bluuebunny/crossref_metadata_2025 huggingface dataset:
P1-01-13: Prognostic Impact of CD8 in Node-Negative Breast Cancer.
Authors: Schmidt M , Chen Z , Hellwig B , Böhm D , Lebrecht A , Gebhard S , Gehrmann M , Koelbl H , Hengstler JG
Published: June 2012

Background: Infiltration of cytotoxic T lymphocytes (CTL) is a common feature in breast cancer. We examined the prognostic impact of CD8 positive CTL in formalin-fixed-paraffin-embedded (FFPE) tissue using immunohistochemistry (IHC) in node-negative breast cancer and validated our findings in previously published cohorts using RNA expression.

Methods: CD8 was evaluated in FFPE tissue of 339 medically untreated node-negative breast cancer patients utilizing IHC. Results were validated utilizing microarray based gene-expression data of four cohorts of medically untreated node-negative breast cancer patients (n=824). Impact of CD8 on metastasis-free survival (MFS) was analyzed with univariate and multivariate Cox regression. Meta-analysis of previously published cohorts was performed using a random effects model. Prognostic significance was examined in the whole cohort and in different molecular subtypes (ER+/HER2−, ER−/HER2−, HER2). Correlation between RNA expression and IHC was analyzed according to Spearman.

Results: Immunohistochemical detection of CD8+ CTL was associated with MFS in univariate (hazard ratio [HR] 0.76, 95% confidence interval [95% CI] 0.59−0.98, P=0.037) as well as in multivatiate analysis (HR 0.61, 95% CI 0.46−0.81, P=0.001). Protein levels of CD8 had a good correlation with RNA expression (r=0.488). Higher RNA expression of CD8 was related to better MFS in a meta-analysis of the whole cohort (HR 0.74, 95% CI 0.60−0.90, P=0.0023). However, prognostic significance was confined to ER+/HER2− (HR 0.71, 95% CI 0.51−0.99, P=0.0201) and HER2+ (HR 0.60, 95% CI 0.38−0.97, P=0.037) but not to ER−/HER2− molecular subtypes (HR 0.77, 95% CI 0.52−1.14, P=0.1965).

Conclusion: CD8 positive CTL have independent prognostic significance in node-negative breast cancer.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-01-13.
P1-02-04: Estrogen Receptor beta Inhibits Breast Cancer EMT by Regulating the Expression of miR-200.
Authors: Thomas C , Gayani R , Nikolos F , Hao R , Katchy A , McCollum C , Williams C , Bondesson M , Krishnamurthy S , Esteva F , Gustafsson J-A
Published: June 2012

Estrogen receptor beta (ERβ) mediates the effects of estrogens in a variety of human tissues and regulates cellular processes involved in initiation and progression of breast cancer such as cell proliferation and migration. Clinical studies produced contradictory data regarding the role for ERβ in prognosis of metastatic breast cancer and the molecular mechanism through which ERβ influences cell migration and invasion has not been fully elucidated. Here we show that induction of ERβ expression inhibits epithelial to mesenchymal transition (EMT) in metastatic breast cancer cells. This correlates with an ERβ-mediated induction in the expression of the epithelial marker E-cadherin and downregulation of its transcriptional repressors ZEB1 and SIP1. ERβ alters the expression of ZEB1 and SIP1 by inducing the expression of the miR-200a, miR200b and miR-429. Downregulation of these miRNAs in ERβ-expressing cells resulted in decreased cell-cell contact and decline of E-cadherin levels. In addition, ERβ was found to inhibit the invasiveness of metastatic breast cancer cells in a zebrafish xenotransplantation model. We are now examining breast cancer specimens derived from ductal carcinomas and metaplastic breast cancers to see whether ERβ levels decline in the mesenchymal regions and ERβ expression is correlated with epithelial markers. These data propose a crucial role for ERβ in the regulation of EMT and in prognosis of invasive and metastatic breast cancer.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-02-04.
P1-02-11: The BCL2 Antagonist of Death, BAD Is Down-Regulated in Breast Cancer and Inhibits Cancer Cell Invasion.
Authors: Cekanova M , Fernando R , Wimalasena J
Published: June 2012

Background: Recent clinical evidence suggest that expression of BCL2 and its antagonist BAD are good prognosticators for survival in breast cancer patients. BAD protein was previously shown by us to inhibit cyclin D1 expression and cJun activation, both may enhance invasiveness of tumor cells(Fernando et al JBC 07). However, the role of BAD and other BCL2 family proteins in invasion/metastasis of breast cancer is poorly characterized.

Methods: By immunohistochemistry nuclear and cytoplasmic staining of several proteins including BAD in normal epithelial was compared to that of cancer cells in human specimens. Western blotting and ELISA methods were used to compare BAD overexpressing MCF7 breast cancer cells with control cells for the expression of variety of signaling molecules, proteins that take part in metastasis and invasion and ability of cells to invade was measured. PCR was used to measure m RNA levels and reporter constructs utilized for transcriptional factor activity studies.

Results: Grade II breast cancer specimens express less total and phosphorylated forms of BAD in nuclei and cytoplasm compared to normals. BAD expression decreased Sp1, β-catenin and STAT proteins, which may increase cyclin D1 in vitro. BAD inhibited activation of the CRE and AP1 elements, and phosphorylation of BAD on S112 and S136 is required for this activity. BAD inhibited the Ras/MEK/ERK pathway and JNK, which may underlie inactivation of c Jun. BAD, like BCL2, may decrease metastasis,therefore, ability of BAD to modulate the expression of metastasis related proteins were measured and MMP10, VEGF, SNAIL, CXCR4, E-cadherin, and ***TlMP2 were regulated by BAD with a concomitant reduction in cell invasion.

Conclusion: Higher expression of BAD in breast cancer and a role in metastasis and proliferation suggests that BAD is valuable prognostic marker in breast cancer and a multi functional protein. Further given the overwhelming clinical evidence that BCL2 prolongs survival, a reevaluation of the role of BCL2 family proteins in metastasis is urgently required.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-02-11.
P1-04-02: In Vitro Qualification and Quantification of Murine Mammary Stem/Progenitor Cells.
Authors: Dong Q , Wang D , Bandyopadhyay A , Liu Z , Yu L , Gao H , Moncada K , Huang C , Walter CA , Sun L-Z
Published: June 2012

Increasing evidence suggest that tumors with a cell origin of more basal or stem-like sources are often highly metastatic and associated with poor prognosis. Identification of the cell of origin thus has important implications for development of new preventive and therapeutic approaches. To identifying the cells of origin of various breast cancers, understanding the normal cellular hierarchy within the breast tissue is an important prerequisite. However, current understanding of normal mammary stem/progenitor cell is limited due to the lack of a robust in vitro assay. The newly developed cell surface markers (CD24+CD49fhi) can only be used for low percent of enrichment. The only gold standard assay for functional evaluation of stem cell property is the in vivo transplantation of cleared mammary fat pad assay, yet this assay is expensive, time consuming, and requires highly trained skills, and thus cannot be used routinely for experiments. The goal of our study is thus to develop a robust in vitro assay to qualify and quantify murine mammary stem/progenitor cells. Our rationale lies on our recent findings of in vitro study of different fractions of murine stem/progenitor cells. When we use isolated primary epithelial cells for in vitro mammosphere formation, we found that stem cell enriched fraction and progenitor cell enriched fraction both formed very small mammospheres (≤ 50 μm) and these spheres appear to be of clonal origin (one cell gives rise to one sphere). When we plated these small spheres into 3D extracellular matrix for colony formation, however, they gave rise to two distinct structures: stem-enriched fraction generated predominately solid structure while progenitor-enriched fraction is dominated by hollow structures. Previous studies have linked the solid structure to the basal/myoepithelial lineage and the hollow structure to the luminal lineage. We thus suspected that the small mammospheres derived from the stem or progenitor enriched fractions could be originated from single stem or progenitor cell, and the number of spheres could indicate the number of original stem/progenitor cells within these enriched fractions. Subsequent in vivo transplantation with single sphere or single solid structure cultured in 3D extracellular matrix derived from green fluorescent protein transgenic mice proved that single sphere or 3D solid structure can repopulate the gland-free mammary fat pad. To conclude, the in vitro mammosphere formation in combination with subsequent differentiation in 3D extracellular matrix can be used as a robust in vitro assay for qualification and quantification of murine mammary stem/progenitor cells. This in vitro assay will greatly facilitate our understanding of genes regulate stem cell self-renewal, proliferation, differentiation as well as mechanisms keeping them at quiescent state within the niche.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-04-02.
P1-04-07: Poly (ADP-Ribose) Polymerase-1 (PARP-1) Is Overexpressed in Human Breast Cancer Stem Cells: Results from a Proteomic-Based Approach.
Authors: Gilabert M , Ginestier C , Audebert S , Pophillat M , Toiron Y , Birnbaum D , Borg J-P , Charafe-Jauffret E , Gonçalves A
Published: June 2012

Introduction: Cancer stem cells (CSC) have been increasingly recognized as playing a major role in various fields of breast tumor biology including carcinogeneis, metastasis and resistance to cytotoxic drugs and radiotherapy. Identification of protein biomarkers associated with breast CSC may help understanding CSC biology as well as identifying novel diagnostics and specific therapeutic targets. 2-D Fluorescence Difference Gel Electrophoresis (2-D DIGE) is a method that labels protein samples with fluorescent dyes before 2-D electrophoresis, enabling accurate analysis of differences in protein abundance between samples.

Methods: Using flow cytometry-based ALDEFLUOR assay (ALD), we isolated CSC-enriched (ALD-positive) and non CSC-enriched (ALD-negative) cell populations of MDA-MB-453, a human breast cancer cell line. Total proteins were extracted from both fractions using urea-based buffer and subjected to 2D-DIGE. Differentially expressed spots were excised, proteins were gel-extracted, digested and identified using MALDI-TOF MS (Ultraflex, Brucker Daltonics, Billerica, USA)

Results: 2D-DIGE revealed differential expression of various protein spots between ALD-positive and ALD-negative MDA-MB-453 cells. MALDI-TOF MS analysis allowed identification of 11 differentially expressed proteins, among which 7 were down-regulated and 4 were up-regulated in ALD-positive MDA-MB-453 cells, including Poly (ADP-ribose) polymerase-1 (PARP-1). Overexpression of PARP-1 in MDA-MB-453 cells was further confirmed by western blot using specific monoclonal antibody and such an observation was extended to 4 additional human breast cancer cell lines including HCC1937, MDA-MB-436, SUM149 and SUM159. These 5 human breast cancer cells were found to display a limited short-term sensitivity (within the micromolar range) to olaparib, a specific PARP-1 inhibitor. However, a negative correlation was found between the level of overexpression and the ability of olaparib to inhibit the growth of ALD-positive cells. Conclusion: A proteomic-based approach revealed that PARP-1 was up-regulated in ALD-positive, CSC-enriched from various human breast cancer cell lines. Such an overexpression may contribute to clinical resistance to PARP-inhibitors.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-04-07.
P1-04-09: Biphasic Effects of Docetaxel and Hedgehog Signaling Antagonists on Breast Cancer Tumor-Initiating Cells In Vivo.
Authors: Zhang X , Moraes RC , Landis MM , Wu M-F , Hilsenbeck SG , Cairo MM , Toftgar R , Chang JC , Lewis MT
Published: June 2012

Recent data suggest the existence of a subset of breast cancer cells variously termed cancer stem cells, tumor-initiating, or tumor-propagating cells, that are capable of self-renewal and of regenerating tumors upon transplantation that are biologically consistent with the tumor of origin. These cells appear to be intrinsically resistant to systemic chemo- and radiation therapies and may therefore be responsible for treatment resistance, and disease recurrence. Several signal transduction networks have been implicated in normal and malignant stem cell self-renewal. These include those of the Hedgehog, Notch, Wnt, EGF, FGF families of ligands. Antagonists of these networks have become attractive targeted therapeutic agents. In this study, we evaluated two different Hedgehog signaling antagonists, one targeting Smoothened (the main membrane effector of activated signaling) and one targeting GLI1/2 (the two transcription factors mediating ligand response), for their ability to shrink patient-derived xenograft tumors and to affect the number or function of breast tumor-initiating cells in vivo. Both antagonists showed little effect as single agents, but could augment tumor shrinkage by docetaxel. However, these effects were only observed in those models showing evidence of canonical SMO/GLI-mediated signaling. In dilutional transplantation assays, both SMO or GLI targeted agents have a biphasic effect on the behavior of the tumor-initiating population such that large cell numbers yielded few tumors while intermediate cell numbers yielded tumors at a comparable rate as controls. These data suggest that combination therapy alters the functional state of stem cells in the context of larger cell numbers rather than targeting the tumor-initiating cell directly. These data therefore have significant relevance for the interpretation of ongoing clinical trials of hedgehog signaling inhibitors in solid tumors, including breast cancer.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-04-09.
P1-07-10: Validation of a Diagnostic Molecular Signature (EHT Dx14) on Fine-Needle Aspirate Samples from Breast Tumors.
Authors: Delaloge S , André F , Fehlbaum P , Sol O , Vielh P
Published: June 2012

Background: A 1228-probeset molecular classifier (EHT Dx14) able to discriminate benign breast lesions from breast adenocarcinoma with a 96% performance was previously identified in a panel of 71 fine-needle aspirate (FNA) samples of nodular breast lesions collected at Institut Gustave Roussy (IGR). The microarray which enabled the signature identification was based on the genomewide RNA analysis using the ExonHit's SpliceArray™ technology. The objective of the current study was to validate the performance of the EHT Dx14 molecular classifier on an independent cohort of FNA samples stored at the Centre de Ressources Biologiques at IGR.

Patients and methods: The samples were collected from women undergoing investigations for suspicious nodular breast lesions classified BIRAD ACR 4 or 5 on mammogram and/or ultrasonography. The morphological cytological analysis of FNA samples classified the lesions as either benign, malignant or indeterminate. All samples were associated with a definite clinico-pathological diagnosis of the breast lesions. The first part of the study aims at confirming the sensitivity, specificity and overall performance of EHT Dx14 among lesions clearly classified by morphological cytology. The second part aims at establishing its performance among lesions with indeterminate cytological results.

Results: In the first part of the study, the EHT Dx14 transcriptomic signature was applied on a set of 94 FNA consecutive samples categorized as benign (n=47) or adenocarcinoma (n=47) on cytology. The benign/malignant status of the lesions was confirmed by subsequent investigations or follow-up. Results of EHT DX-14 assessment of the samples were respectively 97.9% (95%CI: [88.7%-99.9%]) for sensitivity and 91.5% (95%CI: [79.6%-97.6%]) for specificity. Results on indeterminate FNA samples are being investigated in the second part of the study.

Conclusion: This study did confirm the high performance of EHT Dx14 signature in identifying malignant from benign breast lesions in clinically suspect lesions. EHT Dx14 may provide adequate biological diagnosis in the absence of capability of morphological evaluation.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-07-10.
P1-07-13: Efficiency of a Laboratory Developed HER2 FISH Test on Circulating Tumor Cells.
Authors: Mayer JA , Pham T , Wong KL , Scoggin J , Sales EV , Clarin T , Pircher TJ , Mikolajczyk SD , Cotter PD , Bischoff FZ
Published: June 2012

INTRODUCTION: Most circulating tumor cell (CTC) platforms rely on EpCAM for capture and cytokeratin (CK) for detection. However, an important population of cells that are CK-negative (i.e. cells with epithelial-mesenchymal transition (EMT) phenotype) will be missed. We report a new strategy to efficiently isolate a more heterogeneous population of CTCs using an antibody cocktail.

METHODS: In the first prospective study, blood (20 mL) was collected from 89 patients diagnosed with various late stage metastatic/recurrent cancers (breast, CRC, lung, prostate) following IRB approval. PBMCs were incubated with either EpCAM alone or a mixture of 10 capture antibodies to target both epithelial and mesenchymal cells. CTCs were subsequently captured in the OncoCEE™ channels and detected with cytokeratin (CK) and CD45. A second prospective IRB approved study involving 54 patients diagnosed with late stage metastatic/recurrent breast cancer was performed using similar detection strategies (CK cocktail mixture and anti-CD45) with the addition of HER2 FISH to determine amplification status among captured CK+/CD45- and CK-/CD45-cells.

RESULTS: In the first study, overall detection of CK+ cells was 83% with EpCAM alone and 93% with antibody cocktail. In addition, a median of 0.4 CK+ cells/mL and 1.0 CK+ cells/mL was observed using EpCAM and antibody cocktail, respectively. In the second study, CK+/CD45- cells were detected in 43 of 54 cases (80%). Among the 43 cases in which CK+/CD45- cells were detected, high concordance (93%) in HER2 status between primary tumor and CTCs was observed with HER2 amplification noted in both CK+/CD45- (50%) and CK-/CD45- (50%) cells.

CONCLUSIONS: We have developed a novel and robust method for CTC enumeration that utilizes a cocktail of antibodies for the detection of a heterogeneous (CK+ and CK-) population of CTCs. Our findings suggest an important population of CK- cells is being missed by current stain criteria in breast cancer patients. Data also demonstrate that recovery of CTCs from peripheral blood using the OncoCEE™ platform is efficient and suitable for FISH-based laboratory testing.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-07-13.
P1-08-14: Arsenic Methylation Is Associated with Breast Cancer Risk in Northern Mexico.
Authors: López-Carrillo L , Hernández-Ramírez U , Torres-Sánchez L , Gandolfi JA , Ornelas-Aguirre JM , Cebrián ME
Published: June 2012

Background: Arsenic (iAs) exposure has been associated with several types of cancer. However, most studies to date have not yet implicated iAs as a cofactor for breast cancer (BC). BC incidence and mortality are 3-fold higher in Northern Mexico (NM) than in the rest of the country. iAs levels in drinking water are a source of concern since they have been above international standards. We hypothesized that iAs exposure and individual Arsenic-methylation ability are potential cofactors for BC risk.

Material and Methods: We investigated 840 BC incident cases and 973 controls from a population-based case-control study performed in NM. Women were directly interviewed about dietary and reproductive BC covariables. iAs metabolites in urine were determined by HPLC/ICP-MS. Methylation capacity was assessed by calculating the percentage of iAs species and primary [MMA(V)/(As(III)+As(V)] (PMI) and secondary [DMA(V)/MMA(V)] methylation indexes (SMI).

Results: The range of total urinary Arsenic was 0.4−303.9 ug/L. Most women (91%) had values above 50 ugAs/L.%MMA(V) was significantly higher in BC cases while%DMA(V) was significantly lower. PMI was positively associated with BC (OR T3vsT1=1.47; CI95=1.17−1.84; p for trend=0.001), while SMI was negatively associated (OR T3vsT1=0.54; CI95=0.42−0.68; p for trend<0.001). Discussion: As exposure may pose a risk for BC, particularly in women with higher capacity to methylate iAs to MMA(V) and lower capacity to further methylate MMA(V) to DMA(V). This is consistent with studies indicating that the proportion of MMA(V) in urine was positively associated whereas that of DMA(V) was inversely associated with the risk of skin, bladder and lung cancer. Further research is needed about the temporal relationships between disease and methylation capacity and the presence of potential association modifiers, including environmental and genetic factors involved in one carbon metabolism.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-08-14.
P1-08-18: Aspirin Exposure and Nodal Status at Diagnosis in Women with Stage I-III Breast Cancer.
Authors: Barron TI , Sharp L , Bennett K , Visvanathan K
Published: June 2012

Background: The recent Nurses’ Health Study reported an association between aspirin use and a reduction in breast cancer (BC) recurrence and BC-specific mortality. It has been suggested based on preclinical data that these effects may be due to the inhibition of tumor invasion and metastasis by aspirin. To test this hypothesis we examined the association between aspirin exposure prior to BC diagnosis and nodal involvement at diagnosis.

Methods: A retrospective analysis was conducted using linked pharmacy claims and cancer registry data from the National Cancer Registry Ireland. Women with an incident diagnosis of stage I-III BC between 2001–2006 and no prior invasive cancer were included. Women with a prescription for aspirin at any time in the year prior to diagnosis were defined as exposed. Multivariate logistic regression was used to estimate odds ratios (OR) and 95% confidence intervals (CI) for (a) N0 disease; (b) N1 disease (c) N1/2/3 disease at diagnosis in aspirin exposed (overall exposure & exposure quartiles by duration of use) versus unexposed women. All analyses were adjusted for age, tumor size/grade, ER, PR and HER2 receptor status. Analyses were repeated including women with stage IV BC at diagnosis. Survival analyses are ongoing.

Results: 4212 women with stage I-III BC at diagnosis were identified. 1086 (25.8%) had a supply of aspirin available to them in the year prior to diagnosis (median dose 75mg, interquartile range 75mg, 75mg). Women taking aspirin were older than those who did not (74.3 yrs/64.4 yrs p<0.05). There was no difference in tumor sizes, grade, ER, PR or HER2 receptor status between exposed and unexposed women. In the multivariate logistic regression analysis aspirin exposure was associated with significantly increased odds of an N0 tumor (Table 1; OR 1.27 95%CI 1.09, 1.48) and significantly decreased odds of an N1 or N1/2/3 tumor. Additionally, women taking aspirin for >95% of the year prior to diagnosis had the highest odds of an N0 tumor (Table1; OR 1.50 95%CI 1.14, 1.97). There was no difference in the odds of an N0 tumor for women taking aspirin for <40% of the prior year (OR 1.01 95%CI 0.78, 1.31). Inclusion of stage IV BC (n=789) in the analysis did not significantly alter these results.

Discussion: This study demonstrates a significant association between aspirin use in the year prior to BC diagnosis and the degree of nodal involvement at diagnosis. Furthermore there was a clear dose-response relationship between increasing duration of exposure and nodal status. These associations are unlikely to be due to confounding by healthy user/adherer behaviors as tumor sizes did not vary significantly between unexposed and exposed women, or across quartiles of exposure duration.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-08-18.
P1-09-01: Effect of Obesity on Gene Expression in Invasive Breast Tumors.
Authors: Ellsworth RE , Croft DT , Ellsworth DL , Shriver CD
Published: June 2012

Background: Obesity is a risk factor for breast cancer in postmenopausal women and weight gain after diagnosis is associated with decreased survival and less favorable clinical characteristics such as greater tumor burden, higher grade, and poor prognosis, regardless of menopausal status. Despite the negative impact of obesity on clinical outcome, molecular mechanisms influencing breast cancer prognosis remain elusive.

Methods: Postmenopausal women with invasive breast cancer (n=20) in the Clinical Breast Care Project who were obese at diagnosis (BMI ≥30) were matched by age, stage, ER status, HER2 status, and tumor grade to women who maintained a healthy weight (BMI = 18.5−24.9). Sections of pure invasive carcinoma were obtained by laser microdissection from flash-frozen or OCT embedded sections. Gene expression data using U133 2.0 microarrays was generated to compare genome-wide patterns of expression in breast tumors between obese and normal-weight patients. Data were analyzed using Partek Genomics Suite.

Results: Principal Component Analysis accurately clustered the specimens into two groups, healthy weight or obese. Using a false-discovery rate (FDR) p-value <0.05 with a minimum 2.0-fold change of expression, 119 probes from 106 genes were differentially expressed, with 35 genes expressed at significantly higher levels and 71 at significantly lower levels in tumors from obese women compared to those from healthy-weight patients. Hierarchical clustering using these 119 probes reliably partitioned all samples as healthy-weight or obese. Genes differentially expressed include lower expression of several members of the histone cluster 1 family, APOD and IGF1, and higher expression of FABP7, members of the major histocompatibility complex and LTF in tumors from obese women.

Conclusions: Invasive breast tumors from obese women are genetically different from those of healthy-weight women. Altered gene expression may affect tumor cell proliferation and survival, contributing to aggressive phenotypes and altered immune response. These underlying molecular differences may contribute to the less favorable prognosis observed in obese women with breast cancer.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-09-01.
P1-10-01: Utility of Routine Cardiac Ejection Fraction (CEF) Measurement Prior to Anthracycline-Based Chemotherapy (ABC): A Study of 466 Patients with Early-Stage Her2-Negative Invasive Breast Cancer.
Authors: Policepatil SM , Go RS , De Maiffe BM , Gundrum JD , McHugh VL
Published: June 2012

Background: Despite the lack of evidence to support its utility, routine CEF measurement prior to ABC is a common practice in the community, required in most US clinical trials, endorsed by the American Heart Association and American College of Cardiology, and appears on Food Drug Administration-approved labeling guidelines.

Objectives: We determined the frequency of the following events in newly diagnosed invasive breast cancer patients: a) CEF measurement prior to ABC; b) asymptomatic left ventricular dysfunction; c) modification in initial treatment strategy as a result of CEF measurement; d) development of congestive heart failure (CHF) after ABC.

Methods: From our cancer registry, we obtained a list of all female patients with newly diagnosed stage I-III breast cancer who were treated with chemotherapy as part of initial therapy at our institution in 2000–2010 (N=562). We excluded those with prior CHF, ABC, and those who had Her2-positive disease (n=66). CEF <50% was considered low. CHF risk factors including coronary heart disease, cigarette smoking, hypertension, obesity, and diabetes mellitus were collected.

Results: We included 496 patients with the following stage distribution: I (22%), II (54%), and III (24%). The mean age was 53 years (SD±11; range 25–88). In 30 (7%) patients, ABC was considered inappropriate by the physician provider due to poor performance status, co-morbidities, and physician or patient preference. Of the 466 patients considered for ABC, CEF was measured in 241 (52%) patients before chemotherapy. In the latter group, only 1 (0.4%) patient was found to have asymptomatic left ventricular dysfunction with CEF of 48%. Among patients considered for ABC, 10 (2%) did not receive ABC for the following reasons: low or borderline CEF (2), patient preference (3), co-morbidities (2), and randomization to non-ABC in clinical trials (3). After a mean follow up 57 months (SD±34), only 3 of 456 (0.7%) patients who received ABC developed CHF. Two had normal CEF prior to treatment while the other did not have CEF measured. The practice of CEF measurement prior to ABC varied among physician providers, ranging from 23% to 67% (mean/SD 47%±15%). Those who had CEF measured before chemotherapy were older (55 vs. 50 years; P <0.001) and had more CHF risk factors (1.3 vs. 0.9; P <0.001).

Conclusions: At our institution, routine CEF measurement prior to ABC as initial therapy for early-stage Her2-negative invasive breast cancer patients is a common though not uniform practice. Asymptomatic left ventricular dysfunction is rarely detected and generally mild. Our findings do not support the utility of routine CEF measurement in this population and challenge current practice and guidelines. Potential cost implications to our health care system can be substantial.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-10-01.
P1-12-05: Complete Pathological Response of Ductal Carcinoma In Situ after Treatment with Neoadjuvant Herceptin.
Authors: Chalmers CR , Mallon EA , Touqan N , Horgan KJ , MacPherson I , Doughty JC
Published: June 2012

Introduction: The introduction of neo-adjuvant herceptin has led to pathological complete response rates of up to 60% in patients with breast cancer. The traditional (NSABP) definition of pathological complete response includes cases demonstrating residual DCIS and little is known about the effects of neo-adjuvant herceptin in this situation. Current evidence suggests that DCIS is chemo-resistant, but it has also been shown that DCIS is more likely to be HER-2 positive than primary cancer.

Results: We describe six patients who received neo-adjuvant chemotherapy and herceptin for biopsy-proven, node-positive, large breast cancers either with an extensive component of concomitant DCIS or a separate DCIS focus. Neo-adjuvant treatment did not result in any changes in the pattern of mammographic calcification seen pre-operatively (Figure 1 a-c), however histopathology of all six specimens showed ductal spaces containing macrophages and calcification but no residual lining epithelium, features consistent with pathological complete response of the DCIS component (Figure 1d). Figure 1: Pathologically complete response of DCIS after neo-adjuvant treatment with herceptin. (a) Pre-treatment mammogram (b) Post neo-adjuvant chemotherapy and herceptin mammogram showing resolution of the mass effect of the primary cancer and unchanged residual mammographic calcification associated with the extensive DCIS component (c) Wire-localised specimen demonstrating excision of calcification (d) Histopathology of the same specimen showing calcium-filled ducts and no residual lining epithelium. Discussion: In conclusion, neo-adjuvant herceptin is associated with complete pathological response of both breast cancer and DCIS. The presence of unchanged persistent mammographic calcification in known regions of DCIS after treatment complicates any role of mammographic surveillance in this group and raises the question of optimal surgical management in the future management of these individuals.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-12-05.
P1-12-20: The Safety and Tolerability of Vorinostat in Combination with Lapatinib in Advanced Solid Tumors.
Authors: Chumsri S , Tait NS , Medeiros MM , Bauer KS , Betts K-MT , Lewis JC , Bao T , Feigenberg SJ , Kesmodel SB , Stearns V , Edelman MJ , Sausville EA , Tkaczuk KHR
Published: June 2012

Background: Lapatinib has been previously shown to markedly decrease cancer stem cells (CSC) in HER2−positive breast cancer. In preclinical models, we have demonstrated that histone deacetylase inhibitors (HDACi) such as vorinostat can induce differentiation and decrease CSC. The combination of vorinostat and lapatinib is synergistic with a combination index of 0.32 (synergism if CI <1). We therefore undertook a pilot study to evaluate the combination of these two drugs in advanced solid tumors.

Method: Patients were eligible if they were: age ≥ 18 years with incurable solid tumors, ECOG PS 0–2, adequate organ function, and no prior exposure to HDACi. The first 3 patients received lapatinib at the dose of 1,250 mg continuous daily and vorinostat 300 mg 4 days on 3 days off. The second dose level with lapatinib 1,250 mg continuous daily and vorinostat 400 mg 4 days on 3 days off were administered in 6 patients. Cycles were repeated every 21 days until disease progression. Echocardiogram and radiologic evaluation were performed every 12 weeks. During the first cycle, pharmacokinetic (PK) evaluation was performed on days 18 and 21.

Results: Nine consented patients (7 with metastatic breast cancer, 1 with non-small cell lung cancer, and 1 with thyroid cancer) have been enrolled with the median age of 52 (range 25–66). Patients received an average of 6 prior treatments (range 2–10). No dose limiting toxicity or drug related death have been observed. Grade 1–2 toxicities including diarrhea, fatigue, muscle cramps and stomatitis were observed. No grade 3 or 4 hepatic, renal or cardiac toxicity were observed (including no QTc prolongation and no significant reduction in the left ventricular ejection fraction). Patients have received the maximum of 7 cycles (median 3 cycles, range 2–7). Response: as of June 2011, 2 patients are still on treatment. Two patients achieved stable disease (triple negative metastatic breast cancer and HER2−positive breast cancer), 6 patients with progressive disease, and 1 patient is too early to evaluate for response. PK analysis will be presented at the time of the meeting.

Conclusions: The combination of vorinostat and lapatinib is tolerable and has some antitumor activity in heavily pretreated advanced solid tumors. A phase II study in HER2−positive metastatic breast cancer is underway with lapatinib 1,250 mg continuous daily and vorinostat 400 mg 4 days on 3 days off.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-12-20.
P1-12-21: Adjuvant Trastuzumab Treatment without Adjuvant Chemotherapy in Early Breast Cancer.
Authors: Dall P , Koch T , Lenzen G , Kuhn T , Hielscher C , Reichert D , Maasberg M , Ehscheidt P , Eustermann H , Fischer G
Published: June 2012

Background: Trastuzumab (T) is approved in most Western countries for the treatment of early, HER2+ breast cancer (BC) parallel or sequential to adjuvant chemotherapy (CT). Nevertheless, as in metastatic disease, the antibody seems to be used without chemotherapy in a selected group of HER2+ patients (pt). The purpose of this analysis is to characterize this subgroup as well as describe safety and efficacy outcome parameters in this German prospective observation trial.

Methods: At present, 2870 patients (pts) have been enrolled and documented in this ongoing non-interventional study from 270 German centers. At data base closure for this analysis, sufficient documentation was available from 2422 eligible pts.

Results: The proportion of pts receiving T without preceding or concomitant CT (noCT) was 180/2422 (7.4%). This subgroup was characterized by higher age (median 58 vs.56 in the CT group, p=0.0026; ≥70 years: 18%/10%), smaller primaries (pT1 49%/43%, p=0.11), more favorable grading (G3 45%/53%, p=0.045), a higher proportion of positive hormone receptor (67%/61%, p=0.096) and less radiotherapy (64%/79%, p<0.0001). The strong association to radiotherapy is probably due to the fact, that the choice of this additional modality reflects the overall risk assessment of the pt.

In contrast, there was no difference in nodal involvement (pN0: 52%/51%; positive nodes: mean 2.5/2.4). In multivariate analysis (logistic regression model, not incorporating radiotherapy), hormone receptor status is not predictive (p=0.41), while age ≥65 (p=0.0011), grade 1/2 (p=0.046) and pT1 (p=0.089) independently remain at least borderline significant. The mean number of T administrations was 18 and the median duration of T therapy was 12 months in the noCT group as well as the CT group. 57% of the noCT pts received adjuvant endocrine therapy. In the CT group, 81% received anthracyclines and 61% taxanes.

Based on a still low number of only 13 observed events in the noCT group, no differences in relapse-free survival could be detected (p=0.38). Pathological cardiac findings in the pt's history was not predictive of therapy selection (6%/7%) and were also similarly distributed at the end of T therapy (7%/8%). Cardiac function disorders of CTC grade 3/4 were reported in 1% of both groups, across all grades slightly less frequent in the noCT group (2.8%/4.0%).

Conclusions: A small, but distinct group of early BC pts without adjuvant chemotherapy receives T treatment for HER2+ disease. These patients are characterized by higher age and favorable primary tumor staging and grading, but not by cardiac comorbidities.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-12-21.
P1-12-26: Global Patterns of Care for HER2/Neu Overexpressing Breast Cancer.
Authors: Chavarri-Guerra Y , Liedke P , Symecko H , Moy B , Higgins M , St. Louis J , Finkelstein D , Goss P
Published: June 2012

Background: Her2/neu overexpression is an independent adverse prognostic factor present in approximately 25% of invasive breast cancers. HER2−overexpressing breast cancer is particularly common in younger patients and therefore poses a significant public health burden. Anti-Her2/neu adjuvant trastuzumab significantly reduces the risk of recurrence as well as improves survival. In view of the serious potential ramifications of not receiving anti-Her2/neu therapy when appropriate, we undertook this international project to determine clinical access to Her2/neu testing and treatment patterns for women with HER2/neu-positive early breast cancer.

Methods: A web-based survey was sent to 386 physicians from 33 countries who participated in the “TEACH” trial, a double blind placebo-controlled phase III study of a novel anti-Her2/neu therapy, lapatinib, in women with primary Her2/neu-positive breast cancer. The survey contained 27 questions addressing physician and patient demographics, access to Her-2/Neu testing in everyday clinical practice, and anti-Her2/neu treatment options in a variety of clinical scenarios.

Results: One hundred and fifty one participants (39%) from 28 countries answered the survey. Ninety eight percent of the participants reported having Her2/neu tumor expression routinely measured for clinical practice in their institutions by immunohistochemistry (83%), FISH (78%) and other methods (17%). Among Asian physicians, 18% did not have routine testing available and sent primary tumors for central testing for TEACH eligibility. Forty eight percent of physicians surveyed reported instances when they had recommended adjuvant Her2-directed therapy to a patient who eventually did not receive it. The proportion of physicians from developing countries that reported patients not receiving therapy was higher than those from developed countries (68% vs. 38%, respectively). The main reason for not receiving trastuzumab was cost in developing countries, while in more developed countries patient refusal and co-morbidities were the main reasons.

Discussion: This survey reflects availability of HER2 breast tumor testing and anti-Her2/neu therapy among physicians from 28 countries worldwide who participated in an anti-Her2/neu therapy clinical trial of a free anti-Her2/neu therapy. These results indicate that a high proportion of women with Her2/neu-overexpressing breast cancer may not receive standard anti-Her2/neu adjuvant therapy especially in developing countries, the barrier to treatment being cost of therapy. We are extending our access to care survey project to a more unselected diverse group of physicians in developing countries.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-12-26.
P1-14-01: Randomized Phase II Trial of Weekly vs. q 2-Weekly vs. q 3-Weekly Nanoparticle Albumin-Bound Paclitaxel with Bevacizumab as First-Line Therapy for Metastatic Breast Cancer.
Authors: Seidman AD , Conlin AK , Bach A , Forero-Torres A , Wright G , Hackney MH , Clawson A , Schofield D , Iglesias J , Hudis CA
Published: June 2012

Background: Nanoparticle albumin-bound paclitaxel (nab-P) 260 mg/m2 is superior to paclitaxel 175 mg/m2 (P) every 3 weeks (Gradishar et al. JCO 2005) in metastatic breast cancer (MBC), and weekly uninterrupted P is more effective than q3wk P in MBC (Seidman et al. JCO 2008). Bevacizumab (bev) nearly doubles response rate and time to progression (TTP) when added to P as 1st line therapy for MBC (Miller et al. NEJM 2007).

Methods: This open-label, phase II study randomized patients (pts) to nab-P at 260 mg/m2 q3wk (arm A) vs. 260 mg/m2 q 2wk with filgrastim (arm B) vs. 130 mg/m2 weekly uninterrupted (arm C), all with bev (15 mg/kg q 3 weeks arm A, 10 mg/kg q 2 weeks arms B and C). Patients were required to have measurable, HER2 negative MBC and no prior chemotherapy for MBC. The primary endpoints were response rate and toxicity.

Results: Of 212 pts randomized, 208 (75 arm A, 54 arm B, 79 arm C) were treated, with balanced demographics and baseline characteristics. The median age was 57 (range 29–85), 82% were postmenopausal and 89% had visceral disease (64% lung, 50% liver). ECOG PS 0:60%, 1:35%, 2:5%. 62% had prior neo-adjuvant or adjuvant chemotherapy for early stage disease: anthracycline: 54%, taxane: 38%. No significant differences in confirmed complete and partial response rates were noted (A: 40%, B: 44%, C: 46%). Median TTP was longer in Arm C (9.0 months) versus both arms B (6.3 months) and A (8.0 months), overall p=0.065. There were no differences in overall survival (Arm A: 21.3 months, Arm B: 19 months, Arm C: 25.3 months). As per protocol-specified stopping rule, arm B was closed early due to an unacceptable safety profile with significantly more grade ≥ 2 fatigue (B:57%, A: 39%, C:39%, p=0.048) and bone pain (B:19%, A:10%, C:4%, p=0.024). Sensory neuropathy was common; grades 2/3/4: Arm A: 29%/32%/1%, Arm B: 15%/50%/2%, Arm C: 27%/43%/1%). Sensory neuropathy was commonly readily reversible with dose delay and reduction. Febrile neutropenia occurred in <2% of pts in all arms. Arm C patients experienced significantly less arthralgia compared with arms A and B, but dose delays were frequent (86% of pts) on this planned uninterrupted weekly schedule. Bevacizumab-related events were consistent with prior phase III trials of taxane/bev; there were no new safety signals.

Conclusions: Significant and similar antitumor activity was observed in all arms. Weekly nab-P with bev (arm C) resulted in longer TTP. Weekly nab-P with bev (arm C) appears to have the highest therapeutic index, however sensory neuropathy is limiting, suggesting that a 3 week on/1 week off schedule could be preferable and should be studied comparatively.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-14-01.
P1-14-04: Prolonged (≥1 Year) Exposure to First-Line Bevacizumab Combined with Paclitaxel in Patients with HER2−Negative Metastatic Breast Cancer Treated in a Routine Oncology Practice Study.
Authors: Kuemmel S , Schneeweiss A , Foerster FG , Geberth M , Tesch H , Klare P , Schumacher C , Hollburg W , Soeling U , Schmidt M
Published: June 2012

Background: First-line bevacizumab (BEV) combined with weekly paclitaxel (PAC) significantly improves progression-free survival (PFS) and response rate (RR) vs PAC alone in HER2−negative metastatic breast cancer (mBC), as shown in E2100. The benefit of BEV combined with other chemotherapy (CT) agents was demonstrated in AVADO and RIBBON-1. BEV was continued for ≥1 year in 21% of patients in the global ATHENA safety study and in 42% of patients in the JO19901 single-arm Japanese study of BEV-PAC. We analyzed data from the subgroup of patients treated for ≥1 year in a German routine oncology practice study to provide insight into the safety and efficacy of prolonged first-line BEV-PAC.

Methods: Patients who had received no prior CT for their mBC received BEV-PAC per the European label. Efficacy and safety were documented for up to 1 year (or until progression, death, or BEV discontinuation if earlier) with additional long-term follow-up. Data from patients treated with BEV for ≥1 year were extracted for this analysis.

Results: By Jan 2011, data were available for 818 patients, of whom 157 (19%) had already received BEV for ≥1 year. Baseline characteristics of this subset relative to the overall population are summarized in the table.

In 79% of those treated for ≥1 year, BEV was continued as a single agent after discontinuation of CT. The overall RR in patients treated for ≥1 year was 81% (complete response in 20%). Median PFS was 17.9 months (events in 44% of patients) vs 9.4 months in the overall population (events in 68%). Overall survival data are immature, as 81% of those treated for ≥1 year are still alive. The most common grade 3/4 adverse events in patients treated with BEV for ≥1 year were hypertension (11% of patients), pain (10%), and leukopenia (7%). There were no cases of gastrointestinal perforation, arterial thromboembolic event, or reversible posterior leukoencephalopathy syndrome in those treated for≥1 year. Analysis of adverse events according to time of onset is ongoing.

Conclusions: A notable proportion of patients appear to derive benefit from prolonged exposure to first-line BEV-containing therapy. In the present analysis, baseline characteristics appeared more favorable in the subset of patients treated for ≥1 year than in the overall population. Efficacy data clearly have a bias towards improved outcome in those able to continue BEV for ≥1 year, as these patients had sustained disease control for ≥1 year. However, the efficacy of prolonged BEV-containing therapy is of interest and suggests that some patients achieve sustained disease control with continued first-line BEV-PAC with limited side effects.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-14-04.