bluuebunny/crossref_metadata_2025_split huggingface dataset:
Development of a blood based breast cancer test for Indian population.
Authors: Tobin D , Bårdsen K , Lindahl T , Kauczynska M , Punia D , Kumar Y , Desai C , Shroff C , Børresen-Dale A , Sharma P 
Published: December 2010

Abstract
Abstract #5013
Background: The incidence of breast cancer (BC) in India is increasing as the society becomes more westernized. No national screening program exists and there is a high mortality rate which may be linked to the late detection of the disease. Here we report the development of a gene expression based blood test to detect BC. We have previously presented data from European/US cohort with accuracies ranging between 75% - 82% suggesting that test can be developed to early detect breast cancer. We now report the findings from a multi-centre study initiated to test the efficacy of the test for early detection of breast cancer (BC) in an Indian population.&#x2028; Methods: A multicentre-study was initiated where blood samples were collected from women recruited in 5 different groups 1) early stage BC, 2) late stage BC, 3) women at high risk - without BC, 4) benign breast lesions, 5) without abnormal mammographic findings. The women recruited in group 3 is a part of ongoing prospective study. Recruitment was balanced between pre- and post-menopausal women among the remaining groups. Samples were collected in PAXgene tubes and shipped to a central laboratory where RNA extraction and quality control. Gene expression analysis was performed using TaqMan® low density arrays (LDA's) containing a BC-specific gene signature in a 96-gene assay format. A total of 442 samples from groups 1, 2, 4 and 5 were used to develop the India model and estimate its prediction efficacy. The 442 samples were divided between a training cohort (N=292) and a test cohort (N=150). Gene expression data of the training cohort were used to develop the breast cancer specific diagnostic model and its performance was determined by predicting the class of the independent test cohort.&#x2028; Results: The developed model correctly predicted the class of 110/150 test samples, resulting in an overall accuracy of 73%. Prediction performance was similar for early and late stage cancer and for benign and healthy (no mammographic findings). No significant difference in diagnostic performance was seen between pre- (57/75 correctly predicted) and post-menopausal (53/75 correctly predicted) women indicating the clinical value of our test in younger women.&#x2028; Conclusion: The Indian multi-centre study demonstrates the utility of the BC-specific 96 assay signature in detecting BC in an Indian population with similar diagnostic performance seen between women with late stage (stage 2+) and early stage BC. We have previously presented data from European and US cohorts with similar diagnostic performance suggesting the broad applicability of our test. The Breast Cancer test showed the same high level of accuracy in pre-menopausal as in post-menopausal women indicating its clinical applicability in younger women where mammography is of less value due to dense breast tissue.
Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 5013.
Minimal residual disease detection in peripheral blood of primary breast cancer patients – translational research in the SUCCESS-study.
Authors: Jenderek C , Jückstock J , Schindlbeck C , Rack B , Fuchs R , Noeding S , Krabisch P , Ebner F , Lichtenegger W , Beckmann MW , Friese K , Sommer H , Janni W 
Published: December 2010

Abstract
Abstract #5019
Background: The detection of occult tumor cells in the bone marrow of patients with breast cancer has been shown to be an independent predictive and prognostic factor for early disease recurrence and decreased overall survival. However, it is more feasible to monitor treatment efficacy by sampling peripheral blood. The translational research program of the German SUCCESS-trial evaluates Minimal Residual Disease (MRD) in peripheral blood during adjuvant treatment of breast cancer patients. Follow-up results of the detection of MRD at primary diagnosis and after adjuvant chemotherapy (CT) are presented.&#x2028; Patients and Methods: Cells were separated by Oncoquick® (Greiner-bio one, Frickenhausen, Germany) followed by labeling of epithelial cells with an anti-cytokeratine-antibody (directed against cytokeratins 8, 18 and 19) and immunocytochemical staining. Every preparation was screened by two independent persons. MRD was defined as one or more cytokeratine-positive cell.&#x2028; Results: 1237 breast cancer patients were analyzed at primary diagnosis. After completion of adjuvant CT 655 patients returned for a second blood sampling. The majority of the tumors was small (40.1% pT1, 52.7% pT2, 5.9% pT3, 1.1% pT4) but mostly of intermediate or unfavorable grade, (G1 4.6%, G2 48.9%, G3 46.2%). 65% of the patients were node-positive (35% pN0, 45.6% pN1, 13.8% pN2, 5.7% pN3) and a positive hormone receptor status was seen in 69.1%. In 25.4% the Her2 status was positive. In 20.3% of all patients MRD was found at primary diagnosis. After CT MRD was observed in 15,7%. The mean number of detected cells was 2.3 (range: 1-25) before and 2.1 (range 1-23) after chemotherapy. Neither tumor size (p=0.48, p=0.47), lymph node metastases (p=0.34, p=0.81), histopathological grading (p=0.83, p=0.85) nor hormone receptor status (p=0.72, p=0.65) of the primary tumor correlated with the presence of MRD before or after CT respectively. The Her2/neu status of the primary tumor did not correlate before, but after CT with the presence of the MRD (p=0.63, p=0.08).&#x2028; Conclusion: The detection of MRD in peripheral blood can be widely used and is suitable for repeated measurements. Further follow-up of the SUCCESS trial will show this method´s validity for risk stratification and monitoring of treatment efficacy in breast cancer patients.
Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 5019.
HER2+ circulating tumor cells (CTCs): a more sensitive and specific marker for CTCs positivity than total CTCs count in early breast cancer.
Authors: Ignatiadis M , Durbecq V , Markowicz S , Chaboteaux C , Rouas G , Mounia B , Benjamin B , Liebens F , Georgoulias V , Mavrudis D , Piccart M , Sotiriou C 
Published: December 2010

Abstract
Abstract #5023
Background: CTCs have been detected using the CellSearch System in early and metastatic breast cancer (BC). However, no studies have been reported for the detection of HER2+ CTCs using the CellSearch System. In this study we aimed to analyze CTCs for HER2 expression in different stages of BC progression from ductal/lobular carcinoma in situ (DCIS/LCIS) to early and metastatic BC. Patients and Methods: 20ml of peripheral blood per woman was analyzed from healthy women, DCIS/LCIS or early BC patients, whereas 7.5ml of blood was analyzed from metastatic BC patients. The presence of CTCs as well as HER2 expression was assessed with the CellSearchSystem (Veridex, USA). After immunomagnetic enrichment with an anti-Epcam-antibody, cells were labeled with anti-cytokeratin (8,18,19), anti-HER2 and anti-CD45 antibodies. CTCs were defined as Cytokeratin-positive/CD45-negative cells, whereas HER2+ CTCs were defined as Cytokeratin-positive/CD45-negative/HER2-positive cells. Results: The presence of CTCs and HER2+ CTCs in healthy women, DCIS/LCIS, early and metastatic BC is depicted in table.&#x2028; &#x2028; Since 1 and 2 CTCs were detected in 7 (14.8%) and 4 (8.5%) of 47 healthy women, respectively, &gt;1CTC or &gt;2CTCs were selected as cutoffs for further analysis. Using the above cutoffs, 7 (18.9%) and 5 (13.5%) of 37 patients with early BC were considered CTC-positive, respectively. Since no HER2+ CTCs were detected in any of the 47 healthy women, a cutoff of ≥1 HER2+ CTCs was chosen. With this cutoff, more women with early BC [11 (29.7%) of 37 women] were considered CTC-positive. Conclusion: HER2+ CTCs seems to be a more sensitive and specific marker for CTCs' positivity than total CTCs counts in early breast cancer. Clinical relevance of the above cutoff for CTCs positivity using HER2+ CTCs detection will be performed in ongoing clinical trials of HER2 positive early breast cancer.
Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 5023.
Metastatic potential of Her2/neu positive and Her2/neu negative circulating epithelial tumor cells (CETC) released during primary systemic chemotherapy.
Authors: Pachmann K , Camara O , Gajda M , Rabenstein C , Pachmann U , Runnebaum IB 
Published: December 2010

Abstract
Abstract #5031
Background: Most tumor patients do not die from the primary tumor but from metastases in vital organs. To form metastases, cells must exit from the original tumor and reach these organs mainly via blood before they can settle and grow. Peripheral blood can be regarded as a transport system for CETC. Here we report on the influence of trastuzumab on the number of CETC and the impact on metastasis formation.&#x2028; Material and Methods: Tumor cells were quantified with an automated microscope (Laser Scanning Cytometer®, Icis or ScanR) from anticoagulated blood drawn before each new therapy cycle from patients treated with primary systemic chemotherapy and before and after surgery and subsequent maintenance therapy. After red blood cell lysis leucocytes were stained with PE-anti-CD45 and tumor cells with FITC-anti-EpCAM and changes registered as % increase or decrease. The response of CETC to therapy was correlated to outcome.&#x2028; Results: All breast cancer patients scheduled for primary systemic treatment had CETC before therapy. The number correlated with tumor size. The variable initial decrease in cell numbers highly correlated with the final tumor size reduction in her2/neu negative patients predicting for relapse free survival. All patients responded to the subsequent taxane therapy with an increase in CETC. In her2/neu positive patients who received combined taxane/trastuzumab there was only a modest increase in CETCs whereas patients with her2/neu positive disease who received taxane only showed a steep increase in CETC. 4 her2/neu positive patients who did never receive trastuzumab all had increasing CETC numbers during the postoperative observation time and all have suffered relapse. 6/6 patients who received trastuzumab only after surgery all had decreasing CETC numbers and all are in sustained complete remission after up to 4 years. 6/15 patients showed no change or an increase in CETC in spite of trastuzumab treatment and of these 4 have suffered relapse. Most relapses occurred 1-2 years after stopping trastuzumab.&#x2028; Discussion: Monitoring CETC will not only provide the earliest and most reliable indicator of successful neoadjuvant treatment and spare patients unnecessary treatment. In patients with the highly aggressive her2/neu positive tumors such cells may have an increased potential to settle and re-grow. This warrants further therapy studies to control what patients may benefit from additional or modified therapy already before metastases appear.
Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 5031.
Overexpression of LMW cyclin E results in faster M phase progression via Cdc25C interaction leading to chromosome missegragation in breast cancer cells.
Authors: Bagheri-Yarmand R , Keyomarsi K 
Published: December 2010

Abstract
Abstract #5032
Cyclin E is a G (1) cyclin essential for S-phase entry and has a profound role in oncogenesis. Cyclin E is overexpressed and present in low-molecular-weight (LMW) isoforms in breast cancer cells and tumor tissues compared to normal cells and tissues. Such alteration of cyclin E is linked to poor patient outcome. We have previously shown that ectopic expression of low molecular weight of Cyclin E (LMW-E) in breast cancer cell lines induces genomic instability by promoting aneuploidy and the formation of structural chromosomal lesion. To determine the mechanism of induction of genomic instability by ectopic expression of LMW-E, we generated the MCF-7 cells inducibly expressing either the full length (EL) or low molecular weight (LMW-E) form of cyclin E. We observed 5.5 fold higher number of cells with centrosome defects in LMW-E induced versus EL induced cells. Notably, elevated levels of LMW-E led to a significant increase in fragmented nuclei and multinucleated cells. Immunofluorescence analysis also showed a 4-5 fold increase in the number of cells containing lagging chromosomes and anaphase bridges. Karyotype analysis for gross chromosomal aberrations metaphase preparation of the uninduced and EL and LMW induced cells showed a significantly higher percentage of chromosomal defects including dicentric chromosome and chromosomal breaks associated with LMW-E overexpression. We also show that while overexpression of EL leads to accumulation of cells in S and G2/M phases of the cell cycle, the overexpression of LMW of cyclin E causes a faster M phase progression. As a consequence, LMW-cyclin E overexpressing cells are resistant to nocodazole-induced M phase arrest. Using biochemical assays, we show that EL and LMW-E interact with the phosphatase Cdc25C and when either EL or LMW are in complex with CDK2 that they phosphorylate Cdc25C. Such association of LMW-E with Cdc25C results premature inactivation of cyclin B/CDK1 upon treatment of these cells with nocodazole. Additionally, continued induction of LMW-E results in premature inactivation of PLK1 in mitotic cells and deregulated localization of PLK1 such that it is no longer associated to central spindles during anaphase and telophase. Collectively, these results suggest that overexpression of LMW-E interferes with the activation of CDK1 during mitosis. The relatively low activity of CDK1 during the passage through prophase, prometaphase, and metaphase interferes with the regulation of the spindle checkpoint and facilitates the premature transition of dividing cells from metaphase to anaphase resulting in chromosomal instability and aneuploidy. This is the first observation of how mechanistically the overexpression of a proto-oncogene can lead to genomic instability in breast cancer.
Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 5032.
Amplication of the estrogen receptor 1 (ESR1) gene in mastopathy predicts development of breast cancer.
Authors: Kilic E , Rufle A , Holst F , Ihnen M , Simon R , Zlobec I , Terracciano LM , Sauter G 
Published: December 2010

Abstract
Abstract #5037
Background: Mastopathy is a disease with fibrocystic changes (FCC) of the breast tissue commonly seen between the ages 30-50, but rare in postmenopausal women, and therefore suggestive of an association with estrogen stimulation. Whether FCC bears an elevated breast cancer risk is unclear. The role of ESR1 amplification in FCC has not previously been investigated. Material and Methods: Paraffin embedded tissues from 59 women with invasive breast cancer (BrCa) were analyzed. For all women, tissues with FCC taken at least 1.5 years before first diagnosis of BrCa (termed here “FCC old”) were also available. Average latency between FCC and BrCa was 6years (range1.5-17y). Immunohistochemistry (IHC) with antibody against estrogen receptor alpha (ER) and FISH-analyses with a probe for ESR1 (Zytovision, Bremerhafen/Germany) were performed. As a control, tissues from 19 women with FCC without a diagnosis of in situ or invasive breast carcinoma in the observed time frame (12-18y) were taken. Clinical-pathological data, such as histological subtype, tumour stage, tumour diameter, BRE-grade, patient's age and others, were available for statistical analysis. Results: After FISH analyses, 9 of 59 (15%) BrCa were amplified for ESR1. All ESR1-amplified BrCa were strongly positive for ER with IHC (at least 80% of tumour cells labelled). Tumour type or stage and BRE-grade did not correlate with ESR1-status. In all cases of ESR1 amplification in BrCa areas of amplification in FCC adjacent to the tumour were identified. Interestingly, in women with ESR1 amplification in BrCa, an amplification detectable in tissues with FCC prior to the first diagnosis of BrCa (“FCC old”) was noted but was absent in tissues from women with FCC who did not develop BrCa in the observed time frame. Non-neoplastic ESR1 amplified cells within FCC were strongly positive for ER with IHC. In unaltered mammary glands ESR1 amplification was not observed. Discussion: In this pre-selected collective of women with BrCa and history of FCC we found amplification of the ESR1 gene in 15% of BrCa. In these women, ESR1 amplification was also detectable in tissue with FCC taken prior to the first diagnosis of BrCa. This investigation shows that ESR1 amplification is an early event in breast pathology and its occurrence in FCC predicts development of BrCa.
Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 5037.
Lapatinib selectively inhibits HER2 over-expressing breast cancer stem-like cells.
Authors: Johnson RL , Farnie G , Clarke RB , Bundred NJ 
Published: December 2010

Abstract
Abstract #5052
Introduction&#x2028; Standard drug treatment kills proliferating cells but recent studies show the tumourigenic cancer stem cells (CSC) are not affected, leading to recurrence. HER2 over-expression in Ductal Carcinoma In Situ (DCIS) and invasive cancer is associated with poor prognosis. We determined the effect of a dual tyrosine kinase inhibitor (Lapatinib) on CSC growth in mammosphere (non-adherent) and 3D culture in HER2 over-expressing cell lines and primary DCIS samples.&#x2028; Methods&#x2028; MCF7, BT474 (HER2+), DCIS.com and SUM225 (HER2+) cell lines were cultured as mammospheres in the presence or absence of Lapatinib (0.1-5mM). FACS analysis was used to measure proliferation (Ki67) and apoptosis (annexin / PI) rates. Western blotting was used to analyse levels phosphorylated MAPk and AKT in mammospheres. 3D matrigel culture of cell lines and primary DCIS cells was carried out with and without Lapatinib (1mM), the size and number of colonies grown were assessed. Statistical analysis was by unpaired 2-tailed T test.&#x2028; Results&#x2028; Mammosphere forming efficiency (MFE) of HER2 over-expressing but not HER2 negative cell lines was inhibited by Lapatinib (at IC50 p&lt;0.05) and the IC50 for inhibited cell lines determined (Table 1). Lapatinib treatment at the IC50 concentration in both BT474 and SUM225 reduced proliferation by 10% after 24 hours of mammosphere culture (p&lt;0.05), however there were no differences in apoptosis. Lapatinib reduced pMAPk and pAKT in HER2 over-expressing cells during mammosphere culture in a dose dependent manner. Secondary mammosphere culture demonstrated a sustained 50% reduction in mammosphere formation from primary mammospheres cultured with Lapatinib, suggesting self renewal was inhibited in mammosphere culture.&#x2028; In 3D culture of HER2 over-expressing SUM225 and primary DCIS samples Lapatinib (1µM) significantly reduced the size of the colonies but no difference was observed in the number of colonies formed (Table 1).&#x2028; &#x2028; Conclusion&#x2028; Lapatinib selectively inhibits breast cancer stem cell activity by reducing proliferation in HER2 over-expressing lines measured using non-adherent and 3D culture. These findings need to be verified by an in vivo limiting dilution tumourgenicity assay.
Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 5052.
The effect of intermittent versus continuous energy restriction on biomarkers of breast cancer risk.
Authors: Harvie M , Chapman M , Cuzick J , Flyvbjerg A , Jebb S , Mattson M , Howell A 
Published: December 2010

Abstract
Abstract #506
Excess energy, whether it is the form of high energy diets or low energy expenditure (sedentary lifestyle), is a risk factor for breast cancer. Observational studies show that continuous energy restriction (CER) and exercise reduce risk, particularly of postmenopausal breast cancer. Animal data indicate that intermittent energy restriction (IER) may be superior or at least as effective as continuous energy restriction in preventing or reducing the growth rate of mammary tumours. IER may also be more acceptable than CER which has been difficult to implement.&#x2028; We have tested the effectiveness and acceptability of IER and have performed a randomised trial of IER (n=53) versus CER (n=54) in overweight or obese premenopausal women. Energy restriction was 25% in both arms. IER subjects had approximately 550 kcal on 2 days per week and add lib food on other days; CER subjects had approximately 1500 kcal / day over 7 days. Nineteen women (18 %) withdrew from the study before 6 months (IER = 12, CER = 7, main reasons: stress = 4, pregnancy = 3, change in employment = 3, couldn't stick to diet = 3).&#x2028; Change in weight and biomarkers for last observation carried forward are reported.&#x2028; At 6 months. Both IER and CER are effective for loss of weight -7.9 vs. - 6.7% P = 0.26 and waist circumference -6.3 vs. -5% P = 0.13, and for lowering serum levels of leptin -43.3 vs. -39.8% P = 0.53; C – reactive protein -14.8 vs.-20.3% P = 0.15 and free androgen index -9.8 vs.-14.4% P = 0.90. Both IER and CER increased circulating levels of sex hormone binding globulin + 18.1 vs. 10.8% P = 0.51, and plasma adiponectin +11 vs. +1% P = 0.09. IER is better at lowering insulin resistance at 6 months than CER -29.6 vs. -15.3 % P &lt;0.05.&#x2028; These data indicate that 2 days IER gives similar results to CER with respect to weight loss and many of the risk parameters measured and may be an alternative approach to continuous energy restriction. The superior reductions in insulin resistance with IER which may be related to periods of very low calorie intake (approximately 70% restriction). We are waiting data of the effects of IER vs. CER on the IGF-1 axis and oxidative stress. Future studies will investigate the mechanism of IER, its optimal duration and its influence on mammary cell function.
Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 506.