bluuebunny/crossref_metadata_2025 huggingface dataset:
P4-09-16: Factors Affecting the Development of Axillary Lymph Node Metastases in T1a-T1b Breast Cancers.
Authors: Khair TA , Boolbol SK , Boachi-Adjei K , Klein P
Published: June 2012

Background: The presence of axillary nodal metastases is a poor prognostic indicator in patients with breast cancer. In the majority of patients, axillary nodal involvement correlates with tumor size and most commonly in tumors greater that 1 cm. Many series report an incidence of axillary nodal metastases in 3–37% of patients with tumors less than or equal to 2 cm. The presence of axillary nodal metastases will often change the stage of the cancer and role of adjuvant systemic treatment; therefore, it is important to understand what features are associated with these small lymph node positive tumors. The purpose of this study is to identify characteristics of T1a and T1b breast cancers that present with axillary nodal metastasis. Methods: A retrospective review of a prospective database identified 878 patients with T1a and T1b breast cancer. Pathologic features including tumor grade, hormone receptor status, lymphatic and vascular invasion, Her-2 neu status, and tumor histology were examined in patients with and without axillary nodal metastases. Pearson's χ2 test along with stepwise linear regression analysis were used to determine which factors were associated with the development of axillary nodal metastases.

Results: There were 289 T1a lesions and 589 T1b lesions studied. Four independent variables were found to be associated with axillary nodal metastases in T1a-T1b breast cancer. As illustrated in Table 1, a significant difference was seen in nodal positivity among patients with ER negative tumors compared to those with ER positive tumors (p=.0000). An increased incidence of nodal positivity was also observed among patients with LVI compared with those without LVI (p= .0000). A significant difference was observed in nodal positivity among patients with perineural invasion compared to those without (p=.0006). An increased incidence of nodal positivity was also observed among T1a and T1b tumors with a higher histologic grade (p= 0409, p= .0001). Of note, when comparing T1a to T1b breast cancers, there was no significant difference found among the identified exposure variables of interest.

Conclusion: In this series, the overall prevalence of nodal positivity in T1a and T1b breast cancer is 12%. An absence of a relationship between T1a and T1b cancers with nodal positivity was observed. However, the presence of nodal positivity showed a significant correlation with ER negativity, LVI, perineural invasion, and higher histologic grades.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-09-16.
P4-09-17: Leptin and Adiponectin Expression in Breast Cancer.
Authors: Jeong Y-J , Jeong H-Y , Bong J-G , Park S-H , Choi J-H , Oh H-K
Published: June 2012

Background: Several adipocytokines, such as leptin, resistin, and adiponectin, are associated with obesity and breast cancer. Multiple studies have indicated that adipocytokines may significantly influence the growth and proliferation of tumors. However, the relationship between adipocytokines and breast cancer invasiveness has not been investigated in human tissue. The aims of this study were to determine the expression of leptin, leptin receptor (ObR), adiponectin and adiponectin receptor (AdipoR) in human breast cancer, to evaluate their prognostic significance in the breast cancer.

Material and Methods: Specimens from 198 patients with primary breast cancer were enrolled, and representative paraffin tumor blocks were selected for constructing tissue microarrarys (TMA). Immunohistochemical staining for leptin, ObR, adiponectin, and AdipoR was performed using TMA, and the clinicopathologic characteristics were evaluated from the patient's medical records. Results: Stage 0 breast cancer accounted for 41 cases, and 157 cases were invasive cancer. A univariate analysis showed that positive rates of leptin and ObR expression in the ductal carcinoma in situ (DCIS) group were significantly higher than those of the invasive cancer group (97.4% vs. 34.0%, p<0.001; 74.4% vs. 29.8%, p<0.001). However, positive rates of adiponectin and AdipoR expression in the invasive cancer group were significantly higher than those in the DCIS group (53.7% vs. 33.3%, p=0.024; 59.9% vs. 26.3%, p<0.001). High leptin expression was significantly associated with high Ki-67 expression (p=0.016). High adiponectin expression was significantly correlated with smaller tumor size (p=0.001). The disease-free survival (DFS) and overall survival (OS) rate at 5 years were 89.3% and 96.7%, respectively. No difference in DFS and OS was observed based on leptin, ObR, adiponectin, AdipoR expression.

Discussion: Previous studies, using serum of breast cancer patients or breast cancer cell line, revealed that leptin and ObR may affect processes related to cancer initiation and progression, and adiponectin has protective effect on tumor progression. However, the precise effects of leptin, ObR, adiponectin and AdipoR in breast cancer are still unclear. Our results suggest that losses of leptin and ObR expression could be associated with invasive cancer, whereas high adiponectin and AdipoR expression may be associated with breast cancer invasiveness.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-09-17.
P4-09-21: A Novel Prognostic Marker for Triple-Negative Breast Cancers.
Authors: Wallon UM , Sabol JL , Zemba-Palko V , Carp NZ , Ciocca RM , Wojciechowski BS , Prendergast GC
Published: June 2012

BACKGROUND: Triple-negative breast cancers (TNBC) represent a highly aggressive form of this disease with few treatment options available. Currently, even the smallest node negative cancers are considered by many to warrant treatment with chemotherapy (CTX). While many recur early (within 2–3 years), there is a subset of long-term survivors illustrating the heterogeneity within this group. Here we report our ongoing effort to establish tissue inhibitor of metalloproteinase-4 (TIMP-4) as a prognostic marker in all early breast cancers a. While the canonical function of TIMPs is to inhibit tissue degradation, numerous reports have established that TIMPs exert tumor promoting activity. In our prospective study, we evaluated TIMP-4 as prognostic marker for TNBC and its role in disease progression.

METHODS: Specimens from our retrospective and prospective cohorts were assessed by immunohistochemical staining using standard techniques and a monoclonal antibody for TIMP-4b in accordance with the IRB approved protocol. Staining intensity was documented on a scale of 0–3. No data was released to the treating physicians at the time of collection. Outcome data from a total of 240 pts was obtained through tumor registry and clinician practices. Staining intensity was then correlated with outcome to calculate sensitivity and specificity of the marker. To determine the role of TIMP-4 in TNBC cell behavior we have performed microarray analyses. The effects of TIMP-4-induced signaling were tested using invasion and clonogenic survival assays under normal growth conditions and after exposure to gamma radiation.

RESULTS: Elevated TIMP-4 expression identified a high risk of relapse and short survival time with 75% sensitivity and 80% specificity. No discernable differences were noted between retrospective and prospective cohorts. Array analyses revealed activation of the PI3K/AKT pathway in the presence of TIMP-4. Furthermore, elevated TIMP-4 increased the invasive behavior of breast cancer cells in Matrigel™-coated invasion chambers and reduced sensitivity to gamma irradiation. These effects were reversible by addition of either a PI3K inhibitor or an anti-TIMP-4 antibody, suggesting their use as potential therapeutic strategies.

CONCLUSIONS: On the basis of these clinical data we suggest that TIMP-4 may offer a simple prognostic marker for TNBC patients at highest risk. The presence of TIMP-4 identifies a patient population likely to recur quickly despite standard CTX treatment. Our research suggests that targeted therapy of the PI3K/AKT pathway and/or a biological therapeutic approach directed against TIMP-4 may be of benefit in this subset of pts and should be explored. Therefore, TIMP-4 testing of TNBC patients could aid in the selection of a treatment regimen to improve survival outcome.

a Liss, M et.al. Am. J. Pathol. 2009

b Donover, P et.al. J. Cell. Biochem. 2010

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-09-21.
P4-10-11: Genetic and Environmental Predictors, Endogenous Hormones and Growth Factors and Risk of Estrogen Receptor-Positive Breast Cancer in Japanese Women.
Authors: Yoshimoto N , Nishiyama T , Toyama T , Takahashi S , Shiraki N , Sugiura H , Endo Y , Iwasa M , Fujii Y , Yamashita H
Published: June 2012

The incidence of breast cancer in Japanese women has doubled in all age groups over the past two decades. We have recently demonstrated that this marked increase is mostly due to an increase in the estrogen receptor (ER)-positive subtype. It is necessary to establish risk factors capable of predicting the risk of ER-positive breast cancer which will enable the efficient selection of candidates for preventive chemotherapy. We analyzed genetic factors, including 14 single nucleotide polymorphisms (SNPs), environmental risk factors (body-mass index (BMI), age at menarche, pregnancy, age at first birth, breastfeeding, family history of breast cancer, age at menopause, use of hormone replacement therapy, alcohol intake and smoking), serum hormones and growth factors (estradiol, testosterone, prolactin, insulin-like growth factor 1 (IGF1) and IGF binding protein 3 (IGFBP3)) and mammographic density in 913 women with breast cancer and 278 disease-free controls. To identify important risk factors, risk prediction models for ER-positive breast cancer in both pre- and postmenopausal women were created by logistic regression analysis. In premenopausal women, 1 SNP (CYP19A1-rs10046), age, pregnancy, breastfeeding, alcohol intake, serum levels of prolactin, testosterone and IGFBP3 were considered to be risk predictors. In postmenopausal women, 1 SNP (TP53-rs1042522), age, BMI, age at menopause, serum levels of testosterone and IGF1 were identified as risk predictors. Risk factors may differ between women of different menopausal status, and inclusion of common genetic variants and serum hormone measurements as well as environmental factors might improve risk assessment models. Further validation studies will clarify appropriate risk groups for preventive chemotherapy.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-10-11.
P4-13-03: Retrospective Analysis of Palonosetron Compared with Older 5-HT3 Receptor Antagonists (Ondansetron, Dolasetron, and Granisetron) in Preventing Anthracycline-Induced Nausea and Vomiting in Breast Cancer Patients from Four Phase 3 Trials.
Authors: Rugo HS , Schwartzberg L , Morrow GR , Barbour SY , Ballinari G , Thorn MD , Cox D
Published: June 2012

In patients with breast cancer, the risk of developing chemotherapy-induced nausea and vomiting (CINV) is particularly high due to the common use of highly emetogenic chemotherapy coupled with pt-related risk factors including female gender and young age. Despite prophylactic antiemetics, controlling CINV is difficult in this high risk population, particularly in the delayed phase following CT. Palonosetron, a potent 5-HT3 receptor antagonist (RA) with a distinctly different pharmacokinetic and receptor binding profile, has demonstrated improved CINV protection compared to older 5-HT3 RAs in phase 3 and 4 clinical trials in a variety of settings and tumor types.

We conducted a retrospective subset analysis of pts with breast cancer, the majority of whom (89%) received anthracycline combination chemotherapy (doxorubicin or epirubicin and cyclophosphamide, AC/EC) from four phase 3, randomized, double-blind, parallel group studies to assess the comparative effectiveness and safety of single IV doses of palonosetron (0.25 or 0.75 mg) versus older 5HT3-RAs (ondansetron 32 mg, dolasetron 100 mg, and granisetron 3 mg) in preventing CINV in pts receiving this highly emetogenic chemotherapy. The primary efficacy evaluation was for complete response (CR), defined as the proportion of patients with no emetic episodes & no use of rescue medication during 0–24hrs (acute), >24-120hrs (delayed), & 0–120hrs (overall) using a logistic regression model. A comparative descriptive safety assessment was also performed.

A total of 1197 patients with breast cancer were included in this analysis (palonosetron n=714; older 5HT3-RAs n=483). Higher CR rates were seen for palonosetron vs. older 5HT3-RAs during the acute (63.4% vs 59.4%, p= 0.08), delayed (59.2% vs. 44.9%, p<0.0001) and overall (50.7% vs.38.5%, p<0.0001) phases. Rates for CR were similarly higher for palonosetron-treated patients in the large subgroup of patients receiving AC/EC based chemotherapy. Consistent improvements associated with palonosetron were seen for secondary endpoints of complete control (CR + no more than mild nausea), no emesis, and severity of nausea. The adverse event (AE) profile for palonosetron was similar to older 5HT3-RAs, with headache and constipation being the most commonly reported treatment-related AEs with similar incidences for all 5-HT3 RAs.

This retrospective analysis of patients with breast cancer, including a large subgroup receiving anthracycline-based highly emetogenic chemotherapy, demonstrated significantly improved CINV prevention with palonosetron compared to older 5HT3-RAs, particularly in the delayed phase. Safety was comparable. This data has important implications for current CINV prevention guidelines.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-13-03.
P4-14-04: Tolerance of Docetaxel: Is Obesity a Risk Factor for Toxicity? A Pilot Study.
Authors: Parent D , Rallet A , Jovenin N , Yazbek G , Cure H , Rey J-B
Published: June 2012

Background Few data are available in the literature regarding chemotherapy dosage adjustment in obese patients. Docetaxel is a lipophilic substance, highly bound to albumin; obesity might result in an accumulation of docetaxel and may lead to a higher rate of toxicity. The objective of this study was to assess the tolerance to docetaxel in obese and non obese patients, according to their body mass index (BMI).

Methods A retrospective study was carried out. Patients were divided according to their BMI, considering a limit of 30 kg/m2. All patients received docetaxel (100 mg/m2) as a part of adjuvant chemotherapy for breast carcinoma. Toxicity of docetaxel such as neutropenia, hand-foot syndrome, mucositis, and neuropathy were assessed in both groups. Patients receiving concomitant therapies leading to similar adverse events were excluded (e.g. cyclophosphamide, 5-FU, …). The included patients received docetaxel alone or in association with trastuzumab or bevacizumab. Qualitative data analysis was performed using Fisher's exact test. Quantitative data were studied using the Student test.

Results Among the 49 patients, 18 (37%) presented with a BMI > 30 kg/m2. The median age was 53 years-old in the group BMI > 30 and 55 years-old in the group BMI < 30 (p = 0.9383). The median dose intensity delivered was 91% (74-100%) in the group BMI> 30 and 97.7% (75-100%) in the group BMI < 30 (p = 0.06). The median value for albumin was 75 g/L (59-86 g/L) in the BMI > 30 and 71 g/L (56-79 g/L) in the BMI < 30 (p = 0.122). Toxicity in both groups was as follows:

No neuropathy was observed in both groups.

Conclusions Obese patients (BMI > 30 kg/m2) presented with more toxicities (p = 0.013). Hypoalbuminemia did not represent a risk factor for toxicity in this study as patients had normal albumin levels. Obesity is a risk factor for toxicity in patients treated with docetaxel. Dosage calculation according to body surface area is not suitable for these patients. Toxicity in the group BMI < 30 seems to demonstrate greater susceptibility to the molecule for these patients.

Further studies will try to identify the different mechanisms of toxicity of docetaxel involved obese patients, including the interaction between docetaxel and fats. Measures have also to be taken to obtain a favorable effectiveness/toxicity balance in obese patients.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-14-04.
P4-17-01: Trastuzumab Does Not Increase the Incidence of Central Nervous System (CNS) Relapses in HER2−Positive Early Breast Cancer: The HERA Trial Experience.
Authors: Pestalozzi B , Holmes E , Metzger O , de Azambuja E , Hogge L , Scullion M , Gelber R , Piccart-Gebhart M , Cameron D
Published: June 2012

Background: Retrospective studies of HER2−positive metastatic breast cancer (BC) showed an incidence of CNS metastases of 21% to 34%. We investigated the incidence and clinical aspects of CNS relapse (CNS-R) in patients (pts) enrolled in the HERA trial, a prospectively randomized adjuvant trial in node + or high-risk node - HER2−positive early BC pts.

Methods: 3401 pts were randomized into the 1-year trastuzumab (1yT) or the observation (obs) arms of HERA (Piccart-Gebhart et al, 2005, Gianni et al, 2011). The cumulative incidences of first disease-free survival (DFS) events in the CNS vs other sites were estimated using competing risk analysis. The database of the main study had a clinical cut-off date of 9th June 2008. To obtain additional information regarding CNS-R (including occurrence of CNS-R after first DFS event), a specific CNS-directed questionnaire was sent to investigators of pts who were deceased as of July 2009. Information collected included the date of CNS-R, whether it was symptomatic, the type of CNS-R (brain metastases (BM) or meningeal carcinomatosis (MC)), methods of diagnosis, and treatments at the time of CNS-R. Results: 1yT significantly reduced the risk of other DFS events (p=0.000017, Gray's test), but not of CNS-R (p=0.55) as first event (see table). During the first year of follow up, CNS-R accounted for 15 (14.9%) of the 101 first DFS events in the 1yT arm and 15 (7.7%) of the 194 first DFS events in the obs arm. The analysis of baseline patient and tumor characteristics associated with CNS-R as first event confirmed known risk factors such as young age (<35y), T3 tumor, ≥ 4 + LN, ER neg, and G3.

413 of the 481 questionnaires (85.9%) were returned. 217 of the 413 deceased pts had a CNS-R diagnosed prior to death (52.5%), with more events occurring in the 1yT arm (see table). By contrast, the incidence of CNS-R as first DFS event was balanced across the arms.

Based on the survey data, CNS-R was symptomatic in 189 pts (87.1%) with no differences between arms. BM were present in 211 pts (97.2%), absent in 5 (2.3%), and missing information in 1 (0.5%). MC was diagnosed in 25 pts (11.5%), absent in 187 (86.2%), missing information in 5 (2.3%). Frequencies for BM and MC were very similar in both arms.

Conclusion: This retrospective analysis of a prospective large study shows more than 50% incidence of clinically diagnosed CNS-R in HER2−positive BC pts who have died. CNS-R was symptomatic in most pts. CNS-R at any time was less frequent in the 1yT arm (88 vs 129). There is no evidence that adjuvant trastuzumab increases the incidence of CNS-R.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-17-01.
P4-17-09: Comparative Performances of Prognostic Indexes for Breast Cancer Patients Presenting with Brain Metastases.
Authors: Braccini A-L , Azria D , Thezenas S , Ferrero J-M , Romieu G , Jacot W
Published: June 2012

Introduction: Incidence of brain metastases (BM) from breast cancer has increased over the past decade, especially for HER2(+) tumours. Several scores and prognosis systems have been developed in the brain metastases (BM) setting in order to help physicians tailor treatment options depending on patient prognosis and to stratify patients enrolled in clinical studies. The aim of our study was to compare the clinical relevance of the major classifications and existing prognostic scores in a population of breast cancer patients affected by BM.

Methods and Materials: In this retrospective study conducted in Montpellier and Nice Cancer Centres, we retrospectively reviewed the clinical and biological data of 250 patients diagnosed with breast cancer BM between 1995 and 2010. Prognostic value and accuracy of recursive partitioning analysis (RPA), graded prognostic assessment (GPA), basic score for brain metastases (BS-BM) and a clinico-biological score (BS) developed in phase I study and validated in the BM setting were compared.

Results: Median age at BM diagnosis was 55 years-old. Most patients (74%) had good performance status (0-1). Brain was the first metastatic site in one third of patients (33.6%). In 12.4% of cases, no extracerebral metastases were detected at BM diagnosis. Among the 250 patients’ tumors, 43.6% overexpressed HER2 receptor and 25.6% were triple negative (negative estrogen receptor, progesterone receptor and HER2 status). After a median follow up of 4.5 years, median overall survival from BM diagnosis was 8.9 months (CI 95%, 6.9−10.3 months). The four scores were able to discriminate patients according to their survival. Chi2 tests showed a correlation between the different scoring systems. In multivariate analysis, the elimination model identified RPA (p = 0.031; hazard ratio, 0.77; 95% confidence interval, 0.64−0.91) and BS (p = 0.043; hazard ratio, 0.5; 95% confidence interval, 0.31−0.80) as the only two independent predictors of survival. RPA was the most accurate score in order to identify patients with high life expectancy, while RPA and BS were the most accurate scores to classify patients with short life expectancy.

Conclusions: In a general unselected population of breast cancer affected by BM, RPA seems to be the most useful score. Integration of biological parameters in addition of existing clinico-radiological scores is promising in order to improve the prognostic determination accuracy.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-17-09.
P4-17-11: Central Nervous System Involvement and Clinical Outcome, Review of 135 Patients, 9-Year Follow Up.
Authors: Watanabe J , Ogiya A , Tadokoro Y , Tanaka K , Takahashi K , Uematsu T , Mitsuya K
Published: June 2012

Background: Breast cancer infiltrates into central nervous system (CNS) with the incidence of 10–20%. It is commonly accepted CNS screening does not contribute to maintain patients’ survival in metastatic breast cancer (MBC). We review our MBC cases with CNS involvement with the aim of searching prognostic factors and evaluating benefits derived from early detection of CNS lesion. Patients and Methods: From October 2002 to January 2011, 473 MBC patients treated at our hospital were reviewed and assessed. CNS lesion was found in 133 of 473 (28.5%) mostly by magnetic resonance imaging. Receptor statuses in patients with CNS disease were as follows; Estrogen receptor-positive 52.4%, HER2−positive 41.1% and triple-negative 12.7%. Asymptomatic patient at the diagnosis of CNS metastasis, in other words, CNS screening performed case, was 57.8%. Liver metastasis prior to CNS involvement was observed in 51.2% of patients.

Results: Within our 9-year follow-up, 122 patients received irradiation or operation, and 118 died. While major cause of death was lung or liver failure, 23.7% was considered as CNS death. Median overall survival from the diagnosis of primary breast cancer, survival from the MBC development and survival from CNS involvement were 1951, 1027 and 321 days respectively. Patients having HER2−disease showed superior survival after detection of CNS disease than HER2−negative patients. (median 514.0 vs 220.0 days, p=0.001) Asymptomatic patients, excluding underwent best supportive care (BSC), could not receive any survival benefit from early detection of CNS lesion. Liver metastasis prior to CNS involvement did not affect patients’ survival. Nineteen patients (14.1%), 17 of them were asymptomatic, received stereotactic irradiation as an initial treatment for CNS lesion, and showed superior survival after CNS event than patients received whole brain irradiation, surgery or BSC (median 670.0 vs 287.0 days, p=0.01).

Conclusion: Our review disclosed superior survival of HER2−disease with CNS metastasis. Early detection of CNS metastasis is only beneficial to patients when stereotactic irradiation will be suited for.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-17-11.
P4-17-12: Analysis of Predictive and Prognostic Factors for Metastatic Breast Cancer Patients with Brain Metastasis Treated by Whole Brain Radiotherapy.
Authors: Park IH , Lee KS , Shin KH , Ro J
Published: June 2012

Background We addressed the progression free survival of brain metastasis (PFS_BM) in metastatic breast cancer (MBC) patients treated by whole brain radiotherapy (WBRT) for brain metastasis (BM). In addition, we investigated predictive and prognostic factors for the PFS_BM and OS_BM.

Patients and methods A total of 212 patients with BM, treated by WBRT at the National Cancer Center between January 2000 and December 2010 were reviewed. The PFS_BM was defined as the time interval from the date of start of WBRT to the date of a progression of metastatic lesions in the brain or death or last follow-up.

Results The median age of patients was 45 years (range, 22–72 years) and the median time to brain metastasis was 12.7 months (range, 0.0−72.8 months).

Of all patients, 50 (23.6%) patients were hormone receptor (HR) positive and 104 (49.1%) HER2 positive. Six patients received a surgery or a focal radiotherapy for metastatic lesions prior to WBRT. Seventy one (33.4%) patients had well controlled extracranial systemic disease (CR+PR+SD) when brain metastasis was diagnosed. The median survival after BM (OS_BM) was 5.4 months (95% CI, 4.4−6.4 months) and approximately 14.6% of patients died mainly of progressive brain metastasis. The PFS_BM was significantly affected by poor performance status (PS ≥3) (P<0.001), uncontrolled systemic disease status (P=0.029), disease free interval (DFI) < 2 years (P=0.011), no chemotherapy after BM (P<0.001), and HER2 positivity (P=0.002). Of those, poor PS, no chemotherapy after BM, and uncontrolled systemic disease status were remained important factors on a multivariate analysis. In terms of OS_BM, poor PS (P<0.001), older age (P=0.005), number of systemic metastatic sites >3 (P<0.001), uncontrolled systemic disease status (P<0.001), disease free interval (DFI) < 2 years (P=0.014), no chemotherapy after BM (P<0.001), and visceral involvement (P=0.009) were significant factors. Of those, poor PS (P<0.001), uncontrolled systemic disease status (P<0.001), and no chemotherapy after BM (P<0.001) were important factors for shorter OS_BM when adjusted with other factors.

Conclusions Uncontrolled extracranial systemic disease status, no chemotherapy after BM, and poor PS were significant factors for shorter PFS_BM and overall survival after brain metastasis.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-17-12.
P4-18-03: Basal-Like Phenotype of BRCA Associated Ductal Carcinoma In-Situ.
Authors: Kristjansdottir K , Laprise J , Scalia-Wilbur J , Simon R , Steinhoff M , Legare R
Published: June 2012

Background: Molecular profiling has identified different subtypes of invasive carcinoma. The basal-like tumors represent a subgroup that is known for aggressive behavior. The NCCN guidelines recommend genetic testing for patients with triple negative breast cancer as over 80% of breast cancers in BRCA1 mutation carriers are triple negative tumors, and 70% of those fulfill criteria for basal-like tumors. Ductal carcinoma in-situ (DCIS) is thought to represent a non-obligate precursor to invasive breast cancer. Little is known about basal-like precursor lesions and their association with germline BRCA mutations. The aims of this study were to characterize the immunohistochemical phenotype of BRCA associated DCIS and evaluate the clinical importance of such lesions.

Material and Methods: We identified 32 cases of DCIS from 28 women with a known BRCA1/2 mutation, from our genetics database. Tumor samples were reviewed to identify areas of DCIS for staining. Five micron sections were stained on the Dako Autostainer with antibodies to estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER-2), epidermal growth factor receptor (EGFR), and CK 5/6.

Results: 44% of the DCIS tumors were from BRCA1 mutation carriers and 56% from BRCA2 mutation carriers. 53% of the samples were triple negative (staining negative for ER, PR, HER-2) and 69% of the triple negative tumors were basal-like tumors (staining positive for either EGFR or CK5/6). The basal-like DCIS specimens were all high grade, and they all came from BRCA1 positive patients. 71% of BRCA1 positive DCIS had basal-like features.

Conclusions: DCIS in BRCA1 mutation carriers shows basal-like features that are in line with historical control for invasive carcinomas in the same group, suggesting that DCIS is a precursor lesion for BRCA1 positive cancer. Although further investigation is needed, our data suggest that patients with basal-like DCIS should be offered genetic testing.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-18-03.
P4-19-04: Evaluation of Histopathological Parameters in Male Breast Cancer Reveals Differences Compared with Female Breast Cancer.
Authors: Nilsson C , Johansson I , Ahlin C , Thorstenson S , Bergkvist L , Amin R-M , Holmqvist M , Hedenfalk I , Fjällskog M-L
Published: June 2012

Purpose: To characterize male breast cancer (MBC) by evaluating established histopathological parameters and their prognostic value.

Methods: 197 male patients with invasive breast cancer and available paraffin-embedded tumor tissue were retrospectively assessed. Patient files were reviewed for clinicopathological data. Tumors were re-evaluated for histologic grading on conventional sections. Immunohistochemical (IHC) stainings for estrogen receptor (ER), progesterone receptor (PgR), human epidermal growth factor receptor 2 (HER2), silver-enhanced in situ hybridization (SISH), Ki-67, CK5/6 and epidermal growth factor receptor (EGFR) were performed on tissue micro arrays. Data on vital status and cause of death were retrieved from the Cause of Death Registry. Cox proportional regression models were used for uni- and multivariate analyses.

Results: ER and PgR positivity was demonstrated in 93 and 78 % of patients, respectively. Nottingham histologic grade III was seen in 41% and HER2 positivity in 11% of all patients. Defining molecular phenotypes using IHC markers revealed luminal (ER+ and/or PgR+, and HER2−) in 83%, luminal/HER2+ (ER+ and/or PgR+, and HER2+) in 11%, basal-like (ER-,PgR-,HER2−,EGFR+ and/or CK5/6 +) in 1%, but no cases of HER2-like or unclassified. Node-positivity (HR 4.6; 95% CI 1.9−11.4), tumor size > 20 mm (HR 2.3; 95% CI 1.1−5.0) and ER-negativity (HR 5.2; 95% CI 1.9−14.2) were significant risk factors for breast cancer death. Grade, HER2 status, Ki-67, or age did not demonstrate independent prognostic information. No difference in breast cancer deaths was demonstrated between luminal and luminal/HER2.

Conclusion: Male breast cancer tumors seem to be of grade III more often than female breast cancer, whereas HER2 expression appears equally common. In our study, the most important predictors for breast cancer death in male breast cancer were lymph nodes, tumor size and ER status. The most common molecular phenotype was luminal.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-19-04.
P4-20-03: T-Cell Cytokine Production Related to Progression of Breast Cancer Patients.
Authors: Cohen EN , Gao H , Lee B-N , Giordano A , Tin S , Anfossi S , Parker CA , Cristofanilli M , Valero V , Alvarez RH , Hortobagyi GN , Woodward WA , Ueno NT , Reuben JM
Published: June 2012

Background: Impaired immunosurveillance and immune dysregulation contribute to the pathogenesis and progression of breast cancer (BC). Upon activation, T cells synthesize inflammatory cytokines such as TNF-α that can promote or inhibit tumor growth. We therefore investigated T-cell cytokine syntheses as a predictor of disease progression.

Methods: We recruited 115 BC patients [25 with locally advanced breast cancer (LABC), 21 with metastatic breast cancer (MBC), 25 with non-metastatic inflammatory breast cancer (IBC), and 44 with metastatic IBC (mIBC)] and 31 healthy donors (HD) for this ongoing study. The tumor phenotype consisted of 69 hormone receptor (HR) positive (including 26 patients with HER2 positive disease), 16 HR negative but HER2+, 30 triple negative BC (TNBC). To evaluate T cell function, peripheral blood mononuclear cells from patients and HD were stimulated overnight with immobilized anti-CD3 and soluble anti-CD28 antibodies and assessed for the percentage of T cells that synthesized cytokines by multi-parameter flow cytometry. The associations of T cell cytokine production profile with patient progression free survival (PFS) were analyzed by Kaplan Meier Test.

Results: The median follow-up (FU) of 113 evaluable patients was 14.1 months with a median time to relapse of 10.5 months; 54 patients had stable disease (SD) and 59 patients had progression of disease (PD). In the entire cohort, on univariate analysis, metastasis, IBC, stage, and previous treatment predicted for worse PFS (p< 0.05). In non-metastatic patients (LABC+IBC), absolute count of anti-CD3 activated CD8+ T cells producing IL-17 was significantly higher in the SD patients compared with patient with PD (p=0.038), but it did not predict PFS (p=0.073). Similarly in metastatic patients, anti-CD3 activated CD4+ T cells producing TNF-α were significantly higher in patients with SD (p=0.025) and was predictive of longer PFS (p=0.033). Considering all patients with IBC (IBC + mIBC), although patients with PD had significantly fewer (percent and absolute number) anti-CD3 activated T cells capable of producing cytokines, this immune impairment was mostly related to metastasis and previous treatment. However, the percentage of anti-CD3 activated CD8+ T cells producing TNF-α was an independent positive prognostic indicator of PFS (p=0.002).

Conclusion: Higher than average cytokine syntheses by anti-CD3 activated T cells are significantly associated with longer PFS. These data are consistent with the hypothesis that an adaptive immune response can control disease progression.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-20-03.
P4-20-04: Cytokine Synthesis by Activated Dendritic Cells in Relation to Disease Progression in Inflammatory Breast Cancer (IBC).
Authors: Gao H , Cohen EN , Lee B-N , Giordano A , Tin S , Anfossi S , Parker CA , Cristofanilli M , Valero V , Alvarez RH , Hortobagyi GN , Woodward WA , Ueno NT , Reuben JM
Published: June 2012

Background: Deficiencies in innate and adaptive immune responses by plasmacytoid dendritic cells (pDC) and myeloid DC (mDC) have been linked to poor clinical outcome in breast cancer (BC) (Treilleux, Clin Cancer Res, 2004, PMID 15569976). pDC produce IFN-a and pro-inflammatory cytokines that regulate innate and adaptive immunity in breast cancer. mDC present in blood and secondary lymphoid organs secrete IL-12 and induce inflammatory cytokine production by T cells. Therefore, we studied DC activity in the peripheral blood and assessed their function with clinical outcome in breast cancer patients.

Methods: We recruited 115 BC patients [25 with locally advanced non-IBC (LABC), 25 with IBC, 21 with metastatic breast cancer (MBC), and 44 with metastatic IBC (mIBC)] and 31 healthy donors (HD) for this study. Peripheral blood pDC and mDC were activated through toll-like receptor (TLR)-7 to assess IFN-α and IL-10 production whereas mDC were activated through TLR-8 to assess production of IL-12 and TNF-α by multi-parameter flow cytometry. Associations between cytokine production by TLR-activated pDC and mDC with progression free survival (PFS) and overall survival (OS) of patients were analyzed by Kaplan Meier Test.

Results: The median follow-up (FU) of 113 evaluable patients was 14.1 months with a median time to progression of 10.5 months; 54 patients had stable disease (SD) and 59 had progression of disease (PD). Metastasis, previous treatments, and IBC contributed to shorter PFS and OS. Compared to HD, BC patients had significantly fewer total DC (p=0.008), mDC (p=0.008), and pDC (p=0.003) per μL. In general, the number of TLR-7-activated pDC per μL that produced IFN-a(p=0.023) or IL-10 (p=0.027) and the number of TLR-8-activated mDC per μL that produced IL-12 (p<0.001) or TNF-α (p=0,008) were significantly lower in BC patients than in HD. However, patients with DC that produced these cytokines above the median levels of HD had shorter PFS or OS. In IBC patients, higher numbers of TLR-8-activated mDC that produced TNF-α (p=0.025) or IL-12 (p=0.003) predicted shorter OS. In mIBC patients, a higher number of TLR-7-activated pDC producing IFN-α (p=0.024) or IL-10 (p=0.034) predicted shorter PFS.

Conclusion: BC patients had significantly fewer pDC and mDC in peripheral blood than HD. IBC patients with above average numbers of TLR-activated DC capable of producing proinflammatory cytokines had a significantly shorter PFS or OS. Disease progression in IBC is related to an increased number of activated dendritic cells producing inflammatory cytokines.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-20-04.
P5-01-07: Identification of SORBS2 as a Candidate Marker To Predict Metastatic Relapse in Breast Cancer.
Authors: Alsafadi S , Scott V , Pautier P , Goubar A , Lazar V , Dessen P , Lacroix L , Duvillard P , Morice P , André F , Delaloge S
Published: June 2012

Background

Elucidation of promising cancer biomarkers from gene expression data can provide important insight into the relationship between signaling networks and cancer. SORBS2, sorbin and SH3 domain containing 2, is a multi-adapter protein involved in signal transduction associated to the cytoskeleton and was reported to be strongly repressed in pancreatic and cervical cancers.

Methods: With the purpose of identifying genes involved in metastatic process, we compared gene expression profiling of 19 invasive ovarian cancers and 24 borderline tumors. Prognostic value of the selected genes was then tested in a gene expression array database that includes 1659 patients with early breast cancer (Gyorffy B et al. 2010). Upon isolation of SORBS2 as a predictor, its involvement in cell migration and tumor progression was investigated in vitro. Small interfering RNA targeting SORBS2 was used to downregulate its expression in T47D and Hela, two cell lines overexpressing SORBS2. Functional effect of siRNA-induced knockdown of SORBS2 on cell viability was determined by WST-1 assay and Trypan Blue exclusion test. Effect on cell migration was evaluated by wound-healing and transwell assays. Western blot analyses were also performed to examine the expressions of proteins involved in cell survival, death and migration.

Results: High-throughput analyses of genes that are differentially expressed between borderline ovarian tumors and invasive carcinoma demonstrated that SORBS2 is significantly downregulated in invasive carcinoma (FDR<10e-10). Moreover, SORBS2 loss was a strong predictor of relapse in a gene expression array dataset of 1659 patients presenting an early breast cancer (HR=0.62, 0.52−0.72, p=6.4e-10). In vitro studies showed that SORBS2 specific downregulation stimulated cell proliferation, by an average of 40%, in T47D and Hela cell lines. SORBS2 knockdown had no effect on either cell migration rate or protein expressions of p-FAK and p-Paxillin, two focal adhesion proteins which are critical to cell adhesion and migration. No significant variations in protein expressions of Akt/mTOR pro-survival or caspases activation pathways were observed. Gene expression profiling before and after SORBS2 dowregulation is ongoing in the two cell lines.

Conclusion: This study is the first to provide evidence for an antiproliferative activity of SORBS2 with no effect on cell migration in breast cancer cells. Our clinical and in vitro data suggest that SORBS2 is a candidate marker to predict relapse in patients with early breast cancer. Molecular mechanisms mediating the antiproliferative effect of SORBS2 are currently being investigated.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P5-01-07.
P5-01-15: The Functional Role of the Estrogen-Regulated Gene GREB1: Characterization of a Novel GREB1 Knockout Mouse Model.
Authors: Hnatyszyn HJ , Rodriguez C , Herbert L , Olson R , Lippman ME
Published: June 2012

Background: Gene regulated in breast cancer 1 (GREB1) was initially discovered in breast cancers as an estrogen-regulated gene that mediates estrogen-stimulated cell proliferation and is a candidate clinical marker for response to endocrine therapy. However, little is known of the functional role of GREB1 protein in normal breast tissue or breast cancers.

Methods: To address this unknown role, our laboratory designed and created a novel Greb1 Knockout Mouse model (C57/bl MEL Greb1 KO). This constitutive model results in the loss of Greb1 mRNA and protein expression in cells where expression of Cre recombinase promotes the cleavage of exon 1 and intron 1 of the gene encoding Greb1. ROSA26 Cre C57/b1 MEL Greb1 KO mice heterozygous for the floxed Greb1 allele were crossed to generate experimental litters. Initial experiments were designed to evaluate if the complete loss of Greb1 expression in offspring homozygous for the floxed Greb1 allele was lethal during gestation. Experimental litters were tail clipped and genotyped using gDNA and genotype-specific PCR.

Results: Offspring homozygous for the floxed Greb1 allele were identified in expected Mendelian ratios with wild type and heterozygous siblings. Loss of Greb1 expression was confirmed using RT-PCR, in situ hybridization and immunoblotting. Loss of both Greb1 alleles was not observed to be lethal during gestation for either male or female pups. Preliminary gross observation of these homozygous KO mice revealed no overt anatomical differences, however, they were 25–30% smaller than their heterozygous and wild-type siblings. Breeding experiments are underway to determine the fertility of crossbred Greb1 homozygous KO mice. Imaging experiments and necropsy with histochemical analysis of tissues will reveal any alteration in architecture and function. These findings will be summarized in this presentation.

Discussion: As GREB1 has been identified as an estrogen-regulated gene involved in breast cancer cell proliferation and a potential target for new therapeutic strategies, it is important to understand the contribution of GREB1 to the differentiation, development and function of normal tissues as well as in breast cancers. Characterization of this novel Greb1 KO mouse model will provide answers to these functional questions surrounding GREB1.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P5-01-15.
P5-01-17: HER2 Amplification in Primary Tumor: A Potential Marker for Presence of Circulating Tumor Cells in Inflammatory Breast Cancer Patients?
Authors: Bhattacharyya A , Krishnamurthy S , Lodhi A , Hall C , Anderson A , Jackson S , Ueno N , Bedrosian I , Kuerer H , Lucci A
Published: June 2012

Background: Inflammatory breast cancer (IBC) is a rare but aggressive form of invasive breast cancer accounting for 3–6% of all cases and have higher rates of distant recurrence. Circulating tumor cells (CTCs) are known to predict outcome in metastatic breast cancer (BC) patients, but little is known about their prognostic significance in non-metastatic BC. We hypothesized that CTCs can be identified in patients with IBCs and may correlate with primary tumor characteristics. Methods: All patients had blood samples collected at the time of primary surgery. CTCs (per 7.5 ml blood) were detected using the Cell Search™ system (Veridex) and were defined as nucleated cells lacking CD45 but expressing cytokeratins (CK) 8, 18, or 19. The presence of ≥ 1 epithelial cells meeting morphologic criteria for malignancy was considered a positive result. Statistical analyses employed Chi square and Fisher's exact tests using STATA IC 11. Results: We prospectively evaluated 41 IBC patients enrolled in an IRB approved protocol undergoing surgery for stage I-III breast cancer. Median follow-up was 30 months. Mean age was 52 years. Thirty five patients (94%) had positive lymph nodes (LNs) at presentation, 30 (75%) had high-grade tumors and 20 (53%) had lymphovascular invasion. Eleven patients (28%) were ER positive, 11 (27%) were PR positive and 18 (44%) were HER2 positive. IBCs were more likely to be high grade (P<0.0001), ER negative (P<0.0001), PR negative (P<0.0001), HER2 positive (P<0.0001), High Ki-67 (P= 0.005) and had a BMI of more than 25kg/m2 (P=0.04). Eleven (27%) patients were CTC positive. CTCs were more likely be found in HER2 positive (8/18; 44%) vs. HER2 negative primary tumors (3/20; 15%) [OR= 4.53; 95% C.I. = 1.02−19.52; P= 0.04]. We found no statistically significant correlations between primary tumor characteristics (ER, PR, LNs, high grade) and presence of CTCs. Conclusions: About a quarter of IBC patients had CTCs at the time of primary surgery. In these patients HER2 overexpression predicted the presence of CTCs. Studies with longer follow-ups is needed to determine if CTCs predicted survival.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P5-01-17.
P5-06-06: The RNA Binding Protein FUS Is a Potential Marker for Breast Cancer Progression and Therapy Response.
Authors: Brooke GN , Bevan CL , Rudraraju B , Palmieri C
Published: June 2012

Background

The RNA binding protein FUS is a multifunctional protein that has recently been demonstrated to be directed to target genes via non-coding RNA, where it represses transcription via disruption of the transcriptional complex. We have shown that FUS is an important regulator of prostate cancer cell growth; FUS over-expression causes G1 arrest, promotes apoptosis and blocks proliferation in vitro and in vivo, whereas knock-down increases proliferation. This regulation appears to be, at least in part, via direct regulation of cyclin D1. Further, analysis of patient samples demonstrated that FUS levels are inversely correlated with Gleason grade and directly correlated with survival and the presence of bone metastases. We hypothesised that FUS is a predictive and/or prognostic marker for hormone-dependent cancers.

Materials and Methods: FUS expression in breast cancer was analysed in silico and using Western blotting and immunohistochemistry on a tumour microarray. FUS levels were knocked down by siRNA in MCF-7 cells and chemosensitivity analysed using caspase 3/7 assays.

Results: Analysis of the Finak dataset (2008) demonstrated that FUS expression is significantly lower in breast cancer compared to normal breast. Immunohistochemistry revealed that FUS expression is also inversely correlated with grade. Importantly, FUS expression was also correlated with survival (Kaplan-Meier Plotter); significantly longer relapse free survival was seen in patients whose tumours had high levels of FUS. FUS expression also appears to correlate with therapy response since analysis of the Chang dataset (2003) revealed that FUS levels are significantly lower in patients resistant to Docetaxel and we found FUS levels to be lower in the tamoxifen resistant MCF-7 cell line (MCF-7 TAMR) compared to the parental line. We therefore hypothesised that loss of FUS expression may be driving therapy failure. In support of this, reducing FUS expression in the parental MCF7 line resulted in a significant reduction in docetaxel-induced Caspase 3/7 activity, suggesting that FUS is a regulator of chemosensitivity.

Discussion: FUS may be an important regulator of breast cancer progression since its levels are inversely correlated with grade and directly correlated with relapse free survival. Further, lower FUS expression is also correlated with reduced hormone therapy and chemo-sensitivity. Since knock-down of FUS reduced MCF-7 chemosensitivity, we believe that the observed changes in FUS expression are, in part, driving therapy failure and hence propose that FUS is a novel target and biomarker for breast cancer.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P5-06-06.