bluuebunny/crossref_metadata_2025 huggingface dataset:
Immunotherapy following high-dose chemotherapy (HDCT) and peripheral blood progenitor cell transplantation (PBPCT) in the treatment of high-risk early breast cancer (HREBC) and metastatic breast cancer (MBC).
Authors: Recchia F , Candeloro G , Necozione S , Sureda M , Brugarolas A , Rea S
Published: December 2010

Abstract #2159

Background: Meta-analyses have shown that HDCT with PBPCT is not superior to standard dose chemotherapy in the treatment of high risk early breast cancer (HREBC) and metastatic breast cancer (MBC). The reason for the limited efficacy is due to failure to eradicate all cancer stem cells, resistant to chemotherapy, and capable to reproduce the tumor in immunodeficient mice. Lymphopenia has been described after HDCT. We have previously shown that low dose inteterleukin 2 (IL-2) and oral 13-cis- retinoic acid (RA) improved lymphocyte (L) count, and decreased vascular endothelial growth factor (VEGF), of patients with advanced tumors having a clinical benefit from chemotherapy. Primary endpoint of this study was to verify if IL-2 and RA, improving L count, and decreasing VEGF after HDCT with PBPCT of patients with advanced breast cancer, reduced angiogenic switch. Secondary endpoints were the evaluation of progression-free survival (PFS) and overall survival (OS).  Methods: From 02-1998 to 02-2003, 30 patients with HREBC and 18 with MBC were entered into the study. One month after HDCT, patients received, for 1 year, 5 days/week, 3 weeks/month, subcutaneous IL-2 (1.8 x 106 IU daily) plus RA (0.5 mg/kg body weight). Immunotherapy was continued, with intermittent schedules, up to 5 years. Patients with estrogen receptor positive (ER+) tumors received hormonal therapy, while those in premenopause received also a LHRH analogue for 5 years.  Results: Characteristics of HREBC patients: median age, 48 years (range 34-60); median number of positive axillary nodes, 13 (range-11-27); ER+/ER-: 20/10; characteristics of MBC patients: median age, 46 years (range 28-60); median number of disease sites 1.5, (range-1-4); ER+/ER-: 5/13; Major adverse effects from IL-2 were fever, rash and autoimmune reactions; no treatment-related mortality was seen. After a median follow-up of 61 months, a statistically significant improvement of L and decrease of VEGF were observed for both HREBC and MBC. 5 years PFS and OS rate were, for HREBC 76% and 85%, respectively, for MBC, 22% and 35%, respectively.  Conclusions: These data show that IL-2 administration after HDCT and PBPCT is feasible, has a moderate toxicity, gives a statistically significant improvement of L a decrease of VEGF, and seems to give an improvement of expected PFS and OS, both in HREBC and MBC.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2159.
Identification of high-risk lesions through automated natural language processing (NLP) of pathology reports.
Authors: Ozanne EM , Sharko J , Drohan B , Grinstein G , Hughes KS
Published: December 2010

Abstract #3001

Purpose  Pathology reports contain extensive research information that is inaccessible except through costly and time consuming chart reviews. This is due to the fact that pathology reports are recorded as semi-structured prose with critically important descriptive text intended for human interpretation. Key challenges for processing this data include interpreting multiple methods of describing the same finding, and subsequently aggregating the findings of multiple reports into episodes of care. Investigators tested NLP techniques in the processing of pathology reports into structured data and episodes of care, allowing for the rapid identification and epidemiologic modeling of high-risk breast lesions.  Methods  Using state-of-the-art NLP software (ClearForest, A Thomson Reuters Company, Waltham, MA), breast pathology reports stored as text files were processed into a structured electronic database using these steps: 1) identification of diagnosis of interest (i.e. high risk lesions, cancer), 2) use of NLP to identify all terms and phrases used to report each finding (e.g. atypical hyperplasia, hyperplasia with atypia), 3) grouping of relevant terms into categories, 4) identification of categories occurring in each patient report, and 5) grouping of patient reports into episodes of care (defined as all reports within 6 months of an initial diagnosis).  Results  Under IRB approval, 27,931 breast pathology reports from Massachusetts General Hospital in 16,208 patients seen between 1990-2007 were analyzed. The results were compared against manually reviewed pathology reports for quality control. For DCIS diagnoses, the initial error rate for both the NLP process and the manual process was 2%. The NLP process was then re-tuned using the identified discrepancies which reduced the error rate to zero. Using the refined model, we identified 1) patients with atypical lesions (atypical ductal hyperplasia (ADH), severe ADH, atypical lobular hyperplasia (ALH), and lobular carcinoma in situ (LCIS)) without prior or concurrent cancer, and 2) patients who developed cancer greater than 6 months post diagnosis.    Conclusion  This process successfully identified high-risk diagnoses that were otherwise relatively inaccessible, and appears to match the accuracy of a human research associate. The results of this first implementation are promising and will be further validated over time. In the future, this approach can be applied to other medical reports and diseases. NLP has significant potential to decrease the cost of research and for improving patient care.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 3001.
Usefulness of intraoperative touch smear cytology in breast-conserving surgery.
Authors: Sumiyoshi K , Nohara T , Iwamoto M , Tanaka S , Kimura K , Takahashi Y , Tsuji M , Kurisu Y , Tanigawa N
Published: December 2010

Abstract #3011

[Objective] In the treatment of breast-conserving surgery, positive margins are closely related to intramammary recurrence, but methods of assessing resection stumps during breast-conserving surgery have not been standardized. The present study investigated the usefulness of intraoperative touch smear cytology at our department.  [Subjects and Methods] From 2005 to 2007, a total of 308 patients underwent breast cancer surgery. Subjects comprised 160 patients who underwent breast-conserving surgery and touch smear cytology. Results of touch smear cytology were compared to those of histological tissue analysis.  [Results] Touch smear cytology displayed 70% sensitivity (14/20), 97.1% specificity (136/140) and a diagnostic accuracy of 93.8% (150/160). Six false-negative cases and 4 false-positive cases were identified. Of the 6 false-negatives, cancer cells were seen in the mammary duct in 5 cases, and the degree of cancer cell atypia in the stump was low. Residual cancer cells were seen in the stump in 18 cases, and additional resection was performed on 16 cases. Cancer cells were identified histologically in the additionally resected tissue in 8 of these 16 cases (50.0%). The direction of positive cytology was towards the mammary papilla in 16 cases, lateral tissue in 5 cases and contralateral mammary papilla in 2 cases. The tendency was that that the greater the cancer cell volume as assessed by touch smear cytology, the higher the frequency of positive margins as assessed by histological tissue analysis.  [Discussion] Histological tissue analysis and touch smear cytology can be performed to assess resection stumps during breast-conserving surgery. Histological tissue analysis is accurate, but: 1) analyzing all areas of a large resection stump is difficult; 2) preparing high-quality frozen sections is difficult due to the high adipose content of breast tissue; 3) assessing the malignancy of intraductal proliferating lesions is difficult; and 4) part of the tissue sample is damaged. Touch smear cytology is easy to perform, offering a very useful technique yielding comparable results to histological tissue analysis.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 3011.
Simultaneous quantitative assessment of tumor cells in blood and bone marrow in primary breast cancer patients.
Authors: Moore AP , Magbanua MM , Scott J , Moore D , Alvarado MD , Esserman LJ , Park JW
Published: December 2010

Abstract #302

Background: Both circulating tumor cells (CTCs) in peripheral blood and disseminated tumor cells (DTCs) in bone marrow have been detected in primary breast cancer, but typically not quantitatively, simultaneously and using the same methodology. Here, we simultaneously assessed CTC and DTC levels in 390 primary breast cancer patients (Stages I-III), including treatment-naïve and post-neoadjuvant patients.  Materials and Methods: CTCs/DTCs were enumerated via a novel two-step assay (IE/FC) consisting of immunomagnetic enrichment (IE) and flow cytometry. First, samples were enriched for epithelial cells using magnetic beads coated with anti-EpCAM monoclonal antibody (mAb). Next, fluorescently labeled mAb specific for an alternate EpCAM epitope was combined with a mAb against CD45 and a nucleic acid stain in multiparameter flow cytometry. Tumor cells were also isolated by IE followed by FACS sorting.  Results: The assay quantitatively detected tumor cells in either blood or bone marrow obtained at time of breast surgery. Tumor cells were detected with greater frequency and at generally higher levels in bone marrow than in blood. In blood, CTCs ranged from 0 to 33.7 cells/mL (mean = 0.9 ± 2.4 cells/mL) and in bone marrow DTCs ranged from 0 to 501.8 cells/mL (mean = 16.8 ± 38.7 cells/mL). Direct comparisons in each patient revealed complex but significant correlation (Spearman's rank correlation = 0.12, p = 0.02) between CTC and DTC levels. In the subgroup of patients receiving neoadjuvant therapy (N=84), 51/79 (64%) were considered positive for DTC (>5.5 cells/mL, with range 0.3-262.2, mean 34±17.1) while 15/79 (19%) were considered positive for CTC (>0.8 cells/mL, with range 0-38.7, mean 1.4±5.7.) Treatment-naïve patients were more likely to be considered positive in the blood (29% vs 19%, p=0.03.) than neoadjuvant patients, possibly reflecting treatment effect. CTCs or DTCs were isolated from selected patients with high cell burden and subjected to expression or genomic analyses, which confirmed the malignant nature of the cells and indicated the feasibility of molecular profiling of isolated CTCs/DTCs.  Discussion: This approach provides quantitative detection and isolation of CTCs/DTCs in blood and bone marrow, and may be useful for analysis of early events in cancer progression.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 302.
Dual targeting of EphA2 and ER restores tamoxifen sensitivity in ER/EphA2-positive breast cancer.
Authors: Gokmen-Polar Y , Toroni RA , Badve S , Bruckheimer E , Kinch MS , Miller KD
Published: December 2010

Abstract #3020

Background: Overexpression and altered function of EphA2 receptor tyrosine kinase are critical in the progression of breast cancer and provide a novel target for breast cancer therapy. We have previously demonstrated that EphA2 overexpression decreases estrogen dependence and tamoxifen sensitivity both in vitro and in vivo. EA5, a novel monoclonal antibody that mimicks the binding of ephrin A to EphA2, reverses the effect of EphA2 overexpression and restores tamoxifen sensitivity in EphA2-transfected MCF-7 cells (MCF-7EphA2) in vitro. Here we report the impact of EA5 on in vivo tumor growth and its ability to overcome in vivo tamoxifen resistance in MCF-7EphA2 xenografts. Furthermore, we investigated the mechanisms by which EphA2 overexpression decreases the estrogen dependence and contributes to tamoxifen resistance in ER+ breast cancer models.  Material and Methods: MCF-7 cells transfected with vector (MCF-7neo) or EphA2 (MCF-7EphA2) were implanted in the right and left mammary fat pads of athymic mice. Treatment with EA5 (5 mg/kg/i.p./5 days/week), vehicle, tamoxifen (1 mg/oral gavage/5 days/week), or EA5 and tamoxifen in combination was initiated once tumors were established. To explore the role of EphA2 overexpression on ER-dependent mechanisms, we used two different ER+/EphA2-transfected cell line models (MCF-7neo/ MCF-7EphA2 and T47Dneo/ T47DEphA2). We measured ER activity and expression of ER-dependent proteins in response to 17β-estradiol (E2;10-10 M), 4-hydroxy-tamoxifen (10-6 M, and EA5 antibody (3μg/mL). Interaction of ER and EphA2 was investigated using GST-pulldown and co-immunoprecipitation approaches. Gene array studies suggested that EphA2 may impact ER via noncanonical pathways, leading to an investigation of focal adhesion kinase (FAK) signaling.  Results: EA5 inhibits primary tumor growth and restores tamoxifen sensitivity in the MCF-7EphA2 in vivo model;EA5 had no impact on in vivo tumor growth in MCF-7neo xenografts. Using T47DEphA2 in vitro model, we verified that EphA2 decreases ER activation in response to E2 stimulation consistent with our earlier results in MCF-7EphA2 model. We found no direct interaction between ER and EphA2 and no difference in expression of canonical ER-dependent proteins, ER coactivators or corepressors. However, E2 stimulation phosphorylates FAKTyr925 in ER+/EphA2+ cell lines but not in ER+/EphA2- cell lines. Treatment of T47DEphA2 cells with EA5 and tamoxifen leads to dephosphorylation of FAKTyr925 in the presence of E2 stimulation.  Conclusion: Our data demonstrate that dual targeting of EphA2 and ER is a promising approach for delaying resistance to tamoxifen. The data support our hypothesis that EphA2 impacts ER function via a FAK dependent pathway.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 3020.
Proteomics analysis of tissue samples obtained before and during letrozole treatment in postmenopausal patients with locally advanced breast cancer.
Authors: Hans V , Evans DB , Lonning PE , Geisler J
Published: December 2010

Abstract #3030

Introduction  We report on the analysis of protein expression in the tumor tissue of postmenopausal patients before and after 4 months of letrozole treatment in the neoadjuvant setting. All patients had ER and/or PGR positive breast cancer and were enrolled in the “FAST-study”. Letrozole was given orally as 2.5 mg once daily. Tumor tissue was obtained by large, open biopsies prior to treatment and during final surgery (in general: mastectomy).  Methods  We used paired samples from 13 postmenopausal breast cancer patients. Breast cancer tissue and plasma samples were obtained prior to treatment and after 4 months of therapy with letrozole in the neoadjuvant setting. Thus, all in all 26 tumor samples were used from 13 individual patients. Proteins were extracted from 20-50 mg of breast cancer tissue, labeled with fluorescent dyes and separated using 2-dimensional difference gel electrophoresis (2-D DIGE). After image analysis, differentially expressed proteins (p<0.01, paired T- and Wilcoxon tests) were identified using MALDI tandem mass spectrometry.  Results and discussion  The 2-D DIGE based method allowed monitoring of 2000-2500 protein species in each sample. After alignment of all these patterns, around 25 cellular proteins were found to be differentially expressed in the pairwise pre-/post-treatment comparison, along with several plasma proteins. Among these cellular proteins were several reported tumor markers such as stathmin and NDPKA, chaperones such as HSP-60, and members of the annexin family. We are currently investigating the correlation between these proteins and the clinical parameters.  To our knowledge this is the first report on proteomics analysis comparing samples obtained before and during treatment with a third-generation aromatase inhibitor. The study demonstrates the feasibility of performing a high-resolution proteomics analysis on tumor material derived from needle biopsies and yielded a number of potential markers, which will require further confirmation in a larger population. The results might be useful to elucidate adaptive mechanisms to estrogen suppression in vivo.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 3030.
Coassociation of ERα and p160 proteins predicts resistance to endocrine treatment; SRC-1 is an independent predictor of breast cancer recurrence.
Authors: Redmond AM , Bane FT , Stafford AT , McIlroy M , Dillon MF , Hill AD , Young LS
Published: December 2010

Abstract #3032

The p160 coactivators AIB1 and SRC-1 are known to play a critical role in modulating transcription in breast cancer cells in conjunction with ligand-bound estrogen receptor. The interactions of the p160 proteins with this nuclear receptor are also important in the development of resistance to endocrine treatments. Using quantitative coassociation immunofluorescent microscopy, the colocalisation of the p160s and ERα was increased in the LY2 endocrine resistant cell line following treatment with the anti-estrogen tamoxifen in comparison to the endocrine sensitive MCF-7 cell line. In cell cultures derived from patient tumours at the time of primary surgery prior to treatment, there was an increase in association of the coactivators with ERα following treatment with estrogen but dissociation was evident in the presence of tamoxifen. Immunohistochemical staining of a tissue microarray, constructed from 560 breast cancer patients, revealed that SRC-1 was a strong predictor of reduced disease-free survival, both in patients receiving adjuvant tamoxifen treatment and untreated patients (p<0.0001 and p=0.0111 respectively). AIB1 was not a significant independent predictor of disease recurrence. SRC-1 was assigned a hazard ratio of 2.12 when survival analysis using a Cox proportional hazards model was applied. Quantitative coassociation analysis of the p160 coactivators with ERα in the patient TMA revealed significantly stronger colocalisation of SRC-1 and ERα in patients who are known to have relapsed than those patients who did not recur (p=0.00001). This data suggests SRC-1 is pivotal in tumour aggressiveness and is a powerful predictor of progression of disease in breast cancer patients.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 3032.
Amphiregulin is an estrogen-responsive gene which predicts recurrence-free survival in patients with estrogen receptor positive breast tumors.
Authors: Kenny PA
Published: December 2010

Abstract #3039

Estradiol is a potent growth factor in estrogen receptor positive breast disease. Activated estrogen receptors have been reported to activate many hundreds of genes, yet the key effector genes for breast cancer cell proliferation remain poorly understood. Here, we have focused on one such target gene, Amphiregulin, which encodes a ligand for the Epidermal Growth Factor Receptor. We report that Amphiregulin is induced by estrogens and repressed by estrogen receptor antagonists in several breast cancer cell lines (MCF7, ZR75B and T47D), and that it promotes breast epithelial cell proliferation. Using microarray analysis of primary tumors, we find that Amphiregulin and ER-alpha expression are highly positively correlated in human breast cancer cases (n = 295). Additionally, in a cohort of women with ER+ breast tumors (n = 60), high levels of Amphiregulin predicted response to tamoxifen. We further demonstrate that Amphiregulin production can be blocked by inhibiting TACE/ADAM17, a metalloproteinase which regulates the shedding of Amphiregulin from the cell surface.  Collectively, these data suggest that Amphiregulin may be a key transcriptional target of estrogen receptor signaling and a marker for Estrogen Receptor positive tumors that are especially dependent on estrogen and, therefore, particularly sensitive to tamoxifen.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 3039.
A role for GATA-3 in control of estrogen receptor alpha expression.
Authors: Hostetter C , Licata LA , Keen JC
Published: December 2010

Abstract #3050

Expression of the GATA-3 binding protein correlates with estrogen receptor (ER) alpha expression in primary and cultured breast cancer cells. While the significance of this correlation and role of GATA-3 in breast cancer remains unclear, GATA-3 may serve as a good prognostic indicator in breast tumors. Recently, a reciprocal interaction between GATA-3 and ER has been demonstrated. GATA-3 may regulate ER levels through binding to an enhancer region upstream of the ER gene while ER can associate with and regulate GATA-3 expression. Meta-analyses of several cancer microarrays indicate that GATA-3 is co-expressed with ER and other ER responsive genes, further suggesting a role of GATA-3 in the ER signaling pathway. Our laboratory has been investigating the role of the RNA binding protein HuR/ELAV1 in control of mRNA stability in breast cancer cells. We have previously reported that ER mRNA stability is increased through binding of HuR to the 3'UTR. Microarray analysis of those transcripts associated with HuR in MCF7 cells showed that GATA-3 mRNA associates with HuR compared to an IgG control. Biotin pulldown assays using a biotinylated GATA-3 RNA probe confirmed that HuR binds to the GATA-3 3'untranslated region (UTR). Inhibition of HuR using siRNA probes decreased GATA-3 mRNA and protein expression, further indicating a role for HuR in regulation of GATA-3 mRNA stability. siRNA mediated inhibition of GATA-3 expression significantly reduced ER mRNA and protein levels. Surprisingly, we found that GATA-3 bound to the ER mRNA by binding to two out of four regions within the ER 3'UTR, suggesting that while GATA-3 may play a role in upregulation of ER mRNA transcription through the enhancer interaction, GATA-3 may stabilize ER mRNA once transcribed by binding to the ER 3'UTR. Studies are ongoing to fully characterize this role of GATA-3 in control of ER mRNA stability.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 3050.
Targeting the peroxisome proliferator-activated receptor gamma one in the treatment of breast cancer.
Authors: Kilgore MW , Zaytseva YY , Wang X , Southard RC
Published: December 2010

Abstract #3055

Peroxisome proliferator-activated receptor gamma 1 (PPARγ1) is a member of the nuclear hormone receptor superfamily and is over expressed in breast cancer compared to human mammary epithelial cells (HMEC). Although synthetic ligands of PPARγ have been reported to mediate differentiation, inhibit proliferation, and induce apoptosis of cancer cells in culture it remains unclear whether these are mediated through PPARγ or by off target effects. Furthermore, the controversial nature of these studies and the disappointing results from clinical trials raise questions about the efficacy of these drugs in treating breast cancer. Recent work from our lab and others suggests that targeting the expression or endogenous transactivation of PPARγ represents a novel strategy in the treatment of breast cancer. We have previously demonstrated that the increase in PPARγ1 expression in seen in breast cancer cells is driven by a tumor-specific promoter. Furthermore, we have identified the myc-associated zinc finger protein (MAZ) as a critical mediator of PPARγ1 expression in these cells. In this study, we further demonstrated the significance of MAZ in the transcriptional regulation of PPARγ1 and confirmed that high expression of PPARγ1 in MCF-7 breast cancer cells is MAZ dependent. Additionally, using RNA interference (RNAi) techniques to knockdown PPARγ1 or MAZ or driving the expression of a dominant negative form of PPARγ1 in MCF-7 breast cancer cells, we demonstrated that blocking the expression of either protein resulted in reduced cellular proliferation and enhanced apoptosis. These findings suggest that in the absence of any exogenous ligand an increase in PPARγ1 signaling, driven at least in part by MAZ, contributes to an imbalance between proliferation and apoptosis, and may be an important mediator or of breast cancer progression. These results also highlight the need to understand the molecular mechanisms that drives tumor specific expression of PPARγ1 and development pure antagonist as a therapeutic treatment of this devastating disease.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 3055.
Does the recurrence of pregnancy-associated breast cancer during subsequent pregnancy 10 years after mastectomy support the unique hormonal milieu of pregnancy as a cause of tumorigenesis? A case and literature review.
Authors: Schlager A , Unger JG , Schnabel F
Published: December 2010

Abstract #3058

We report a case of pregnancy-associated chest wall recurrence of breast adenocarcinoma 10 years following mastectomy and reconstruction for initial pregnancy-associated breast cancer (PABC). This represents the first reported case in the literature of a pregnancy-associated recurrence of pregnancy-associated breast cancer. The patient had no episodes of recurrent breast cancer during the interim 10 years, and only experienced a recurrence with a rapidly growing left chest mass during the subsequent pregnancy. Thus, the possibility that the malignancy may have been triggered by the pregnancy cannot be ignored.  Although prior studies following breast cancer patients and their subsequent pregnancy have not shown an increase risk of recurrence, these studies were performed in “all comers” with breast cancer, and would not have had the statistical power to distinguish a small subset of patients with an increased risk, such as PABC patients. As a result, these patients have been assumed to be at no greater risk for recurrence with pregnancy and have been advised as such.  It is reasonable to assume that PABC patients may, in fact, represent a unique subset of breast cancer patients with the unique hormonal milieu of pregnancy playing a mechanistic role in the propagation and proliferation of the disease. In this case a patient with prior PABC had a rapidly growing progesterone receptor positive breast cancer during a subsequent pregnancy. Further, studies have indicated that there is a possibility that the hormone status of women during pregnancy may in fact lead to a change in tumor characteristics, and therefore their risks. This instance would support the concern and represent the sentinel report describing a new subset of breast cancer with a unique spectrum of risk factors.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 3058.
Blocking internalization of pBP1, a secreted homeoprotein, kills breast cancer cells but not normal breast epithelial cells.
Authors: Rheey J , Bivona L , Berg P
Published: December 2010

Abstract #3068

Background: BP1 is a member of the homeobox gene family, a highly conserved family of transcription factor genes. pBP1 is a homeotic protein that is upregulated in 80% of invasive ductal breast tumors. We have previously shown that the percentage of pBP1 positive breast cancer cases increases with the extent of cellular proliferation and carcinogenesis. BP1 expression is associated with aggressive tumors: 100% of ER negative tumors were BP1 positive, compared with 73% of ER positive tumors. pBP1 is secreted by MCF-7 cells, T47D cells, and MDA-MB-231 breast cancer cells, but not by MCF10A or H16N2 normal breast epithelial cells. In all cell lines tested, secreted pBP1 is internalized, resulting in stimulation of growth. Moreover, incubation of MCF-7 or MCF10A cells in recombinant pBP1 results in stimulation of several oncogenes, including bcl-2, Twist, and Met. These results led us to ask whether we could abrogate mitogenic stimulation by addition of anti-pBP1 antibody.  Materials and Methods. Cells were incubated in 10 µg/ml anti-BP1 Ab (Novus) or 10 µg/ml IgG for three days. Growth was measured using MTT assays and apoptosis was assessed by flow cytometry after staining with Annexin V (Trevigen).  Results. Addition of anti-BP1 Ab to the growth medium of MCF-7 cells, T47D cells, and MDA-MB-231 cells caused significant growth inhibition and apoptosis. In contrast, addition of anti-BP1 Ab to the growth medium of MCF10A and H16N2 cells did not lead to significant growth inhibition or apoptosis.  Discussion. Our novel finding that pBP1 is secreted by breast cancer cells but not normal breast epithelial cells was the impetus to test the effect of adding anti-BP1 Ab to the growth media of both types of cells. Remarkably, anti-BP1 Ab kills only the breast cancer cells, suggesting they may be BP1-dependent. Our data indicate it may be possible to specifically target BP1 positive breast cancer cells in the 80% of women with BP1 positive tumors.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 3068.
Protein tyrosine kinase 6 (PTK6): a new/potential therapy target in breast cancer?.
Authors: Aubele M , Walch AK , Braselmann H , Ludyga N , Atkinson MJ , Luber B , Auer G , Bartlett JM
Published: December 2010

Abstract #3073

Background: Elevated expression of the cytoplasmic tyrosine kinase PTK6 (breast tumor kinase, BRK) occurs in approximately two-thirds of primary breast tumors, where it is implicated in EGF receptor-dependent signaling and epithelial tumorigenesis. The role of PTK6 in breast cancer development and prognosis is unclear, its function in-vivo remains undefined.  Material and Methods: We performed a retrospective study of PTK6 expression in 426 archival breast cancer samples from patients with long term (≥240 months) follow up. Immunohistochemical analysis of the following proteins was determined: PTK6, all 4 HER receptors, Sam68 (a substrate of PTK6), and the signalling proteins MAP kinase (MAPK), phosphorylated MAPK (P-MAPK), and PTEN. Correlations between markers were determined, and the prognostic value of parameters by univariate and multivariate survival analysis. We compared PTK6 gene copy number with the protein expression levels using FISH analysis.  Results: The disease-free survival of patients ( ≥240 months) was positively correlated with PTK6 protein expression, and was inversely correlated with nodal status and tumor size (p≤0.01). A time dependence for the prognostic value of key markers was evident, with HER2/neu only showing prognostic significance at early time points (60 months).  PTK6 expression in tumor tissue significantly correlated with the expression of PTEN, MAPK, P-MAPK, and Sam68 (p≤0.05). To investigate whether these correlations may be due to molecular interactions, we used the T47D cell line for immunoprecipitation and Western Blot analysis. Co-precipitation-type protein-protein interactions could be demonstrated between PTK6 and MAPK, P-MAPK, HER2/neu, HER3, HER4, PTEN, and Sam68.  Quantitative analysis revealed an increased PTK6 gene copy number in 75% of the tumors. Polysomy of chromosome 20 was responsible for most cases (47%) whilst gene amplifications were less frequent (28%).  Discussion: We show that PTK6 protein expression is of significant prognostic value in predicting the long term outcome of breast carcinomas. Further, several potential binding partners were identified, indicating the involvement of PTK6 in three main signal transduction pathways.  Supported by: Deutsche Krebshilfe # 10379 / EU LSHC-CT-2004-503426.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 3073.
Synergisitic inhibition of DNA repair by combining ß-lapachone with Nu7026 as a strategy for killing primary and metastatic breast cancers.
Authors: Bey EA , Kessinger C , Blanco E , Bachoo R , Williams N , Boothman DA
Published: December 2010

Abstract #3078

The primary tumors of most breast cancer patients are removed via resection or complete mastectomy. However, years after having their primary tumors removed many patients will need chemotherapy for metastatic disease that arises in distal sites such as lung and brain. It has been reported that many breast cancers overexpress the 2 electron oxidoreducase NQO1, a marker of tumorigenesis for a variety of cancers. Since NQO1 expression levels are low in normal tissue but elevated 10-100 fold in breast cancers, we hypothesized that NQO1 levels could be exploited for chemotherapy with ß-lapachone. ß-lapachone is a naturally derived plant product that is bioactivated by NQO1. In this study we show that ß-lapachone (a DNA repair inhibitor) synergizes with the DNA-PK inhibitor Nu7026 to kill human breast cancer cell lines that express NQO1. Normal human breast tissues and breast cancer cell lines that did not express NQO1 were not affected by ß-lapachone. In addition by using in vivo breast cancer models we demonstrated that ß-lapachone effectively inhibited breast tumor growth in athymic mice. We also showed that ß-lap was efficacious against lung tumors derived from MDA-MB-231 breast cancer cell lines. Finally, our simulated brain metastatic model showed that ß-lap accumulated in brain tumors derived from MDA-MB-2331 cells. Together these data suggest that ß-lap and ß-lap in combination Nu7026 may be a potent combination for treating primary and metastatic breast cancer disease.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 3078.
Prophylactic surgical uptake in a cohort of BRCA carriers: an Irish perspective.
Authors: Carroll PA , Kennedy J , Boyle T , Connolly EM
Published: December 2010

Abstract #3095

Introduction: Women with BRCA 1/2 mutations have a lifetime risk of approximately 45-87%1 for developing breast cancer and a 20-60% risk for ovarian cancer. The managment of these patients including prophylactic surgery is contorversial and presents difficult choices for patients. Uptake of prophylactic surgery in BRCA carriers in an Irish setting has not previously been investigated.  Methods: St. James hospital is a tertiary referral hospital for BRCA counselling and testing. All information was gathered and analysed from the Cancer genetics department database in our institution which provides a counselling service for a number of hereditary diseases.  Results: Deleterious BRCA 1/2 mutations have been identified in 57 families since testing began in 1992. There are a total of 109 women in this cohort.  54% of these women were diagnosed with primary breast cancer. 23 (47.9%) women were symptomatic on presentation. Only 3 (6.3%) were detected during screening. Inital surgery and prophylactic surgery uptake in this group is displayed in table 1.  31% (n=34) of our BRCA carrier cohort remain unaffected. Uptake of surgical prohylaxis is outlined in table 2. 17 women in this group opted for no form of prophylaxis. 88% of the women will require regular screening for breast cancer.  Age at bilateral prophylactic masectomy was 30.5 years. Of the 39 women in both groups who elected for bilateral salpingo oophorectomy, only 28% (11) women were under the age of 40 years.    CPM Contralateral Prophylactic Mastectomy  TAH/BSO Total Abd Hysterectomy/Bilat Salpingo oopherectomy    TAH/BSO Total Abd Hysterectomy/Bilat Salpingo oopherectomy  BPM Bilateral Prophylactic Mastectomy  Discussion: Prophylactic mastectomy offers a 95% risk reduction for breast cancer of 95%, despite this, a survival benefit has not been proven. Prophylactic oophorectomy before the age of 40 years has been associated with a reduction in breast cancer of 50% and up to 80% reduction in ovarian/peritoneal malignancy.There is also data to support that oophorectomy provides a survival benefit in BRCA carriers. The rates of uptake of prophylactic surgery in both the affected and unaffected groups in our cohort are lower than that of international standards2. The reasons for this are currently under investigation.  References:  1. Ford D et al. Genetic heterogeneity and penetrance analysis of the BRCA1/2 genes in breast cancer families. Am J Hum Genet 1998;62(3):676-89.  2. Metcalfe KA, et al.International variation in rates of uptake of preventive options in BRCA1/2 mutation carriers. Int J Cancer. 2008 May 1;122(9):2017-22.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 3095.
BRCA mutation in Chinese population: results from the Hong Kong hereditary and high risk breast cancer programme (HRBCP) and the Hong Kong hereditary and high risk breast cancer family registry.
Authors: Kwong A , Wong CL , Ma ES , West DW , Ford JM
Published: December 2010

Abstract #3097

Background: Little is known about the frequency and spectrum of BRCA 1/2 mutations among Chinese population. Previous research suggests possible differences in the prevalence and penetrance of inherited mutations in these genes among Asians and the earlier presentation of breast cancer in Asian population may have more relevance to genetic causes. The access to genetic counseling and testing in Asia have been limited. The HRBCP (www.HRBCP.org) was established in 2007. This is first in Hong Kong and provides genetic testing, counseling and research on the spectrum of the disease in Asia and in particular Chinese population. The data from this programme will be entered in the newly established The Hong Kong Hereditary Breast Cancer Family Registry.  Methods: Probands who were diagnosed to have breast cancer age 50 or below, have a family history or personal history of breast and/or ovarian cancer, bilateral breast cancer, male breast cancer, triple negative breast carcinoma were recruited. Genetic counseling were given and consent for testing were obtained. Blood and tumor samples were collected. The entire coding regions of the extracted DNA/RNA and flanking introns of BRCA1 and 2 were screened for germline mutations using full gene sequencing and MLPA.  Results: A total of 119 Chinese have been sequenced and analysed. 21 (18%) deleterious mutations were identified of which 4 were novel mutations. 8 (38%) were BRCA 1 mutations and 13 (62%) were BRCA 2 mutations. 66.6% had first degree relatives with breast cancer. 49 (41.2%) were under the age of 40 years old of which 11 (22%) were found to carry a BRCA mutation. 7 (64%) of this younger age group with mutation have family history of breast cancer. For the other 4, 1 had triple negative cancer, 1 had a family history of ovarian cancer, 1 unknown and 1 with no risk except for young age. Of the 37 family members tested, 21 (57%) have BRCA mutations and only 9 of them have breast or ovarian cancers. 1 only had stomach cancer. 6 male family members were found to carry the BRCA mutation with no history of cancer.  Conclusions: Approximately 18% of the tested blood samples from clinically high risk Chinese women carry a deleterious mutation in BRCA1 or 2. In contrast to Caucasian data, there was a comparative higher rate of BRCA2 mutations. Further research on the spectrum of the mutations in Chinese and other Asian population will allow the tailor-making of an ethnic-based risk assessment model and also screening, prophylactic and preventative measures to be taken.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 3097.
The benefit using an integrated electronic trial platform for a neoadjuvant therapy trial (HEDON) in primary breast cancer – results of an evaluation study.
Authors: Schwarz-Boeger U , Zahlmann G , Paepke S , Schmidt M , Jacobs VR , Kiechle M , Harbeck N
Published: December 2010

Abstract #3114

Background: At present, academic medical centers conducting investigator initiated trials (IIT) have the choice of relying on paper or going electronic regarding electronic data capture and data management solutions. Many of the electronic systems have originally been developed for requirements of the pharmaceutical industry and all of the currently available systems would thus be isolated in the existing IT setting of the medical centers.  We here suggest an approach to integrate a data management system into existing hospital information systems (HIS) and to provide the end users (e.g. physicians, investigators, study nurses) with consolidated user interfaces and integrated systems.  Methods: In 2005, a collaboration between the Department of Obstetrics and Gynecology, Klinikum rechts der Isar, Technical University of Munich and Siemens Medical Solutions was initiated as a pilot project.  An integrated platform for clinical trials was developed and implemented to support the HEDON trial (Herceptin-Docetaxel-Neoadjuvant), an investigator initiated clinical phase II trial concerning primary systemic therapy of breast cancer. The HEDON trial started in February 2006 and will have recruited all of a total of 90 planned patients in the end of this year. In addition, a translational research trial (TransHEDON) evaluating therapy response markers is an essential component of the clinical trial.  Using the integrated platform, clinical data as well as laboratory data are automatically populated from the HIS into the electronical study CRFs, after data confirmation by investigators or study nurses (depending on their defined role) using the validation buffer of the system. The existing PACS and HIS are made accessible from the platform which has special capabilities to greatly improve user interactions with all relevant systems. To quantify the benefit of such an integrated system, an integrated evaluation project was set up including the quantification time savings for all involved personnel. For a pre-defined time period, data capture and data management processes are performed paper based and electronically. This allows thorough and critical analysis of all involved processes  Resukts; Errors that occur specially due to manually entering laboratory data can be dramatically minimized using such an electronical data capture system. Another important benefit is time savings. The data will be presented.  Conclusion:This single-sign-on electronic platform for clinical trials has been validated in its whole and fulfils the regulatory requirements as specified by GCP or 21 CFR part 11 compliance for trials of the FDA. We consider this integrated system to be very helpful for implementation of clinical trials into a routine hospital setting.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 3114.
Phase II trial of dasatinib in triple-negative breast cancer: results of study CA180059.
Authors: Finn RS , Bengala C , Ibrahim N , Strauss LC , Fairchild J , Sy O , Roche H , Sparano J , Goldstein LJ
Published: December 2010

Abstract #3118

Background: Dasatinib (SprycelR; BMS-354825) is a potent orally-available inhibitor of Src-family kinases and other kinases with anti-proliferative, anti-osteoclastic and anti-metastatic activity demonstrated pre-clinically. Expression profiling suggested that basal-like cancers may be preferentially sensitive to dasatinib. Methods: A Phase II single-agent trial of dasatinib, using a continuous schedule, was performed in patients with advanced triple-negative (as proxy for basal-like) breast cancers. Subjects were required to have measurable locally-advanced or metastatic triple-negative (ER/PR-negative, Her2-normal) disease and prior anthracycline and/or taxane therapy. A 2-stage Gehan design was adopted, with RECIST-defined response as primary endpoint; subjects discontinued for toxicity were considered non-responders. The original dasatinib dose of 100 mg BID (n=21) was reduced to 70 mg BID (n=23) to improve tolerability. Biomarkers were analyzed in tumor and plasma samples obtained for PK analysis. Results: From 12/06 through 12/07, 44 subjects were treated at 14 institutions: median age 55 yrs, median time from diagnosis 30 mo, prior therapy for advanced disease in 29 (66%). Of 43 response-evaluable subjects, 7 discontinued for toxicity prior to on-study assessment. Of 36 subjects with radiographic assessment, there were 2 confirmed PR [1 continues >1 year + 1 discontinued for intolerance at week 16] plus 2 SD lasting >16 weeks. Four additional subjects had transient clinical benefit reflected by improvement in bone pain (anecdotal) or short-term tumor shrinkage (reductions of 11 - 29%). Tolerability was improved at a dose of 70 mg compared with 100 mg BID. In preliminary analysis, fewer subjects experienced any serious adverse event (13% at 70 mg BID vs 48% at 100 mg BID), fewer reported Grade 3 toxicity, including gastrointestinal (10% vs 26%), pleural effusion (4% vs 9%), generalized edema (0% vs 9%) or pericardial effusion (0% vs 9%), and fewer had dasatinib dose reduction (24% vs 61%). Fatigue, myalgia/arthralgia and headache were comparable at the two doses. No Grade 4 drug-related events occurred. Grade 3-4 abnormal laboratory values were uncommon. Biomarker and PK data will be presented. Conclusions: Modest but encouraging single-agent activity was observed with dasatinib in patients with advanced triple-negative breast cancers, with clinical benefit rate of 9.3% (4/43). Future studies are warranted to address optimal dose and schedule of dasatinib in combination with chemotherapy for this challenging tumor type.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 3118.
RAD001 (everolimus) in combination with weekly paclitaxel and trastuzumab in patients with HER-2-overexpressing metastatic breast cancer with prior resistance to trastuzumab: a multicenter phase I clinical trial.
Authors: O'Regan R , Andre F , Campone M , Naughton M , Manlius C , Pylvaenaeinen I , Sahmoud T , Hurvitz S
Published: December 2010

Abstract #3119

Background: Resistance to trastuzumab (H) is associated with loss of PTEN and AKT/mTOR activation. Preclinical models demonstrate that RAD001, an oral inhibitor of mTOR (a downstream component of the PI3K/AKT signaling pathway), enhances the efficacy of trastuzumab and suggest that RAD001 could reverse resistance to trastuzumab.  Methods: Paclitaxel (T, 80 mg/m2) IV was administered over 60 min on D 1, 8 and 15 q4w. Trastuzumab 4 mg/kg loading dose was administered IV over 90 min on Day 1 (if patient (pt) was not already receiving H), followed by weekly 2 mg/kg IV over 30 min. RAD001 was administered as either a daily (5 and 10 mg) or weekly (30, 50 and 70mg) cohorts. Treatment continues until progression or unacceptable toxicity.  Results: As of June 9 2007, 22 heavily pretreated pts were enrolled; 6 to the 5mg daily, 6 to the 10mg daily and 10 to the 30mg weekly cohorts. All pts were resistant to H and all patients, except one, received prior taxane. Median number of prior regimens was 6 (range: 1-26). Two DLTs were observed during the first cycle; febrile neutropenia on the daily 5mg arm and confusional state on the weekly 30mg arm. To date, 17 pts were evaluable for efficacy (Table 1). Among the 7 pts who were resistant to both T and H, 5 had PR (4/5 pts and 1/2 on the 5mg daily and 30mg weekly cohorts, respectively) and 2 had SD >16 weeks. Fifteen pts remain on treatment. The most commonly reported AEs are neutropenia and stomatitis. Recruitment continues.  Conclusions: RAD001 is well tolerated in combination with weekly TH regimen. Preliminary efficacy shows a high rate of tumor responses in heavily pre-treated pts, including pts resistant to both T and H. Mature results and biomarkers data will be presented.   

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 3119.