics were collected. A self-administered questionnaire queried use of any of the following CAM therapies: herbal teas, megavitamins, acupuncture, religious services, nonreligious services, aspirin, and/or non-prescription pain medicines. Chi-square test followed by Cox proportional hazards regression were used to compare overall survival between U.S.-born and foreign-born breast cancer cases as well as CAM ever- versus never-users.Results: Univariate analyses revealed that 59.5% of foreign-born breast cancer cases, versus 55.7% of U.S.-born reported having ever used CAM (Chi-square=0.76; p=0.38). In multivariate analyses, foreign-born women with breast cancer had roughly 3 times greater risk for all-cause cause mortality (HR=3.38; 95% CI: 2.14, 5.33) compared with U.S.-born breast cancer cases, adjusting for history of CAM use, education, number of existing co-morbidities, stage of tumor at diagnosis, and age at time of survey. Stratified analyses revealed that CAM use was not significantly associated with all-cause mortality using the above multivariate model among foreign-born (HR=1.35; 95% CI: 0.77, 2.37), or U.S.-born breast cancer cases (HR= 2.36; 95% CI: 0.99, 5.64).Discussion: These findings suggest that although foreign-born breast cancer cases are at greater risk for all-cause mortality, history of CAM use does not significantly protect nor increase the risk of death for this population.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 3069.