bluuebunny/crossref_metadata_2025 huggingface dataset:
UCSC cancer genomics browser.
Authors: Zhu J , Sanborn JZ , Wang T , Hsu F , Benz S , Szeto C , Esserman L , Haussler D
Published: December 2010

Abstract #2022

As experimental techniques for a comprehensive survey of the cancer landscape mature, there is a great demand in the cancer research field to develop advanced analysis and visualization tools for the characterization and integrative analysis of the large, complex genomic datasets arising from different technology platforms.  The UCSC Cancer Genomics Browser is a suite of web-based tools designed to integrate, visualize and analyze genomic and clinical data. The secured-access browser, available at https://cancer.cse.ucsc.edu/, consists of three major components: hgHeatmap, hgFeatureSorter, and hgPathSorter. The main panel, hgHeatmap, displays a whole-genome-oriented view of genome-wide experimental measurements for individual and sets of samples/patients alongside their clinical information. hgFeatureSorter and hgPathSorter together enable investigators to order, filter, aggregate and display data interactively based on any given feature set ranging from clinical features to annotated biological pathways to user-edited collections of genes. Standard and advanced statistical tools are available to provide quantitative analysis of whole genomic data or any of its subsets. The UCSC Cancer Genomics Browser is an extension of the UCSC Genome Browser; thus it inherits and integrates the Genome Browser's existing rich set of human biology and genetics data to enhance the interpretability of cancer genomics data.  We demonstrate the UCSC Cancer Genomics Browser by integrating several independent studies on breast cancer including the I-SPY chemotherapy clinical trial and other studies focused on chemotherapeutic response or long-term survival. The types of data that are visualized and analyzed by the browser include microarray measurements of gene expression, copy number variation and phosphoprotein expression, MRI imaging measurements, and clinical parameters.  Collectively, these tools facilitate a synergistic interaction among clinicians, experimental biologists, and bioinformaticians. They enable cancer researchers to better explore the breadth and depth of the cancer genomics data resources, and to further characterize molecular pathways that influence cellular dynamics and stability in cancer. Ultimately, insights gained by applying these tools may advance our knowledge of human cancer biology and stimulate the discovery of new prognostic and diagnostic markers, as well as the development of therapeutic and prevention strategies.  Funding sources: CALGB CA31964 and CA33601, ACRIN U01 CA079778 and CA080098, NCI SPORE CA58207, California Institute for Quantitative Biosciences, NHGRI.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2022.
Validation and comparison of nomograms and scores to predict non-sentinel lymph node status in breast cancer patients with metastatic sentinel lymph node: a prospective multicentric study.
Authors: Coutant C , Houvenaeghel G , Olivier C , Fondrinier E , Marchal F , Guillemin F , Seince N , Barranger E , Thomas V , Lambaudie E , Daraï E , Uzan S , Levêque J , Rouzier R
Published: December 2010

Abstract #203

Background  Five tools have been developed to predict non-SN status in breast cancer patients with SN metastasis: two nomograms (the Memorial Sloan-Kettering Cancer Center nomogram (MSKCC nomogram) and the nomogram developed by Degnim) and three scoring systems (the Tenon score, the score from the M.D. Anderson Cancer Center (MDA score), and the score developed by Saidi). Our purpose was to assess the validity of these tools in a prospective multicentric study of 561 SN-positive patients.  Methods  For 561 SN-positive patients who underwent ALND, we tested the accuracy of the five mathematical models. We calculated the false-negative (FN) rates and the areas under the receiver operating characteristics curves (AUC) as a measure of discriminatory capacity. We also tested the accuracy of each model in the subgroup of 246 patients with micrometastasis or isolated tumor cells in the SN; and in the subgroup of 165 patients with histological tumor size measuring more than 2 cm. Results were compared to the optimal logistic regression model that could be developed on the 561 patient cohort.  Results  At least one non-SN was positive in 147 patients (26.2%). The Tenon score, MSKCC nomogram, MDA score, nomogram of Degnim and score of Saidi applied to all 561 patients achieved AUCs of 0.81, 0.78, 0.73, 0.73, and 0.65 respectively. The FN rate were 4.4% (12/273), 6.5% (13/201), 5.7% (8/140), 12.5% (2/16) and 15% (21/140) respectively. In the subgroup of patients with only SN micrometastases, the FN rates were 2.9% (6/209) and 3.5% (5/143) for the Tenon score and the MSKCC nomogram respectively. The AUCs were 0.81 and 0.72 respectively  In the subgroup of patients with histological tumor size measuring more than 2 cm, the FN rates were 0% (0/10) and 28.6% (4/14) for the Tenon score and the MSKCC nomogram respectively. The AUCs were 0.69 and 0.66 respectively. For comparison, a logistic regression model developed on this multicentric cohort yielded AUCs of 0.83, 0.82, and 0.75 for the whole cohort, patients with micrometastasis or isolated tumor cells in the SN and in patients with tumor > 2 cm, respectively.  Conclusion  In this multicenter study, the Tenon score outperformed other scoring system. It was also more discriminant than other scores in patients with micrometastasis or isolated tumor cells in the SN and in patients with tumor > 2 cm.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 203.
Phosphoproteomic study of breast cancer: high-throughput SH2 screening.
Authors: Machida K , Claffey KP , Thompson CM , Shin D , Mayer BJ
Published: December 2010

Abstract #2031

Background: Protein tyrosine phosphorylation plays an important role in breast cancer cells. Tyrosine kinases such as Src and HER-2 have been shown to be overexpressed in invasive breast cancer tissues, and thus are used as prognostic markers and therapeutic targets. However, substrate proteins for tyrosine kinases in breast cancer tissues have not been extensively studied due to technical difficulties including low sensitivity and large sample requirement. Recently we established a high-throughput method using Src homology 2 (SH2) domain probes, termed SH2 profiling, to profile proteomes based on interaction between SH2 domains and tyrosine-phosphorylated proteins. In this study we aimed to optimize the assay for breast cancer tissues and evaluate SH2 profiling based classification. Material and Methods: Protein lysates from 10 breast cancer patient tissues as well as cancer cell lines were prepared and spotted in multiple duplicates on a nitrocellulose membrane. The blocked membrane was incubated with HRP-labeled GST-SH2 domain probes using a 96 well chamber apparatus. Bound probes were detected by chemiluminescence and signals were quantified using densitometry. Quantitative SH2 binding profiles of breast cancer samples were compared using a hierarchical clustering program. Results and discussion: 1. Although signal-to-noise ratio of breast cancer tissues was relatively low compared to cell lines, possibly due to non-cancerous material in clinical samples, specific signal was detectable using as little as 100 micrograms of sample protein for the entire assay. 2. Unsupervised clustering analysis based on SH2 binding patterns correctly separated HER-2 positive from HER-2 negative tumors, indicating the diagnostic potential of this method. Further validation studies with more samples are needed. 3. Although phosphoproteomics has not yet been adopted in the clinic because of issues related to sample requirements, reproducibility, and sensitivity, this pilot study strongly suggests that clinical applications of phosphoproteomics are feasible. This study is supported by the Breast Cancer Alliance.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2031.
Identification of robust, clinically relevant phenotypes of breast cancer from genome scale molecular profiling.
Authors: Eklund AC , Qiyuan L , Juul N , Richardson AL , Szallasi Z
Published: December 2010

Abstract #2036

Genome scale analysis by microarrays or array CGH holds the promise to identify novel subclasses of breast cancer or characterize functional modules of clinical relevance in a robust fashion. However, the success of these endeavors is highly dependent on several factors such as the general noise structure and noise level of high throughput measurements or the strength of association between biomarkers/gene modules and clinical outcome. We will address two issues deeply rooted in the high throughput nature of genome scale profiling, which are relevant for the accurate analysis of clinical microarray data sets: systematic bias in clinical microarray data and establishing strategies that allow extracting robust, convergent information/classification from genome scale molecular profiling of breast cancer.  As will be demonstrated, clinical microarray data are burdened with a high level of systematic bias. We identified sources of technical bias affecting many genes in concert, thus causing spurious correlations in clinical data sets and false associations between genes and clinical variables. A method will be presented to correct for technical bias in clinical microarray data, which increased concordance with known biological relationships in multiple data sets.  Gene expression profiling based classification of breast cancer and prognostic or clinical response associated gene expression signatures are usually derived from a single data set. However, any result extracted from a single data set will reflect to a large extent the technical (which genes are measured reliably on the microarray) and biological (such as cohort selection) bias of the given cohort. An alternative approach uses multiple, in this case 5 different, analogous clinical data sets and determines the robust, convergent information emerging from the cross data-set analysis. We will present such a method, which will reduce the impact of data set specific bias and outline robust functional modules in breast cancer, ultimately leading to the reevaluation of gene expression profiling based subtyping and diagnostic gene expression signatures in breast cancer.  Finally, we will present evidence for the existence of at least two fundamentally different types of genome instability in breast cancer with direct implications for response to chemotherapy.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2036.
Concentrations of plasminogen activator inhibitor type1 (PAI-I) and urokinase-type plasminogen aktivator (uPA) in primary breast cancer (BC) tissue and corresponding axillary lymph node metastasis.
Authors: Artmann A , Celik K , Kiechle M , Rummeny EJ , Schmitt M , Harbeck N
Published: December 2010

Abstract #2046

Background: Components of the plasminogen activator system play a key role in tumor invasion and metastasis. Tumor tissue levels of urokinase-type plasminogen activator (uPA) and its inhibitor PAI-1 showed a strong prognostic impact in node-negative and node-positive primary BC. uPA/PAI-1 were the first novel prognostic factors in primary BC reaching the highest level of evidence for their clinical utility and they have recently been recommended by ASCO for clinical decision making.  Material and Methods: For the first time, correlations between uPA and PAI-1 concentrations in 73 primary tumor tissues and corresponding axillary lymphnodes (ALN) in node-negative (pN0) and node-positive (pN+) BC (1991-98) and their prognostic impact on disease-free (DFS) and overall survival (OS) were evaluated.  Results: Median follow-up was 56 months. Overall, PAI-I (p=0,014) and uPA concentrations (p<0,001) were significantly higher in tumour tissue than in the corresponding ALN. There was no significant difference of PAI-1 in primary tumor tissue between pN0 and pN+ disease. In involved ALN (pN+), concentrations of PAI-1 but not of uPA, were significantly higher than in pN0 (p<0.001). Moreover, patients with higher PAI-1 concentrations in the corresponding ALN than in their primary tumor had a significantly shorter DFS (p<0,001) and OS (p=0.005).  Discussion: Our clinical findings support the basic research data implicating uPA and PAI-1 as key proteolytic factors in metastasis-asociated processes such as adhesion, angiogenesis, invasion, proliferation and migration. For the first time, we are able to demonstrate that particularly PAI-1 seems to be involved in tumor metastasis in primary breast cancer with its concentrations being higher in involved ALN than in the primary tumor. Besides uPA/PAI-1 being validated and clinically useful prognostic factors, interference with the uPA/PAI-1 system thus promises to be an interesting therapeutic concept in BC.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2046.
Prognostic implications of isolated tumour cells and micrometastasis in lymph nodes in non-adjuvant treated breast cancer patients.
Authors: Park D , Kåresen R , Naume B , Sauer T
Published: December 2010

Abstract #205

Introduction : The status of axillary lymph nodes (ALNs) is the most powerful predictor of survival and has clinical implications for adjuvant treatment of breast cancer patients. Occult metastases in ALNs of patients initially staged as negative have been detected up to 40%, using the various methods with serial sectioning and/or immunohistochemical (IHC) staining. However, there is no consensus concerning the prognostic significance for occult metastases. The aim of this study was to estimate the prognostic value of isolated tumor cells (ITC, ≤0.2 mm) and micrometastases (MM, 0.2-2.0 mm) in invasive breast cancer.  Materials and Methods: The ALNs of 295 patients with axillary node-negative invasive breast cancer treated from 1995 to 1998 was evaluated with serial sectioning and cytokeratin IHC (a single section from each of 10 levels) and hematoxylin-eosin (one section from 3rd and 10th level). Disseminated tumor cell status in bone marrow was also known. Of 295 patients, 232 were offered no systemic adjuvant therapy. Patients were categorized by largest cluster size (≤0,2mm, 0.2-2.0mm), and then their survival rates, i.e. disease-free survival (DFS), distant disease-free survival (DDFS) and breast cancer specific survival (BCSS) were compared with those of node-negative patients. We also investigated the correlation between occult metastases in ALNs and bone marrow (BM).  Results: The median patient age was 59 years and the median tumour size was 1.5 cm. At a median observation time of 98 months (9-127 months), 33 patients had developed relapse,2 local and 31 systemic. 17 patients had died of breast cancer. Occult metastases were found in 92 of 295 patients (31%); ITC – 58% (53/92), MM -34% (31/92) and macrometastases (>2.0mm)- 8% (8/92). The cases with macrometastases were excluded from survival analysis. The results presented in the table below demonstrate significant prognostic impact of MM on BCSS, borderline significance on DFS and DDFS. This was not shown for ITC. No significant correlation was found between occult metastases in ALN and BM in both total patients (Χ2-test, p=0.680).    Conclusions: Our observation of a prognostic impact of MM in non-adjuvant treated patients indicates that adjuvant therapy might be warranted in this group. Lack of a similar finding in patients with ITC questions the need of adjuvant therapy in this group. The lack of correlation between occult metastases in ALN and BM indicates different “homing” mechanisms for the two compartments.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 205.
JNK2 enhances mammary carcinoma cell migration by regulating EPS8 expression and localization.
Authors: Mitra S , Van Den Berg C
Published: December 2010

Abstract #2050

Cell migration is a tightly regulated event critical for normal development and adult wound healing, but it is also a key step in the tumor metastasis cascade. Growth factors initiate intra-cellular signaling pathways that stimulate cell movement by virtue of remodeling the actin cytoskeleton. Fine regulation of this process renders a degree of polarity to the cells, altering their cytoskeleton and providing the requisite mechanical torsion to move forward. Breast cancer cells extravasate from their primary site and travel to other organs forming metastatic colonies.Various signaling molecules including members of the Mitogen Activated Protein Kinase (MAPK) such as c-Jun N-Terminal Kinase (JNK) have been implicated in regulating cancer cell migration. How growth factors transmit signals to form dorsal ruffles and filopodia for cell migration is not precisely understood. Additionally, JNK is encoded by three genes, resulting in 10 isoforms, hence its role in metastasis is complicated. Here, we used the Polyoma Virus Middle T Antigen (PyVMT) transgenic mouse model where wildtype jnk2 was expressed or knocked out. In tumors we found that PyVMT jnk2-/- tumors express higher Epidermal Growth Factor Receptor Substrate 8 (EPS8) mRNA and protein. EPS8 was originally identified as a substrate for Epidermal Growth Factor Receptor (EGFR), and was recently reported to have an actin capping function. Hence, EPS8 is a prime candidate for connecting the EGFR signaling to actin cytoskeletion remodeling, and therefore mediate cell migration. In vitro using Boyden Chamber based migration assays, we found that the PyV MT jnk2+/+ cells migrated fivefold more than the PyV MT/jnk2-/- cells. Pan inhibition of JNK using a small peptide inhibitor TAT-JIP VII significantly reduced migration of the PyV MT/jnk2+/+ cells but was ineffective at further reducing PyVMT jnk2-/- cell migration, suggesting that the JNK2 is the major isoform to convey this function. Furthermore, shRNA mediated knock down of EPS8 in the PYVMT jnk2-/- cells, increased migration of these cells to levels comparable with the wild type counterparts. We subsequently undertook immunocytochemistry studies to evaluate EPS8 localization and cytoskeletal changes upon serum stimulation. Cells expressing JNK2 showed more dorsal ruffles upon serum stimulation, and EPS8 localized to the plasma membrane and in dorsal ruffles. In contrast, EPS8 was predominantly localized in focal adhesions in the PyVMT jnk2-/- cells, which may explain their reduced migration potential. Together our model supports an integral role for JNK2 regulation of EPS8 expression and localization to facilitate cell migration. Our current studies are aimed at further deciphering the JNK/EPS8 pathway to understand the exact molecular mechanism by which these proteins regulate a critical step in mammary cancer cell metastasis.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2050.
Identification of non-catalytic matrix metalloproteinase 1 residues that participate in collagenolysis.
Authors: Lauer-Fields JL , Fields GB
Published: December 2010

Abstract #2063

Collagen serves as a structural scaffold and a barrier between tissues, and thus collagen catabolism (collagenolysis) is required to be a tightly regulated process in normal physiology. In turn, the destruction or damage of collagen during pathological states plays a role in tumor growth and invasion, cartilage degradation, or atherosclerotic plaque formation. Only a small number of proteases have been identified capable of efficient processing of triple-helical regions of collagens. Several members of the zinc metalloenzyme family, specifically matrix metalloproteinases (MMPs), possess collagenolytic activity, and at least one of these (MMP-1) has been strongly implicated in the progression of breast cancer. The present study has utilized triple-helical peptide (THP) substrates and inhibitors to dissect MMP-1 collagenolytic behavior. Analysis of MMP-1/THP interactions by hydrogen/deuterium exchange mass spectrometry (HDX MS) in combination with site-specific mutagenesis and enzyme kinetic assays have allowed us to identify non-catalytic sites in MMP-1 that participate in collagenolysis. As we have recently demonstrated [J. Am. Chem. Soc. 129, 10408 (2007)], non-catalytic sites can be utilized to create THP inhibitors that target proteases implicated in cancer progression while sparing proteases with host-beneficial functions.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2063.
STAT1 and STAT3 synergistically suppress EGF-induced MMP-9 activity in SKBR3 human breast cancer cell line.
Authors: Kim S , Choi J , Lim H , Lee S , Kim W , Yang J , Nam S , Lee J
Published: December 2010

Abstract #2064

Background: EGF is a cytokine associated with cell proliferation, migration, and tumor invasion through EGFR activation in various cancer cells. EGFR activates its downstream molecules such as protein kinase C-δ, ERK, PI3-kinase, STAT-3. It has been shown that EGFR induces matrix metalloproteinase-9 (MMP-9), leading to invasion and migration of cancer cells. However, it has not been fully elucidated the roles of STAT-1 and STAT-3 in breast cancer cells. We investigated the regulatory mechanism of STAT-1 and STAT-3 on EGF-induced MMP-9 activity in SKBR3 cells.  Material and Methods: EGF-induced MMP-9 activity regarding to ERK, JNK, PI-3 kinase and STAT-3 were evaluated by Western blot. Then, we transfected SKBR-3 cells with adenoviral vectors containing STAT-1 and STAT-3 to make SKBR-3 cells overexpressing each gene. We also treated SKBR-3 cells with STAT-3 siRNA. Overexpression and silencing of genes were identified with Western blot. With the treatment of EGF, MMP-9 activity was measured with gelatinase Zymography.  Results: We confirmed that EGF increases MMP-9 activity and induces the phosphorylation of ERK, JNK, PI-3 kinase and STAT-3 in dose-dependent manner. We found that EGF-induced MMP-9 activity was significantly decreased by MEK1/2 inhibitor, UO126, and PI3-kinase inhibitor, LY294002, respectively, but not by JNK inhibitor, SP600125. In contrast to previous study, EGF-induced MMP-9 activity was inhibited by STAT-3 overexpression. We also found that it was increased by STAT-3 siRNA transfection. Moreover, we found that STAT-1 and STAT-3 co-overexpression synergistically decreased EGF-induced MMP-9 activity in STAT-1 and STAT-3 co-overexpressing SKBR-3 cells.  Discussion: We demonstrated that EGF-induced STAT-3 activation suppresses tumor invasion through down-regulation of MMP-9 in SKBR-3 cells. Moreover, we found that there is a synergistic relationship between STAT-1 and STAT-3, inhibiting EGF-induced MMP-9 activity.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2064.
The use of proximity ligation assay to identify HER2:HER3 heterodimers in early breast cancers.
Authors: Spears M , Munro AF , Taylor KJ , Mallon EA , Bartlett JM
Published: December 2010

Abstract #2068

Background: The HER receptor family includes four members: HER1, HER2, HER3 and HER4. Expression of these proteins is linked to breast cancer development and progression. HER2 gene amplification is found in 20-30% of breast cancers. Overexpression of HER2 has been linked with high grade tumours and poor prognosis. Clinical studies have demonstrated a strong relationship between HER2 and HER3 with 70% of HER3-positive tumours also being positive for HER2. Co-expression of HER2/HER3 has been suggested to drive endocrine and chemotherapy resistance. However such resistance is driven not by co-expression but by signalling via HER2/HER3 dimers which have previously been impossible to detect in situ. Proximity ligation assays (PLAs) can detect homo- and heterodimers in situ at a single-molecule level and allow study of signalling pathways. Paired antibodies are used to detect protein complexes in tissues.  Methods: Using tissue microarrays containing 100 breast cancer samples, we analysed HER2 homodimers and HER2:HER3 heterodimers by PLA. HER2 amplification was detected using FISH analysis.  Results: PLA was used to detect HER2 homo and heterodimers in vivo and in vitro. HER2 homodimers were detected in amplified and non-amplified cases. A significant association (p<0.00001) was identified between HER2 homodimerisation and amplification. HER2 homodimerisation was not associated with any clinicopathological data.  HER2:HER3 heterodimers were detected in vivo and in vitro through PLA and western blot analysis. The median number of HER2:HER3 heterodimers (7.8 signals per cell) was greater than HER2:HER2 homodimers (2.0 signals per cell) in this series indicating HER2:HER3 may be the preferred dimer. HER2:HER3 was significantly associated with nodal status (p=0.003) and grade (p=0.02).  Conclusions: Using an in situ PLA we demonstrated the in vivo expression of HER2 homodimers in human breast cancer and increased HER2 homodimerisation linked to HER2 gene amplification. We have demonstrated HER2:HER3 heterodimers to be linked to nodal status and grade. Our observations suggest that PLA is an invaluable method to detect homo and heterodimers in situ and could potentially be used a diagnostic tool in the future.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2068.
Prognosis in BRCA1, BRCA2 associated breast cancer (BC): a prospective Breast Cancer Family Registry (BCFR) international population-based cohort study.
Authors: Goodwin P , Phillips K , West D , Ennis M , Hopper J , John EM , O'Malley F , Milne R , Andrulis I , Friedlander M , Longacre T
Published: December 2010

Abstract #2072

Hereditary BC occurs at a younger age and is associated with more adverse tumor-related features than sporadic breast cancer (BC) (defined here as BC in those with no 1st or 2nd degree family history of breast or ovarian cancer). Using pre-specified criteria, we assembled a population-based cohort of newly diagnosed BC at 3 centers: Ontario, Canada (1996-98), San Francisco Bay area, USA (1995-2000), Melbourne/Sydney, Australia (1991-1998). Medical information was obtained from medical records; women were followed prospectively for recurrence, new cancers and death. Pathology data were obtained from central review or pathology reports. BRCA1 and BRCA2 mutation testing was performed on 77% and 70% of cases, respectively (sporadic BC cases were not tested at 2 centers). Hereditary and sporadic BC cases were compared using Cox proportional hazards (stratified by center). 3215 eligible cases were enrolled in the BCFR, with a mean age at diagnosis of 46.9 years. Median follow-up was 7.61 years; 565 women had distant recurrences and 547 died. There were 92 cases with BRCA1 and 72 with BRCA2 mutations; 1549 (48.2%) had sporadic BC; the remainder had familial BC as defined above. BRCA1 mutations were associated with young age, estrogen and progesterone receptor (ER and PgR) negativity and high grade; BRCA2 mutations were associated with node positivity and high grade. Distant disease-free survival (DDFS) and overall survival (OS) did not differ significantly between BRCA1 carriers and sporadic cases in univariate or multivariate analyses. DDFS and OS were worse in BRCA2 carriers than in sporadic cases (HR 1.6, p=0.04 and HR 1.8, p=0.01, respectively) in univariate analyses but not in multivariate analyses (DDFS HR 1.0, p=0.98; OS HR 1.13, p=0.61). The small group of BRCA2 carriers who did not receive adjuvant chemotherapy had a significantly worse OS (multivariate HR 3.63, p = 0.005). Furthermore, BRCA2 carriers who received adjuvant tamoxifen had significantly worse OS than women with sporadic BC (HR=2.0, p=0.03). We conclude that BRCA1 and BRCA2 mutations do not independently impact DDFS or OS. Significantly worse outcomes were seen in BRCA2 carrier subgroups defined by adjuvant treatment; this requires further investigation and may have implications for clinical practice.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2072.
Pharmacokinetics and tolerability of lapatinib administered once daily in combination with tamoxifen.
Authors: Cardoso F , Koch KM , Awada A , Lokiec F , Brain E , Bogaerts J , Marreaud S , Chung S , Campello V , Fumoleau P
Published: December 2010

Abstract #2073

Background: The pharmacokinetics (PK) and tolerability of daily oral dosing with lapatinib (LAP) and tamoxifen (TAM) in patients (pts) with advanced or metastatic breast cancer (MBC) are examined in this study. Optimal dose selection requires characterization of PK to accommodate the potential for a bi-directional PK interaction involving LAP inhibition and TAM induction of CYP3A4 and P-glycoprotein (Pgp).  Methods: Twenty three previously treated pts with ER and/or PR positive MBC, not ErbB selected, were randomly assigned to one of two groups, to separately assess each direction of the potential PK interaction at steady state during daily dosing of 1250mg LAP and 20mg TAM. In one group, the PK of LAP is assessed on the 14th day of dosing LAP alone, and again on the 28th day dosing of LAP and TAM together. In the other group, the PK of TAM is assessed on the 28th day of dosing TAM alone, and on the 7th day of dosing TAM and LAP together. LAP and TAM are dosed together throughout subsequent 28 day cycles.  Results: To date 23 pts have been enrolled with a median age of 58 (38-83) and a median PS of 1. Preliminary PK data for LAP from six patients summarized in the table below showing geometric median (95% confidence limits) parameters indicate that LAP plasma concentrations are decreased by TAM. Final PK analysis will be presented at the meeting. To date, the most common adverse events at cycle 1 were: fatigue gr1/2 (100%), diarrhoea gr 1/2 (53%), rash gr 1 (40%), anorexia gr 1 (40 %) and nausea gr1/2 (40 %). Transaminase elevation (SGPT/ALT) increased from 3 pts at baseline to 6 pts at cycle 1 including one gr3.    Conclusions: The preliminary PK results are consistent with data indicating TAM induces hepatic CYP3A, the primary route of LAP elimination. This important information may impact in ongoing adjuvant lapatinib studies.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2073.
Survival differences among women with newly diagnosed stage IV and recurrent breast cancer.
Authors: Dawood S , Kristine B , Hortobagyi GN , Giordano S
Published: December 2010

Abstract #2077

Background   The objective of this retrospective study was to determine whether differences in survival exist between women with newly diagnosed stage IV disease and those with recurrent breast cancer.  Methods  Women with newly diagnosed stage IV disease or recurrent breast cancer with distant metastases diagnosed between 1992 and 2007 were identified from the Breast Cancer Management System Database at UTMDACC. Among women with recurrent breast cancer, distant disease-free interval (DDFI) was defined as the time from diagnosis of primary breast cancer to the date of the first distant metastases. For all patients, overall survival (OS) was measured from the date of first distant metastases to the date of death from any cause or last follow-up. The Kaplan-Meier product limit method was used to estimate OS, and OS was compared across groups using log-rank statistic. Cox-proportional hazards models were fit to determine the association between stage IV breast cancer (recurrent versus newly diagnosed) and OS after controlling for other patient and tumor characteristics.  Results  The analysis included 3524 patients of whom 643 (18.2%) had newly diagnosed disease and 2881 (81.8%) had recurrent disease. Median follow-up was 19 months. Median OS among patients with newly diagnosed stage IV disease and recurrent disease was 39.2 and 27.2 months, respectively (p<0.0001). Patients with recurrent disease with DDFI < 5 years had worse OS compared to those with newly diagnosed disease (p<0.0001). Patients with DDFS >5years had better OS compared to those with newly diagnosed stage IV disease (p<0.005). In the multivariable model patients with recurrent disease had an increased risk of death compared to patients with newly diagnosed stage IV disease (HR=1.75, 95% CI 1.47-2.08, p<0.0001). When the multivariable model was stratified by DDFI, patients with recurrent disease with DDFI <5 years had 1.74 times the risk of death compare to those with newly diagnosed stage IV disease (95% CI 1.44-2.10, p<0.0001). The risk of death was not statistically different among patients with recurrent disease with DDFI >5 years compared to those with newly diagnosed stage IV disease (HR=1.11, 95% CI 0.88-1.41, p=0.38).  ConclusionThis large cohort study provides further insight into the natural history of recurrent and newly diagnosed stage IV breast cancer. Compared to women with newly diagnosed stage IV disease, those with recurrent disease whose DDFI was >5 years and <5 years had similar and worse OS respectively.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2077.
Male breast cancer in Chinese population – a ten year review.
Authors: Kwong A , Suen DT
Published: December 2010

Abstract #2094

Objective: Breast cancer is uncommon in men, occurring in 1% of the male population based on Western data. There is limited knowledge about the natural history and prognosis of male breast cancer in Chinese population. This study aims to be the first to perform a 10 year review of the clinical presentation and outcome of Chinese male breast patients.  Methods: A retrospective study of patients with male breast cancer treated in Hong Kong from January 1995 to December 2005 was performed.  Results: A total of 83/124 medical records of male breast cancers treated in 16 hospitals during this 10 year study period were available and reviewed. The mean age at diagnosis was 65 years old. Of those where family history was known, only 3% had a family history of breast cancer. Only 7 patients had gynaecomastia. The mean time of presentation from time of symptoms occurrence was 23.2 months (range 0.06- 216). The most common presentation was presence of a breast lump. 75 (97%) of these patients underwent mastectomy, half of whom also had axillary dissection. 4 patients had LD flap reconstruction in addition to the mastectomy. 67 (87%) of these were invasive carcinoma of which 1 was invasive lobular carcinoma, 2 were mucinous carcinoma, 8 were invasive papillary carcinoma and the rest were invasive ductal carcinoma. 9 (11.7%) were in-situ carcinomas, where 6 were intraductal papillary carcinoma and the rest were ductal carcinoma in-situ. One patient had breast sarcoma. Of the invasive tumors, 74% were classified as Grade II and above. 29.8% of the patients had nodal involvement and the most common stage of presentation were either Stage 1 (33) or Stage 2 (27%). Majority of the tumors were oestrogen (95.8%) and progesterone (81%) receptor positive and 50% were cerbB2 positive. Where treatment data was available, all patients with ER positive tumors except for 6 patients received tamoxifen. 2.7% received neoadjuvant chemotherapy, 19.2% adjuvant chemotherapy and 47.3% received radiation therapy. The mean follow up was 54.5 months (1-178) months. The 5-year disease free and overall survival were 88% and 71.2% respectively. Interestingly 15 patients also had a second primary cancer not of breast origin.  Discussions: Male breast cancer in Chinese men is rare and present at an older age but at an early stage. Disease free survival was better than overall survival likely due to death cause by disease other than breast cancer due to the older age of presentation. Although a majority of our cohort did not have family history of breast cancer, there was a high incidence of second primary cancer not of breast origin. Further investigation with genetic study in this group of patients is likely to be of relevance.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2094.
Dose-dense (DD) doxorubicin and cyclophosphamide (AC) followed by weekly paclitaxel (P) with trastuzumab (T) and lapatinib (L) in HER2/neu-positive breast cancer is not feasible due to excessive diarrhea: updated results.
Authors: Dang C , Lin N , Moy B , Come S , Lake D , Theodoulou M , Troso-Sandoval T , Dickler M , Gorsky M , D'Andrea G , Modi S , Seidman A , Drullinsky P , Partridge A , Schapira L , Wulf G , Gilewski T , Atieh D , Mayer E , Isakoff S , Sugarman S , Fornier M , Traina T , Bromberg J , Currie V , Robson M , Burstein H , Overmoyer B , Ryan P , Kuter I , Younger J , Schumer S , Tung N , Zarwan C , Schnipper L , Chen C , Winer E , Norton L , Hudis C
Published: December 2010

Abstract #2108

Background: DD q 2 weekly (w) AC → P + T x 1 year (y) has an acceptable safely profile w/ congestive heart failure (CHF) rate of 1/70 pts (Dang, JCO 2008). Lapatinib (L) is effective in HER2 (+) BC. We conducted a pilot study of dd AC → w P + T + L to determine its feasibility and cardiac safety.  Methods: Enrolled pts had HER2 (+) BC; LVEF > 50%. Rx consisted of AC at 60/600 mg/m2 x 4 q 2 w (w/ pegfilgrastim 6 mg day 2) → P at 80 mg/m2 x 12 q w + T x 1 y; L (1000 mg daily beginning w/ P + T and continued x 1 y). MUGA is obtained at baseline and at months (mo) 2, 6, 9, and 18. Rx is considered feasible if 1) > 80% pts can complete the PTL phase without a dose delay or reduction and 2) the cardiac event rate (CHF or cardiac death) is < 4%. Pts can remain on-Rx w/ one dose reduction of L (1000 mg → 750 mg) for a G 3 event or < G 3 toxicity (unacceptable).  Results: From March 2007 to April 2008, we enrolled 95 pts. Median (med) age was 45 years (range, 28-73). At a med follow-up of 7 months, 90 are evaluable. Of the 90 pts, 34 (37%) withdrew from study during the PTL phase; 29 for a 2nd event of G 3 or unacceptable < G 3 toxicities (15 G 3 diarrhea, 4 G 1/2 diarrhea, 1 G 3 rash, 2 G 2 rash, 1 G 3 dyspnea and also had G 3 diarrhea, 1 G 3 ↑QTc also had G 3 diarrhea, 1 G 3 ↑ALT also had G 3 diarrhea, 1 G 3 paronychia, 1 G 3 pneumonitis, 1 asymptomatic LVEF ↓, 1 myocarditis) and 5 for other reasons (2 personal reason, 1 PCP pneumonia, 1 progression, 1 P hypersensitivity). Overall, 25/90 (27%) pts had G 3 diarrhea and 31/90 (34%) pts required a dose reduction of lapatinib. Med LVEF at baseline is 67% (N=95), at mo 2 is 68% (N=90), at mo 6 is 65% (N=53), and mo 9 is 65% (N=28). To date there are no patient drop-outs due to significant LVEF declines after dd AC; one patient dropped during PTL out due to an asymptomatic LVEF decline.  Discussion: L at 1000 mg/day is not feasible combined w/ weekly P and T by protocol stipulation (> 20% pts required L dose reduction) primarily due to excessive G 3 diarrhea. These results have led to the modification of Design 2 (Arm D) of ALTTO. We will report updated results.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2108.
Role of the prostaglandin receptor EP3 in regulating vasculogenic mimicry of inflammatory breast cancer.
Authors: Robertson FM , Simeone A , Lucci A , Woodward W , Reuben J , Ueno N , Cristofanilli M
Published: December 2010

Abstract #2127

Background: Inflammatory breast cancer is an aggressively angiogenic, metastatic and lethal variant of breast cancer. One of the characteristics of aggressive tumors types such as IBC is the ability of tumor cells to undergo vasculogenic mimicry (VM), which is the formation of organized capillary like structures in vitro and in vivo much like endothelial cells, providing a conduit for oxygen and nutrients to reach hypoxic and nutrient deprived tumor cells that have outgrown their adjacent vascular supply. Using genomic and proteomic profiling as well as Western blotting, immunohistochemistry, invasion assays using the modified Boyden chamber assay as well as digital imaging, we evaluated the signal transduction pathways involved in regulating proliferation, invasion, and VM by IBC tumor cells.  Results: We found that cyclooxygenase-2 (Cox-2), which produces the bioactive lipid, prostaglandin E2 (PGE2) is one gene that is highly expressed IBC tumor cell lines, SUM149 and SUM190. The biological activity of PGE2 occurs via binding of PGE2 to one of 4 members of a G protein coupled receptor family, designated as the EP receptors, EP1, EP2, EP3 and EP4. Western blot analysis and immunochemistry revealed that EP3 receptor protein is produced at very high levels by both IBC tumor cell lines to a significantly (p<0.05) greater extent than either human MCF-7 or MDA-MB-231 non-IBC breast tumor cells. Since EP3 is known to be an inhibitory prostanoid receptor, we evaluated the dose dependent effects of the EP3 agonist, sulprostone, on proliferation, invasion, and VM. Sulprostone [0.1, 1, 10 and 100 μm] inhibited proliferation of SUM149 and SUM190 IBC cells in a dose and time dependent manner to a significantly greater extent (p<0.01) than MCF-7 or MDA-MB-231 breast tumor cells. Sulprostone inhibited invasion of SUM149 IBC tumor cells, with no effect on invasion of a basement membrane by the MDA-MB-231 human non-IBC breast tumor cells. SUM149 IBC tumor cells undergo VM, which was completely inhibited by 10 μM sulprostone at 24 hrs, as visualized by Periodic Acid Schiff (PAS) staining and digital imaging. The ability of sulprostone to completely inhibit VM occurred in conjunction with inhibition of production of laminin 5-γ2 fragments and matrix metalloproteinase-2 (MMP-2) activity. Studies are currently underway to determine the in vivo effect of sulprostone on IBC tumor growth, VM, and metastasis of SUM149 IBC xenografts.  Discussion: These results suggest that targeting EP3 may provide a selective advantage for abrogating not only proliferation, and invasion but also vasculogenic mimicry that is a key component of the rapid angiogenic program observed in IBC.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2127.
A phase II, randomized, open-label, dose-finding study of TAS-108, a novel steroidal antiestrogen, administered orally in postmenopausal women with advanced or metastatic breast cancer.
Authors: Iwata H , Inaji H , Nakayama T , Yamamoto N , Noguchi S , Ikeda T
Published: December 2010

Abstract #2132

Background: A phase II study was conducted to identify the recommended dose of TAS-108, a novel steroidal antiestrogen with full ERα antagonist, mainly metabolized by CYP3A4, in post menopausal Japanese women with advanced or metastatic breast cancer. This study was prepared referring to the multi-center phase II trial that performed in four countries (USA, Russia, Mexico and Chile), and performed in Japan. Both studies are not the same designs.  Patients and Methods: Patients with ER and/or PgR positive were eligible if they had one or two standard endocrine therapies, with or without one prior chemotherapy, for advanced disease. TAS-108 was administered orally at three different doses (40mg/80mg/120mg per day) for 24 weeks. The treatment was terminated if PD was observed. In this clinical study, clinical benefit (CB, CR+PR+SD for at least 24 weeks) rate as the primary end point was evaluated. The secondary endpoints were overall response rate and best overall response using RECIST; time to progression; adverse events (AEs) graded by CTC-AE (ver.3.0). Endometrial thickness (ET) and bone mineral density (BMD) as specific items of hormonal therapy were evaluated by transvaginal or transabdominal ultrasound, and dual-energy X-Ray absorptiometry method, respectively.  Results: Ninety eight patients were enrolled and randomized. One ineligible patient in the 120mg dose group dropped out without administration; therefore a total of ninety seven patients were administered and assessed for efficacy and safety. Thirty three patients received 40mg, 32 received 80mg and 120mg. The three treatment groups were well balanced. CB was observed in 10 patients (30.3%) in the 40mg group, and 8 patients (25.0%) in both the 80mg and 120mg groups. Retrospective analysis showed TAS-108 had effectiveness to patients who had failed on prior tamoxifen (TAM) or aromatase inhibitors (AI) treatment, including relapse during the adjuvant therapy or within 6 months from its completion. CB rate at the three doses respectively were 25.0/33.3/25.0% in 14 (4/6/4) patients failed on TAM, and 30.8/31.6/20.0% in 70 (26/19/25) patients failed on AI.  TAS-108 was well tolerated and most of the AEs were Grade1 or 2. The most frequent drug-related AEs were hot flush (22 patients, 22.7%, 7/8/7 patients in 40/80/120mg), perspiration (11 patients, 11.3%, 2/3/6), nausea (9 patients, 9.3%, 1/3/5), arthralgia (6 patients, 6.2%, 0/3/3). Neither remarkable change nor apparent dose-related trend among the three groups was observed in ET or BMD.  Conclusion: According to the protocol, 40mg was chosen as the recommended dose of TAS-108 for breast cancer. It was suggested that TAS-108 has clinical benefit for patients with breast cancer who have failed on prior TAM or AI treatments.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2132.
Internalization and retention of nanodiamonds by MCF-7 cells in vitro: a potential drug delivery platform.
Authors: Khan SA , Ivancic DZ , Zaichuk T , Huang H , Chen M , Lam R , Zhang M , Chatterton RT , Ho D
Published: December 2010

Abstract #2134

Background: Selective uptake and prolonged retention of drugs will facilitate pharmacotherapy to the breast. This may be accomplished by complexing of drugs to carbon-based nanodiamond (ND) particles, 2-8nm size. ND particles are attractive for this purpose because they are inert and non-toxic.We tested the uptake and retention of ND particles by the estrogen receptor positive breast cancer cell line MCF-7.  Methods: Polylysine coated ND were labeled with fluorescein isothiocyanate (FITC-ND), and examined by UV-visible spectra and found to be stable. MCF-7 cells were cultured on Lab-Tek II chamber slides using DMEM with 10% FCS, insulin, sodium pyruvate, L-glutamine, and Pen/Strep. To assess the efficiency of ND internalization, we added 10ug/ml ND-FITC in complete media to the chamber slides and incubated for 30 min, 1 hr, 3 hr, 4 hr, 17 hr, and 24 hr. FITC-ND was withdrawn from the medium after an initial exposure of 17 hr by washing of cells, and retention of fluorescence was measured at intervals, up to 3 days after withdrawal. To assess the ability of MCF-7 cells to take up NDs under low metabolic conditions MCF-7 cells were incubated in serum-free medium. As a control soluble FITC alone was incubated at 10 ug/ml with MCF-7 cells. Cells were imaged on the laser confocal microscope and FITC detection at 488nm was analyzed by three dimensional Z stack imaging.  Results: Fluorescence was initially associated with the membrane surface, and later was visualized in the cytoplasm and in the perinuclear zone. Cellular uptake of FITC and ND-FITC occurred at 30 minutes and reached a maximum at 17 hr of incubation. After washing of cells, the retention studies showed little decrease in the intensity of the ND-FITC signal within the cells up to 96 hours. The FITC molecule was readily released from the cells by passive diffusion within 24 hours while NDs appeared to be compartmentalized in cytoplasmic vesicles.    Conclusions: Internalization of ND particles within the cell cytoplasm occurs within a relatively short time, and the ND particles are retained for at least 96 hours following removal from the culture medium. This prolonged retention is likely to be an advantage when considering novel routes of drug delivery to the breast, such as transdermal or intraductal approaches.  Funded by Avon.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2134.