bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P1-02-01: Vasohibin-1 in Human Breast Carcinoma: A Potential Negative Feedback Regulator of Angiogenesis
Authors: Tamaki K , Ishida T , Miyashita M , Sato Y , Tamaki N , Ohuchi N , Sasano H.
Published: April 2011

Background: Vasohibin-1 is a recently identified negative feedback regulator of angiogenesis induced by vascular endothelial growth factor (VEGF)-A. This is the first study to evaluate immunoreactivity and mRNA expression of Vasohibin-1 in human breast disorders. Material and Methods: We examined 251 breast specimens including invasive ductal carcinoma (IDC: 148), ductal carcinoma in situ (DCIS:62), fibroadenoma (FA:16), inflammatory lesion (6), fibrocystic change(9) and non-pathological breast tissue(7). We immunolocalized vasohibin-1 and compared its immunoreactivity to that of VEGF-A, VEGF receptor 2 (Flk-1), CD31 and Ki-67. The correlation of vasohibin-1 immunoreactibity with overall survival (OS), and disease free survival (DFS) of the breast carcinoma patients was also evaluated. In addition, we evaluated Ki-67 and CD31, and Ki-67 and vasohibin-1 double-immunostaining for further characterization of neovascularization. We also evaluated mRNA expression of Vasohibin-1 using a real-time quantitative RT-PCR with LightCycler system.

Results: Vasohibin-1 was detected in endothelial cells of human breast and its immunodensity was significantly higher in IDC and inflammatory lesion than the other types (P<0.001). As for DCIS, there was a significant positive correlation between vasohibin-1 expression level and nuclear grade or Van Nuys classification of carcinoma cells (P<0.001). In addition, a significant positive correlation was detected between vasohibin-1 and VEGF-A, Flk-1 or Ki-67 (P<0.001). There was also positive associations between vasohibin-1 and OS (P=0.004) and DFS (P<0.001). Results of double-immunostaining demonstrated the ratio of Ki-67-positive cells among vasohibin-1-positive endothelial cells (46.5%) was significantly higher than those among CD31-positive cells (23.5%). In addition, there was a significant positive correlation between Vasohibin-1 mRNA level and Ki-67 or high nuclear grade(P<0.001).

Discussion: This is the first study demonstrating the status of vasohibin-1 in human breast lesions, which indicates that vasohibin-1 is associated with neovascularization and may especially play important roles in the regulation of intratumoral angiogenesis in human breast cancer. Vasohibin-1 is supposed to be induced in the downstream of VEGF-A/Flk-1 signaling pathway and considered an appropriate biomarker for intratumoral neovascularization. In addition, these results clearly indicated that vasohibin-1 expression may become one of the prognostic markers for metastasis and prognosis, and also become one of the appropriate biomarker of the potential of subsequent stromal invasion of carcinoma cells.

Figures available in online version.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P1-02-01.
Abstract P1-02-04: The Influence of Matriptase-2 on Angiogenesis and Tumour Growth In Vivo.
Authors: Webb SL , Sanders AJ , Mason MD , Jiang WG.
Published: April 2011

Background: Metastasis is a key part of cancer progression and angiogenesis plays a large role in the ability of cancer cells to spread around the body and in the progressive growth of the primary tumour. Matriptase-2 is a cell surface serine protease believed to have implications in cancer metastasis. The current study aims to assess the importance of matriptase-2 in angiogenesis and tumour growth, using in vitro and in vivo models. Methods: Matriptase-2 was over-expressed in the HECV endothelial cell line, previously displaying minimal matriptase-2 expression, through the transfection of a mammalian expression construct containing the full coding sequence for matriptase-2. Following verification of forced expression, the cellular effects of matriptase-2 were examined using a number of in vitro cell models. The effect of matriptase-2 over-expression was also examined in in vivo mouse models by co-delivery of endothelial cells and tumour cells.

Results: Matriptase-2 significantly reduced the motility (46.47±16.18 vs 10.93±2.80 p=<0.01) of the HECV cells and their ability to form tubule structures (23841.60±744.75 vs 9817.80±933.85 p=0.01) in an in vitro angiogenesis Matrigel tubule formation model. Matriptase-2 was found to have no significant effect on the growth (691.52±100.17 vs 763.50±91.43 p=0.110) and cell-matrix adhesion of the HECV cells (64.20±8.70 vs 57.42±4.50 p=0.530). However, the in vivo studies showed a reduction in tumour growth and development following co-injection of cancer cells with endothelial cells over-expressing matriptase-2 compared to co-injection with control endothelial cells. The sizes of tumours in two tumour models were: 70.99±19.52mm3 for tumour cells with control endothelial cells vs 0.17±0.10 mm3 for tumours with Matriptase-2 over-expression endothelial cells, p=<0.01; and 80.41±18.31 mm3 vs 43.12±20.33 mm3 p=0.08, in the other tumour model.

Conclusion: Matriptase-2 appears to negatively influence angiogenesis through its inhibitory effect on endothelial cell motility and tubule formation. This is corroborated by the reduction of tumour growth, in vivo. Together this data suggests that matriptase-2 has an important role in the control of angiogenesis and tumour development.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P1-02-04.
Abstract P1-02-05: EPLIN? Can Negatively Impact on Angiogenesis and Is Associated with ERK Signalling
Authors: Sanders AJ , Ye L , Jiang WG , Mason MD.
Published: April 2011

Background: EPLIN (epithelial protein lost in neoplasm) is a cytoskeletal associated protein involved in regulating actin dynamics and cell motility. EPLIN expression is frequently lost in cells as they progress to a cancer state and this loss may account for enhanced cancer cell motility. EPLIN has also been shown to be lost in clinical breast cancer, which is associated with patients clinical outcome. Recently, EPLIN has been linked to ERK signalling. This study examined the potential of EPLIN to influence the angiogenesis process and its links to ERK in endothelial cells Methods: Over-expression of EPLIN α was achieved through transfection of the vascular endothelial cell, HECV, with a mammalian expression construct containing the full coding sequence of EPLINα. Enhanced levels of EPLINα were verified through RT-PCR, Q-PCR and Western blot analysis before assessing the angiogenic potential of control and transfected cells using angiogenesis and cell migration models. Treatment of transfected and control cells with ERK inhibitor was used to assess the potential interaction between EPLINα and ERK in endothelial cells. Results: Substantial enhancement in EPLINα expression was seen in transfected cells compared to control cells at both transcript and protein level. Over-expression of EPLINα in HECV endothelial cells resulted in a reduced capacity, compared to control, of these cells to form tubule like structures in an in vitro angiogenesis tubule formation assay (6566 +/− 856µm tubule perimeter of control cells vs 2881 +/− 546µm EPLINα overexpressing HECV cells, p = 0.007). This was seen together with a reduced cell migration following overexpression of EPLINα in the cells. Treatment of control cells with ERK inhibitor significantly reduced tubule formation (3537 +/− 277µm tubule perimeter of ERK inhibitor treated control cells vs 6566 +/− 856µm total perimeter of untreated control cells, p = 0.01), however treatment of HECV cells transfected with EPLINα expression construct caused tubule formation levels to return to control levels (6566 +/− 856µm tubule perimeter of untreated control cells vs 5326 +/− 784µm tubule perimeter of ERK inhibitor treated EPLINα overexpressing HECV cells, p = 0.317).

Discussion: EPLINα can negatively influence tubule formation in vitro and thus may impact on the angiogenesis process. Furthermore, its impact on tubule formation appears to be linked to ERK signalling. Together, this study suggests a role for EPLINα in angiogenesis, which may be linked with ERK signalling.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P1-02-05.
Abstract P1-03-09: Combination of nab-Paclitaxel and Bevacizumab Inhibited Tumor Growth and Metastasis of New Inflammatory Breast Cancer Models
Authors: Volk LD , Wilber A , Trieu V , Desai N , Ran S.
Published: April 2011

Background: Inflammatory breast cancer (IBC) is a highly lethal form of breast cancer which is characterized by skin redness, irritation, swelling, pain, as well as extensive lymph nodes (LN) and hematogenous metastasis. Animal models of IBC are highly desirable for studying its pathology and for designing effective therapies. In this study, we established two new luciferase-and fluorescently-labeled IBC models and tested the efficacy of the novel drug combination nab-paclitaxel and bevacizumab. Methods: Inflammatory SUM149 breast cancer cells were stably transfected with Red Fluorescent Protein (RFP) and Renilla luciferase to establish SUM149-RR, or infected with lentivirus encoding Green Fluorescent Protein (GFP) and Firefly luciferase to establish SUM149- GL. The new cell lines were characterized both in vitro and in vivo. Immunodeficient mice bearing SUM149-RR tumors of 150mm3 in size were treated with saline (control), bevacizumab (4 mg/kg, i.p., twice a week, for 10 weeks), nab-paclitaxel (10 mg/kg, i.v., qdx5), or the combination. Metastasis was analyzed by measuring luciferase activity in the lymph nodes (LN) and lungs.

Results: Luciferase measurements and in vivo imaging showed that both SUM149-RR and-GL clones were highly metastatic to LN, lungs, liver, brain, and spleen. SUM149-RR tumors in control mice displayed ulcerations, edema and redness similar to the clinical disease, while tumors in mice treated with bevacizumab or combination therapy showed no signs of inflammation. Bevacizumab alone decreased tumor growth at later but not early stages of tumor growth, whereas nab-paclitaxel alone inhibited tumor growth by 73%. Combination therapy increased inhibition to 96%, and resulted in 22% (2/9) complete responses. Histologically, tumors from bevacizumab treated groups were more morphologically intact with reduced vascular abnormalities than tumors from control or nab-paclitaxel treated mice. LN and lung metastasis was significantly reduced in all treated groups as compared with control.

Conclusions: The SUM149-RR and SUM149-GL lines are new double-tagged models of human IBC that allow organ visualization and accurate quantification of metastasis. These models can be used to study the biology and treatment of IBC. Combination of nab-paclitaxel with bevacizumab was highly effective against SUM149-RR, suggesting the potential usefulness of this regimen for treatment of IBC patients.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P1-03-09.
Abstract P1-03-10: A Model To Prevent Pregnancy Associated Breast Cancer: The Effect of HCG on Tumorigenesis in MMTVneu Mice
Authors: Rouzier R , Vercambre-Donck M , Wafo E , Uzan S , Aractingi S.
Published: April 2011

Background: HER2 expression is associated with increased parity-induced risk of breast cancer in women, and women that have been pregnant and have breastfed acquire a 4.2-fold increased risk of developing HER2- positive breast cancers than their counterparts that never breastfed. The mechanisms responsible for the increased risk of developing HER2- positive breast cancer following pregnancy and lactation remain to be elucidated. Animal and ‘in vitro’ experiences show that human chorionic gonadotropin (hCG) is capable to protect from breast cancer. The aim of this study was to assess the effect of HCG and pregnancy on tumorigenesis in MMTVneu mice and to study the mechanisms in terms of differentiation and stem cells depletion.

Methods: Mice were intraperitoneally injected BrdU then intraperitoneally injected HCG. Mice were sacrificed for the detection of label retaining cells. The two inguinal groups of mammary gland were dissected. For each gland, ducts and lobules were identified and drawn manually. We used Mercatorâ software for morphometry: ductal and lobular surface were measured and reported to the surface of the selected region. Cells retaining the label after 8-10 weeks (adulthood) were identified as LRCs. A mammary gland tumor free survival was conducted. Results: We observed that the differentiation and maturation of the mammary gland caused by the HCG was different in MMTVneu mice and controls: whereas WT littermates presented with sleek ductal appearance before parity, increased lateral side branching and budding was observed in mammary gland ducts of the MMTV-neu females and WT receiving HCG. The effect of HCG was mediated by the ovary. The effect of HCG on the stem cell pool was limited. Early HCG exposition did not modify ErbB2/Neu-induced mammary tumorigenesis but had an independent tumorigenic effect.

Conclusion: Early stimulation with HCG does not mimic early parity. The tumor development was not different between MMTV-neu mice that received or not HCG. We observed an independent ovary mediated tumorigenic effect of early administration of HCG.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P1-03-10.
Abstract P1-03-12: The First Reproducible Model of Primary Human Ductal Carcinoma In Situ (DCIS) Using the Mouse Intraductal (MIND) Method
Authors: Valdez K , Smith W , Fan F , Medina D , Behbod F.
Published: April 2011

Past studies of the molecular mechanisms regulating human DCIS progression to invasion have been limited by a lack of suitable human models. The current models have not utilized primary human DCIS placed in their natural microenvironment, i.e., inside the mammary ducts, and do not mimic the heterogeneity of human DCIS. Previously, we reported the development of a mouse xenograft model using established breast DCIS cell lines, SUM-225 and DCIS.COM by the mouse intraductal (MIND) method. MIND involves injection of human DCIS cells into the primary ducts of immunodeficient mouse mammary glands via the nipple. In this study we demonstrate the reproducible growth of human primary DCIS cells derived from patient biopsy samples using the MIND model. Core biopsies from consenting patients are processed to isolate the epithelial cell component and injected. To date, 69 patients have consented to provide core biopsy tissue and we have received tissue from 38. Of these, 14 samples have been confirmed as positive for DCIS, and 2 as atypical hyperplasia, based on the pathology report of adjacent biopsies. The biopsy samples were heterogeneous with respect to ER, PR, Ki67, Her-2 and histology. A total of 24 mammary glands from immunodeficient mice have been injected with these cells and analyzed for growth by immunofluorescent (IF) with antibodies directed at human-specific cytokeratin-5 and/or 19 and smooth muscle actin.

Table 1. Summary of grewlh of human primary biopsies in (tie MIND xenograft

The average number of cells recovered per mg of tissue are shown in Table 1. Biopsies diagnosed as normal yielded only 91±34 cells/mg compared with most DCIS cases that showed an average cell number of 971 per mg tissue. Our results indicate that human DCIS and hyperplasia cells from biopsy samples are capable of growing within the NOD-SCID IL2Rgammanull (NSG) mouse mammary ducts. NSG mice mammary ducts are more suitable hosts for the growth of human primary DCIS cells, compared to SCID-Beige. Furthermore, the higher number of cells recovered following digestion predicted the positive DCIS growth rate. DCIS cells formed single and multi-layered epithelium inside the ducts and were heterogeneous with respect to the expression of human specific cytokeratins. The MIND xenografts recapitulated the patient's original DCIS as evidenced by IF staining for the biomarkers ER and Her-2 in the primary human biopsies and MIND xenografts. These results provide the first reproducible model of primary human DCIS for studying the temporal processes of early breast cancer progression. Future experiments will characterize the cellular basis for the subtypes of DCIS and delineate the distinct molecular and cellular mechanisms of early breast cancer invasion, and cancer stem cell growth and self-renewal.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P1-03-12.
Abstract P1-08-02: Short-Term Pregnancy Levels of Estradiol Delays Mammary Tumor Development by Altering PI3K/AKT Pathway in Her2/neu Mice
Authors: Parada JA , Arumugam A , Agullo P , Myers S , Delgado A , Boopalan T , Lakshmanaswamy R. , Paul L.
Published: April 2011

Approximately 20-30% of all breast cancers overexpress the human epithelial growth factor receptor 2 (Her2). Breast cancers that overexpress Her2 have been associated with a more aggressive phenotype and decreased survival. Currently, the FDA has only approved trastuzumab, a recombinant monoclonal antibody against the Her2 receptor, for treatment of Her2- overexpressing breast cancer. Even though trastuzumab has been effective, many breast cancers do not respond to treatment or they eventually progress to a resistant state. Hence, several research investigations are underway to identify therapies besides trastuzumab that could be used for the treatment of Her2-positive breast cancer. It has been earlier demonstrated that uniparous Her2/neu transgenic mice developed smaller mammary tumors and these tumors had significantly decreased metastatic potential. Also, we had earlier demonstrated that short-term treatment with pregnancy levels of estradiol was very effective in delaying tumor progression and decreasing tumor burden. In this present investigation, we have attempted to understand the molecular mechanisms underlying this anti-tumor effect of short-term pregnancy levels of estradiol treatment against Her2/neu driven mammary carcinogenesis. Seven week old activated Her2/neu (murine-mammary-tumor virus (MMTV)-c-neu) FVB transgenic mice were treated for 3 weeks with 100mg of estradiol in silastic capsules. We have shown that this dose of estradiol treatment mimics pregnancy levels of the hormone in circulation. Control animals received empty silastic capsules for the same duration. Mice were palpated once every week for nine months to monitor the development of mammary cancer. Histopathological examination was performed to confirm the carcinomatous nature of the palpable tumors. A group (n=3) of mice were terminated from control and estradiol treated groups immediately after three weeks of treatment, and also, at 6 and 12 weeks post withdrawal of the treatment. Mammary tumor and normal mammary tissue were excised, snap-frozen in liquid nitrogen and stored at −80°C. Protein and total RNA were extracted from these tissues. Pathway focused microarray analysis was done on the PI3K/AKT pathway. Results demonstrate that, in parous and estrogen treated mice, proteins and genes involved in tumor-suppression (Mtcp1,Rbl2,Srf and Ywhah) and immune response (Cd14,Tlr4) were upregulated; which have been shown to play a role in reducing tumors by induction of apoptosis. This trend in gene expression alteration is present in both normal and mammary tumor tissue in the estradiol treated and parous groups and demonstrates that short-term pregnancy levels estrogen is altering the PI3K/AKT pathway; which correlates with our previous findings that short-term estradiol treatment is effective in tumor delay.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P1-08-02.
Abstract P1-09-07: Survival, Health Care Utilization and Cost after Recurrence among Patients with Triple Negative Breast Cancer in a U.S. Managed Care Setting
Authors: Baser O , Wei W , Henk HJ , Xie L , Teitelbaum A.
Published: April 2011

Background: Triple negative breast cancer (TNBC) is characterized by the lack of expression of the genes for estrogen receptor, progesterone receptor or human epidermal growth factor recepter-2 and therefore it has a shorter recurrence-free interval. We compared the survival rates, health care costs and utilization of breast cancer recurrent patients who have triple negative versus no triple negative breast cancer.

Methods: Analyses were conducted using Impact Intelligence Oncology Management (IIOM) (2002-2009) linked with United Health administrative claims. Patients were followed from the time of initial breast cancer diagnosis until death, disenrollment or end of the observation period. Continuous enrollment was required during the observation period. Staging and estrogen and progesterone receptors (ER/PR) and HER2 status were obtained at time of initial diagnosis. Patients’ age, region, cancer stage, and comorbid conditions were controlled for risk-adjusted comparison. A stratified log-rank test was used to determine the equality of survival functions. Cox regression was used for the risk-adjusted survival analysis. Health care resource use during the follow-up period after recurrence was modeled with negative binomial regression. Inverse probability weight adjusted generalized linear models were used to estimate risk-adjusted total health care costs.

Results: The study included 289 recurrent breast cancer patients, out of which 87 (30%) had triple negative breast cancer. At time of recurrence, TNBC patients had similar age, plan, region, and cancer stage distribution relative to non-TNBC patients. During follow-up, TNBC patients had significantly lower survival rate (log-rank test P<0.001). After controlling for baseline differences, the overall mortality risk rates for TNBC recurrent patients were almost three times higher than non-TNBC recurrent patients (HR:2.84, p=0.056). The risk-adjusted annual number of hospitalizations was also higher for TNBC patients with a recurrent event (1.65 vs. 1.13, p=0.05). There were no differences in health care costs after recurrence between TNBC and non-TNBC patients.

Conclusion: Although similar in health care costs, triple negative breast cancer is associated with higher mortality and number of hospitalizations after the recurrence of breast cancer.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P1-09-07.
Abstract P1-11-01: ANZAC: A Randomised Neoadjuvant Biomarker Study Investigating the Anti-Tumour Activity of the Addition of Zoledronic Acid to Chemotherapy in Breast Cancer
Authors: Winter MC , Syddall SP , Cross SS , Evans A , Ingram CE , Jolley IJ , Hatton MQ , Mori S , Holen I , Coleman RE.
Published: April 2011

Background: Pre-clinical studies have demonstrated sequence-dependent synergistic anti-tumour effects of chemotherapy (CT) and zoledronic acid (Z) in bone and soft tissue tumours. We have also reported that the addition of Z to neoadjuvant CT appears to improve pathological response in the surgical resection specimen. The ANZAC study aimed to investigate the short-term anti-tumour effects of neoadjuvant CT +/− Z in patients with invasive breast cancer, evaluating biological end-points including apoptosis, proliferation and angiogenesis.

Methods: Forty patients were randomised to receive a single 4mg infusion of Z 24 hours after the first cycle of FE100C chemotherapy, as per the most effective schedule demonstrated pre-clinically, or CT alone (CT n=20, CT+Z n=20). Randomisation was stratified for tumour (T) stage, ER, HER2 status and menopausal status to minimise biological differences in the two treatment groups that may influence chemosensitivity. All patients had repeat breast core biopsy at Day 5 (D5) +/− Day 21 (D21). Apoptotic index (AI) and proliferation were measured on core biopsy specimens using TUNEL and Ki67 immunohistochemistry, counting a total of 2000 and 1000 tumour cells respectively. Serum VEGF was also measured at baseline prior to CT +/− Z, and on D5 and D21. Differences between the groups in change from baseline to subsequent timepoints were investigated using the Mann-Whitney U test.

Results: Baseline clinico-pathological characteristics were well balanced between the two groups. For the different biological end-points, percentage change from baseline to D5 and from baseline to D21 are shown in Table 1.

At D5, a greater reduction in serum VEGF occurred in patients treated with CT+Z compared to CT: median percentage change -23.8% (IQR -32.9, -15.8) vs. -8.4% (IQR -27.3, +8.9); p=0.02), but these effetcs were lost by D21. Cell turnover index fell at D5 in both groups (increased apoptosis and reduced proliferation) but recovered much more rapidly with CT+Z than CT alone by D21 to levels above baseline (p=0.006). Conclusions: No definite evidence of a direct anti-tumour effect of the addition of Z to CT was observed, but potentially relevant biological effects were seen in this small study. The clinical relevance of recovery of cell turnover by D21 with CT+Z is unclear. Studies with more frequent dosing of Z are warranted to exploit any potential anti-angiogenic effect of Z.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P1-11-01.
Abstract P1-11-03: Pathological Complete Response after Neoadjuvant Chemotherapy + Trastuzumab Treatment Predicts Survival and Detects a Patient Subgroup at High Need for Improvement of Anti-HER2 Therapy. Three Year Median Follow-Up Data of the TECHNO Trial
Authors: Untch M , Fasching PA , Konecny GE , Hasmueller S , Lebeau A , Kreienberg R , Camara O , Müller V , du Bois A , Kühn T , Stickeler E , Harbeck N , Höss C , Kahlert S , Beck T , Fett W , Mehta K , von Minckwitz G , Loibl S.
Published: April 2011

Purpose: To evaluate the efficacy and safety of epirubicin/cyclophosphamide followed by paclitaxel/trastuzumab as neoadjuvant treatment in patients with HER2-overexpressing primary breast cancer (BC). Patients and Methods: Patients with HER2-overexpressing primary BC (≥2cm or inflammatory BC) received preoperative 4 cycles epirubicin/cyclophosphamide (90/600 mg/m2), q3w) followed by 4 cycles paclitaxel 175 mg/m2 q3w with trastuzumab 6 mg/kg body weight q3w (8 mg/kg as loading dose) followed by surgery. Trastuzumab was continued after surgery until completion of 12 months treatment. The primary endpoint was pathological complete response (pCR) defined as no invasive tumor in the breast and axillary lymph nodes.

Results: Two hundred seventeen patients were enrolled. 39% achieved a pCR. Breast conserving surgery was possible in 138/217 (64%) of the patients. Median follow-up was 41 months and 3-year disease-free survival (DFS) was 88% in patients with pCR compared to 73% in patients without pCR (p=0.01). Three year overall survival (OS) was 96% in patients with pCR compared to 86% without pCR (p=0.025). In multivariate analysis pCR remained a significant prognostic factor for DFS and OS (DFS hazard ratio (HR) 2.5 [95% CI 1.2-5.1] p=0.013; OS HR 4.9 [95% CI 1.4-17.4] p=0.012). A cardiac event was reported in 8/217 (3.7%) patients; 6 had an LVEF decrease and two developed 2 (0.9%) clinical congestive heart failure.

Conclusion: The neoadjuvant combination of trastuzumab and chemotherapy results in a high chance for a pCR. However, those patients without a pCR have an increased risk for relapse and death and are therefore candidates for further improvement of anti-HER2 directed therapy.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P1-11-03.
Abstract P1-11-10: Sequential Versus Upfront Intensified Neoadjuvant Chemotherapy in Patients with Large Resectable or Locally Advanced Breast Cancer (INTENS), First Results from a Phase III Study of the Dutch Breast Cancer Trialists’ Group (BOOG)
Authors: Vriens BE , Van de Vijver KK , Boetes C , van Gastel SM , Wals J , Smilde TJ , van Warmerdam LJ , van Laarhoven HW , van Spronsen DJJ , Borm GF , Tjan-Heijnen VC.
Published: April 2011

Background: Taxanes have an established role as (neo)-adjuvant treatment of breast cancer. In the NSABP-B30 and the BCIRG 005, adriamycin and cyclophosphamide followed by docetaxel (AC-T) was compared with docetaxel in combination with adriamycin and cyclophosphamide (TAC) in the adjuvant setting. AC-T outperformed TAC if TAC was given for 4 cycles (B-30). However, AC-T was not significantly different from TAC if TAC was given for 6 cycles (BCIRG 005). In the present study, we compared AC-T with 6 cycles of TAC in the neo-adjuvant setting.

Methods: Women presenting with breast cancer, cT2≥3cm, cT3, cT4 and/or cN+, with measurable disease and no prior treatment, age ≥18 and ≥70 years and Karnofsky-Score ≥70% were eligible. Patients were randomized to AC (60/600 mg/m2 q3wk x 4 cycles) followed by T (100 mg/m2 q3wk x 4 cycles) or TAC (75/50/500 mg/m2 q3wk x 6 cycles). If indicated, trastuzumab and/or endocrine therapy were given as adjuvant treatment. The primary endpoint of this study was the pathologic complete response (pCR) rate to neoadjuvant chemotherapy at surgery. Safety data will be presented separately at a later date.

Results: In total, 201 patients (n=100 AC-T, n=101 TAC) were included between February 2006 and April 2009. Baseline characteristics (ACT/TAC) were well balanced for age (median 49/49y), T-status (cT1-2 51/49%), N-status (cN0 37/40%), hormone receptor status (positive in 65/66%) and HER2 (positive in 28/18%). AC-T resulted in pCR of the breast in 28% of patients and TAC in 19% of patients (Odds ratio (OR) 1.61; 95% CI 0.79-3.33). In patients with positive lymph nodes in situ at start of chemotherapy (no sentinel node procedure), combined pCR of the breast and axilla was 19 % (n=42) and 13% (n=38). Conclusion: AC followed by T resulted in a higher pCR rate, though not statistically significant.

Support: Unrestricted grants from sanofi-aventis NL BV and Amgen BV

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P1-11-10.
Abstract P1-11-14: Neoadjuvant Chemotherapy in Lobular and Ductal Carcinoma: Comparison of Clinical, Pathological Response Rates and Survival
Authors: Coussy F , Cuvier C , Hamy AS , Giacchetti S , De Roquancourt A , Espié M.
Published: April 2011

Introduction: Neoadjuvant chemotherapy is a standard of care in locally advanced breast carcinomas. The major purpose of neoadjuvant chemotherapy is breast conserving surgery. Advantages in survival have only been shown in patients undergoing achieving pathological complete response (pCR). Lobular carcinomas seem less chemosensitive than ductal carcinomas to neoadjuvant chemotherapy (NCT).

Purpose: To compare the clinical and pathological response rate and the outcome of lobular (ILC) versus ductal (IDC) invasive breast carcinomas after NCT.

Patients and methods: Between 1985 and 2010, 385 patients with locally advanced and/or inflammatory breast carcinomas from Saint Louis hospital received neoadjuvant chemotherapy. Forty-four (11, 4%) ILC and 341 (88, 6%) IDC were diagnosed by surgical or core needle biopsy before CT. All patients received anthracyclines based CT,181 (47%) additionally received a taxane (four cycles of epirubicin 75 mg/m2 and cyclophosphamide 750 mg/m2 then four cycles of taxotere 100mg/m2 or six cycles of a dose dense regimen of 75 mg/m2 epirubicin and 1200mg/m2 cyclophosphamide, every 14 days),and underwent breast surgical excision (lumpectomy or mastectomy) and axillary node dissection.Clinical response was defined by the lack of palpable tumor in the breast before surgery.Pathological complete response was defined by no residual invasive tumor in breast. Radiotherapy and hormonotherapy were delivered to patients, when appliable.

Results: Clinical response to NCT was higher for ILC (27, 3%) than IDC (13,4%). ILC with clinical response tended to have higher histological grade 2 (91,6 % versus 30,4% for IDC),more estrogens receptors (RE) positivity (91,6% vs 45,6%), p53 wild type (66,6% vs 30,4%) and HER2- negative tumors (91,6% vs 76%) Pathologic response (pCR) rate was lower for ILC than for IDC (2, 7% vs 9, 3%). Only one ILC (grade 2, RE+, HER2 negative, p53 wild type) underwent pCR. Thirty two (9, 3%) IDC haved pCR: 15,6% haved histological grade 2, 15,6% RE positivity,9,3% p53 wild type, 62,5% HER2 negative. Thirteen percent of ILD and 9 % of ILC haved breast conservating surgery .At a median follow up of 60 months, ILC patients tended to have longer overall survival (55% vs 48%) and recurrence free survival (44% vs 36,8%) than IDC.

Conclusions: ILC was characterized by better clinical response rates but lower pathologic response and breast conservating surgery rates .Despite the low pCR rate, patients with ILC tended to have better outcomes than did patients with IDC. Pathological complete response to NCT in ILC did not seem to have prognostic significance. Further data are warranted to help clarify the characteristics genomics and proteomics of ILC which explains this better outcome.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P1-11-14.
Abstract P1-11-17: Neglected and Non-Neglected Locally Advanced Breast Cancer: Effects of Neoadjuvant Chemotherapy and Survival
Authors: El Saghir NS , El Asmar NS , Abboud M , Nachef ZK , Mikdashi HF , Hjeij MA , Abbas J , Abi Saad G.
Published: April 2011

Background: Locally advanced breast cancer is a frequent clinical entity that compromises a biologically heterogeneous group that can also be divided chronologically by the duration between onset of symptoms and presentation for medical care.

Methods: Retrospective review of all patients with locally advanced breast cancer (LABC) treated in a general oncology practice between 1997 and 2007. Patients with LABC at presentation and have available follow-up were included. Patients were staged or restaged according to AJCC 7th edition. Patients were divided between two groups: those who presented within 3 months of onset of symptoms and signs of disease (non-neglected) and those who presented after more than 3 months (neglected). DFS was calculated from time of diagnosis till time of distant recurrence or last follow-up. Overall survival was calculated from time of diagnosis till time of last follow-up or death. Study was approved by IRB of the American University of Beirut. Data was entered and analyzed using SPSS. Survival was calculated and estimated according to Kaplan Meier method.

Results: Total number of patients with LABC was 109 pts. Median age at diagnosis was 48 yrs (Range: 21-78 y). Hormone receptors were positive in 71% of cases. Number of neglected cases at presentation were 61 patients and non-neglected 48. Pre-operative therapy, adjuvant chemotherapy, hormonal therapy and radiation therapy were conformal with standard current guidelines. Neoadjuvant chemotherapy was given in 24% (27/109) of all cases. Adjuvant trastuzumab was used starting June 2005. Patients were followed closely every three months and compliance with therapy was excellent.

Overall Survival (OS) of all patients was 68% at 5 years and 34% at 10 years. Median Survival was 100 months for non-neglected and 62 months for neglected disease (P: 0.06)

Survival data

Conclusions: Locally advanced disease confers a bad prognosis and poor outcome in patients with breast cancer. Patients who present within 3 months of onset of symptoms have a better survival than those who present with neglected disease. Neglected disease, even though not necessarily biologically aggressive, continues to have an adverse effect on survival. Survival continues to decline between 5 and 10 years. Additional patient demographics and tumor characteristics as well as shorter delays at presentation will be presented.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P1-11-17.
Abstract P1-11-18: Neoadjuvant Use of 5-Fluorouracil/Epirubicin/Cyclophosphamide (FEC) Followed by Docetaxel (T) with Concurrent COX-2 Inhibitor in Women with Stage II/III Breast Cancer: A Sub-Analysis of Cardiac Function
Authors: Chow LWC , Yip AYS , Chan RHW , Lam Y-M , Toi M.
Published: April 2011

BACKGROUND: Anthracycline and taxane have been widely used and studied in neoadjuvant setting for treatment of locally advanced breast cancer. Although addition of COX-2 inhibitors might enhance the anticancer effect of chemotherapy, many studies were halted with the concern of possible cardiotoxicity. This sub-study is to evaluate the change in cardiac function of patients during treatment with pre-operative FEC-T with concurrent celecoxib until and after surgery.

METHODS: A group of 47 patients, age ranged from 33 to 58 years (mean age 46±6 years), who received four cycles of 3-weekly FEC followed by four cycles of 3-weekly T with concurrent oral celecoxib before surgery (F: 500mg/m2; E: 100mg/m2; C: 500mg/m2; T: 100mg/m2; Celecoxib: 200mg bid), were studied. Echocardiography was performed at baseline, after 4 cycles of FEC and after 4 cycles of T before surgery, and 6 months, 12 months and 18 months after surgery to evaluate changes in left ventricular ejection fraction (LVEF) across time-points. Paired sample statistics was conducted.

RESULTS: Patients tolerated well with this regimen. Neither life-threatening toxicity nor clinical symptom of cardiac toxicity was observed. Of 47 evaluable cases, LVEF was assessed in 47 patients pre-operatively and in 32 patients post-operatively. During concurrent celecoxib treatment, the mean LVEF increased from 68.5%±6.6% at baseline to 72.3%±3.6% after FEC and to 73.0%±4.1% after T. When compared to baseline, the changes were statistically significant after FEC (p=0.002) and after T (P<0.0001) in 47 patients. LVEF however did not change at 6 months (72.8%±3.8%), 12 months (72.7%±3.1%) and 18 months (72.4%±3.3%) after surgery in 32 patients. The remaining 15 patients are still under follow up for cardiotoxicity until 1 year after surgery.

CONLCUSIONS: Neoadjuvant FEC-T with concurrent celecoxib is well-tolerated. The short-term treatment increased LVEF during the course of neoadjuvant chemotherapy and remains good LVEF after surgery. This is an interesting finding which warrants further investigation of the possible protective effect on cardiac function.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P1-11-18.
Abstract P1-11-23: Cannabinoid Receptor 2 Compounds in the Attenuation of Breast Cancer Cell Proliferation: Mechanisms of Action
Authors: Hanlon KE , Lozano-Ondoua AN , Chandramouli A , Largent-Milnes TM , Umaretiya PJ , Stapleton DJ , Nelson MA , Mantyh PW , Vanderah TW.
Published: April 2011

Cannabinoids have been well established as mediators of tumor cell proliferation in several cancer models. The activity of cannabinoid receptor 1 (CB1) agonists as well as cannabinoid receptor 2 (CB2) agonists have been extensively studied over the last decade, though their mechanisms of action have yet to be defined in the context of attenuating tumor proliferation. CB1 is abundant in the central nervous system with a low level of expression in the periphery, while CB2 exists primarily on cells of the immune system. Due to the lack of neuronal expression of CB2 receptors, compounds acting at CB2 receptors do not produce the psychotropic effects associated with CB1 receptor compounds. Although CB1 and CB2 compounds have yielded similar antiproliferative results in some tumor cells in vitro, CB2 receptors are markedly upregulated in many tumor cell lines for unknown reasons. Therefore, we focused on compounds selective for the CB2 receptor, including the CB2 selective agonists JWH-015 and AM1241. We show here that CB2 agonists and antagonists alike attenuate the proliferation of mouse and human breast cancer cells in a concentration dependent manner. CB2 agonist induced breast cancer anti-proliferation is not blocked by pretreatment with pertussis toxin, the CB1 cannabinoid receptor antagonist SR141716, or the transient receptor potential cation channel subfamily V member 1 (TRPV1) antagonist capsazepine. The CB1 and CB2 receptors are classified as G-protein coupled receptors (GPCRs) that are generally linked to Gai or Gaq. Agonists and antagonists of G protein coupled receptors are typically defined based on their Ga mediated effect on intracellular cAMP levels. This method of classification is not entirely effective: it ignores the possibility of differential effects on alternative kinases and the bg subunit activity. The anti-proliferative activity of both JWH-015 and SR144528 may be explained by alternative receptor coupling pathways, changes in constitutive activity, or activity at a separate receptor. Here, we show a dose and time dependent decrease in phosphorylated ERK in cells treated with either JWH-015 (CB2 agonist) or SR144528 (CB2 antagonist). Together, these data along with the absence of a pertussis toxin effect suggest that the CB2 compounds are acting in a non-Gai coupled manner, and are attenuating a pro-survival pathway. Further studies are necessary to identify the binding partner responsible for the antiproliferative effects of CB2 compounds. We advance the idea that CB2 receptors on breast cancer cells constitutively activate the MAP/ERK pro-survival pathway and that by preventing constituitive activity of the CB2 receptor, CB2 compounds downregulate phosphorylation of the MAPK/ERK pathway to promote apoptosis of breast cancer cells.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P1-11-23.
Abstract P1-11-24: Concurrent and Sequential Bevacizumab and Preoperative Chemotherapy in the Treatment of Locally Advanced Breast Cancer: UAB 0493
Authors: Carpenter JT , Forero A , Falkson CI , Nabell LM , De Los Santos JF , Krontiras H , Li Y.
Published: April 2011

From 3/08 through 12/09, 32 patients with HER-2 negative locally advanced breast cancer (LABC) received preoperative chemotherapy (CT) with sequential dose-dense chemotherapy consisting of pegylated liposomal doxorubicin 25 mg/M2 every 2 weeks X 3, paclitaxel 175 mg/M2 every 2 weeks X 3, then cyclophosphamide 600 mg/M2 every 2 weeks X 3, all with concurrent bevacizumab 10 mg/kg every 2 weeks. One patient had progression of disease during treatment and a second developed bone metastasis by the completion of CT. 30/32 had at least 30% shrinkage of tumor clinically. No patient developed cardiac failure. All 31 patients who completed CT had > 55% Left ventricular ejection fraction on echocardiogram before and after CT. The only toxicities greater than grade 2 were skin—grade 3 palmar plantar erythrodysesthesia in 1 and grade 3 hypertension with BP >150/90 in 7 patients. 12 doses of bevacizumab were held due to a blood pressure > 150/90. No proteinuria was seen. 5 patients experienced delayed wound healing after operation. 8/32 pathologic complete responses (pCR) were seen, 2/21 in ER and/or PR positive disease and 6/11 in ER and PR negative disease. 4 additional patients had no invasive tumor remaining in the breast but had ≥1 involved axillary node. Hormonal therapy but no further CT was given postoperatively. 20 patients who had residual invasive tumor and/or involved axillary nodes at operation will receive an additional year of Avastin 15 mg/kg every 3 weeks. 1 patient received only 2 doses, 13 have completed or are receiving bevacizumab, 6 patients have not yet started. None of these 20 has relapsed, with a median followup of 15 months (range 6 to 27 months). 1 who did not receive bevacizumab due to delayed wound healing had progression 6 months after CT. Concurrent bevacizumab with preoperative CT may not influence the rate of pCR; postoperative bevacizumab may decrease recurrence after operation in patients with LABC.

Supported through grants from Centocor Ortho Biotech Services & from Genentech

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P1-11-24.
Abstract P1-12-01: Preoperative Endocrine Therapy with Tamoxifen and Goserelin Acetate for Hormone Receptor-Positive Premenopausal Patients
Authors: Shimizu D , Ishikawa T , Yamada A , Tanaabe M , Chishima T , Sasaki T , Endo I.
Published: April 2011

Background: Clinical trials have consistently shown that the response rates of hormone receptor-positive breast cancer patients to neoadjuvant chemotherapy are low. Preoperative endocrine therapy has therefore become a logical alternative for such patients. Several researchers have reported favorable response rates in postmenopausal women. However, the effectiveness of preoperative endocrine therapy for premenopausal women remains unknown. The aim of this study was to evaluate the potential benefits of preoperative endocrine therapy for premenopausal women.

Materials and methods: Forty premenopausal patients with operable estrogen receptor (ER)-positive breast cancer were consecutively treated with neoadjuvant therapy with tamoxifen and Goserelin acetate for 3 months. The patients eligible for this study included clinically node-negative patients with tumors that were less than 2 cm in diameter and negative for human epidermal growth factor receptor 2 (HER2). All the patients were clinically evaluated using ultrasonograpy before and after the endocrine therapy. Further, core-needle biopsy specimens obtained before administration of the endocrine therapy and surgical specimens were pathologically evaluated for ER, progesterone receptor (PgR), HER2, p53, and Ki67 expressions. This study was approved by our institutional IRB.

Results: The patients included in this study were 30-52 years old (median age, 43 years). The size of the tumor was 0.9-2.0 cm (median, 1.5 cm).

The clinical response rate was 20% (8/40) and disease progression was not observed. In the case of 39 of the 40 patients, grade 0 or grade 1 pathological response was observed. Clinical responses were not correlated with pathological responses. The ER expression level before the endocrine therapy differed from that after the therapy in the case of 8% (2/40) of the patients (increased in 4% and decreased in 4%). Further, PgR expression was altered in the case of 50% (20/40) of the patients (increased in 8% and decreased in 42%). Moreover, Ki67 expression was decreased in 90% (36/40) of the patients after the endocrine therapy.

Conclusions: This is the first report on neoadjuvant endocrine therapy for premenopausal patients. Disease progression 3 months after the endocrine therapy was not observed. Because the expressions of Ki67 and PgR decreased, these molecules might be candidate markers of endocrine responsiveness in premenopausal patients. To confirm this assumption, further evaluations involving tumors of a larger size, longer observational period, and more number of eligible patients are planned to be conducted.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P1-12-01.