bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract PD03-09: Metformin Induces Apoptosis in Triple Negative Breast Cancer Cells Via Inhibition of Stat3 Activity
Authors: Deng XS , Wang S , Deng A , Liu B , Edgerton SM , Thor AD.
Published: April 2011

Background: Metformin inhibits breast cancer cell proliferation and colony formation, with S phase arrest and the induction of apoptosis in triple negative breast cancer cells in vitro. In these cells, metformin down-regulates Cyclin D1 and Cyclin E, as well as EGFR, p-EGFR, p-AKT, p-MAPK, p-Src and p-mTOR, whereas it up-regulates p-AMPK (Liu, et al. Cell Cycle, 2009). Non-triple negative breast cancer cells are resistant to metformin induced apoptosis, although they show similar changes in p-AMPK induction with metformin (Alimova, et al. Cell Cycle, 2009). We hypothesized that there are unique signaling intermediates associated with metformin responsivity in triple negative cells and have identified the signal transducer and activator of transcription 3 (Stat3) as a potential target. Stat3 is constitutively activated in a wide range of tumors, including breast cancer (up to 60%). It reportedly promotes cancer cell proliferation and survival.

Methods: Human triple negative breast cancer cell lines (MDA-468, MDA-231, BT20 and HCC70) were used to evaluate interactions between metformin and Stat3 signaling. Activation of Stat3 was examined by Western blot analysis with phosphorylation-specific antibodies. Cell proliferation was determined by MTS assay. Apoptosis was quantitated by an apoptosis ELISA assay and Western blots for PARP and caspase cleavage. In these 4 cell lines, Stat3-over-expressing clones were obtained via transfection of a constitutive active (CA) construct of Stat3. Specific knock-down of Stat3 expression was achieved by using a lentiviral system containing Stat3 small hairpin RNA.

Results: In a dose and time dependent manner, metformin inhibits Stat3 phosphorylation at sites Tyr705 and Ser727, Cyclin D1 and E protein expression, cleavage of PARP and the pro-caspases 3, 8, and 9. Overexpression of the CA-Stat3 attenuates the aforementioned meformin-associated PARP and caspase cleavage as well as apoptosis, and it suppresses metformin induced cell cycle arrest and changes in cyclin D1 and E. In contrast, specific knock-down of Stat3 expression sensitizes the triple negative breast cancer cells to metformin-mediated apoptosis and cell cycle arrest, enhancing the signaling changes we describe above. Conclusion: These data indicate that Stat3 is a critical intermediary for metformin action in triple negative breast cancer cells. Our studies suggest that targeting Stat3 activation may be a useful strategy to treat breast cancer patients with triple negative phenotype.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr PD03-09.
Abstract PD04-01: Does Analysis of Disseminated Tumor Cells in Bone Marrow Give Additional Prognostic Information in Primary Breast Cancer? Analysis of Disseminated Tumor Cells in Bone Marrow — Report of a Prospective Study with 5 Years Follow-Up
Authors: Falck A-K , Bendahl P-O , Ingvar C , Lindblom P , Lövgren K , Rennstam K , Fernö M , Rydén L.
Published: April 2011

Introduction: Disseminated tumor cells (DTC) in bone marrow of patients with breast cancer has, beyond the formerly known and clinically used prognostic markers, in recent publications been identified as an independent predictor of poor prognosis in patients with non-metastatic disease. Detection of DTC could therefore be a way to identify patients with an increased risk of recurrence among patients with a “good” standard prognostic profile. The present prospective study aimed to evaluate the presence and prognostic value of DTC in bone marrow in women undergoing surgery for primary breast cancer.

Material and methods: From 1999 to 2003 bone marrow aspirates were obtained from the sternal crest in 569 patients during surgery for primary invasive breast cancer. Tumor cells in bone marrow were identified with cytokeratins (4,5,6,7,8,10,13,14,15,16,18,19) against epithelial cells and analyzed with immunofluorescence and immunohistochemistry. The results were related to clinical follow up for 5 years using distant disease free survival (DDFS) and breast cancer specific survival (BCSS) as end-points by uni-and multivariate analysis including standard prognostic factors.

Results: Analysis of DTC were performed in 445 patients and cytokeratin positive cells were found in 159 of these (36%). The presence of DTC in bone marrow was not related to lymph node metastases (N+ 38% vs N-35%, p= 0.4), ER+/ER-(38% vs 30%, p= 0.2) or histological grade (40% grade 1; 37% grade 2; 30% grade 3, p= 0.18). BCSS was related to: lymph node engagement (+ vs -; HR 3.5, p< 0.001), ER (+ vs -; HR 0.55, p= 0.046), histological grade (2 vs 1: HR 4.9, p= 0.03; 3 vs 1: HR 7.4, p= 0.008), age (HR 1.02/year, p= 0.04) and tumor size (>20 mm vs <20 mm: HR 1.8, p= 0.05) in Cox multivariate analysis. Presence of DTC did not correlate either with DDFS (p= 0.97) or BCSS (p= 0.75). In the subgroup N0/ER-, a significantly higher rate of distant metastases was observed in patients with micrometastases in bone marrow (2/12 vs 0/25; p= 0.04; log rank test) and BCSS in the same subgroup was significantly related to the presence of DTC in bone marrow (2/12 vs 0/25; p= 0.04).

Conclusion: Detection of micrometastases in bone marrow in primary breast cancer has previously been shown to be a significant independent predictor of poor prognosis in different studies. In a consecutive cohort including unselected patients we were not able to confirm these results. Our data suggest that it is too early to implement the method of bone marrow aspiration and diagnosis of DTC in clinical routine.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr PD04-01.
Abstract PD04-04: Cytokeratin-19 mRNA Positive Circulating Tumor Cells during Follow-Up of Patients with Early Breast Cancer: Prognostic Relevance for Late Relapse
Authors: Saloustros ES , Perraki M , Apostolaki S , Politaki E , Agelaki S , Kallergi G , Xyrafas A , Georgoulias V , Mavroudis DA.
Published: April 2011

PURPOSE: We reported previously that the detection of CK-19 mRNA-positive circulating tumor cells (CTCs) after adjuvant chemotherapy in patients with early breast cancer is associated with poor clinical outcome. Reliable prognostic markers for late disease relapse are not available. In this study we investigated the prognostic value of CTCs’ detection during the first 5 years of follow-up in predicting late disease relapse. PATIENTS AND METHODS: Blood was obtained from 281 patients with early breast cancer who had not experienced disease relapse during the first two years of follow up. CTCs were detected using a real-time reverse transcriptase polymerase chain reaction for CK 19 mRNA after the completion of adjuvant chemotherapy and every six months thereafter for a 5-year follow up period. Patients were classified into four groups based on the CTCs’ status during the first two years and the subsequent three years of follow up (persistently negative, persistently positive, negative turned to positive and the opposite).

RESULTS: The vast majority of patients (92.9%) received some kind of hormonal therapy in addition to chemotherapy. Sixty-nine (24.6%) patients remained CTCs-free throughout the 5-year period. A change in CK-19 mRNA status was observed in 126 (44.9%) patients; 60 (21.4%) patients with initially CK-19 mRNA-positive CTCs during the first 24 months turned CTC-negative afterwards while 66 (23.5%) patients who were initially CTC-negative became CTC-positive. Eighty-six (30.6%) patients remained persistently CK-19 mRNA-positive for the entire 5 years. After 94 months of median follow up (range: 51-147), the relapse rate for each of the above groups of patients is illustrated in the following table.

Table 1: CTCs’ detection status and risk of late disease repapse.

Compared to the persistently negative, only the group of CK-19 mRNA-persistently positive patients had a statistically significant higher risk of relapse (38.4% versus 7.2%, p< 0.001), shorter progression-free survival (not reached vs 135 months, P<0.001) and significant difference in overall survival (not reached vs 146 months, p=0.045).

CONCLUSION: Persistent detection of CK-19 mRNA-positive CTCs in the blood during the first 5 years of follow up after adjuvant chemotherapy is associated with an increased risk of late disease relapse indicating the presence of chemotherapy-and hormonetherapy-resistant residual disease. This prognostic evaluation may be useful when deciding on subsequent adjuvant systemic therapy.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr PD04-04.
Abstract PD04-05: Prognostic Value of Circulating Tumor Cells According to Immunohistochemical Defined Molecular Subtype in Advanced Breast Cancer Patients
Authors: Munzone E , Botteri E , Nolè F , Esposito A , Adamoli L , Zorzino L , Sciandivasci A , Cassatella MC , Goldhirsch A , Rotmensz N , Sandri MT.
Published: April 2011

Background: Breast cancer (BC) is not a single disease with variable morphologic features and biomarkers but, rather, a group of molecularly distinct neoplastic disorders. Four main molecular classes of BC have been distinguished by gene expression profiling, called luminal-A, luminal-B, HER2-positive and triple-negative (TN) cancers. These subgroups correspond reasonably well to clinical characterization on the basis of estrogen receptor (ER) and HER2 status, as well as proliferation markers or histologic grade. Correlating circulating tumor cells (CTCs) with molecular subtypes might identify subgroup of patients (pts) at different risks and groups that might benefit more from prognostic evaluation by CTCs. Patients and methods: We retrospectively analyzed 187 consecutive metastatic BC pts who had baseline CTCs enumeration performed between March 2005 and December 2009 by CellSearch. Pts were categorized in 3 prognostic groups in accordance to the number of CTCs, 0 CTC, <5 CTCs and ≥5 CTCs, and in 4 categories based on the tumour biology: luminal-A with ER >1%, any progesterone receptor (PgR), G1/G2, Ki67<30%; luminal-B with ER >1%, any PgR, G3, Ki67>30%; HER-2 positive with HER-2 overexpressed or FISH amplified; TN with both ER and PgR 0% and HER-2 not overexpressed/FISH not amplified. Kaplan-Meier method for survival analysis was used.

Results: Median age at baseline was 56 years (range 31-78). CTCs were a strong predictive factor for overall survival (OS) in all pts (Log-Rank P<0.001), in pts with bone metastases (MTS) only (n=43; p=0.045), visceral MTS only (n=42; borderline significant p=0.08) and both bone and visceral MTS (n=74; P<0.001). The same figure was not observed in the few pts with only soft tissue MTS (n=28; p=0.315). Sixty-one (33%) pts had Luminal-A tumour, 58 (31%) Luminal-B, 53 (28%) HER-2 positive and 15 (8%) TN. CTCs were a significant prognostic factor in all molecular subtypes as shown in Figure 1. Conclusion: This large series confirms CTCs as an important prognostic factor for metastatic BC in all different molecular subtypes, but pts with 0 CTCs at baseline and TN or luminal B tumors seem to perform better as compared to other categories. Furthermore, pts with metastatic disease including bone and/or visceral sites benefited most from CTCs prognostic evaluation. Further studies with larger series and longer follow-up time could help identify metastatic BC subgroups in which CTCs analysis would be particularly useful and should be recommended. Figure available in online version.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr PD04-05.
Abstract PD04-10: Genome-Wide Copy Number Analysis of Circulating Tumor Cells (CTCs) from Metastatic Breast Cancer Patients
Authors: Magbanua MJM , Sosa EV , Scott JH , Pinkel D , Rugo HS , Park JW.
Published: April 2011

Background: Clinical studies have demonstrated the prognostic and predictive value of the number of CTCs in metastatic breast cancer. However, it is the molecular characterization of CTCs that offers insight into the biology of these tumor cells in the context of personalized treatment.

Methods: Ten to twenty mLs of blood from 32 metastatic breast cancer patients with poor prognosis (≥5 CTCs in 7.5mLs blood) were subjected to immunomagnetic enrichment. Pools of ∼20 CTCs [EpCAM+, CD45-, nucleated] were isolated via fluorescence activated cell sorting, FACS. Genomic DNA was subjected to whole genome amplification followed by array comparative genomic hybridization (CGH) analysis. Serial genomic profiling was performed in three patients. Genomic profiles from archival primary tumor available from 4 patients were compared to matched CTCs. Copy number analysis was performed using the Nexus® Copy Number. Genomic alterations with p ≥0.05 were considered significant. Results: Genomic profiling of CTCs revealed significant recurrent (≥35%) genomic alterations including gains in 8q12.1, 8q24 and losses in 1p36, 4p16, 5q21, 10q22, 11q24-25, and 13q34. Good concordance of CGH profiles obtained from CTCs isolated from serial blood samples attested to the reproducibility of the assay. Furthermore, comparisons of CTCs with matched archival primary tumors confirmed shared lineage with some divergence.

Conclusions: Copy number analysis revealed common genomic alterations in CTCs as well as shared aberrations with matched primary tumor. In addition, we demonstrated the feasibility of serial genomic profiling of CTCs. Future work will determine associations between genomic aberrations in CTCs and clinicopathological parameters. Uncovering genomic alterations in CTCs may lead to the discovery of therapeutic biomarkers to specifically target these cells in breast cancer patients who respond poorly to current aggressive therapies.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr PD04-10.
Abstract PD06-01: A Single Treatment with Targeted Intraoperative Radiotherapy (TARGIT) Is Similar to Several Weeks of External Beam Radiotherapy (EBRT) with Respect to Efficacy and Safety, and Has Obvious Advantages to the Patient and the Economy
Authors: Vaidya JS , Joseph DJ , Tobias JS , Wenz FK , Bulsara M , Alvarado M , Keshtgar MR , Eiermann W , Williams NR , Baum M,
Published: April 2011

Background: After breast-conserving surgery, 90% of local recurrences occur within the index quadrant despite the presence of multicentric cancers elsewhere in the breast. Thus, restriction of radiation therapy to the tumour bed during surgery might be adequate for selected patients. Materials and methods: Having safely piloted the new technique of single-dose targeted intraoperative radiotherapy (TARGIT) with Intrabeam, we launched the TARGIT-A trial on March 24, 2000. In this prospective, randomised, non-inferiority trial, women aged 45 years or older with invasive ductal breast carcinoma undergoing breast-conserving surgery were enrolled to compare TARGIT with whole breast external beam radiotherapy (EBRT). The primary outcome was local recurrence in the conserved breast with a predefined absolute non-inferiority margin of 2-5%. Analysis was by intention-to-treat.

Results: 1113 patients were randomly allocated to TARGIT and 1119 were allocated to EBRT. The Kaplan-Meier estimate of local recurrence in the conserved breast at 4 years was 1-20% (95% CI 0-53-2-71) in the TARGIT and 0-95% (0.39-2-31) in the EBRT group (difference between groups 0.25%, −1.04 to 1.54; p=0.41). The frequency of any complications and major toxicity (TARGIT 3.3% vs. EBRT 3.9% p=0·44) was similar. Radiotherapy toxicity was lower in the TARGIT group (0.5% vs. 2.1%, p=0.002). 14% of patients who received TARGIT also received EBRT as per the protocol, i.e., the remaining 86% patients could safely avoid 3-6 weeks of daily radiotherapy treatments and its obvious associated costs in economic and human terms.

Discussion: For selected patients with early breast cancer, a single-dose targeted intraoperative radiotherapy should be considered as an alternative to whole breast EBRT delivered over several weeks. It could save time, effort, and money for the patient and the healthcare system, which is particularly relevant in the present times of healthcare reform. NB the main trial results have been presented in ASCO 2010 and published in the Lancet Online First.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr PD06-01.
Abstract PD06-08: Size Matters: Incorporation of Sentinel Lymph Node (SLN) Metastasis Size into a Nomogram Predicting NonSLN Involvement in SLN Positive Breast Cancer Patients
Authors: Mittendorf EA , Boughey JC , Degnim AC , Harrell R , Bassett R , Yi M , Meric-Bernstam F , Ross MI , Babiera GV , Kuerer HM , Lucci A , Bedrosian I , Hunt KK , Hwang RF.
Published: April 2011

Purpose: Emerging data from cooperative group trials suggests that not all patients with a positive SLN require completion axillary lymph node dissection (ALND). The controversy regarding who will benefit from additional nodal clearance continues, suggesting a need for improved tools to predict non-SLN involvement. We hypothesized that the size of a SLN metastasis evaluated as a continuous variable would be an important predictor of non-SLN involvement. The goal of this study was to determine clinicopathologic factors including SLN metastasis size that predicted non-SLN involvement and to use these factors to construct an improved predictive nomogram.

Methods: Using a prospective database, we identified 509 patients with invasive breast cancer from 1996 - 2007 with a positive SLN who underwent ALND. Patients receiving neoadjuvant therapy were excluded. Clinicopathologic data including age, primary tumor size, presence of multifocal disease, histology (invasive ductal vs. invasive lobular vs. mixed invasive), tumor grade, estrogen receptor, progesterone receptor, and HER2/neu status, presence of lymphovascular invasion (LVI), the number of SLNs identified, number of positive SLNs, the maximum SLN metastasis size and the presence of extranodal extension were recorded. For SLNs described only as containing isolated tumor cells, size was recorded as 0.1 mm. Univariate and multivariate logistic regression analyses were performed to identify factors predictive of positive non-SLNs. Using these variables, a nomogram was constructed and subsequently validated using an external cohort of 464 patients.

Results: On univariate analysis, the following factors were predictive of positive non-SLNs: number of SLN identified (p<.001), number of positive SLN (p<.001), maximum SLN metastasis size (p<.001), extranodal extension (p<.001), primary tumor size (p=.001), LVI (p=.019) and histology (p=.034). On multivariate analysis, all factors remained significant at the 5% level except LVI. A nomogram was created using these variables. When applied to an external cohort, the nomogram was accurate and discriminating with an area under the receiver operating characteristic curve of .73 (95% CI: .68 - .77). Conclusions: SLN metastasis size is predictive for identifying additional disease in the axilla. These data have been incorporated into a nomogram that accurately predicts the likelihood of having additional, non-SLN metastasis suggesting that routine reporting of the SLN metastasis size would assist with clinical decision making regarding the need to perform a completion ALND. Figures available in online version.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr PD06-08.
Abstract PD07-06: Basal-Like Molecular Subtype Predicts Response to Neoadjuvant Chemotherapy with Epirubicine, Cyclophosphamide, and Docetaxel in Patients with Primary Breast Cancer
Authors: Stickeler E , Klar M , Pils D , Orlowska-Volk M , zur Hausen A , Gitsch G , Tempfer C.
Published: April 2011

Background: Gene expression profiling provides an opportunity to predict response to specific regimens of neo-adjuvant chemotherapy (NAC) in breast cancer patients.

Materials and Methods: In a prospective cohort study of 32 women with primary invasive breast cancer with a measurable lesion, we obtained a tumor specimen by high speed core biopsy before and after 4 cycles of NAC with epirubicine 90mg/m2 and cyclophosphamide 600mg/m2 every 3 weeks, followed by 4 cycles of docetaxel 100mg/m2. Total RNA was extracted from tumor specimens and the whole transcriptome was quantified with Agilent's 44K single color microarray interrogating 44000 unique human genes. Tumor lesions were ultrasonographically measured to assess response using Sinn criteria. Data analysis was performed by GeneSpring v11 and IBM SPSS v18.

Results: Using three single sample predictors, 10 tumors were classified as basal-like and 22 tumors were classified as non basal-like. We found that gene expression-based molecular subtype (basal-like vs. non basal-like) (p=0.003), but not tumor grade (p=0.07), estrogen receptor (p=0.1), progesterone receptor (p=0.6), and HER2 status (p=0.4) predicted response to NAC. Specifically, 7/10 basal-like tumors responded to NAC, whereas 19/22 non basal-like tumors did not respond. Comparing gene expression signatures before and after 4 cycles of NAC, we found that all patients with an initial non basal-like tumor retained this tumor type, whereas 5/7 basal-like tumors, including all responders, lost this molecular subtype. Using regression models based on centroid predicition gene sets, complete prediction of response to NAC based on the initial tumor biopsy as well as on the change of gene expression between two tumor biopsies was achieved with a 21 gene list (p=0.000008) and a 23 gene list (p=0.000007), respectively. Of note, both the expression and upregulation of a single gene, ie HER4, predicted response to NAC in 26/32 (81%; p=0.002) and in 23/25 (92%; P<0.001) patients, respectively.

Conclusions: Basal-like molecular subtype and therapy-induced HER4 gene upregulation predict response to NAC with epirubicine, cyclophosphamide, and docetaxel.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr PD07-06.
Abstract PD07-07: Indicators of Homologous Recombination Deficiency in Breast Cancer and Association with Response to Neoadjuvant Chemotherapy
Authors: Lips EH , Mulder L , Hannemann J , Laddach N , Vrancken-Peeters M-JFTD , van de Vijver MJ , Wesseling J , Nederlof PM , Rodenhuis S.
Published: April 2011

Background: Tumors with homologous recombination (HR) deficiency are highly sensitive to DNA double strand break (DSB) inducing agents, such as alkylating agents and poly (ADP-ribose) polymerase (PARP)-inhibitors. BRCA1 or BRCA2- mutated tumors, which are HR deficient, have characteristic DNA gains and losses that can be assessed by an array Comparative Genomic Hybridization (aCGH) classifier, one for BRCA1 mutations and one for BRCA2 mutations. We have studied these aCGH profiles together with several other HR deficiency indicators in sporadic breast cancers and we have correlated their presence to neoadjuvant chemotherapy response.

Material and Methods: A total of 163 HER2-negative pre-treatment biopsies were examined, procured from sporadic breast cancer patients scheduled to receive neoadjuvant therapy with doxorubicin and cyclophosphamide. Triple negative (TN) and estrogen receptor positive (ER+/HER2-) tumors were analyzed separately. aCGH was performed to assess BRCA1-like and BRCA2-like profiles. In addition, BRCA1 promoter methylation, BRCA1 mRNA expression and amplification of the BRCA2-inhibiting gene EMSY were analyzed. Response to neoadjuvant treatment was assessed by measuring pathological complete remission (pCR) and near pCR at the time of surgery.

Results: Inactivation of BRCA1 was frequent in TN tumors: 57% of these tumors showed a BRCA1-like profile at aCGH. BRCA 1 promoter methylation and reduced BRCA1 mRNA expression were observed in 25% and 36% of the TN tumors, respectively. The BRCA1-like aCGH profile was not clearly associated with a better neoadjuvant treatment response (58% vs. 48%, p=0.47). In ER+ tumors, a BRCA2-like aCGH profile and the amplification of the BRCA2 inhibiting gene EMSY were frequently observed (43% and 13% respectively). A BRCA2-like aCGH profile was associated with a significantly higher response rate (35% vs. 14%, p=0.014). EMSY amplification and a BRCA2-like aCGH profile occurred together in only one case, suggesting mutual exclusivity. EMSY was not associated with treatment response, questioning the role of EMSY in HR deficiency.

Conclusion: Alterations associated with BRCA1 inactivation are present in about half of the TN breast cancers, but were not predictive of chemotherapy response. In ER+/HER2- tumors, the BRCA2-like aCGH profile predicts sensitivity to DSB-inducing chemotherapy, and possibly as well to new targeted agents, such as the PARP inhibitors. After validation in independent series the aCGH classifiers may lead to a diagnostic test that could assist in neoadjuvant treatment selection.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr PD07-07.
Abstract PD07-09: Hypoxia (VEGF-S) Signature and CRYAB Predict Response to Neoadjuvant Anthracycline/Taxane Containing Chemotherapy within Triple Negative and Unselected Breast Tumors
Authors: Cheang MCU , Parker JS , He X , Zhao W , Fan C , Prat A , Carey L , I-SPY Investigators , Perou CM.
Published: April 2011

Background: Aggressive breast tumors are a clinically challenge to manage. A VEGF-signature (VEGF-s) developed in vivo (Hu et al. BMC Med. 09) and the profile of the anti-apoptotic gene and regulator of tumor angiogenesis, CRYAB (Moyano et al. J Clin Invest 06; Dimberg et al. Blood 08), are both associated with distant metastasis. In this study, we sought to test if these two mechanistically related biomarkers predict pathological complete response(pCR) to anthracyline/taxane-based neoadjuvant chemotherapy.

Methods: Gene expression data of pre-treatment fresh frozen tumors were combined, 149 from the ISPY trial (Agilent 44K) and 225 from MD Anderson Cancer Center(MDACC) (Affymetrix HG U133A). Patients from I-SPY were treated with doxorubicin/cyclophosphamide followed by a taxane. Patients from MDACC were treated with paclitaxel followed by sequential fluorouracil-AC. Tumors were classified into intrinsic subtypes using PAM50(Parker et al. J Clin Oncol 09) and Claudin-low(CL) predictor(Prat et al., Submitted). Odds ratios of biomarkers to pCR were determined using logistic regression. Predictive values of multivariable models were estimated using receiver operating characteristic curve. Results: 319 patients had complete data; the pCR rate was similar between the datasets (ISPY 22%, MDACC 20%). Both the VEGF-s and CRYAB were highly expressed within Basal-like(BL) and CL (P<0.0001); these two biomarkers were positively correlated with each other (r: 0.4, P<0.001). In univariate analysis, both VEGF-s (P<0.001) and CRYAB (P<0.001) were significantly associated with pCR. A multivariable test including tumor size, grade, ER, Her2, intrinsic subtypes, VEGF-s and CRYAB, had an AUC value of 0.86 to predict pCR(Table 1). T stage, Her2, intrinsic subtypes and CRYABwere significant.

82 patients with triple-negative tumors within this combined cohort. In univariate analysis, T stage, grade or classifying tumors into BL vs not did not predict pCR; but VEGF-s (p= 0.04) and CRYAB (p = 0.02) were associated with pCR. Multivariable model, including VEGF-s, CRYAB and the clinical variables, had a predictive value of 0.71 to pCR(Table 1), which identified CRYAB as the most significant factor by likelihood ratio test.

Table 1: Multivariable logistic regression models to predict pCR.

Conclusion: VEGF-s and CRYAB expression are associated with aggressive subtypes and may possibly help to predict response of anthracycline/taxane-based neoadjuvant chemotherapy beyond ER, Her2, and intrinsic subtypes.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr PD07-09.
Abstract PD08-03: Competing Causes of Mortality vs. Breast Cancer Mortality at 5-Years among 9766 Postmenopausal Women with Hormone Receptor Positive Early Breast Cancer Treated on the TEAM Study of Adjuvant Hormonal Therapy
Authors: Jones SE , Putter H , Hasenburg A , van de Velde CJH , Rea D , Vannetzel J-M , Paridaens R , Markopoulos C , Hozumi Y , Seynaeve C.
Published: April 2011

Background: The TEAM (Tamoxifen Exemestane Adjuvant Multinational) trial is a contemporary multinational study of the effects of 5 years of adjuvant hormonal therapy, either exemestane (E) or tamoxifen switching to E (TàE). Five-year results were presented at SABCS 2009 showing no difference in outcome between E vs TàE (Rea D, SABCS Abstract 11), thus giving us the opportunity to analyze the causes of mortality in the entire population of 9766 women.

Methods: Postmenopausal women with hormone receptor positive (HR+) early breast cancer (BC) were randomized to E or TàE for 5 years after completion of surgery, adjuvant chemotherapy (when indicated), and radiation therapy based on local guidelines. All data were collected and analyzed at the Central Data Center in Leiden. The primary outcome of DFS as well as safety and other secondary outcomes were analyzed in 2009 after 1285 events had occurred to trigger the analysis. Results: Between 2001 and January 2006, 9766 women were randomized; 100% were HR+, 50% had node negative disease, 68% received adjuvant radiotherapy, and 36% received adjuvant chemotherapy. The median follow-up was 5.1 years with over 60% of women having been followed for 5 or more years. Among 961 deaths, 568 (59%) were caused by BC and 393 (41%) by causes other than BC. The cumulative incidence of death from BC vs. other causes at 5-years for all women and by nodal status is as follows (see table):

There was a general relationship with non-breast cancer deaths and increasing age at diagnosis (≥70 years), as expected. Non-breast cancer causes of mortality included second malignancy (105); cardiac (71), cerebral (33), or pulmonary (28) events; miscellaneous other causes (105); and unknown (51).

Conclusions: Among 9766 postmenopausal women with HR+ BC treated with contemporary hormonal strategies following standard primary treatment, the probability of death from causes other than BC was nearly as high as the probability of dying from BC.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr PD08-03.
Abstract PD08-09: Impact of a Telephone-Based Exercise Intervention on Physical Activity Behaviors and Fitness in a Cooperative Group Setting
Authors: Ligibel JA , Meyerhardt J , Najita J , Shockro L , Campbell N , Pierce J , Paskett E , Shapiro C.
Published: April 2011

Observational studies have demonstrated a 30-50% lower risk of disease-specific and overall mortality in physically active breast and colorectal cancer patients as compared to sedentary individuals. However, there have been no randomized trials looking at the impact of physical activity on cancer outcomes, and the optimal design of such a trial is not yet well-defined. The Active After Cancer Trial (AACT) is a multicenter feasibility study designed to evaluate the ability of a telephone-based intervention to increase physical activity in patients with breast and colorectal cancer. Methods: Sedentary (reporting less than 60 minutes/week of recreational activity) individuals with stage I-III breast or colorectal cancer were eligible for enrollment after completion of all adjuvant chemotherapy and radiation. Participants were randomized 1:1 to a centralized telephone coaching intervention, with a target goal of 180 minutes/week of physical activity, or to a usual care control group. Intervention participants received an average of 10 telephone contacts over 16-weeks. Initial calls focused upon building self-efficacy and later calls concentrated upon relapse prevention and maintenance of exercise behaviors. Participants underwent assessment of physical activity behaviors (7-Day Physical Activity Recall), fitness (6-Minute Walk Test), physical functioning (EORTC QLQ C-30), fatigue (FACIT) and exercise self-efficacy at baseline and 16 weeks after enrollment.

Results: One hundred and twenty-one patients were enrolled through 10 Cancer and Leukemia Group B (CALGB) institutions; 100 patients had breast cancer and 21 had colorectal cancer. Average age was 54.3, 74% of patients had received chemotherapy and mean time since completion of adjuvant treatment was 24 months. Participants randomized to the exercise group experienced significant improvements in fitness and physical functioning as compared to controls.

Table. Baseline measures and change (post-minus pre) scores in intervention and control patients (±SD)

Intervention participants also reported a doubling in minutes of weekly physical activity, but this was not a significant increase compared to controls.

Conclusions: Sedentary breast and colorectal cancer survivors can be enrolled in a physical activity intervention. A centralized telephone coaching intervention successfully increased fitness and physical functioning, although self-reported exercise time was not significantly changed. A large-scale clinical trial within the co-operative groups is feasible.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr PD08-09.
Abstract PD09-02: Prognostic Effect of Obesity on Breast Cancer
Authors: Tryfonopoulos D , Christakou H , Rapti V , Kontou N , Panopoulos C , Koumakis G , Demiri S , Papadimitriou C , Missitzis I , Apostolikas N , Efremidis A.
Published: April 2011

Background: Body mass index (BMI), prior estrogen use, and family history have been described as risk factors for invasive breast cancer. Our purpose was to investigate the influence of obesity on established prognostic pathologic and molecular characteristics of breast cancer. Material and methods: Five-hundred and twelve female consecutive breast cancer patients of mean age 53±11 years were included in this study. Data were recorded in a prospective electronic databank upon first presentation. Risk factors (BMI, family history, prior estrogen use, menopausal status, parity, metabolic syndrome, smoking, alcohol, hypertension, hyperlipidaemia and diabetes) as well as histologic characteristics (TNM, nodal status, grade, LVI) and tumor phenotype (ER, PR, Cerb-B2) were obtained. Patients with a BMI<27 were considered of normal weight whereas patients with a BMI≥27 were considered as obese. Univariate and logistic regression analysis were performed. Results: The 2 groups did not differ for age (p=ns). Patients with a BMI≥27 were more frequently node positive, had a higher incidence of grade 3, LVI positive tumors and had more often a positive family history of breast cancer (P<0.05). On the contrary, patients with a BMI<27 reported more frequently prior estrogen use and were more commonly nulliparous (P<0.05). Regression analysis confirmed that in patients with BMI<27 age and menopausal status were associated with node positive disease, independently of tumor phenotype, grade, LVI, family history, estrogen use and parity (0.951, 95%CI 0.91-0.99 and 3.232, 95%CI 1.2-8.5). Using the same analysis, in patients with BMI≥27 LVI was associated with node positive disease, independently of all other factors (5.253, 95%CI 1.1-26.9).

Discussion: Different risk factors appear to influence lean and obese breast cancer patients. Patients with BMI<27 more frequently have a history of estrogen use and nulliparity. Age and menopausal status are independently associated with node + disease in lean patients. Obese patients (BMI>27) have more frequently positive family history and more aggressive histology (node +, grade 3, LVI+). LVI in obese patients appears to be independently associated with node + disease.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr PD09-02.
Abstract PD09-04: Weight Loss in Postmenopausal Women Is Associated with Modulation of Serum and Tissue Based Risk Biomarkers
Authors: Fabian CJ , Kimler BF , Phillips TA , Zalles CM , Klemp JR , Malone LM , Hursting SD.
Published: April 2011

Background

In a pilot study, a structured program of reduced energy diet, physical activity, and weekly group behavioral intervention with other high risk women was successful at producing a median 11% weight loss with at least a 5% weight loss in 88% of subjects. We evaluated the association of weight loss with changes in serum and breast tissue risk and mechanisms of action biomarkers. Methods

High risk postmenopausal women with BMI >25 kg/m2 had breast tissue harvested by random periareolar fine needle aspiration (RPFNA) before and after a 6-month energy balance intervention. Specimens were evaluated for biomarkers including cytomorphology, proliferation (immunocytochemical Ki-67), gene expression by RT-qPCR, and expression of cytokines and adpokines by Luminex assay. Fasting serum was assayed for insulin, glucose, adiponectin, leptin, high sensitivity CRP, IL-6, prolactin, SHBG, estradiol and testosterone using ELISA or Luminex.

Results

For 24 biomarker evaluable subjects, 21 had >5% weight loss (median = 11%). Cytologic atypia was present in 10/24 at baseline and 4/24 at 6 months (p=0.034). For 20 subjects with sufficient cells for assessment of Ki-67 at both times, median baseline Ki-67 was 0.7% and off study 0.3%, with a median change of -0.2% (p=0.19). Statistically significant changes (≥0.003; Wilcoxon) were observed for serum levels of adiponectin, adiponectin:leptin ratio, and SHBG (increases); and leptin, bioavailable estradiol and hsCRP (decreases). Reduction was also observed for insulin (p=0.018) and bioavailable testosterone (p=0.033). These results were duplicated (p≥0.014) by Luminex for adiponectin, leptin, adiponectin: leptin ratio, and insulin; plus hepatocyte growth factor (HGF, decrease). Also, an increase in the adiponectin:leptin ratio was observed for the RPFNA specimens (p=0.012). Gene expression (RT-qPCR) of pS2 was significantly modified (decrease, p=0.035). Further, the weight loss (expressed as relative change) was highly statistically correlated with change (relative) in serum leptin, adiponectin:leptin ratio, SHGB, and free estradiol; as well as with relative change in adiponectin:leptin ratio in RPFNA specimens. Conclusion

Weight loss in high risk postmenopausal women is accompanied by significant modulation of numerous serum and breast tissue-based biomarkers. For several risk and response biomarkers there is a significant correlation between change in the biomarker and the weight loss achieved. This suggests the possibility of identifying mechanisms of action and signaling pathways for dietary/energy balance interventions that may reduce risk for development of breast cancer.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr PD09-04.
Abstract PD09-06: Obesity Promotes Breast Cancer Progression and Tamoxifen Resistance Via Cross-Talk between Growth Factor and Estrogen Signaling Pathways
Authors: Bowers LW , Cavazos DA , De Angel RE , Price RS , Hursting SD , Brenner AJ , deGraffenried LA.
Published: April 2011

Epidemiological studies indicate that obesity increases the risk of breast cancer by approximately 50% in postmenopausal women. This population most commonly develops estrogen receptor (ER) alpha positive breast cancer, suggesting that elevated estrogen synthesis by the adipose tissue and local mammary epithelia is the primary mediator of breast tumorigenesis in obese postmenopausal women. However, the specific mechanisms by which obesity promotes the development of postmenopausal breast cancer remain unclear. In vitro and animal studies have indicated that obesity-associated growth factor and cytokine signaling also play a role in breast tumorigenesis. Specifically, obesity is accompanied by elevated levels of insulin, insulin-like growth factor 1 (IGF-1), and leptin, and these all have significant tumorigenic effects in the breast.

In the current study, we utilize an in vitro model of obesity to examine the effects of elevated circulating hormones and growth factors on breast cancer cells. C57BL/6 mice were randomized at six weeks of age to either a 60% fat diet to induce obesity or a 10% fat diet to produce a lean comparison and maintained on these diets for 12 weeks prior to sacrifice. Sera was obtained from the mice at the time of euthanasia and used to treat ER alpha positive MCF-7 breast cancer cells at a concentration of 2% in serum-free media. Preliminary data indicates that sera from the obese mice induces 31% greater proliferation of MCF-7 cells in comparison to sera from lean animals (p=0.0025). This proliferation is mediated primarily via a phosphoinositide 3-kinase (PI3K) pathway, as evidenced by the ability of LY294002, a PI3K inhibitor, to neutralize the proliferative effect of the obese sera. The addition of LY294002 to MCF-7 cells treated with 2% obese sera decreased proliferation by 43% (p=0.0083). Intriguingly, tamoxifen did not significantly affect proliferation in the cells treated with obese sera (p=0.2425), suggesting that obesity may induce tamoxifen resistance.

Consequently, we hypothesize that obesity promotes breast cancer progression and tamoxifen resistance via cross-talk between growth factor and ER alpha signaling pathways. Further studies will be conducted to examine the pathways through which circulating factors mediate obesity-induced breast cancer progression. These will include measurement of the expression and activation of ER alpha, Akt, Erk1/2, IGF-1 receptor, and other proteins in MCF-7 cells in response to treatment with sera from obese and lean mice. By improving our understanding of the mechanisms linking obesity and post-menopausal breast cancer risk, we ultimately hope to develop a rational and effective chemopreventive regimen for this population.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr PD09-06.
Abstract PD09-08: Obesity in Long-Term Disease Free Survivors of Invasive Breast Cancer
Authors: Bahadur S , Roques-Davis R , Shi R , Burton G , Quispe D.
Published: April 2011

Background: As the number of breast cancer survivors increases due to improvements in screening and treatment for breast cancer, there is a particular interest in identifying lifestyle factors associated with long-term survival. Obesity may be one of the modifiable risk factors that can play a role in the outcome of breast cancer patients however studies shown controversial results. Besides, little is known about the role of obesity in long-term survivors. The objective of this study is to examine the role of obesity in breast cancer patients who remained disease free at least 5 years after diagnosis.

Methods: We reviewed the medical records of all women with invasive breast cancer diagnosed at LSUHSC between 1980 and 2005. The patients with stage IV breast cancer at diagnosis were excluded from the analysis. For this study we selected patients whom were disease free at least five years after the breast cancer diagnosis. We classified our patients by body mass index (BMI) at the time of breast cancer diagnosis into: normal weight (BMI <25 kg/m2), overweight (BMI 25-29 kg/m2), and obese (BMI >30 kg/m2). Data on demographics clinical characteristics, tumor characteristic, breast cancer therapy, and relapse were analyzed within BMI classes. Results: A total of 967 patients were diagnosed with invasive breast cancer between 1980 and 2005. Out of those 49% were African American, 29% were overweight, and 49% were obese. From this population 597 (62%) patients were disease free more than 5y: 217 (49%) obese, 113 (26%) overweight, 112 (25%) normal weight. Obese breast cancer survivors in comparison to overweight and normal weight survivors were more likely to: be African American (61%, P<0.01), received Medicaid coverage (14.2%, p=0.05) and to have lower rates of mastectomy (p=0.017). There was not difference in tumor characteristics (ER status, Nodal status, Tumor size) and adjuvant treatment (chemotherapy, radiotherapy and hormonal therapy) between obese and non-obese patients. One hundred and ten patients (19%) developed a recurrence 5 years after diagnosis (48 obese and 33 overweight): 65 local (48 obese and 33 overweight) and 45 systemic (21 obese and 10 overweight).

Discussion: Our study has some limitations typical of retrospective studies and the generalization of the results may be limited. Our analysis suggests that in long-term disease free survivors obesity and race might be associated. To further clarify the role of obesity in long-term breast cancer survivors prospective studies with long term follow up are warranted.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr PD09-08.
Abstract PL3-1: The Upside of Irrationality
Authors: D Ariely
Published: April 2011

Dan will discuss some of the challenges and underlying benefits of irrationalities in our day-to-day life. Using behavioral economic principles as a starting point, Dan will look at how these irrationalities affect our day-to-day behaviors both in the workplace and in our personal lives.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr PL3-1.
Abstract S2-6: Aromatase Inhibitors and Risk of Myocardial Infarction, Stroke and Fracture
Authors: Ligibel JA , O'Malley A , Fisher M , Daniel G , Winer EP , Keating NL.
Published: April 2011

Randomized trials have demonstrated that aromatase inhibitors (AI's) are more effective than tamoxifen in preventing breast cancer recurrence. Since many women with early stage disease have a modest risk of recurrence, it is important to consider carefully the potential side effects of treatments in this setting. Trials have suggested increased risk of fracture is the only serious side effect of AI's, but study participants are often healthier than the general population, and little is known about toxicities of AI's in nontrial populations. We sought to evaluate the association between use of AI's and the incidence of myocardial infarction (MI), stroke, and fracture in community-based populations.

Methods: Using encounter and pharmacy data from WellPoint plans in the HealthCore Integrated Research Database, 44,463 women aged ≥50 years with at least 2 diagnosis codes for breast cancer between 2001 and 2007 were identified and followed through the end of 2008. We used propensity score methods to identify matched controls from a cohort of 88,101 women aged ≥50 years enrolled in WellPoint plans who had no diagnosis codes for breast cancer. We identified receipt of aromatase inhibitors and tamoxifen and used Cox proportional hazards models with time-varying treatment variables to assess whether treatment with aromatase inhibitors or tamoxifen was associated with MI, stroke, and fracture among women with breast cancer and to assess the association of breast cancer with the outcomes of interest.

Results: Among 44,049 breast cancer patients and 44,049 matched controls, the average age was 67 and 87% of cases and controls had no baseline comorbidities. After a median follow-up of 899 days for cases and 1004 for controls, 1.3% of women had an MI, 2.9% an ischemic stroke, 1.5% a hip fracture and 9.5% any fracture. Breast cancer patients treated with AI's had a significantly lower adjusted hazard of ischemic stroke and a significantly higher adjusted hazard of fracture compared with breast cancer patients not receiving an AI or tamoxifen.

Table. Adjusted hazard ratio (95% CI) associated with breast cancer and hormonal therapy

Treatment with AI or tamoxifen was not associated with MI or any fracture. Patients treated with tamoxifen had a significantly lower adjusted hazard of stroke and hip fracture compared with breast cancer patients not receiving hormonal therapy.

Conclusion: The side effect profile of AI's in this community-based population was similar to that seen in clinical trials and suggests that these drugs are associated with few serious side effects.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr S2-6.