bluuebunny/crossref_metadata_2025 huggingface dataset:
P1-07-09: Estrogen Receptor (ER) mRNA and ER-Related Gene Expression in Breast Cancers That Are 1%-10% ER-Positive by Immunohistochemistry.
Authors: Iwamoto T , Booser D , Valero V , Murray JL , Koenig K , Esteva FJ , Ueno NT , Zhang J , Shi W , Qi Y , Matsuoka J , Hortobagyi GN , Hatzis C , Symmans WF , Pusztai L
Published: June 2012

Purpose: Our goal was to examine whether borderline estrogen receptor (ER)-positive cancers, defined as 1–10% positivity by immunohistochemistry (IHC), show the same global gene expression pattern and high ESR1 mRNA expression as ER-positive cancers or are more similar to ER-negative cancers.

Patients and methods: ER status was determined by IHC in 465 primary breast cancers and with Affymetrix U133A gene chip (ESR1 mRNA gene expression: Probe set = 205225_at). We compared expressions of ESR1 mRNA and a 106-probe set ER-associated gene signature score between ER-negative (n=183), 1–9% (n=25), exactly 10% (n=6), and > 10% ER-positive (n=251) cancers. We also assessed the molecular class of the borderline ER-positive cases using the PAM-50 classifier.

Results: Among the 1–9%, 10% and > 10% IHC positive cases, 24%, 67% and 92% were also ER-positive by ESR1 mRNA expression. The average ESR1 expression was significantly higher in the > 10% IHC-positive cohorts compared to the 1–9% or completely negative cases but in these latter two cohorts ER expression was similarly low. The average ER gene signature scores were similar for the ER-negative and 1–9% IHC-positive cases, but significantly lower than in > 10% ER-positive cases. None of the 1–9% ER-positive cases were classified as Luminal A, 2 were Luminal B and 12 were Basal-like. Among the 10% ER-positive cases, 2 were Luminal A and 1 was Luminal B. Conclusion: Overall, 24% of the 1–9% and 67% of the 10% ER-positive cancers show ESR1 mRNA levels and gene signatures that are consistent with ER-positive, potentially endocrine sensitive tumors.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-07-09.
P1-07-17: Multiplex Plasma Biomarkers Associated with Bone Metastasis from Breast Cancer.
Authors: Leitzel K , Ali S , Bisen A , Lomakin A , Simonyan V , Lipton A
Published: June 2012

Background: Over two-thirds of breast cancer patients who relapse will develop bone metastasis, and there are currently no effective biomarkers to predict for this event. Multiplex immunoassay technology for 60 bone metastasis-associated biomarkers was conducted on pretreatment plasma from 40 patients enrolled in a 2nd-line phase III hormone therapy trial of metastatic breast cancer, to discover effective circulating biomarkers predictive of bone metastasis.

Methods: Pretreatment EDTA plasma from 20 patients with bone metastasis, and 20 patients with non-bone metastasis from the letrozole-megace 2nd-line phase III hormone therapy trial of metastatic breast cancer was evaluated using Searchlight multiplex immunoassay technology (Aushon Biosciences, Billerica, MA) on 60 biomarkers custom-selected for known association with bone metastasis. Multiplex data analysis for maximal discrimination of bone metastasis vs. no bone metastasis cohorts was performed by two independent statisticians: one evaluated the predictive capacity of individual biomarkers as well as two-marker combinations constructed by Monte Carlo optimization using Metropolis algorithm (MMC), and the other used a machine learning technique called hybrid bootstrapped RLDA +PCA (regularized linear discriminant analysis with principal component analysis, with dimensionality reduction used in the bootstrapping procedure).

Results: One of the 60 analytes (IL-17) was undetectable at the 1.6 pg/ml lower limit of detection, and was not included in the multiplex statistical analyses. Of the remaining 59 analytes, MMC multiplex analysis yielded 9 biomarkers that predicted for bone metastasis, whereas RLDA + PCA multiplex analysis yielded 20 predictive biomarkers. Common to both analyses, which were limited in this small 40 patient study, were 5 biomarkers, with varying functions including growth factors, cytokines, inflammatory factors, protease inhibitors, and cell adhesion molecules.

Conclusions: Multiplex analysis of 60 biomarkers custom-selected for known association with breast cancer metastasis to bone yielded 5 pretreatment plasma biomarkers classified as predictive of bone metastasis by 2 different multiplex statistical analysis methods. Evaluation of these biomarkers is now warranted in a larger validation cohort of breast cancer patients with and without bone metastasis.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-07-17.
P1-08-15: Pattern of Cardiac Monitoring and Risk of Trastuzumab Associated-Cardiac Dysfunction in a Clinical Practice: A Population Based Study.
Authors: Lee-Ying R , Ubhi C , Roberts S , Lim H , Bhatt H , Gesy K , Ahmed S
Published: June 2012

Background: Adjuvant trastuzumab therapy (ATT) in women with early-stage HER2+ breast cancer has been associated with significant reduction in the disease recurrence and mortality. Cardiac dysfunction (CD) is a known serious adverse effect of ATT. Although periodic cardiac monitoring is recommended during ATT, little is known about pattern of cardiac monitoring and incidence of CD in a clinical setting. The study aimed to determine extent of cardiac monitoring and rate of CD during ATT and to identify factors correlated with CD.

Methods: Medical records of women with localized HER+ breast cancer diagnosed between the years 2005–2007 in the province of Saskatchewan was reviewed. Women with advanced or recurrent disease or if they were treated in the setting of an adjuvant trastuzumab trial were excluded. A logistic regression analysis was performed to determine various clinical variables correlated with CD.

Results: A total 116 eligible women with median age of 54 yrs (range: 27–74) and median BMI of 27 (range: 44–17) were indentified. 40% had a cardiac risk factor & 30% were premenopausal. 51% had node positive & 53% had ER or PR+ breast cancer. 92% received anthracycline-based chemotherapy and 23% received sequential ATT. Of 62 patients with ER/PR+ breast cancer, 61% received adjuvant aromatase inhibitors. Baseline cardiac assessment was performed in 93% women. 98% women underwent periodic cardiac monitoring during ATT, 55% had monitoring performed at the interval of 3–4 months & 82% women had monitoring performed at the interval of 3–6 months. Mean baseline cardiac ejection fraction (EF%) prior to the commencement of chemotherapy and ATT were 65% & 63.9% respectively (p=NS). CD was observed in 32 (28%) women and only 4% were symptomatic. Trastuzumab was interrupted in 34%, and was discontinued in 20% women. Of 32 women with CD, 59% were referred to a cardiologist and 53% were treated with medication. CD was reversible in 84% cases. On multivariate analysis adjuvant aromatase inhibitor therapy was significantly correlated with cardiac dysfunction (Odd ratio 6.9 [95% CI: 1.4−33.0]). During the follow up period 18% women developed recurrent disease and 16% were died.

Conclusions: Our results confirm high compliance with cardiac monitoring, though not as frequently as recommended in the clinical trial setting. Overall the rate of symptomatic decline in cardiac function was similar to the rate reported in the clinical trials, however, a relatively higher incidence of asymptomatic decline in the left ventricle EF% was noted. Among various variables examined, adjuvant aromatase inhibitor therapy was associated with an increased risk of CD.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-08-15.
P1-10-04: Cost-Utility of the 21-Gene Breast Cancer Assay (Oncotype DX®) in the Irish Healthcare Setting.
Authors: Lacey L , Chien R , Hornberger J
Published: June 2012

Objective: The Oncotype DX® Breast Cancer Test is a validated 21-gene assay that predicts the likelihood of benefit from adjuvant chemotherapy in early-stage, node-negative and up to 3-node-positive, ER+ breast cancer as well as the likelihood of distant recurrence at 10-years. Study objective was to investigate the cost-utility (incremental cost per QALY gained) of using Oncotype DX® in early stage, node-negative, ER+ breast cancer patients, from the perspective of the Irish Healthcare system.

Methods: A cost-utility analysis was performed over the lifetime of the patient using a state transition model (Markov), which simulated the costs and quality-adjusted survival associated with and without the use of Oncotype DX® to inform adjuvant chemotherapy decisions. The model included 3 health states: no recurrence, recurrence and death. Transition probabilities between health states, utility scores associated with each of the health state, patient cohort characteristics and impact of Oncotype DX® on chemotherapy decisions data were derived from published sources. Direct medical costs associated with chemotherapy were collected from a representative panel of 5 hospitals in Ireland. Costs and outcomes were discounted at 4% per annum and the analysis was conducted over 30 years (lifetime horizon). Probabilistic and one-way sensitivity analyses were conducted. Results: Costs associated with adjuvant chemotherapy (4 cycles of TC) included the following costs: drug (€1,002); administration and monitoring (€1,646); adverse event prevention (€3,561) and adverse event management (€756). The average total cost of chemotherapy across hospitals therefore summed up to €6,965. The base case incremental cost per QALY gained with ODX testing was estimated to be €9,462. There was a 74% chance for ODX to be cost-effective at a willingness to pay threshold of €20,000. Conclusions: Using Oncotype DX® as a decision tool to inform adjuvant chemotherapy in node-negative, ER+ breast cancer patients, is likely to be cost-effective in the Irish healthcare setting. This estimate is probably conservative as the cost and disutility associated with long-term impact of chemotherapy related adverse events (i.e. such as cardiotoxicity, secondary leukaemia and potential cognitive impairment) have not been considered.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-10-04.
P1-11-06: Barriers to Enrollment in Cancer Therapeutic Clinical Trials: A Comprehensive Cancer Center Experience.
Authors: Bourdeanu L , Niland J , Stiller T , Swain-Cabriales S , Somlo G
Published: June 2012

PURPOSE: Well-conducted cancer therapeutic clinical trials are essential for improving patient outcomes. Unfortunately, less than 5% of new cancer patients participate in clinical trials on average nationwide. Failure to recruit eligible patients represents a major impediment to the success of clinical trials. Barriers to patient accrual in cancer clinical trials must be identified and overcome to increase patient participation.

MATERIALS AND METHODS: A retrospective analysis of 418 patients with a diagnosis of breast cancer seen at City of Hope Medical Center in 2009 was performed. Along with descriptive analysis of patient demographic data, logistic regression analyses were performed to evaluate predictors of enrollment.

RESULTS: Of the 418 new breast cancer patients in 2009, we determined that 138 (33%) were eligible for available therapeutic clinical trials at City of Hope. At the initial visits, physicians did not consider clinical trials in 32% (44/138) of these patients. Of the 138 eligible patients, 58% (80/138) of patients participated in clinical trials. The remainder (58/138, 42%) either declined trial participation despite meeting eligibility criteria (14/58, 24%), or were not considered for clinical trials by their respective physician (44/58, 76%). By logistic regression analysis, patients with Stages II and III disease were significantly more likely to enroll in clinical trials compared to those with stage 0 or I (OR=2.89, 95% CI, 1.17−7.12, P = .02; OR=9.17, 95% Cl, 2.65−31.76, P=0.0005;, respectively). Age, preferred language, marital status, family history of breast cancer, and race were not found to be significant predictors of enrollment in therapeutic clinical trials.

CONCLUSION: While enrollment of eligible breast cancer patients onto therapeutic clinical trials at City of Hope is high (58%), overall the proportion going onto clinical trials remains low (80/418, 19%), though higher than national averages. We found that the majority of patients did not enroll due to lack of availability of a suitable trial, ineligibility, or lack of offering of trial enrollment by the treating MD. Barriers to cancer clinical trial accrual should be prospectively identified and addressed in future studies, with the hope of leading to more vigorous enrollment.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-11-06.
P1-11-08: Breast Cancer Screening Resources and Stage at Diagnosis in Appalachia: A Geospatial Perspective.
Authors: Yao N , Hillemeier MM , Anderson RT
Published: June 2012

Background: National Cancer Institute has designated Appalachia as a priority area characterized by significant disparities in cancer outcomes. However, little research focuses on the availability of cancer care resources in this region and how resource availability may relate to cancer outcomes. This paper will describe the distribution of breast cancer screening resources in Appalachia and examine the relationship between the screening resources and breast cancer stage at diagnosis at the population level.

Method: Percentages of early stage breast cancer incidence are computed based on the county-level data from the four Appalachian state cancer registry data from 2000 to 2008. Per capita breast cancer screening providers or facilities are computed for these Appalachian states. Descriptive analysis, exploratory spatial data analysis, and spatial regression were conducted.

Results: Appalachian counties had significantly fewer primary physicians, OB/GYN specialists, and diagnostic radiologists per capita than non-Appalachian counties. Spatial analysis demonstrates moderate clustering of scarce breast cancer screening resource and low percentage of early stage breast cancer incidence in West Virginia and Appalachian Kentucky. Those diagnosed at early stages were 67.92% of Appalachian vs. 68.34% of non-Appalachian breast cancer patients. The number of diagnostic radiologists per capita is significantly associated with the percentage of early stage breast cancer incidence when controlling for covariates such as county level poverty rates, uninsurance rate, percentage of adults with college degree, and per capita primary care resources.

Conclusions: Fewer diagnostic radiologists in West Virginia and Appalachia Kentucky are associated with lower percentage of early stage breast cancer incidence in the Central Appalachia region. Public health intervention could include policies and regulations to improve breast cancer screening resources in this region.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-11-08.
P1-11-09: Effects of a Multidisciplinary Breast Cancer Clinic in an Appalachian Based Medical Center.
Authors: Ingham JA , Cremeans DK , Daugherty SL , Myhand RC , Sever WE
Published: June 2012

Background: Adena Health System is a not for profit 250 bed institution that provides health care for a 13 county region in southeast, Appalachia Ohio. Access to health care in rural setting is an ongoing challenge. Several barriers to health care access affect this region. Socioeconomic burden and lack of higher education are two of the most dominant barriers. The age-adjusted beast cancer rates are 5.3% higher in the Appalachian region and the stage at presentation is more advanced. In December of 2009, we held our first Multidisciplinary Breast Cancer Clinic (MDBCC). The goal of the MDBCC is to facilitate prompt diagnosis and effective treatment. The MDBCC is staffed by a team of physicians consisting of surgeons, medical oncologists, and radiation oncologists. The team meets with the patient in a one on one setting. That same day the patients are discussed by the team with input from pathology and radiology in order to tailor a treatment plan specifically for each patient. After the conference, the recommendations are discussed with the patient before treatment is initiated.

Material and Methods: In a retrospective review, we evaluated the effect of a MDBCC on days to diagnosis, days from diagnosis to treatment, percentage of patients receiving neoadjuvant chemotherapy, and stage at diagnosis. Since December 2009, there have been over 90 breast cancer patients evaluated in the MDBCC. We randomly selected 60 breast cancer patients treated prior to MDBCC between 2008–09 and 66 patients evaluated at the MDBCC between December 2009–2011 from the tumor registry database.

Results: The average number of days to diagnosis pre-MDBCC was 15.5 days as compared to 4 days post-MDBCC. In 2008–09, the average days to treatment (pathology result to a-port placement, surgery or chemotherapy) was 18 days as compared to 12 days post-MDBCC. The percentage of patients receiving neoadjuvant chemotherapy has not changed significantly, however, there is an upward trend approaching statistical significance (10% per-MDBCC vs. 12% post-MDBCC). The stage at presentation has not changed significantly since the start of the MDBCC.

Discussion: The development of the MDBCC has proven to be a successful team oriented approach to breast cancer diagnosis and treatment. Time to diagnosis and initiation of treatment has improved. Providing patients with access to multiple specialists and forming a treatment plan the same day improves quality of care and lessens economic burden. The MDBCC is clearly feasible and particularly suited to the rural hospital setting. System wide, we hope to see a decrease in the stage at presentation and are confident based upon our current data that this in a realistic goal.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-11-09.
P1-11-13: Improvement in the Quality of Care for Patients with Locally Advanced Breast Cancer through Implementation of an Integrated Electronic Care Pathway.
Authors: Hogeveen S , Han D , George RL , Sweet-Goldstein M , Dinniwell RE , Muradali D , Brezden CB , Haq R , Simmons CE
Published: June 2012

Background: Locally advanced breast cancer (LABC) refers to the most advanced stage of non-metastatic tumours with an incidence of approximately 10% in newly diagnosed breast cancers. Currently, for optimal care, patients with LABC require a multidisciplinary approach including coordinated planning with medical, surgical and radiation oncologists. Many barriers may exist in co-ordinating timely management of these patients. Care pathways are important tools to streamline care and data collection and feedback allows for quality improvement in clinical practice. We created an interactive electronic care pathway and self populating quality assurance database at St. Michael's Hospital (SMH) to facilitate multidisciplinary teams to track LABC patient histories and patient treatments in order to coordinate therapy effectively and expedite care (LABC E-PATH).

Methods: This is an observational before-and-after cohort study of patients with LABC with a retrospective review pre-implementation and prospective collection of clinical data post-implementation. The completeness of workup and the timeliness of treatment pre- and post-implementation were assessed.

Results: With the implementation of the LABC E-PATH in May 2010, the time between the referral of an LABC patient to a medical oncologist and consultation has decreased from 7 days (range 0 to 493) pre-implementation to 2 days (range 0 to 29) post-implementation. The time between referral to medical oncologist and the start of their chemotherapy treatment decreased significantly from a median of 19 days to 8 days (pre-implementation: range = 0 to 493, postimplementation: range = 0 to 128) (p=0.0028). All pre-treatment staging was completed faster post-implementation of the LABC e-path than pre-implementation, expediting time to initiation of chemotherapy. The number of referrals for LABC to the SMH program increased from < 1 patient per month to 4 patients per month post-implementation.

Conclusions: The LABC E-PATH has achieved its goal of expediting care and overcoming barriers to timely treatment in this patient population. It has also ensured timely and appropriate resource allocation. This unique system may also be applied to other disease sites where coordination of a multi-disciplinary team is critical for appropriate patient management.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-11-13.
P1-12-12: Evaluation of Ile655Val HER2 Polymorphism Associated with Cardiac Toxicity Following the Administration of Trastuzumab in Women with Non-Metastatic Breast Cancer.
Authors: Diorio C , Lemieux J , Côté M-A , Provencher L , Nadeau-Larochelle C , Jacob S , Demers É , Tremblay-Lemay R , Saint-Pierre C , Beauchemin M , Barabé F , Laflamme C
Published: June 2012

Background: Trastuzumab is well tolerated without major side effects except for cardiac toxicity. Although a number of clinical parameters have been associated with trastuzumab-associated cardiac toxicity (TACT), there is some indication that genetic variation of the HER2 gene may be associated with TACT in a population of metastatic breast cancer patients. However, this finding needs confirmation and we looked at a population of non-metastatic breast cancer.

Purpose: This study aimed to evaluate the association between cardiac toxicity and HER2 [Ile655Val] polymorphism in non-metastatic breast cancer patients treated with trastuzumab.

Method: The Ile655Val HER2 polymorphism was assessed in 41 women using TaqMan technology. For this study, the genotyping was performed using DNA extracted from normal breast tissue located at more than 1 cm of any other lesions. Charts review was used to collect information on TACT which was defined as a decrease in the left ventricular ejection fraction (LVEF) of ≥15% or ≥10% with a resulting LVEF < 50% or any follow-up LVEF of < 45%. The Fisher exact test was used to evaluate the association between cardiac toxicity and HER2 polymorphism.

Results: No deviation from the Hardy-Weinberg equilibrium has been observed for the allele and genotype frequencies. The distribution of HER2 polymorphism was 10 Ile/Val (24%) and 31 Ile/Ile (76%). In this population, 22% (9/41) developed a cardiac toxicity. The Val655Ile genotype was associated with cardiac toxicity (0.03). In this preliminary study, 50% (5/10) of Ile/Val carriers compared to 13% (4/31) of Ile/Ile carriers showed TACT.

Conclusion: HER2 Ile655Val polymorphism may be an efficient marker of TACT considering this tendency with this small cohort of patients. Larger sample is needed to strengthen this conclusion, since this result may influence on prescribing decision for adjuvant chemotherapy and anti-HER2 therapy in HER2 positive patients.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-12-12.
P1-12-13: Comparative Pharmacokinetics (PK) of Trastuzumab Emtansine (T-DM1) in Patients Who Have or Who Have Not Received Prior Treatment for Human Epidermal Growth Factor 2 (HER2)-Positive Metastatic Breast Cancer (MBC).
Authors: Wang B , Conte P , Casanova LA , da Fonseca Vinholes JJ , Saad OM , Yi J-H , Gupta M , Song C , Olsen SR , Perez EA , Girish S
Published: June 2012

Background: T-DM1, an antibody-drug conjugate composed of trastuzumab, a stable linker, and the cytotoxic agent DM1, is in development for the treatment of HER2−positive cancers. Single-agent T-DM1 3.6 mg/kg every 3 weeks (q3w) has demonstrated clinical activity in 2 phase II studies (TDM4258g and TDM4374g) in patients with pretreated HER2−positive MBC. The efficacy and safety of T-DM1 vs trastuzumab + docetaxel was investigated in patients with no prior MBC treatment in the randomized phase II study TDM4450g/BO21976. Here we report the PK of T-DM1 from that study and compare these data with those from studies that enrolled pretreated patients.

Methods: In all 3 studies, PK parameters, including maximum concentration (Cmax), area under the concentration-time curve (AUC), terminal half-life (t½), steady-state volume of distribution (Vss), and clearance (CL) were estimated by noncompartmental analysis (NCA) for serum T-DM1, serum total trastuzumab (conjugated and unconjugated), and plasma DM1. The effects of baseline trastuzumab and HER2 extracellular domain (ECD) concentration on T-DM1 exposure were explored and the relationship between T-DM1 exposure and clinical response (objective response rate [ORR] and progression-free survival [PFS]) was modeled.

Results: T-DM1 PK from evaluable patients enrolled in 3 studies are shown in Table 1. No significant correlations were observed between efficacy (as measured by ORR) and T-DM1 exposure (AUC, Cmax) after administration of T-DM1 to pretreated patients; efficacy-exposure analyses (ORR and PFS) for previously untreated patients will be presented. Patients with measurable concentrations of trastuzumab at baseline had a greater AUC during cycle 1; however, this did not impact ORR. Baseline circulating HER2 ECD concentrations also had no effect on ORR for pretreated patients. The impact of baseline trastuzumab and HER2 ECD concentrations on ORR and PFS in previously untreated patients will be presented.

Conclusions: Single-agent T-DM1 has similar PK in patients who have received prior therapy for MBC and in those who have not. The PK of T-DM1 was not affected by prior trastuzumab treatment or by circulating HER2 ECD, and no significant correlations were observed between efficacy (ORR) and T-DM1 exposure (AUC, Cmax) or HER2 ECD for pretreated patients. The relationships between efficacy and T-DM1 exposure and HER2 ECD concentrations will be presented for patients without prior MBC treatment.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-12-13.
P1-12-22: Impact on Survival of the Level of HER2/neu Gene Amplification in Patients with HER2−Positive (HER2+) Advanced Breast Cancer (AdvBrCa) Treated with Trastuzumab (H).
Authors: Gullo G , Bettio D , Zuradelli M , Masci G , Giordano L , Bareggi C , Salvini P , Runza L , Santoro A
Published: June 2012

Background The level of HER2/neu (HER2) amplification, defined as HER2/centromeric region of chromosome 17 (CEP17) ratio at dual-color fluorescent in-situ hybridization (FISH) test presents wide variations in clinical practice but its clinical significance is still undefined. We designed this retrospective study to investigate the correlations between level of HER2 amplification in primary (T) and/or metastases (M) and outcome in a cohort of HER2+ AdvBrCa pts treated with a H-containing therapy.

Methods Retrospective multicentric study designed according to REporting recommendations for tumor MARKer prognostic studies (REMARK). To be included pts must have all the following: metastatic or locally advanced (not amenable of curative surgery) BrCa, HER2+ tumour defined as score 3+ at immunohistochemistry or FISH positive, treatment with a H-containing regimen (no H in neo- or adjuvant setting was allowed), at least one tumour sample from T and/or M available, measurable or evaluable disease, adequate follow up (FU) information.Outcome parameters included event-free survival (EFS) and overall survival (OS). All FISH tests were prospectively performed in a central laboratory of cytogenetic specifically for this study and in accordance with the international guidelines on FISH testing.

Results Ninety-one females were identified and 63 included in the final analysis. Forty-seven pts had one specimen available from T or M, 16 pts had two specimens from either T and a corresponding M. All M samples were obtained before treatment with H. Median FU time is 19.2 months (range 1.2−94.6). In 11 out of 16 cases (69%) with two tumour specimens HER2/CEP17 ratio was higher in M than in T with a statistically significant difference in the median HER2/CEP17 ratio between T (8.3, range: 3.1−18.4) and the corresponding M (10.9, range: 4.6−20.8) (p=0.004). The incremental gain in HER2/CEP17 ratio was associated with significantly shorter OS after trastuzumab-based therapy (p=0.023). A trend towards a correlation between increase in level of HER2 amplification assessed in M and shorter EFS and OS was observed. No statistically significant correlation was found between level of HER2 amplification assessed in T and EFS and OS.

Conclusions Level of HER2 amplification reported as HER2/CEP17 ratio is a dynamic parameter in HER2+ AdvBrCa as it increases from T to the matched M in a significant proportion of pts. The gain in HER2 gene copies may represent a prognostic factor for shorter OS when pts with HER2+ AdvBrCa are treated with a H-based therapy. Further studies on the level of HER2 amplification in BrCa metastases are strongly warranted to better understand the biology of HER2−positive breast cancer and to better identify patients with a poorer prognosis after treatment with H.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-12-22.
P1-14-02: Correlation of Circulating Tumor Cells (CTC) and Circulating Endothelial Cells (CEC) with Pathological Complete Response (pCR) during Neoadjuvant Chemotherapy (CT) Combined with Bevacizumab in HER2 Negative Inflammatory Breast Cancer (IBC): Ancillary Study of Phase II Trial BEVERLY 1.
Authors: Pierga J-Y , Bidard F-C , Andre F , Petit T , Dalenc F , Delozier T , Romieu G , Bonneterre J , Ferrero J-M , Kerbrat P , Lemonnier J , Viens P
Published: June 2012

Background: Prognostic value of CTC detection in blood has been reported in primary breast cancer with a rate around 20% in the neoadjuvant setting. No correlation between CTC and pCR to neoadjuvant CT in operable breast cancer has been reported (Pierga CCR 2008, Riethdorf CCR 2010). Predictive value of CEC for response to anti-angiogenic agents is unclear.

Methods: CTC and CEC were detected in 7.5 ml and 4 ml of blood respectively with CellSearch™ System in the neoadjuvant setting of HER2 negative IBC (T4d) patients (pts) enrolled in the phase II multicentre trial BEVERLY 1, evaluating bevacizumab (15mg/kg q3w) in combination with sequential neoadjuvant CT of 4 cycles of FEC followed by 4 cycles of Docetaxel. Patients received postoperatively 10 cycles of bevacizumab and hormonotherapy if tumor was ER+.

Results: From 12/08 to 09/10, 101 pts were included, 96 were evaluable for pCR and 92 for CTC and CEC. Out of 96 pts, 51 (53%) had triple negative breast cancer (TNBC). At baseline, 37 pts out of 92 had ≥ 1 detectable CTC (40%, 95%CI 30–50%).

At baseline, CTC level was not correlated with CEC level, neither with TNBC nor pCR. A drop in CTC incidence was observed from baseline to the 1st follow-up analysis after 4 cycles (p<0.0001). Out of 6 pts with detectable CTC at the end of adjuvant bevacizumab (8 months post surgery) none had pCR (p=0.05 Yates test). There was a significant increase of CEC from baseline to presurgery sample (p <0.001) and a decrease (p=0.04) after tumor removal and end of CT. A higher level of CEC (>20/4ml) before C5 could be associated with a higher probability of pCR (Khi2 test, p=0.003).

Conclusions: We observed a high CTC detection rate of 40% in HER2− IBC, including TNBC. CEC levels increased progressively during neoadjuvant treatment and decreased after its interruption. Baseline CTC and CEC levels were not predictive of pCR. Detection of CTC at 8 months of follow-up was associated with the absence of response to neoadjuvant chemotherapy.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-14-02.
P1-14-03: AVALUZ Study: First Line with Bevacizumab in Combination with Paclitaxel (P) and Gemcitabine (G) in Patients with HER-2 Negative Recurrent or Metastatic BC: PFS Analysis.
Authors: Salvador J , Ciruelos E , Codes de Villena M , Jaen A , Gil M , Galan A , Murias A , Jara C , de la Haba J , Baena JM , Villanueva MJ , Bayo J , Blancas I , Gonzalez E , Perez D , Mel JR , Manso L
Published: June 2012

Background The combination of bevacizumab (B) with taxanes, capecitabine or anthracyclines has shown increased PFS in pts with mBC. The combination with G and P has been evaluated and has demonstrated high response rate and good toxicity profile. The aim of this study is to evaluate the efficacy and toxicity profile of the combination of B with G and P, providing an update of PFS and the toxicity experienced in all patients treated.

Methods A phase II multicenter, national, open-label study in pts diagnosed of recurrent or metastatic HER-2 negative BC, treated with first line B 10 mg/kg, P 150 mg/m2 and G 2000 mg/m2 day 1 and 15 c/28 d until progression disease, unacceptable toxicity or medical decision. This abstract evaluates efficacy by PFS as primary endpoint and as secondary endpoints: response rate and toxicity profile (NCI CTC v3.0 criteria).

Results From January 2009 to December 2009, 82 evaluable patients were recruited in 23 sites. The characteristic of the all of patients included (90) are: median age 51.5 (26-81), ER + 68%, PR + 59%, triple negative pts 19%, previous chemotherapy (neoadj or adj) 64.4% and ≥ 3 metastatic sites 21% of patients (19). The median cycles administered per pts was 7 (1-26) and the relative median dose intensity was 0.93 for B, 0.89 for P and 0.91 for G.

Of the 75 patients with response assessment, the overall response rate obtained was 72% with a clinical benefit of 89.3% (80.6−95.28%) (CR 14.7% (11), PR 57.3% (43), SD 17.3% and PD 10.7%). Of the 82 patients with a median follow up of 16.5 m (0.16- 26.38), preliminary median PFS (43% events) for these pts was 11.48 m (8.98−15.72), the median duration of first response (MDR) was 12.4 m (7.47-NA) and the median OS was 21.35 m (20.72- NA). The one year OS% was 84.15% (74.27 - 90.47).

Toxicity was generally manageable. Related AEs have been reported in 67/82 patients which were mostly mild or moderate. Safety analysis revealed 24 SAEs in 21 patients.

Conclusions Bevacizumab in combination with paclitaxel and gemcitabine showed significant PFS compared with the published data with paclitaxel and gemcitabine and high clinical benefit with manageable safety profile.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-14-03.
P1-17-04: Phase I Study of PARP Inhibitor ABT-888 (Veliparib) in Combination with Cisplatin and Vinorelbine for Patients with Advanced Triple Negative Breast Cancer and/or BRCA-Mutation Associated Breast Cancer.
Authors: Rodler ET , Specht JM , Gadi VK , Kurland BF , Griffin MJ , Hammond JJ , Gralow JR
Published: June 2012

Background: Inhibitors of poly(ADP-ribose) polymerase (PARP) have shown preclinical and clinical activity in targeting tumors with pre-existing DNA repair defects, in particular BRCA1 and BRCA2 deficient tumors. Cisplatin (CP) has demonstrated synergy with ABT-888 (veliparib) in breast cancer xenograft models and has anti-tumor activity in triple negative (TN) and BRCA-1 deficient breast cancer. Vinorelbine (V) combined with CP has shown safety and efficacy in patients with pretreated metastatic breast cancer.

Methods: We are conducting a phase I study to determine the maximum tolerated dose (MTD), pharmacokinetic (PK) and pharmacodynamic profiles (PD), and preliminary anti-tumor activity of veliparib in combination with CP and V in patients (pts) with metastatic breast cancer who are either TN or who have BRCA1 or BRCA2 mutation associated breast cancer. Cohorts receive escalating doses of ABT-888 orally BID days 1–14, CP 75 mg/m2 intravenously (IV) day 1 and V 25 mg/m2 IV days 1,8 every 21 days, in a 3+3 design.

Results: As of 6/14/11, 18 eligible female pts have been enrolled. The median age at registration was 50 years (range 34–78 years). Sixteen pts received at least one prior metastatic regimen (range 0–8). Three pts had previously treated brain metastases. BRCA mutation status was as follows: BRCA1+ (3 pts); BRCA2+ (2 pts); confirmed mutation negative (8 pts); unknown (5 pts). Four pts received the 20 mg BID veliparib dose (one patient in the cohort died of progressive disease early in cycle 1 and was replaced), 3 received 30 mg BID, 6 received 40 mg BID, and 5 have been enrolled at dose level 4 (60 mg BID). Dose limiting toxicities occurred in one patient at the 40 mg BID dose (grade 4 thrombocytopenia), and one patient at the 60 mg BID dose (grade 3 neutropenic fever). Adverse events are typical for a platinum-based chemotherapy regimen and include nausea, fatigue, thrombocytopenia, and neutropenia. MTD has not been reached. Of 11 pts evaluable for radiographic response to date, 6 (55%) had a PR (3 of whom have a BRCA mutation) and 5 (45%) had stable disease. Correlative studies will use immunohistochemistry and gene expression array to evaluate the profile of TN breast cancer and predictors of response to treatment.

Conclusion: Veliparib in combination with CP and V has been generally well tolerated to date. Objective anti-tumor activity was seen in BRCA mutation carriers and in pts with sporadic TN breast cancer. PK, PD, and biomarker analysis is underway. Enrollment continues in the dose escalation cohorts.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-17-04.
P2-01-09: Gene Expression Alterations in the Lymph Node Microenvironment in Response to Successful Metastatic Colonization.
Authors: Ellsworth RE , Valente AL , Kane JL , Ellsworth DL , Shriver CD
Published: June 2012

Background: Breast stroma is known to play an active role in tumorigenesis, undergoing both phenotypic and molecular changes to facilitate and promote tumor development and growth. The metastatic microenvironment also plays a role in successful colonization; however, the genetic changes in these secondary microenvironments associated with metastasis are not well described.

Methods: Women with invasive breast cancer with at least one lymph node with macrometastases and one lymph node with no detectable metastases were identified from the Clinical Breast Care Project. Lymph node tissue was microdissected from both the metastatic lymph node microenvironment and negative nodes and hybridized to U133A 2.0 gene expression arrays. Differential expression was detected using Partek® Genomics Suite™6.5 using a cutoff of P<0.001, >2-fold change.

Results: Nineteen genes were differentially expressed between negative lymph nodes and lymph node tissue microdissected from lymph nodes with metastatic tumors. Eleven genes, including EPCAM, KRT19 and MUC1 were expressed at significantly higher levels in lymph node tissue from metastatic lymph nodes while eight genes, such as CXCL2 and CXCL5, were expressed at significantly higher levels in negative lymph nodes. Results have been validated in external sample sets for AZGP1, CLEC4M, CXCL2, EPCAM, MUC1, PIP and TFPI.

Conclusions: Lymph node tissue differs in gene expression between those harboring metastatic tumors and those without metastasis. Genes expressed at higher levels in lymph nodes with macrometastases are involved in tumorigenesis, suggesting that like the breast stromal microenvironment, the metastatic microenvironment undergoes crosstalk with the tumor cells. In addition, a cluster of genes involved in immune function are expressed at lower levels in metastatic lymph nodes, suggesting that suppression of proper immune response may be required for successful metastatic colonization.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-01-09.
P2-01-19: Expression of Receptor Like Protein Tyrosine Phosphatases mu (PTPRM) in Breast Cancer and the Biological Effects of PTPRM on Breast Cancer Cells.
Authors: Sun P-H , Ye L , Mansel RE , Jiang WG
Published: June 2012

Introduction: Protein tyrosine phosphatases (PTPs) play a major role in many cellular functions including cell survival, proliferation, differentiation and motility. The receptor-like protein tyrosine phosphatase mu (PTPRM) has a similar structure to cell-cell adhesion molecules and has been shown to exhibit homophilic binding and confer cell-cell adhesion in cells including epithelial and cancer cells. However, the role played by PTPRM in breast cancer remains unclear. The present study investigated the expression of PTPRM in breast cancer and the biological impact of PTPRM on breast cancer cells.

Methods: Expression of PTPRM protein and gene transcript was examined in a cohort of primary breast cancer tumours. The association of PTPRM transcripts level and pathological and clinical aspects was then analysed. Expression of PTPRM transcript in breast cancer cells were knocked down using a specific anti-PTPRM transgene. The impact of PTPRM knockdown on breast cancer was evaluated using a series of in vitro cell models and in vivo tumour model.

Result: PTPRM protein was seen generally distributed in the breast tissues, including tumours and normal background tissues. A stronger staining was revealed in cytoplasm of both mammary epithelia and cancer cells in comparison with the surrounding stromal cells. A significantly decrease was seen in poorly differentiated tumours (92.5±279), and moderately differentiated tumours (288±110), compared with that of well differentiated (472±114), p=0.011 and p=0.031 respectively. PTPRM transcript levels were decreased in patients died from breast cancer. The expression of PTPRM transcripts was significantly associated with disease free survival. Patients with lower expression of PTPRM had shorter survival compared with those with high level of PTPRM expression (p=0.029). Knocking down PTPRM from breast cancer cells significantly increased cell adhesion compared with control cells (p<0.001). Likewise, the knockdown also marked increase the rate of cell migration (102.9±11.3μm for control vs 115.3±10.5μm for PTPRM knockdown, p<0.05).

Conclusion: The expression of PTPRM transcripts is decreased in poorly differentiated breast tumours and is correlated with poorer long term survival. This is likely to be contributed to by the biological impact of PTPRM on the aggressiveness of breast cancer cells.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-01-19.
P2-01-20: Reduced Plakoglobin Expression Increases the Metastatic Potential of Breast Cancer Cells.
Authors: Ottewell PD , Whitworth J , Cross SS , Nutter FH , Holen I
Published: June 2012

The majority of deaths from breast cancer are a result of metastases, however, little is understood about the genetic alterations underlying their onset. A recent study of human breast cancer biopsies showed alterations of expression of 9 genes in primary tumours that had metastasized compared to benign tumours. Of these, plakoglobin was the most altered, with metastatic tumours showing a 3x decrease in expression compared to benign. Furthermore, expression of plakoglobin is reduced upon autocrine production of human growth hormone, which is associated with increased breast tumour cell invasion in vivo. Plakoglobin codes for the adhesion protein gamma catenin, an integral part of the cadherin-catenin complex involved in cell-to-cell adhesion. The aim of the current study is to investigate the effects of plakoglobin knockdown on cell proliferation, migration, adhesion and invasion in vitro and in vivo.

The non-metastatic human breast cancer cell lines MCF7 and T47D that express high levels of plakoglobin and gamma-catenin were stably transfected with miRNA to two different regions of the plakoglobin gene, or scramble vector, to produce plakoglobin knockdown and controls for each cell line. Plakoglobin gene expression was monitored by qPCR and gamma-catenin expression by Western blot. Cell proliferation was assessed 24, 48, 72 and 96 hours post seeding; cell migration and invasion were measured over 24 and 48 hours using scratch and modified Boyden Chamber assays. Cell-cell adhesion was assessed by spheroid formation in agar matrix. For in vivo experiments, MCF7 plakoglobin knockdown and control cells were inoculated into the 5th and 10th mammary fat pads of 12-week old female balb/c mice (n=10 per group). Tumour growth was monitored by caliper measurements and local invasion was investigated 6 weeks after tumour inoculation on histological sections. Shedding of tumour cells into the blood stream and evidence of metastatic bone lesions was monitored by flow cytometry and μCT, respectively.

Plakoglobin and gamma catenin expression were reduced by more than 80% in all knockdown cell lines used but were unaltered following transfection with the scrambled vector. Furthermore, knockdown of plakoglobin did not effect expression of its binding partner, e-cadherin. Reduced expression of plakoglobin resulted in a more than 3-fold increase in MCF7 and T47D cell proliferation in vitro (p<0.005) and a 2-fold increase in MCF-7 proliferation in vivo (P<0.05), compared with control. In addition plakoglobin knockdown cells showed a 6 - 15 fold increase in invasion through basement membrane (p<0.005), a 2-fold increase in migration (p<0.005) and exhibited reduced cell-to-cell adhesion compared with control cells. Data show that decreased expression of plakoglobin increases the early pro-metastatic behaviour of cells including increased cell proliferation, migration and invasion.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-01-20.
P2-03-02: An IGF1R Antibody Does Not Inhibit Growth of Tamoxifen Resistant MCF-7 Cells.
Authors: Fagan DH , Yee D
Published: June 2012

The role of the insulin-like growth factor (IGF) system in breast cancer has been well defined, and inhibitors of this pathway are currently in clinical trials. The majority of anti-IGF1R clinical trials are in estrogen receptor-positive patients who have progressed on prior endocrine therapy with some early reports showing no benefit for addition of IGF1R inhibitors to endocrine therapy. The effect in endocrine-resistant models, mimicking the clinical trial scenario, has not been adequately investigated. To examine the effectiveness of IGF1R inhibitors in vitro, tamoxifen-resistant (TamR) cells were generated by culturing MCF-7L cells in the presence of 4-hydroxy-tamoxifen for more than 1 year. TamR cells had diminished levels of IGF1R with unchanged levels of insulin receptor (IR), and enhanced phosphorylation of Akt. Further, TamR cells failed to further respond to IGF-I-induced Akt activation while retaining responsiveness to both insulin and IGF-II. Additionally, IGF-I failed to enhance the proliferation and anchorage-independent growth of TamR cells; however, both insulin and IGF-II stimulated proliferation and anchorage-independent growth. An IGF1R antibody (dalotuzumab) was able to inhibit IGF-I-mediated Akt phosphorylation, proliferation, and anchorage-independent growth in parental MCF-7L cells and had minimal effect on insulin and IGF-II stimulation. In TamR cells, dalotuzumab failed to inhibit proliferation or anchorage-independent growth in response to either insulin or IGF-II. An IGF1R tyrosine kinase inhibitor, (AEW 541) with equal potency for the IGF1R and IR, inhibited IGF-I-, IGF-II-, and insulin-stimulated Akt phosphorylation, proliferation, and anchorage-independent growth in MCF-7L parental cells. Interestingly, AEW 541 also inhibited insulin- and IGF-II-stimulated Akt phosphorylation, proliferation, and anchorage-independent growth in MCF-7L TamR cells. We conclude that cells selected for tamoxifen resistance in vitro have downregulated IGF1R making antibodies directed against this receptor ineffective. Preliminary studies in vivo, suggest that initial co-treatment with tamoxifen plus dalotuzumab was more effective than either drug alone. In contrast, acquired tamoxifen resistance might require dual inhibition of IGF1R and PI3K targets to completely suppress IGF system signaling as currently under study in the clinic. Alternatively, IGF1R tyrosine kinase inhibitors may be effective by inhibiting both IGF1R and IR signaling in cells selected for tamoxifen resistance.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-03-02.