bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P6-08-14: TRPS-1, a New GATA Family Transcription Factor, Regulates Epithelial-Mesenchymal Transition and Maintains an Estrogen Responsive, Claudin-Positive Phenotype in Breast Cancer Cells
Authors: Bao Y , Chen JQ , Wu Y , Leng X-H , Wang E , Marincola F , Radvanyi LG.
Published: April 2011

We recently reported that TRPS-1 (Trichorhinophalangeal Syndrome-1), a new GATA family transcription factor, is a ductal epithelial cell-specific gene expressed in normal breast and over-expressed in the majority of human breast cancer (BC), especially in ER+ BC. We also found that TRPS-1 is a marker for better prognosis when expressed over a critical threshold level using a new quantitative IHC method applied to over 150 primary BC cases of all subtypes. Here, we studied the molecular function of TRPS-1 using a lentiviral shRNA knockdown approach in an ER+ ductal epithelial BC cell line (T47D; luminal B type) normally expressing high levels of TRPS-1. Strikingly, knockdown of TRPS-1 in T47D cells resulted in a complete loss of epithelial markers, including E-cadherin and CK8/18 protein together with the loss of ER and a host of ER-associated genes, such as GATA-3 and FOXA1. In contrast, N-cadherin, vimentin, slug, and other genes associated with a mesenchymal phenotype were induced. Gene expression profiling using microarray found a number of additional differentially expressed genes following TRPS-1 knockdown, including a large decrease in the Prolactin receptor (PRLR), Claudin-1 and 6, EpCAM, PGR, and CD24, while AKT3 and MET (HGF-R) were increased. The Claudin-1 and PRLR gene were down-modulated 149-fold and 167-fold, respectively. This was verified at the protein level, with a total loss of cell surface Claudin-1 and PRLR expression in the TRPS-1 knockdown cells. Overall, the gene expression profile of TRPS-1 knockdown T47D cells resembled the recently described “Claudin-low” BC subset. TRPS-1 knockdown also had dramatic functional consequences, with the cells acquiring a mesenchymal, spindle-like morphology and high resistance to growth arrest and apoptosis under conditions of serum and estrogen deprivation. Moreover, these cells formed aggressive tumors in nude mice in an estrogen-independent manner after orthotopic injection into mammary fat pads, while control cells produced only small non-palpable tumors. In addition, transient induction of TRPS-1 expression in ER/TRPS-1-negative MDA-MB-231 cells led to a significant expression of ER together with other ER-associated genes. Our data suggests that TRPS-1 is a critical new GATA family transcription factor in the mammary gland that is at the hub of a cell lineage-determination pathway regulating epithelial-mesenchymal transition. In BC it may function in maintaining a more highly differentiated and less aggressive Claudin-positive ductal epithelial phenotype.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P6-08-14.
Abstract P6-09-05: Insulin Resistance, Diabetes Mellitus and Breast Cancer Risk in Pre-and Postmenopausal Women: Modification Effect by Moderate Physical Activity
Authors: Torres-Mejía G , Ángeles-Llerenas A , Ortega-Olvera C , Lazcano-Ponce E , Romieu I.
Published: April 2011

Background

It has been hypothesized that diabetes may influence the risk of breast cancer (BC) by impaired regulation of the insulin and insulin-like growth factors (IGF) pathways. Current evidence supports a reduction in BC risk by doing regular physical activity due, in part, by reducing insulin resistance and improving glucose control. Material and Methods

A population based case control study was conducted to evaluate the association between insulin resistance and diabetes mellitus on the risk of BC in Mexican women. A total of 1000 incident cases and 1074 matched controls to cases by five years of age, site and health institution were analyzed. Women provided information on health, diet and physical activity by means of a face to face interview; information about the family history of diabetes mellitus and whether or not women were diagnosed with this disease. Anthropometric measurements and blood samples were obtained from all women. A conditional logistic regression model was used to assess this association. Results

In postmenopausal women, there was an interaction between physical activity and diabetes mellitus on the risk of BC. The effect of diabetes mellitus type 2 decreased with increasing moderate physical activity (OR = 3.67, 95% CI 1.78 — 7.58; OR = 2.86, 95% CI 1.37 — 5.96; OR = 0.82, 95% CI 0.19 — 3.59, by increasing tertiles of daily hours of moderate physical activity respectively); in premenopausal women, this association was not statistically significant (OR = 1.55, 95% CI 0.86-2.79), however in a subsample of premenopausal women, an increase was observed in the risk of BC per unit change in the increase of C-peptide, an indirect marker of insulin resistance (OR = 1.98, 95% CI 1.41 - 2.76). Discussion

The results of this study are consistent with numerous mechanisms linking physical activity directly to reduce BC risk probably, in part, through reducing insulin resistance and improving glucose control.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P6-09-05.
Abstract P6-12-01: A Meta Analysis of Risk of Cardiovascular Events in Patients with Metastatic Breast Cancer (MBC) Treated with Anti Vascular Endothelial Growth Factor (VEGF) Therapy — Bevacizumab
Authors: Nasim S , Sousa BA , Dent RA , Pritchard KI.
Published: April 2011

Background and objectives: Bevacizumab, used in combination with chemotherapy has demonstrated efficacy in randomized trials in MBC. Hypertension, congestive heart failure and cardiomyopathy have been reported in trials of bevacizumab in breast cancer. The aim of this systematic review and meta analysis is to determine the overall risk of grade 3-4 hypertension (HTN), left ventricular dysfunction (LVD) and venous (VT) and arterial thromboembolic events (ATE) related to Bevacizumab in patients with MBC.

Methods: We selected randomized phase III trials which compared chemotherapy with or without bevacizumab in MBC as first or second line therapy. Data extraction was carried out from results published in the literature or from conference proceedings of the selected studies. For the analysis, we used a fixed-effects and random effects models to calculate the pooled event-based relative risk ratios (RR) with 95% confidence interval (CI). The Cochran's Q statistic and I2 statistics were first calculated to assess the heterogeneity among the proportions of the included trials. We collected all grade 3-4 events reported in these trials in relation to cardiovascular events: HTN, LVD, congestive heart failure, cardiomyopathy, VT and ATE.

Results: Five trials were included in the meta-analysis: E2100, AVADO, RIBBON-1, RIBBON-2 and Miller et al study (capecitabine alone or plus bevacizumab in previously treated MBC). In total 2126 patients received chemotherapy in combination with bevacizumab and 1444 chemotherapy alone. When there was tendency for heterogeneity, random-effects models were used. Thromboembolic events were pooled together as VT and ATE as information was not clear in some of the reported studies. CHF and cardiomyopathy were considered for LVD events in the Miller study as LVD was not reported. Patients treated with bevacizumab-containing regimens had a RR for HTN of 10.32 (95% CI, 4.23- 24.79; p<.0001), RR for LVD 2.58 (95% CI, 1.06 - 6.32; p=0.04) and RR for VT and ATE of 1.71 (95% CI, 0.81- 3.60; p=0.16) as shown in figure 1.

Conclusion: HTN is a recognized side effect of bevacizumab therapy. The risk of LVD is also significantly higher with bevacizumab therapy (RR 2.58) as shown in this pooled analysis, but the risk of thrombotic events is not increased.

Figure available in online version.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P6-12-01.
Abstract P6-13-01: Impact of the Number of Prior Chemotherapy Regimens on Overall Survival (OS) among Subjects with Locally Recurrent or Metastatic Breast Cancer Treated with Eribulin Mesylate: Results from the Phase III EMBRACE Study
Authors: Blum JL , Twelves CJ , Akerele C , Seegobin S , Wanders J , Cortes J.
Published: April 2011

Background: The randomized, controlled, open-label, phase III EMBRACE study compared the efficacy and safety of eribulin mesylate (E7389) vs. treatment of the physician's choice (TPC) in heavily pretreated subjects with late-stage breast cancer. The study met its primary endpoint by showing that eribulin significantly prolonged OS vs. TPC (HR: 0.809; 95% CI: 0.660-0.991; p=0.041). This exploratory subgroup analysis aimed to determine the influence of the number of prior regimens on OS among eribulin-treated subjects.

Methods: Eligible subjects had locally recurrent or metastatic breast cancer and had received 2-5 prior chemotherapy regimens, including an anthracycline and a taxane, with ≥2 of the regimens given for locally recurrent or metastatic disease. Subjects had to show progression on or within 6 months of the last chemotherapy regimen. Subjects were stratified by geographic region, HER2/neu status, and prior capecitabine treatment and then randomized 2:1 to receive eribulin 1.4 mg/m2 intravenously over 2-5 minutes on Days 1 and 8 every 21 days or an approved TPC.

Results: A total of 762 subjects in 135 centers worldwide were randomized to receive either eribulin (n=508) or TPC (n=254). Subjects had a median age of 55 years, 92% were white, 76% were postmenopausal, and 81 % had taxane-refractory disease. Overall, 47% and 53% of subjects had received ≥3 and >3 prior chemotherapy regimens in total, respectively. Seventy-five percent and 25% of subjects received ≥3 and >3 prior chemotherapy regimens in total for locally recurrent or metastatic disease, respectively. Subgroup analysis identified consistently longer median OS with eribulin vs. TPC in subjects who received ≥3 prior chemotherapy regimens, both in total and specifically for locally recurrent or metastatic disease (Table). Lesser improvements in OS with eribulin vs. TPC were observed in subjects who received >3 prior chemotherapy regimens. The study was not powered to show statistical significance for this subgroup analysis. Conclusion: In this retrospective subgroup analysis, the OS benefit with eribulin appears greater for subjects with locally recurrent or metastatic breast cancer who received fewer previous therapies. Additional studies are underway to confirm these findings in less heavily pretreated patients.

Table. Overall survival by treatment history vs. TPC

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P6-13-01.
Abstract P6-13-04: Thymidylate Synthase (TS) and Thymidine Phosphorylase (TP) Expression as Predictive Markers of Capecitabine Mono-Chemotherapy for Patients with Anthracyclin-Taxane Pretreated Metastatic Breast Cancer (MBC)
Authors: Lee SJ , Choi YL , Cho EY , Kim ST , Park YH , Ahn JS , Im Y-H.
Published: April 2011

Background: Capecitabine, one of the widely used cytotoxic agents for breast cancer, achieved high tumor control rate with low toxicity even in heavily-pretreated patients with metastatic breast cancer (MBC). Capecitabine (Xeloda®) is an oral fluoropyrimidine pro-drug that is transformed to fluorouracil (FU) in several steps, the last of which is conversion of 5'-deoxy-5-fluorouridine to FU by TP. And TS is a target enzyme of 5-FU. The primary purpose of this study was to evaluate the role of TS and TP as biomarkers to predict clinical outcomes of capecitabine monotherapy for MBC.

Method: Of Three hundreds fifty-three patients with MBC treated with capecitabine at Samsung Medical Center from 2000 to 2008, ninety patients who were previously treated with anthracycline and taxanes containing regimens and had available tissues for immunohistochemical stainings with estrogen (ER), progesterone (PgR), and human epidermal growth factor receptor-2 (HER2), TS, and TP were included in this analysis. All patients had received capecitabine 2000-2500mg/m2/day for 14days q 3 weeks.

Results: The median age of these 90 patients was 49 (26-76) years. The response rate (RR) was 25.6% and the disease control rate (DCR) was 65.6%. The median progression-free survival (PFS) and overall survival (OS) from the 1st day of capecitabine monotherapy were 4.8 (95% C.I.; 3.1-6.5) and 26.7 (95% C.I.; 17.1-36.3) months, respectively. High TS expression (TS score ≥100) was more common in patients with triple negative (TN) subtype than in those with other subtypes (16% for HR+, 22% for HER2+, 44% for TN, p=0.023). Median PFS was significantly lower in high TS expression (6.6 vs. 3.0 months; p=0.017) and low TP expression (TP<50) patients (6.0 vs. 3.3 months; p=0.013). High TS score and low TP score were identified as unfavorable independent risk factors for PFS to capecitabine monotherapy in Cox regression multivariate analysis (Hazard Ratio (HR) 1.7, p=0.037 for high TS score; HR 1.8, p=0.014 for low TP score). And, high TS and low TP score were associated with poorer overall survival.(29.9 vs. 14.1 months; p=0.008 for TS /29.9 vs. 20.4 months; p=0.043 for TP)

Conclusion: Our data suggests that high TS score and low TP score correlated with shorter PFS for capecitabine monotherapy of patients with anthracycline and taxane pretreated MBC. Potential role of TS, TP expression as predictive markers of capecitabine monotherapy in patient with MBC should be confirmed by prospective studies.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P6-13-04.
Abstract P6-14-06: Prospective Testing of Three Different Gene-Signatures for Patient Selection for Dasatinib Therapy in Metastatic Breast Cancer
Authors: Pusztai L , Moulder S , Litton J , Valero V , Ueno N , Melhem-Bertrandt A , Morrow PK , Dotter K , Mattair D , Strauss L , Hortobagyi GN , Qi Y , Symmans WF.
Published: April 2011

Background: Several gene signature-based predictors of response to targeted drugs have been proposed in the literature but none has been prospectively tested as patient selection tools in the clinic. The goal of this trial is to assess the positive predictive value of 3 conceptually different multi-gene signatures as predictors of response to the multitargeted kinase inhibitor dasatinib.

Methods: This clinical trial requires biopsy of a metastatic lesion for gene expression profiling and employs a parallel, multi-arm, two-step, phase II design. Three markers are assessed including a (i) cell-line derived dasatinib-sensitivity signature, (ii) a src-pathway activity signature and (iii) a dasatinib target index calculated as the weighted average expression of all known dasatinib targets. Only markerpositive patients are treated with dasatinib 100 mg po daily and each marker arm is considered as a separate study with early stopping rules for futility (minimum sample size 9, maximum sample size 40/marker arm). A predictor is considered worthy of further study if the clinical benefit rate (i.e. positive predictive value) is ≥25%.

Results: Forty seven patients were accrued from July 2009 through June, 2010, 49 biopsies were performed (soft tissues n=31, liver n=8, bone n=3, lung n=1, adrenal gland n=1), 6 samples had poor cellularity and 3 failed array QC. There was no patient recall, hospitalization or emergency room visit due to biopsy procedure. The median time from biopsy to genomic prediction result was 5 days (range 3-7). Twenty three (57%) patients had positive result for at least 1 predictor (5 were positive for 2) and 20 are receiving therapy (3 withdraw or progressed before therapy began). Responses as of June 2010; Target index arm (n=9): 5 PD (progressive disease), 4 SD (3 stable disease at 8 weeks 1 SD at 16 weeks); SRC Pathway arm (n=5): 3 PD, 2 SD at 8 weeks; Cell line predictor arm (n=6): 2 PD, 1 SD at 8 weeks, 3 not yet reached response evaluation. None of the 3 predictive marker arms have met early stopping yet and accrual is ongoing. Conclusion: Gene-expression signature based patient selection for targeted therapy is feasible and FNA biopsies of metastatic lesions for genomic testing are safe. Updated efficacy results will be reported.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P6-14-06.
Abstract P6-14-11: ß-Catenin Inhibitor SST-024 Can Sensitize Breast Cancer Tumor Initiation Cells (TICs) to Ionized Radiotherapy
Authors: Xu W , Debeb BG , Alvarez de Lacerda LC , Woodward WA.
Published: April 2011

Recent studies have shown that tumor initiation cells (TICs) are more resistant to ionized radiotherapy (IR) than differentiated cancer cells. In our lab, we have previously shown that aberrant Wnt signaling can contribute to radio-resistance of TICs in breast cancer. Herein, we use SST-024 (StemSynergy), an inhibitor of ß-catenin and WNT signaling, to test if pharmacologic inhibition of ß-catenin can sensitize breast cancer TICs to IR. Sum-149, an inflammatory breast cancer cell line, Sum-159, a non-inflammatory basal-type breast cancer cell line and IBC-3, a short-term cultured inflammatory breast cancer sample were used in these studies. We found that SST-024 inhibited Sum-149 mammosphere formation, a surrogate for TIC growth in vitro, at IC-50 0.3 nM while IC-50 for Sum-159 was 3 nM and IC-50 for IBC-3 was 4 nM. Similar efficacy against TICs was seen in assessments of alternative TIC surrogates including aldefluor staining by flow cytometry and CD44+CD24- marked cells. On immunoblot analysis SST-024 down-regulated unphosphorylated, active B-catenin and downstream target c-myc as well as EMT marks, including N-cadherin, Snail, Twist and Slug, and DNA repair regulator ATM. Moreover, standard monolayer and 3D mammospheres clonogenic assays demonstrate significant radiosensitization of SUM-149 TICs (mammospheres) as well as monolayer colony formation. In summary,

SST-024 demonstrates significant efficacy against inflammatory breast cancer TICs and sensitize both TICs and non-TICs to IR implying targeting Wnt β-catenin signaling may be a feasible strategy to radiosensitize breast cancer TICs.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P6-14-11.
Abstract P6-14-16: A Systematic Review of the Evidence for Treatment of Metastatic Breast Cancer beyond 2nd Line
Authors: Lalla D , Sedelnikova M , Wieffer H , Brammer M , Modha R.
Published: April 2011

Background: Optimal treatment of metastatic breast cancer (MBC) post second-line is not well defined. Therapy options are limited by previous exposure, while HER2 status may remain relevant to decision-making. A systematic review was conducted to identify trials reporting efficacy data, and review the current treatment landscape for HER2+ and - patients for third and later line treatment of MBC.

Methods: A systematic search strategy from Medline, EMBASE and Cochrane (CCTR) databases retrieved citations for two-stage review. Citations that reported 3rd or later line treatment of MBC patients were included. Selected conference proceedings from the last 3-5 years were searched for relevant abstracts and posters. Included studies were restricted to those with data reported for 10 or more patients.

Results: 39 citations from 29 trials, investigating 22 treatment regimens, were identified which reported efficacy data specific for a 3rd or later line population: in 22 trials, this was a sub-group analysis of a mixed line population. 76% of included studies were single arm: only 1 RCT was identified. 21% of included studies investigated the treatment of HER2+ populations; no studies reported outcomes specific for the HER2- population. Overall response rate (ORR) was the most reported outcome. The majority of responses were partial, complete response was rarely observed beyond 2nd line. Overall and progression-free survival were rarely reported. The highest ORR was reported for mitomycin C and capecitabine in HER2 mixed patients. Table 1 presents response rates reported in HER2+ patients.

Conclusions: There does not appear to be a standard regimen used in the treatment of patients beyond the second line. In patients with HER2+ breast cancer, T-DM1 had the highest reported response rate.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P6-14-16.
Abstract P6-15-07: A Dose-Escalation Study with a Special Drug Delivery System (SDS) of BEZ235, a Novel Dual PI3K/mTOR Inhibitor, in Patients with Metastatic/Advanced Solid Tumors
Authors: Rodon J , Infante J , Burris H , Tabernero J , Ranson MR , Rouyrre N , Duval V , Silva A , Hackl W , Baselga J.
Published: April 2011

Background: The PI3K pathway plays a major role in cancer cell growth and survival and is the most frequently altered pathway in cancer. BEZ235 is a potent and highly specific oral PI3K/mTOR inhibitor. Administered as hard-gelatine capsule, BEZ235 was well tolerated with a favorable safety profile. Available data showed that BEZ235 led to PRs (1 patient (pt) with NSCLC [PTENnull], 1 pt with ER+ breast cancer [mutation unknown]) and prolonged disease stabilization (4 to >24 months) in pts with PI3K pathway dysregulated tumors (N=9/51), 5/9 with ER+, 1/9 with HER2+ breast cancer. Here, we present data from patients (pts) receiving treatment with a special drug delivery system of BEZ235 (SDS) with improved PK properties (Cao SABCS 2010).

Results: 22 pts have been treated with BEZ235 SDS capsule at 3 dose levels (qd): 400 mg (5); 800 mg (6), 1000 mg (11). Median age was 56 years. The most common tumor types among pts enrolled were: breast cancer (4), CRC (4) and NSCLC (3). 2/22 pts were treated for >3 cycles, 16/22 pts progressed within the first three cycles of treatment. The maximum tolerated dose (MTD) is 1000 mg/d. Dose-limiting toxicities (DLTs) included: 2 Grade (G)3 fatigue (800 mg and 1000 mg) and 1 G3 skin rash (at 1000 mg). The most common treatment-related AEs included: fatigue, diarrhea, nausea, vomiting (G1-3). Preliminary PK data showed a slow absorption rate, Cmax at ∼6 h post-dose (range 2-8 h). The elimination halflife is ∼6 h across doses and visits. Steady state was reached after 8 days of treatment; the mean (SD) exposures at steady state (AUC, rss) for the dose of 400, 800, and 1000 mg/d were 8864 (12210), 19060 (19860), 46010 (36150) ng*h/mL, respectively. The exposure across dose levels (AUC0- 24, SS) was above the exposure required for tumor stasis in PI3K pathway dysregulated tumors, estimated based on CT measurements in patients(Bottino SABCS 2010). Currently, the efficacy of BEZ235 SDS, alone and in combination with trastuzumab, in patients with PIK3CA mutated breast cancer is investigated. As of today 1 clinical PR was observed with BEZ235+trastuzumab in a patient with breast cancer (HER2+, PIK3CA mutated, trastuzumab refractory) with brain and peripheral metastases.

Conclusions: BEZ235 SDS treatment is well tolerated. The MTD is 1000 mg/d. Available PD and efficacy data show that BEZ235 is active in breast cancer (especially PI3K pathway dysregulated tumors). Phase II studies in breast cancer combining BEZ235 SDS with other agents will start soon.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P6-15-07.
Abstract P6-15-19: PI3K-MTOR Inhibitor BEZ235 Shows Antitumor Activity Against a Panel of Basal-Like Cells and Potentiates the Antitumor Activity of Taxanes and Vinorelbine
Authors: Montero JC , Esparis-Ogando A , Pandiella A , Ocana A.
Published: April 2011

Background: Intracellular signalling pathways associated with the oncogenesis of basal-like breast cancer are relatively unknown. Basal-like breast cancer constitutes 15% of all breast tumors. Although chemotherapy is the only treatment, no data support the use of one agent over others. Furthermore, combinations of chemotherapies with targeted agents against activated signalling pathways have not been explored. Here we have analyzed the action of the dual PI3K-AKT-mTOR pathway inhibitor BEZ235 on a panel of basal-like breast cancer cell lines. Methods: We used several triple negative breast cancer cell lines, and analyzed the action of BEZ235 on their proliferation and cell survival using MTT metabolization and other apoptosis assays. Biochemical experiments were performed to investigate the molecular bases of the action of BEZ235. Experiments in combination with chemotherapies were also performed to identify synergic combinations.

Results: PI3K-AKT pathway was activated in most of the cell lines studied. Administration of the dual PI3K-mTOR inhibitor BEZ235 decreased AKT phosphorylation in a dose dependent manner. It also reduced signalling through the mTORC1 and mTORC2 complexes. BEZ235 also decreased proliferation of most cell lines with IC50 values <1 μM. Administration of BEZ 235 with taxanes and vinorelbine showed synergy. Conclusions: The PI3K-AKT-mTOR signalling axis is frequently activated in basal-like cells. The efficient antitumoral effect of the inhibition of this route with BEZ235 indicates that targeting this pathway may be a relevant therapeutic strategy in triple negative breast cancer.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P6-15-19.
Abstract PD01-02: Integration of Capecitabine into Anthracycline-and Taxane-Based Adjuvant Therapy for Triple-Negative Early Breast Cancer: Final Subgroup Analysis of the FinXX Study
Authors: Lindman H , Kellokumpu-Lehtinen P-L , Huovinen R , Jukkola-Vuorinen A , Tanner M , Kokko R , Ahlgren J , Auvinen P , Bono P , Joensuu H
Published: April 2011

Background: Standard adjuvant chemotherapy regimens for patients with moderate-to-high risk early breast cancer typically contain a taxane, an anthracycline and cyclophosphamide. The randomised, phase III FinXX study aimed to investigate whether the integration of capecitabine into such a regimen enhances outcome. Here we present final 5-year exploratory analyses from the subgroup of patients with triple-negative early breast cancer (TNBC).

Methods: Patients aged 18-65 years, with histologically confirmed invasive node positive breast cancer or node negative progesterone receptor negative breast cancer > 20 mm, and with a WHO performance status of 0 or 1, with no previous neoadjuvant chemotherapy were randomised. Patients received 3 x XT (capecitabine 900mg/m2 bid d1-15 + docetaxel 60mg/m2 d1)≥3 x CEX (cyclophosphamide 600mg/m2 d1, + epirubicin 75mg/m2 d1 + X 900mg/m2 bid d1-15, q3w) or 3 x T (80mg/m2 d1)≥3 x CEF (C 600mg/m2 d1, E 75mg/m2 d1, 5-FU 600mg/m2 d1, q3w). The primary endpoint was RFS, defined as the time from randomisation to the first time the patient is recorded as having disease recurrence or the date of death if the patient dies due to causes other than disease recurrence. Secondary endpoints included overall survival and safety.

Results: Between January 2004 and May 2007, 1,500 women from Finland and Sweden were randomised (XT→CEX n=753, T→CEF n=747). Of these, 202 patients (13.5%) had TNBC (XT→CEX n=93, T→CEF n=109). After a median follow-up of 59 months, RFS was significantly improved in the TNBC XT→CEX arm versus control (HR 0.48, 95% CI 0.26-0.88; p=0.018). The following endpoints were also significant: 5-year RFS in patients with TNBC versus those without TNBC (HR 0.50, 95% CI 0.36-0.69; P<0.001); 5-year distant disease-free survival in TNBC patients receiving XT→CEX versus control (HR 0.51, p=0.035); overall survival in TNBC patients receiving XT→CEX versus control (HR 0.42, p=0.019). Conclusions: The FinXX trial is the first to report the efficacy of capecitabine in combination with anthracycline-/taxane-containing therapy in the adjuvant treatment of early breast cancer. The exploratory subgroup analysis shows that XT→CEX is more effective than T→CEF in triple-negative BC, a population with a high unmet need; XT→CEX versus control resulted in a significant improvement in RFS and OS after 5 years.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr PD01-02.
Abstract PD01-07: Transcriptome Analysis of Triple Negative Breast Cancers Identifies Six Distinct Biological Subgroups and Reveals Therapeutic Strategies
Authors: Lehmann BD , Bauer JA , Chen X , Sanders ME , Shyr Y , Pietenpol JA.
Published: April 2011

Background: Currently, there are targeted therapies for estrogen receptor (ER) positive and HER2-amplified breast cancers. However, treatment options for patients with triple negative breast cancers (TNBCs), tumors that are hormone receptor negative (ER and progesterone, PR) and lack HER2 amplification, has been limited by the heterogeneity and lack of moleclular understanding of the subtypes of this disease. Use of genomic information (gene expression, copy number analysis, methylation arrays, and next-generation sequencing) will unravel the complexity of the disease and identify novel targeted therapies.

Methods and Findings: In order to identify biologically distinct TNBC subgroups and associated molecular drivers, we compiled an extensive TNBC transcriptome dataset from 21 independent breast cancer studies. We performed reiterative k-means clustering analysis on a 386 TNBC gene expression profile training set and identified six stable clusters that display unique gene expression patterns and gene ontologies. An independent set of 201 TNBC gene expression profiles was used to validate the six TNBC subgroups. We identified two basal-like subgroups (BL1 and BL2) characterized by cell cycle and DNA damage response genes, two mesenchymal-like subgroups (M and MSL) enriched in cell differentiation, epithelial-mesenchymal transition and growth factor pathways, an immunomodulatory subgroup (IM) defined by immune cell surface antigens, receptors and signal transduction genes, and a luminal subgroup (LAR) driven by androgen receptor signaling. Using tumor gene expression signatures, we identified representative cell lines to model each of the TNBC subgroups and also pharmacologically targeted prominent signaling pathways.

Conclusions: Through transciptome analysis we subclassified TNBC into six biological subgroups and identified potential therapeutic targets for future exploration. The data generated in this study have significant value for target selection in drug discovery, clinical trial design and selection of biomarkers that will enable the alignment of TNBC patients to the appropriate targeted therapy.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr PD01-07.
Abstract PD02-07: Impact of Mesenchymal Stem Cells on Radiation Therapy Response
Authors: Lacerda L , Gupta A , Debeb BG , Xu W , Meyn R , Marini F , Andreeff M , Woodward WA , Klopp AH.
Published: April 2011

Background: Radiation increases engraftment of human bone-marrow derived mesenchymal stem cells (MSC) in tumors and this effect is augmented following repeated doses of radiation. We have recently demonstrated that MSC increase formation of mammospheres, which are enriched with radiation resistant tumor-initiating cells. Thus, we hypothesized that MSC may impact radiation therapy response through enhanced survival of tumor-initiating cells.

Materials & Methods: To evaluate the impact of MSC on radiation response we performed in vitro clonogenic survival assays using 2D and 3D clonogenic assays. Colony (2D) and sphere (3D) formation was quantified following irradiation of different cell lines (breast, lung, head and neck and cervix) at 2, 4 and 6 Gy in the presence of MSC conditioned media. In vivo, MSC were co-injected subcutaneously at increasing percentages with 4T1 murine breast cancer cells in BALB/c mice. The effects of MSC on radiation response were evaluated by examination of tumor size and weight following tumor irradiation at 2 and 5 Gy. Results: In vitro 2D clonogenic cell survival assays demonstrated that MSC conditioned media increased survival of breast (MDA231), lung (H460), head and neck (HN5), and cervical (SiHa) cancer cells following irradiation with 2 and 4 Gy as compared to control fibroblast conditioned media. Moreover, MSC conditioned media increased the number of HMLE (breast normal ephitelial cells) and SUM149 (breast cancer cells) mammospheres formed (55 vs 195and 263 vs 1054, respectively) and increased clonogenic survival of mammospheres after 2, 4 and 6 Gy compared to serum-free media. In vivo, irradiation with 5 Gy reduced murine breast 4T1tumors weight by 75% as compared to control untreated tumors. However, tumors co-injected with MSC did not significantly respond to 5 Gy. Discussion: MSC increase tumor resistance to radiation therapy in vivo and in vitro in 2D and 3D cultures. This effect is mediated at least in part by paracrine factors secreted by MSC. Our results here described suggest that MSC secrete factors which activate survival pathways on cancer cells exposed to sub-lethal radiation doses.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr PD02-07.
Abstract PD02-08: Development of DNA Aptamers That Inhibit Invasion, Migration, and Growth of Breast Cancer Cells
Authors: Kopco RL , Iida J , Lehman JR , Somiari S , Somiari R , Mural RJ , Shriver CD.
Published: April 2011

Triple negative (TN) breast cancers are defined by a lack of expression of estrogen, progesterone, and her-2/neu receptors. It is widely recognized that TN breast cancers have a poorer prognosis than other subtypes of breast caner. Given the lack of effective targeted therapies for TN breast cancer patients, understanding of the mechanisms of growth and invasion of these tumors will provide insight into developing novel approaches to lower the mortality from TN breast cancer.

We utilized three TN breast cancer cell lines (HCC38, HCC1806, HCC1937) as model systems to characterize mechanisms of growth, migration, and invasion. FACS analyses demonstrated that these TN cell lines showed a phenotype of CD44+CD24-/lowepithelial specific antigen (ESA)+. An inhibitory antibody against exon v10 of CD44 significantly inhibited three-dimensional (3D) growth of tumor cells. Importantly, this antibody also inhibited alpha2beta1/alpha3beta1 integrins-mediated migration and invasion into matrigel. These results suggest that CD44 harboring exon 10 facilitates progression and metastasis of TN breast cancer cells in vivo by enhancing growth, migration, and invasion. We also demonstrated that this exon interacts with a cell surface tyrosine kinase,

EphA2. Thus, we hypothesize that CD44 would form a molecular complex with EphA2 through exon v10 on TN breast cancer cells that plays a key role in facilitating invasion and metastasis.

In order to develop novel reagents to inhibit TN breast cancer invasion and metastasis, we have utilized SELEX (systematic evolution of ligands by exponential enrichment) to isolate DNA aptamers that specifically recognize exon v10 of CD44. DNA aptamers are functional molecules with the appropriate sequence and structure to form a complex with a target molecule. Typically, dissociation constants for these aptamer-target complexes are in the high pico-molar to low nano-molar range, which is comparable to antibodies. Given the fact of the ease of chemical synthesis and modifications of oligonucleotides, DNA aptamers that recognize cancer cells will be easier to develop and have a great impact for the next generation of strategies for cancer detection, diagnosis, and therapy. We describe the development of “function-blocking” DNA aptamers specific for exon v10 of CD44 that inhibits TN breast cancer invasion and metastasis.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr PD02-08.
Abstract PD04-07: Monitoring Circulating Tumor Cells (CTC) and Circulating Endothelial Cells (CEC) during Neoadjuvant Combination of Trastuzumab and Bevacizumab with Chemotherapy in HER2 Overexpressing Inflammatory Breast Cancer (IBC): An Ancillary Study of BEVERLY 2 Multicenter Phase II Trial
Authors: Pierga J-Y , Bidard F-C , Petit T , Delozier T , Ferrero J-M , Campone M , Gligorov J , Lerebours F , Roche H , Kraemer S , Mathiot C , Viens P.
Published: April 2011

Introduction: CTC detection in peripheral blood is an independent prognostic factor in early breast cancer but with a low detection rate of 10% to 23% of the patients (B. Rack, ASCO 2010, FC Bidard, Ann Oncol 2010). Changes in the HER2 status of CTC compared to the primary tumor have been reported (Riethdorf S CCR 2010). Predictive value of CEC for response to anti-angiogenic agents is debated.

Material and methods: CTC and CEC were detected in 7.5 ml and 4 ml of blood respectively in IBC (T4d) patients enrolled in the phase II multicenter trial, BEVERLY 2. This study is evaluating the efficacy of bevacizumab (15mg/kg q3w given concurrently) in combination with sequential neoadjuvant chemotherapy of 4 cycles of FEC followed by 4 cycles of Docetaxel-Trastuzumab. Bevacizumab was stopped 4 weeks before and reintroduced 4 weeks (w) after mastectomy. All patients had non metastatic IBC and over expressed HER2 (3+ in IHC or FISH +). The CellSearch™ System, combining EpCAM immunomagnetic selection (IMS) followed by anti-cytokeratin (A45B/B3) and anti-HER2 fluorescently staining for CTC and CD146 IMS and CD105 staining for CEC, was used at baseline, before cycle 5, before and after surgery.

Results: From Oct 2008 to Oct 2009, 52 patients were included in this study and 51 were evaluable for CTC. At baseline, 18 patients out of 51 had ≥ one detectable CTC (35.3%, 95%CI 22-48%, range 1 to 92).

Pathological complete response rate according to local review was 33/47 (70%) or 15/22 (68%) for centrally reviewed cases. At baseline, CTC level was not correlated with CEC level, neither with other patients and tumor characteristics’ (age, nodal status, PeV) nor pCR (centrally reviewed in

24). All positive cases for CTC detection had HER2 positive CTC. Five pts out of 18 (28%) had both HER2+ and HER2 negative CTCs in their blood. A lower level of CEC (< 20/4ml) before C5 could be associated with a higher probability of pCR (Khi2 test, p=0.053). Conclusion: We observed a high CTC detection rate of 35% in this population of patients with HER2+ IBC and a dramatic drop in CTC level during treatment in concordance with the high efficiency of this combination of chemotherapy and targeted therapy. We observed heterogeneity in the HER2 status of CTC in some patients. CEC levels increased progressively during neoadjuvant treatment and decreased after its interruption. Longer follow-up will show if CTC and CEC variations are early predictive factors for this highly efficient combination in HER2+ IBC.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr PD04-07.
Abstract PD05-07: Use of Natural ER Beta Modulators in Treating Hormonal Therapy Resistant Breast Tumors
Authors: Ganapathy M , Hareesh NB , Kirma NB , Tagliaferri M , Cohen I , Krishnegowda N , Vadalamudi RK , Tekmal RR.
Published: April 2011

Although systemic hormone therapies that either block local estrogen production by aromatase inhibitors (AI) such as letrozole or block actions of estrogen/ER actions by anti-estrogens (AEs) like tamoxifen are well tolerated, unfortunately, essentially all breast cancers in women with advanced breast cancer develop resistance to these two classes of agents, and the benefit of adjuvant therapy (where micrometastatic disease is present but cannot be directly observed) is also limited by the development of resistance. In addition, side effects associated with the systemic inhibition of aromatase with current compounds limits their long-term utility of them as chemopreventative agents. We have investigated whether treatment of letrozole resistant breast cancer cells with selective plant-derived ER beta agonists (Liquiritigenin; LIQ or Nyasol; NYA) alone or in combination with letrozole affects the growth of AI resistant breast cancer cells, thereby restores sensitivity to AI. To test whether combination therapy restores sensitivity to letrozole, we have treated letrozole resistant breast cancer cells [LTLTca, derived from MCF-7-aromatase tumors resistant to letrozole] with either letrozole alone or with a combination of letrozole and LIQ/NYA. In addition, effects of these compounds on cell growth was also tested using different breast cancer cells including MCF-7, MCF-7-aromatase, ZR-75-1, ZR-75-aromatase, MDA-MB-231 along with tamoxifen resistant MCF-7 or MCF-7- PELP/ZR-75-PELP1. Compared to single agents, combination treatment not only restored sensitivity to letrozole but also resulted in decreased cell proliferation and increased apoptosis as well as increased ERα and decreased ERα levels in resistant cells. Combination of LIQ or NYA not only diminished cell growth in MCF-7-HER2, MCF-7-PELP1, and SKBR-3 breast cancer cells (that express endogenous aromatase) but also affected the expression of aromatase. Detailed investigations indicated that LIQ/NYA affects the induction of aromatase by specifically inhibiting breast cancer specific aromatase promoter 1.3 as well as its activity through inflammatory cytokine-mediated actions. These studies suggest the therapeutic benefit of LIQ or NYA ERα agonist to resensitize breast cancer cells that are resistant to AEs or AI and their ability to selectively inhibit tumor induction of aromatase. Our study suggests a novel role of natural ER beta agonists as valuable therapeutic modulators to treat endocrine sensitive as well as resistant tumors. (Supported by V-Foundation & P30 CA 54174, RRT)

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr PD05-07.
Abstract PD05-08: Lack of Significant Association between CYP2D6 Polymorphisms and Clinical Outcomes of Adjuvnt Tamoxifen Therapy
Authors: Kim H , Shin H-C , Yom CK , Ahn S , Moon H-G , Han W , Noh D-Y.
Published: April 2011

Purpose: CYP2D6 variant alleles have been suspected to influence on the clinical efficacy of tamoxifen because CYP2D6 is an enzyme which metabolizes tamoxifen into its active forms, endoxifen and 4-hydroxytomoxifen. This study was performed to investigate the correlation between the polymorphisms in CYP2D6 and the clinical outcome of patients receiving tamoxifen. Methods: Between April 2001 and February 2010, 481 patients with hormone receptor-positive, invasive breast cancer receiving tamoxifen monotherapy in Seoul National University Hospital were studied. we analyzed the patients’ CYP2D6 alleles with the technique of PCR and SNaPshot using the blood samples cultured just before the every surgery and investigated the relationships of allelic variants of CYP2D6 to the disease-free survival by the Kaplan-Meier method and Log rank test Results: Each part of all patients’ alleles was genotyped into CYP2D6*1, CYP2D6*2, CYP2D6*5, CYP2D6*10, CYP2D6*41 and All the patients were categorized into 3 types, i.e. 51 homogenous wild type patients, 217 heterogenous variant type ones and 213 homogenous variant type ones according to their allelic types. There was no difference in age, pT, pN, follow up period among these patients. The rate of events which is defined as locoreginal metastasis, distant metastasis, contralateral breast cancer was 7.1%(34 patients totally). Only 2 patients were died due to pulmonary complication with distant metastasis. The number of variant alleles were not associated with the disease free survival.(P = .162; 95% CI in patients with one variant allele vs patients without variant allele and P = .215; 95% CI in patients with two variant alleles vs patients without variant allele) And the number of non-functional alleles were not associated with the disease free survival either.(P = .171; 95% CI in patients with one non-functional allele(CYP2D6*5, CYP2D6*10, CYP2D6*41) vs patients with two functional alleles(CYP2D6*1, CYP2D6*2) and P = .187; 95% CI in patients with two non-functional alleles vs patients with two functional alleles) Even in the subgroups of stage I, stage II or more, premenopause, and postmenopause, there was no difference found in disease free survival. Conclusion: Out results suggest the clinical outcome of the patients who are receiving tamoxifen therapy is independent of the patients’ polymorphisms in CYP2D6 and need more sophisticated investigation into the relationship of the process metabolizing tomoxifen to the clinical effect in the patient's body. Key Words: tomoxifen, CYP2D6, polymorphism

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr PD05-08.
Abstract PD06-06: Novel Surgical Technique To Assess the Response of Metastatic Axillary Lymph Nodes to Neoadjuvant Chemotherapy in Breast Cancer Patients
Authors: Vrancken Peeters M-JTFD , Straver M , Donker M , Loo C , Wesseling J , Holtkamp M , Rutgers ETJ , Rodenhuis S.
Published: April 2011

Background: An important benefit of neoadjuvant chemotherapy (NAC) is the increase in breast-conserving surgery. At present the response of axillary lymph node metastases to chemotherapy is not assessed accurately, since SN biopsy either before or after NAC will not provide an accurate answer. Therefore a reliable axilla-conserving therapy is not yet a benefit. We developed a new surgical technique to evaluate the axillary nodal response by Marking of the Axillary lymph node with Radioactive Iodine seeds (= the MARI procedure)

Method: Prior to NAC, tumor positive axillary lymph nodes were marked with a Iodine-125 seed under ultrasound guidance (the MARI node). After NAC, the marked lymph node was selectively removed with the use of a gamma-detection probe. A complementary axillary lymph node dissection was performed to assess whether the pathological response in the marked node was indicative for the pathological response in the additional lymph nodes.

Results: In 44 patients who were scheduled for NAC the MARI procedure was attempted. In 42 patients the tumor-positive axillary lymph node was successfully marked with a radioactive Iodine-125 seed. Two patients appeared to have overt distant metastasis and did not undergo complete axillary dissection. Thus, in 40 patients the MARI node was selectively removed after NAC. In all these patients a complementary axillary lymph node dissection was performed. In 24 patients the MARI node contained a macrometastasis after NAC. In 4 patients only isolated tumor cells (ITC) were present in the MARI node. In 24 of these 28 patients further nodal involvement was found. More importantly: in 12 patients (=30%) no residual tumor was present in the MARI node (pathological complete response= pCR) and 10 of them also showed a pCR in the complementary axillary lymph node dissection. In one patient a macrometastasis, and in one, ITC were found in one of the additional nodes. Conclusion: In this study we present the first results of a new surgical technique to assess the response of metastatic lymph nodes in the context of NAC for breast cancer; the MARI procedure. We conclude that marking tumor-positive lymph nodes before NAC and selectively removing them after NAC is feasible. Moreover we report that this MARI node is indicative for the response to NAC in the other axillary nodes: in 30% of the treated patients an axillary pCR was found which was correctly indicated by a negative MARI node in all but one patient. Thus the MARI procedure will enable us to select patients in which axillary lymph node dissection can be avoided after NAC.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr PD06-06.
Abstract PD06-10: Clinical Score for Deciding a Sentinel Node Biopsy(SNB) after Neo Adjuvant Treatment(NAC) for Breast Cancer
Authors: Cagnat J , Alran S , Savignioni A , Pierga J-Y , Belichard C , Kirova Y , Salmon RJ.
Published: April 2011

Rationale: Selection of the best candidates for SNB after NAC remains under discussion. A retrospective study identified predictive factors for negative axillary dissection. A score was developed to select patients at high and low risk for metastatic SNB after NAC.

Patients and methods: In 776 patients receiving NAC, clinico biologic patterns were evaluated by a multivariate logistic regression model in order to to predict the negativity of axillary dissection after surgical treatment.

A 4 class score was created using the three significant factors: Initial clinical node status, oestrogen receptors positivity and clinical response to treatment. This model was then prospectively applied tto 180 patients between 2006-8

Results: The probability of having a negative axillary dissection in patients with a score 0, 0-2 and over 2 was respectively 80%, 56% and less than 50%.

Conclusion: We developed a simple and reproducible score to predict the likelihood of having a negative axilla after NAC and consequently to better select the good candidates for SNB after NAC

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr PD06-10.