bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P5-17-03: A new strategy for testing therapies to be effective against seemingly untreatable cancers
Authors: Singh B , Raythatha MR , Shamsnia A , Lucci A
Published: August 2014

Background: Despite a tremendous progress in understanding the genome and biology of cancer, there has been only a slow and limited progress in treating advanced disease and metastases. One main reason for this lack of progress is that preclinical testing is performed with little attention to the true nature of the disease. Most therapy-resistant cancers are highly heterogeneous and they constantly evolve. Methods: Here we describe a strategy for getting to the roots of an aggressive cancer and then evaluating therapies (single agent and combination therapies) for their ability to eradicate such roots. To model the roots of often untreatable disease, i.e., triple-negative inflammatory breast cancer, we started with a well-characterized SUM149 cell line (harbors BRCA1 mutation, oncogenic p53 mutation, has defective RB pathway due to p16 deletion, and lacks of PTEN protein expression). The roots can be selected by applying metabolic challenges (e.g., lack of glucose or glutamine in culture medium). The cell-based model based on most adaptable subpopulation of cancer cells is ideally suited for evaluating combination therapies that are required for combating a heterogeneous and evolving disease. Results: The rare metabolically adaptable (MA) variants (0.01% cells in SUM149 cell line), which can survive and grow without glutamine indefinitely, are structurally adaptable as well (highly enriched in mesenchymal phenotype), resistant to chemotherapeutic drugs, and highly metastatic to multiple organs- lungs, liver, brain, and skin from fat pad xenografts in nude mice (Singh et al., PLoS ONE, 2012). Our function-based selection is more powerful than the biomarker-based selections for isolating the type of rare cells that drive the disease. Our recent CGH array and gene expression microarray data provided evidence of widespread chromosomal imbalances associated with the selection and evolution of MA variants. Interestingly, these analyses revealed that the molecular networks that drive obesity (thus important in cancer) are also important in the survival of MA variants. We are now investigating a variety of experimental therapies (safe sensitizers, inhibitors of drivers, and chemotherapies) to identify those which may eradicate most adaptable breast cancer cells in patients with triple-negative breast cancer. It is noteworthy that our assessment of response versus resistance to therapies is based on long-term assays (1-2 months). Short-term assays, which are often used to assess proliferation and apoptosis, are not good at predicting response in the clinic. Conclusions: Our studies suggest that the MA cells are resistant to most drugs tested thus far as single agent, supporting the validity of our cell-based model. The strategy described here to realistically model therapy resistance and to overcome it has a potential of big clinical impact if it can be aligned with clinical trials.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P5-17-03.
Abstract P6-02-02: mRNA expression of death associated protein 3 (DAP3) and human breast cancer: Clinical correlations and in vitro evidence
Authors: Wazir U , Ye L , Sanders AJ , Kasem A , Jiang WG , Sharma AK , Mokbel K
Published: August 2014

BACKGROUND: To our knowledge, this study is the first to focus on the potential role for death-associated protein 3 (DAP3) in human breast cancer.

MATERIAL AND METHODS: mRNA expression of DAP3 in breast cancer tissues (n = 127) and normal background tissues (n = 33) were determined using quantitative polymerase chain reaction (qPCR) and were correlated with clinico-pathological data accumulated over a 10-year follow-up period.

Furthermore the effects of DAP3 knock down in breast cancer cell lines (MCF-7 and MDA-MB-231) were investigated. The cells were subjected to conventional growth, adhesion and invasion assays.

In addition to the above, electric cell-substrate impedance sensing (ECIS) assay and annexin V/propidium iodide binding apoptosis assay were performed. For the apoptosis assay, the cells were subjected to 48 to 72 hours of serum hunger (depending on cell line) before being analyzed using a flow cytometer.

RESULTS: The expression of DAP3 mRNA was demonstrated to decrease with increasing Nottingham Prognostic Index (NPI2 vs. NPI3, p = 0.036), TNM stage (TNM1 vs. 3, p = 0.07), and tumour grade (grade 1 vs. 3, p = 0.08). Lower DAP3 expression levels were significantly associated with local recurrence (p = 0.013), distant metastasis (p = 0.0057) and mortality (p = 0.019).

Kaplan-Meier plot analysis suggests that patient with higher levels of DAP3 expression had better overall survival compared to patients with lower levels of DAP3 expression (p = 0.075).

DAP3 knock down strains in both cell lines demonstrated increased growth and migration compared to controls during ECIS.

In addition, DAP3 knock down strains of MCF-7 appeared more resistant to serum hunger when compared to controls during the apoptosis assay.

CONCLUSIONS: This study demonstrates an inverse association between DAP3 mRNA levels and tumour stage and clinical stage in breast cancer. This is in keeping with the role of DAP3 as a pro-apoptotic protein. The in vitro evidence lends further credence to this hypothesis. The role of DAP3 may require further investigation to better understand the role of apoptosis in breast carcinogenesis, and may potentially serve as a prognostic marker for human breast cancer.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P6-02-02.
Abstract P6-02-03: TGF-b1-inducible protein BIGH3 (transforming growth factor beta-induced gene human clone 3): A new mechanism inducing breast cancer cell apoptosis
Authors: RezaPoor F , Moritz R , Phelix CF , LeBaron RG
Published: August 2014

Cancer development and progression is a complex process involving tumor cell mechanisms as well as tumor-host cell interactions. Using autocrine and paracrine mechanisms the cytokine TGF-b1 can influence cancer progression, in part through promoting the synthesis and secretion of extracellular matrix molecules. TGF-β1 strongly upregulates the gene TGFBI, which encodes a secretory protein named BIGH3. BIGH3 is an extracellular matrix, adhesion-class proapoptotic protein characterized by four fasciclin-1 (FAS1)-like domains, a cysteine-rich region and several different integrin-binding sequences. The C-terminus of BIGH3 encodes the integrin-binding peptides EPDIM and RGD. These sequences, and the integrin α3β1, have been implicated as important components of BIGH3-induced apoptosis. Breast cancer cells express the integrin α3β1, which is reported to play various roles including signaling, invasion and metastasis. Breast cancer cells and tumor-host cells express BIGH3 protein, which is expected to accumulate in the presence of TGF-β1. The above findings highlight the molecules TGF-β1 and α3β1 in tumor progression and are the basis for our hypothesis that BIGH3 protein itself is sufficient to induce an increase in breast cancer cell apoptosis. In order to test this hypothesis we utilized human breast cancer MCF-7 cells, human recombinant BIGH3 and TUNEL assays. Here, we show for the first time that exogenous human recombinant BIGH3 in cancer cell growth medium evokes a statistically significant increase in breast cancer cell apoptosis. A concentration curve shows that there is a direct relationship between the quantity of BIGH3 in the cells’ milieu and the number of cells undergoing apoptosis. This finding is consistent with receptor-ligand binding and indicates that BIGH3 is a potential target for breast cancer treatment. Importantly, in-house developed anti-BIGH3 antibody completely blocked apoptosis when compared to apoptotic cells in negative control conditions. Stimulation of BIGH3 gene expression in breast cancer cells, and providing exogenous BIGH3 to breast cancer cells, are expected to diminish breast cancer progression. Along this line, BIGH3 may be valuable as an agent for neoadjuvant or adjuvant therapy for human breast cancer patients.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P6-02-03.
Abstract P6-04-17: The irreversible c-Jun N-terminal kinase (JNK) inhibitor, JNK-IN-8, sensitizes basal-like breast cancer cells to lapatinib
Authors: Ebelt ND , Van Den Berg CL
Published: August 2014

Basal-like breast cancers represent 15-20% of breast cancers diagnosed each year. These tumors appear earlier in life and have the worst prognosis due to high prevalence of metastasis. Endocrine and molecularly targeted therapies such as trastuzumab or lapatinib are ineffective against this tumor subtype. A high frequency of p53 mutations, low BRCA1 expression along with high epidermal growth factor receptor (EGFR) expression provide some insight into what types of targeted therapies may be useful for basal-like tumors in the future. We have recently shown that JNK2 mediates the expression of p53 and BRCA1 in response to EGFR activation to promote epithelial to mesenchymal transition and metastasis. Further, high JNK2 expression is associated with poorer survival in patients with basal-like breast cancer. These findings suggest that the JNK pathway may be a promising target for basal-type breast cancer treatment.

Commonly used competitive ATP inhibitors of JNK have suffered from lack of specificity for JNK. However, the Gray laboratory recently developed highly specific covalent inhibitors of JNK (Zhang, et al. 2012). The agent showing the best properties is JNK-IN-8. Our data show that treatment of human and mouse basal-like, breast cancer cell lines with JNK-IN-8 sensitizes them to lapatinib. Single agents had minor affects on cell viability and cell cycle regulation, but combination treatment led to G2/M arrest and endoreduplication along with synergistic apoptotic responses. Further, the expression and electrophoretic mobility of mitotic arrest deficient 2 (MAD2) (a protein mediating chromosomal segregation) decreased after combination treatment which could explain the chromosomal duplication abnormalities observed. Combination treatment using lapatinib and JNK-IN-8 modulates extracellular signal-regulated kinase (ERK) phosphorylation unlike either drug alone suggesting that both JNK and ERK target G2/M check point proteins, and when perturbed, sensitize basal-type breast cancer cells to targeted therapies.

We conclude that use of specific JNK inhibitors in tumors that are resistant to lapatinib, or perhaps trastuzumab, may sensitize them to treatments with these drugs, and may also prevent resistance to these drugs in tumors that were initially responsive. Considering that JNKs are nodes in receptor tyrosine kinase and many other signaling pathways, JNK-IN-8 treatment may improve the efficacy of other targeted therapies as well.

Zhang, T. et al. “Discovery of potent and selective covalent inhibitors of JNK”. Chem Biol. 2012 Jan 27;19(1):140-54.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P6-04-17.
Abstract P6-04-21: Novel therapy of liver metastases of cancer using abraxane loaded into multi-stage nanovector
Authors: Tanei T , Liu X , Ferrari M , Godin B , Yokoi K
Published: August 2014

Purpose; To investigate whether the Abraxane loaded within Multi-Stage Nanovector (MSV) system (MSV-ABX) is more effective in therapy of liver metastases of breast and lung cancer as compared to Abraxane (ABX).

Experimental Design; MTT assay was performed to evaluate sensitivity of 4T1, murine breast cancer cells and 3LL, murine lung cancer cells to ABX or MSV-ABX in vitro. Experimental liver metastases were produced by intra-splenic injection of 4T1 or 3LL cells in Balb/C mice. The mice bearing liver metastases were treated with intravenous injection of (1) control (PBS), (2) ABX or (3) MSV-ABX. Then the mice were sacrificed and liver weight was measured to evaluate the therapeutic efficacy. To evaluate the biological the effects of therapeutics, we performed histological and immunohistochemical analysis of the tumor specimens. Apoptosis of 4T1 and 3LL tumor cells were evaluated by TUNEL staining, and proliferation of the tumor cells were stained using antibody to Ki67. Tumor diameter was measured in H&E stained sections.

Results; There was no significant difference in the sensitivity of 4T1 and 3LL cancer cells to ABX and MSV-ABX, in vitro. All the mice injected with 4T1 or C3LL cells developed multiple liver metastases. The liver weight was significantly lower in the mice treated with MSV-ABX as compared to those in the other groups. The tumor diameters in the liver were significantly smaller in the mice treated with MSV-ABX as compared to those in the other two groups. Immunofluorescent analysis revealed that MSV-ABX inhibited proliferation and increased apoptosis of 4T1 and 3LL tumor cells in the liver significantly more than ABX.Conclusions; These studies demonstrate that MSV-ABX is more effective than ABX in therapy of the liver metastasis of breast and colorectal cancer in our mouse models. These data can be translated into the clinic for planning novel clinical trials using MSV-ABX.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P6-04-21.
Abstract P6-05-01: Basal and induced receptor profiles cluster cell lines into subtypes and predict drug response in a panel of breast cancer lines
Authors: Schoeberl BM , Pace EA , Niepel M , Hafner M , Chai DH , Sorger PK
Published: August 2014

Molecular subtype is a critical determinant of therapeutic approaches in breast cancer. The subtype is based on the expression of three receptors: The Her2/ErbB2 receptor tyrosine kinase (RTK), whose over-expression defines the HER2amp subtype, and the estrogen or progesterone nuclear hormone receptors, whose over-expression defines the HR+ subtype. Triple negative breast cancers (TNBCs) express low levels of all three receptors. HER2amp status serves as a biomarker for therapy with anti-Her2/ErbB2 antibodies such as trastuzumab or pertuzumab and HR+ status is a biomarker for therapy with hormone receptor antagonists such as tamoxifen. TNBCs are usually treated with cytotoxic chemotherapy. However, breast cancer subtypes are heterogeneous, clinical and mRNA subtypes are not identical, and even the best available biomarker, HER2amp status, correctly predicts response to trastuzumab in only a subset of patients.

Projects such as the Cancer Cell Line Encyclopedia aim to identify drug response determinants by relating dose-response profiles for multiple compounds to genomic features across cancer cell collections. Recent studies have suggested that measures of target activity such as phosphorylation are more correlated with drug response than mutation status or gene expression, and steady-state protein data is becoming available for tumors and cell lines. However, few studies have tried to relate the biochemistry of signal transduction to drug response on a large scale, mainly because the measurements are demanding and the utility of the approach is unproven.

In this work, we ask whether measurement of the basal and perturbed states of immediate-to-early signaling proteins in serum-starved cells, a robust measurement that can be collected with relatively little biological variability, is predictive of tumor subtype as well as of drug response. We measured the abundance and basal phosphorylation state of nuclear and cell surface receptors and of downstream signaling kinases in the standardized NCI-ICBP43 cell line collection. Because perturbation reveals features of signal transduction that are not apparent by steady state profiling, and because biological ligands present in the microenvironment can alter drug sensitivity, we also collected response profiles by exposing cells to a diverse set of growth factors and cytokines and measuring the activities of downstream signaling kinases before and after ligand addition. We present how the resulting set of ∼3×105 receptor and cell response measurements segregated with clinical subtype and how these proteomic measurements predict the sensitivity of cells to a range of targeted anti-cancer drugs.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P6-05-01.
Abstract P6-05-02: Molecular genetic grading in breast cancer: Development and validation of a prognostic marker
Authors: Matuschek C , Boelke E , Prisack HB , Budach W , Nestle-Krämling C , Gerber PA , Pagouras C , Bojar H
Published: August 2014

Background: Histopathological grading is an important part for the characterization of breast cancer with regard to prognosis and treatment options. 30-60% of breast cancer patients have a grade 2 (G2) malignancy resulting in an intermediate prognosis and the challenge of finding individualized adequate treatment options. Moreover, the lack of objectivity of the grading system is criticized. With the genetic analysis of breast cancer, the deciphering of its genes and gene expression profiling using microarray technology is a new way of classifying breast cancer

Patients, Materials, and Methods. Tumor tissue from 229 breast cancer patients collected from January 2009 - June 2010 was examined. A histological grading of the tumors was determined and gene expression profiling was performed by microarray technology (Agilent Technologies). Gene expression data were reduced and a novel molecular grade index (MGI-9) was defined as a G1 versus G3 classifier by SAM (Significance Analysis of Microarray) and PAM (Prediction Analysis of Microarray). The validity of the newly defined MGI-9 as a prognostic factor was tested by applying it to three published data sets with external follow-up information for a total of 760 patients

Results: The significance analysis of gene expression profiles of 229 breast cancer patients using SAM showed more than 11,000 significantly differentially expressed genes according to the grading classes G1-2-3. The number of differentially expressed genes was reduced to 228 genes by SAM analysis using only the G1 vs. G3 cases (63 patients). PAM analysis resulted in a more reduced classifier of 44 molecular probes and we could show that a subset of 9 genes gave similar classification results and could reliably used in FFPE Material. The new MGI-9 index was tested in thee external data sets (GSE1456, GSE2034 and GSE3494) and could subdivide the histological G2 group in two molecular subgroups G2A and G2B with significant impact on survival in all test scenarios.

Conclusion: The results show that MGI 9-gene molecular grade index is more informative than the conventional histopathological grading system. In order to validate MGI-9 a prospective, randomized study with long-term follow-up for local recurrence rate and distant metastases is urgently needed.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P6-05-02.
Abstract P6-05-15: Analysis of Ki-67 index on normal breast tissue of premenopausal women exposed to tamoxifen and raloxifene: A randomized, double-blind study
Authors: Da Silva BB , Lopes-Costa PV , Rosal MA , Gebrim LH
Published: August 2014

Background: Raloxifene and tamoxifen were approved for breast cancer chemoprevention in women at high risk for the disease, when only clinical and epidemiological studies were considered. Nevertheless, experimental studies that compare the effects of tamoxifen and raloxifene on proliferative activity in normal breast tissue and adopt a more precise method using monoclonal antibodies (particularly Ki-67, because of its sensitivity) are scarce. Methods: A randomized, double-blind study was conducted in 57 ovulatory, premenopausal women aged 18-40 years, who had been diagnosed with fibroadenoma of the breast. The study was approved by the Internal Review Board of the Federal University of Piauí. All patients signed a written informed consent term prior to admission. The patients were divided into three statistically homogeneous groups with respect to age, tumor size, parity, and age at menarche: Group A: placebo, n = 20; Group B: tamoxifen 20 mg/daily, n = 21; and Group C: raloxifene 60mg/daily, n = 16. The study medication was administered for 22 days starting at the first day of the menstrual cycle. On the 23rd day, the fibroadenoma was removed and a sample of non-neoplastic breast tissue was collected for immunohistochemical evaluation of Ki-67 expression. A minimum of 1000 cells either stained or not with anti-Ki-67 (MIB-1) antibody were counted in each slide. For statistical data analysis (p<0.05), a logistic model was used for stained cell count in relation to total cells (binomial distribution). Results: The mean percentages of Ki-67-stained nuclei in groups A, B and C were 22.16±1.91, 2.37±0.21 and 2.16±0.18, respectively (p<0.001). Conclusion: Exposure to tamoxifen or raloxifene resulted in a significant and similar reduction in proliferative activity of non-neoplastic breast tissue of women of reproductive age evaluated by Ki-67 expression. Thus, further studies must be developed with the objective to validate the prophylactic effect of raloxifene in premenopausal women.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P6-05-15.
Abstract P6-05-20: Designing biomarker panels for pre- and post-menopausal breast cancer diagnostic testing using MSD technology
Authors: Henderson MC , Yeh T-F , Reese DE , Borman S , Hollingsworth A
Published: August 2014

At Provista Diagnostics, we have developed a breast cancer diagnostic test (dtectDx Breast) that analyzes serum levels of five protein biomarkers (IL-8, IL-12, CEA, HGF, and VEGF), which are then used in a proprietary algorithm to generate a score that can be characterized as either “normal” or “elevated”. The dtectDx Breast test has demonstrated a sensitivity/specificity of 90%/95% in a full retrospective training and validation set and 86%/87% in a recent post-market analysis. Though this test performs well in women under age 50, sensitivity and specificity both decrease when analyzing serum from women over age 50. We hypothesize that mean biomarker concentrations differ between pre-menopausal and post-menopausal women, in such a way that an ideal biomarker panel for breast cancer diagnosis in post-menopausal women may involve different analytes and a novel algorithm. To test this, we have partnered with the Mercy Women's Center in Oklahoma City to utilize prospectively-collected serum from women undergoing biopsy for a suspicious mass (all serum was drawn pre-biopsy) and from women with no evidence of a mass (to serve as a control). All patients self-reported menopausal status, thus allowing us to categorize samples as pre- or post-menopausal and determine how analyte levels vary between the populations. We determined mean concentrations for 10 protein biomarkers, all of which have previously been shown to be associated with breast cancer. All analytes were measured using Meso Scale Discovery (MSD) technology, which is highly sensitive and reliably demonstrates a low coefficient of variance between replicates. We have determined that mean serum biomarker concentrations are significantly different between pre- and post-menopausal populations in several analytes, thus suggesting that breast cancer diagnostic biomarker panels perform best when designed specifically for a target population.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P6-05-20.
Abstract P6-05-24: Expression of aquaporin 5 and its polymorphisms predict survival in patients with early breast cancer
Authors: Lee SJ , Chae YS , Park HY , Jung JH , Kim WW , Park J-Y , Jeong JY
Published: August 2014

Background. Our previous study showed the association of AQP5 up-regulation with cancer proliferation and migration in hormone-responsive breast cancer cell lines (MCF-7) and with unfavorable prognosis in a small number of patients with breast cancer. Accordingly, we analyzed the prognostic impact of AQP5 expression and polymorphisms in a large number of patients with early breast cancer (EBC).

Methods. AQP5 expression was investigated based on the immunohistochemistry of tissue microarray specimens from 609 EBC patients who underwent surgery between 2003 and 2008. We scored the staining intensity (IS) and percentage of positive tumor cells (PC). The genomic DNA was extracted from paraffin-embedded tumor-free tissue and then genotyped for 3 polymorphisms (rs3736309, rs1964676, and rs74091167) using the Sequenom Mass array system.

Results. Among the 3 polymorphisms, AQP5 overexpression (IS + PC ≥6) was correlated with AQP5 rs74091167 GG genotype. AQP5 overexpression and AQP5 rs74091167 was significantly associated with disease-free survival (DFS; P < 0.001 and P = 0.021, respectively). Moreover, a multivariate survival analysis revealed that AQP5 overexpression and the GG genotype of AQP5 rs74091167 were significantly associated with DFS (HR = 2.026, 95% CI 1.058-3.881, P = 0.030; HR = 0.377, 95% CI 0.179-0.793, P = 0.010, respectively) adjusted to clinicopathological variables, which was prominent in patients with an ER/PgR-positive tumor.

Conclusions. Consistent with our previous study of breast cancer cell lines, AQP5 expression and AQP5 rs74091167 variant can be considered as a prognostic marker in patients with EBC after curative surgery. In the future, functional relevance of this variant needs to be clarified.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P6-05-24.
Abstract P6-06-08: Impact of initial surgical treatment delay on survival according to hormone responsiveness in breast cancer
Authors: Yoo T-K , Han W , Moon H-G , Kim J , Lee JW , Kim MK , Lee E , Kim J , Noh D-Y
Published: August 2014

Previous studies on the issue of the prognostic importance of treatment delay in breast cancer have shown inconsistent results. Furthermore, the association between the impact of treatment delay and molecular characteristics of tumors has not been adequately addressed. The purpose of this study is to examine the prognostic impact of initial surgical treatment delay after biopsy-proven cancer diagnosis in breast cancer patients.

A total of 1,393 consecutive invasive breast cancer patients, treated in Seoul National University Hospital, Seoul, Korea, between July 2006 and June 2008, were included in this study. Patients with in situ or metastatic carcinoma at the time of diagnosis and patients in whom the dates of the initial pathologic diagnosis were unknown were excluded.

The median time from biopsy-confirmed cancer diagnosis to initial surgical treatment was 25 days (range 0-134). When the patients were classified according to their treatment delay days (0 to 29, 30 to 59, and ≥60days), there was no difference in survival between ‘0-29days’ group and ‘30-59days’ group. However, for patients who experienced more than 60 days of initial delay in surgical treatment, the survival was significantly worse when compared to other groups (p = 0.034). The association between the treatment delay and poor outcome was only seen in patients with ER and PR negative tumors (p = 0.018) while patients with hormone-responsive tumors showed no such association. Patients with ER and PR negative tumors developed more recurrence and had shorter disease-free survival if they had treatment delay of more than 60 days. The prognostic importance of treatment delay of more than 60 days remained significant in predicting disease-free survival after adjusting for other known prognostic factors such as age, tumor size, nodal status, cancer stage, nuclear grade and Ki-67 expression (HR 5.127; 95% CI, 1.350 to 19.472; p = 0.016).

Our results suggest that having treatment delay of more than 60 days is associated with poor treatment outcome in patients with ER and PR negative breast cancer.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P6-06-08.
Abstract P6-06-19: Impact of hospital surgical volume on breast cancer mortality: A population based study in the Netherlands
Authors: Siesling S , Tjan-Heijnen VCG , de Roos M , Snel YEM , van Dalen T , Wouters MW , Struikmans H , Maduro JH , van der Hoeven KJM , Visser O
Published: August 2014

Background: For some low-volume tumour types hospital surgical volume is associated with better survival. For breast cancer there is still much debate. The aim of this study is to determine to what extent the yearly surgical hospital volume determines the mortality in invasive non-metastatic breast cancer patients.

Method: Women diagnosed with primary invasive breast cancer in the period 2001-2005 were selected from the Netherlands Cancer Registry. Hospitals were grouped by their annual volume of surgery for invasive breast cancer. Cox proportional hazard models were performed including all patients with primary non-metastatic breast cancer who underwent breast surgery. Gender, age at diagnosis, morphology, grade, size (pT), number of positive lymph nodes, year of diagnosis and socio economic status (SES) were included as covariates. Follow-up was complete until the 1th of February 2013.

Results: In total 58.982 patients with invasive non-metastatic breast cancer were diagnosed during the period 2001-2005. Hospitals were grouped by volume of surgery: less than 75 (n = 19), 76-100 (n = 30), 101-150 (n = 29), 150-199 (n = 9) and 200 or more (n = 14) surgeries per year. Non-metastatic patients had a 8% higher risk of death in a low volume hospitals than a hospital with >200 surgeries per year (HR 1.08, 95%CI 1.02-1.14).

Cox regression multivariable analyses: the relation of the number of surgical treated invasive breast cancer per hospital per year and the mortality of patients with non-metastatic breast cancer in the Netherlands, 2001-2005Variable HR95%CINumber of operated invasive breast cancers per yearmore than 200reference  150-1991.030.98-1.09 100-1491.010.97-1.05 75-991.040.99-1.08 less than 751.081.02-1.14Corrected for age, gender, morphology, grade, pT, number of positive lymph nodes, year of diagnosis, socioeconomic status

Patient and tumour characteristics like age (HR 1.05, 95%CI 1.05-1.05) and SES (lowest vs highest; HR 1.12, 95%CI 1.07-1.16), grade (low vs high, HR 1.72, 95%CI 1.63-1.82), tumour size (1-2 cm vs 2-5 cm; HR 1.46, 95%CI 1.40-1.51), and a higher number of positive lymph nodes (0 vs 1-3; HR 1.40, 95%CI 1.34-1.46 and 0 vs >10; HR 3.19, 95%CI 3.00-3.39) influenced death to a larger extend than surgical volume.

Conclusion: In the Netherlands, surgical hospital volume influences risk of death marginally, and far less than patient and tumour characteristics. No differences in mortality between hospitals with a surgical volume of more than 75 were revealed for invasive non-metastatic breast cancer patients compared to hospitals with more than 200 operations. Over the more recent period of 2007-2012, only 3 hospitals had less than 75 operations on average per year, resulting in an even more comparable mortality between hospitals in future.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P6-06-19.
Abstract P6-06-25: Usefulness of pretreatment NLR as prognostic factor of late recurrence, 5-year after primary treatment
Authors: Noh H , Lee J , Choi E , Han A
Published: August 2014

Background

Prognostic factor and predictive factors are well established in early-stage breast cancer, but less is known about recurrence, especially 5-year after primary treatment. While mortality of breast cancer is from organic dysfunction with metastatic disease, not from the diseased breast itself, still we do not have adequate information about metastatic disease as we do about primary breast cancer itself.

The aim of this study is to investigate of usefulness of pretreatment NLR(neutrophil to lymphocyte ratio) as prognostic factor that can tell risk of recurrence 5- year after primary treatment.

Patients and methods

A retrospective review of invasive breast cancer patients surgically resected between January 2001 and December 2007. Exclusion criteria were as followings: patients with recurrence or death within 5yrs.

Results

A total of 301 patients were included and 13 patients experienced with recurrence after 5 years.

Table 1. Patients' characteristics Total (n = 301) N (%)Age*48.8(±11.4)Age ≤3525(8.3)35< ≤50165(54.8)50<111(36.9)Histology Ductal270(89.7)Lobular7(2.3)Others24(8.0)Tumor size (cm)*2.73(±1.64)T stage T1126(41.9)T2158(52.5)T317(5.6)N stage N0199(66.1)N167(22.3)N220(6.6)N311(3.7)unknown4(1.3)Histologic grade I72(23.9)II110(36.5)III70 (23.3)unknown49(16.3)Estrogen receptor Negative111(36.9)Positive185(61.5)unknown5(1.6)Progesterone receptor Negative129(42.9)Positive167(55.5)Unknown5(1.6)HER-2 Negative196(65.1)Positive98(32.6)unknown7(2.3)Molecular subtype Luminal A136(45.2)Luminal B66(21.9)HER-2 enriched32(10.6)Triple negative60(20.0)unknown7(2.3)Late recur Yes13(4.3)No288(95.7)*mean±SD

Overall average of NLR value was 2.16, median value was 1.83. The average NLR were 2.80±1.68 in patients with recurrences occurred after 5 years and 2.13±1.15 in patients without recurrence. The NLR cutoff value of 1.83 was determined by median value.

In 1.83≤NLR group has a higher incidence of recurrence after 5 years compared with NLR<1.83 group (8yr DFS 99.2% vs. 93.3%, p = 0.021). Especially in luminal A(ER (+) or PR (+), HER-2(-)), 1.83≤NLR group has a higher incidence of recurrence after 5 years compared with NLR<1.83 group (8yr DFS 100% vs. 87.0%, p = 0.015).

Conclusion

We found that pretreatment NLR is associated with late recurrence that occurs after five years. And we suggest that NLR could be used as one of the prognostic factor that can tell about late recurrence, 5-year after primary treatment.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P6-06-25.
Abstract P6-06-30: Are synchronous and metachronous bilateral breast cancers different? An immunohistochemical analysis aimed at intrinsic tumor phenotype
Authors: Senkus E , Szade J , Pieczynska B , Zaczek A , Pikiel J , Sosinska-Mielcarek K , Karpinska A , Jassem J
Published: August 2014

Background: The biology and pathomechanism of bilateral breast cancers is not fully understood: the risk of developing second (metachronous) breast cancer is 4-6 times higher than that of first primary in general population, whereas for synchronous breast cancers the number of cases is about 100 times higher than could be expected by chance alone. We compared the morphological and immunohistochemical characteristics of primary synchronous (sBBC) vs. metachronous (mBBC) bilateral breast cancers), with special focus on intrinsic tumor phenotype.

Methods: Tumor morphology and expression of 8 immunohistochemical markers were assessed in tissue microarrays containing primary breast tumor cores from 113 mBBC and 61 sBBC. Analyzed markers included hormone receptors (estrogen receptor, progesterone receptor), HER2, Ki67, cytokeratin 5/6, E-cadherin, vimentin and epidermal growth factor receptor. Surrogate intrinsic phenotypes were determined according to St Gallen 2011 criteria.

Results: mBBChad higher incidence of ductal histology (p = 0.037), lower estrogen receptor expression (p = 0.047), and higher expression of cytokeratin 5/6 (p = 0.017) and of vimentin (p = 0.008); in multivariate analysis only vimentin retained the significance (p = 0.01). Ten (13%) and 11 (26%) of mBBCand sBBC, respectively, had luminal A phenotype, 39 (50%) and 15 (36%) were luminal B HER2-negative, 13 (17%) and 12 (28%) - luminal B HER2-positive, 3 (4%) and 2 (5%) - HER2-positive (not luminal), and 12 (16%) and 2 (5%) had triple negative phenotype (p = 0.07).

Characteristics of mBBC and sBBCFeaturemBBC (%)sBBC (%)p valueDuctal histology98 (87)47 (77)0.037Grade 346 (41)17 (28)0.09Extensive intraductal component26 (23)18 (29)0.42Strong ER expression73 (67)48 (81)0.047Strong PR expression55 (49)38 (63)0.07HER2 positivity (2+, 3+)51 (46)29 (50)0.6Ki76 ≤14%24 (22)21 (35)0.06Surrogate intrinsic phenotype   luminal A10 (13)11 (26) luminal B HER2-39 (50)15 (36) luminal B HER2+13 (17)12 (28)0.07triple negative12 (16)2 (5) HER2+ (non luminal)3 (4)2 (5) CK5/6 >10%28 (26)6 (11)0.017Vimentin >10%25 (23)4 (7)0.008E-cadherin (+)88 (83)49 (83)0.99EGFR (+)35 (32)11 (19)0.07

Conclusion: mBBC, compared to sBBC, are characterized by more aggressive phenotype, expressed by lower expression of estrogen receptor and stronger expression of cytokeratin /6 and vimentin; this does not, however, translate into substantial differences in the proportion of intrinsic tumor phenotypes.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P6-06-30.
Abstract P6-06-32: 4EVER: Does mTOR inhibition have a major clinical impact on bone health in postmenopausal women with hormone receptor positive (HR+) locally advanced breast cancer treated with everolimus (EVE) in combination with exemestane (EXE)?
Authors: Hadji P , Tesch H , Fasching PA , Lueftner DI , Janni W , Lux MP , Schneeweiss A , Decker T , Belleville E , Kreuzder J , Muth M , Wallwiener D
Published: August 2014

Introduction:

BOLERO-2, a multi-national, double-blind, placebo-controlled, phase 3 study in postmenopausal women with HR+, HER2- advanced breast cancer (BC) progressing after non-steroidal aromatase inhibitors (NSAI), showed significant benefit in PFS with the addition of the mTOR inhibitor EVE to EXE compared to EXE alone (Baselga J 2012). The BOLERO-2 bone sub study indicated an influence of EVE on bone health after 6 and 12 weeks of treatment (Gnant M 2012). To further investigate the long term influence of mTOR inhibition on bone health, we initiated the phase IIIb, multi-center, open label study 4EVER for postmenopausal women with HR+ locally advanced or metastatic breast cancer treated with EVE +EXE. The primary endpoint is the overall response rate (ORR) after 6 months. Secondary endpoints include progression free survival after 6 and 12 months, ORR after 12 months, overall survival, quality of life, health utilities and health care resources. The exploratory objectives focus amongst others on the assessment of biomarkers of bone turnover.

Methods:

Main inclusion criteria were postmenopausal women with metastatic or locally advanced, ER+, HER2- breast cancer refractory to NSAI. Markers of bone turnover (P-I-NP, CTX, OPG, RANKL, vitamin D, FSH and estradiol) are being assessed at baseline and after 4, 12 and 24 weeks on treatment.

Results:

Baseline characteristics: 334 postmenopausal patients were screened and 297 were enrolled in 5 months. The hormone receptor assessment was performed in 71.0% from primary tumor and in 29% from the metastases (ER+/PgR+ 77.5%, ER+/PgR- 21.5%). The mean time since initial diagnosis was 9.2 years. Mean time since first recurrence/ metastasis was 48.1 months and mean time since most recent recurrence / metastasis 2.4 months. The stage at primary diagnosis was M0 74.2% and M1 22.0%. 68.7% of the patients had bone lesions. Most recent antineoplastic medications were in the adjuvant (24.8%), neoadjuvant (1.1%) and palliative setting (71.0%). 25.6% of the patients had one, 16.4% two and 57.9% three and more prior therapies in metastatic setting.

The results of the baseline bone and hormone maker analysis for baseline and after 4, 12 and 24 weeks on treatment will be presented.

Conclusion:

This exploratory bone study will provide further insights into the influence of mTOR inhibition on bone turnover and overall bone health in women with advanced breast cancer.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P6-06-32.
Abstract P6-06-34: Comparing recurrence risk scores derived from 4 immunohistochemistry stains (IHC4) versus oncoType DX
Authors: Miller DV , Jensen TA , Covington AD , Rosenthal R , Avent JM , Hansen J , Peck L , Wise PD , Leavitt MO , Wallentine JC , Rowley B
Published: August 2014

Background: Prognostically relevant biomarkers in breast cancers can be assayed using nucleic acid (e.g. OncoType DX) or protein based (e.g. immunohistochemistry (IHC)) detection stratagies. Nucleic acid tests measure many more markers compared to immunohistochemistry, but are weighted heavily for ER, PR, and HER2 in their prognostic scoring algorithms. Recent data suggest that a panel of 4 IHC stains (IHC4) may have equivalent prognostic ability to multigene assays. The aim of this study was to retrospectively compare IHC4 scores to OncoType DX recurrence scores in a large cohort of patients in our statewide health system.

Methods: Formalin-fixed paraffin-embedded breast cancer samples previously submitted for OncoType DX testing between 2008 and 2011 were obtained and sections stained for ER, PR, HER2, and Ki67 (the IHC4 staining panel). Stained slides were scanned digitally and scored using customized pathologist assisted image analysis algorithms (Leica Biosystems Aperio, Vista CA). The resulting data were used in the modified “Magee Equation 3” (Klein et al. Mod Pathol. 2013. PMID: 23503643) to calculate an IHC4 recurrence score on a numerically comparable scale to that used in the OncoType score. The IHC4 and Oncotype DX scores were directly compared for concordance and correlation.

Results: 280 breast cancer samples comprised the study cohort. The mean (median) IHC4 score was 20.13 (19), range: 11-42. The mean (median) OncoType score was 18.48 (16), range: 0-75. Using cut-offs of 0-17 for low risk, 18-30 for intermediate risk, and >30 for high risk, the concordance data is shown in Table 1.

Comparison of scores from IHC4 and OncoType by Risk Category  OncoType RS  LowIntermedHigh Low80201IHC4 RSIntermed816819 High029RS = Recurrence Score

Pearson's correlation coefficient for the entire set was 0.6669. The overall concordance was 56.1%; one-step discordance 43.6%; and two-step discordance: 0.3%. Eliminating the intermediate scores, the concordance was 98.8%.

Conclusions: Reasonable agreement was seen between risk scores derived by IHC4 and OncoType DX. Only one discrepancy among 280 scores would have resulted in a different treatment recommendation (low risk by IHC4 and high risk by OncoType). If the IHC4 score is clearly high or low, one would not expect a dramatically different result from Oncotype DX, and the Oncotype DX test may not be needed.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P6-06-34.
Abstract P6-06-39: Chest wall invasion is a more significant prognostic factor than nodal spread, triple-negative or Her2 receptor status in stage III breast cancer: A large, single institution retrospective analysis
Authors: Ansley KC , Batten KG , Singel SM
Published: August 2014

Background

There is a significant variation in response to systemic treatment in stage III breast cancer that is not explained by hormone/Her2 receptor status alone. Identification of additional prognostic factors is needed to further stratify patients for additional treatment. According to the National Cancer Data Base, patients with stage IIIB breast cancer, characterized by having direct chest wall involvement regardless of lymph node spread, have a worse prognosis than patients with stage IIIC breast cancer, generally with significant lymph node involvement (41% vs. 49% 5-year overall survival). We hypothesized that chest wall invasion is an independent prognostic factor and may be more significant than regional nodal spread, triple-negative or Her2 -receptor status, or ethnicity.

Methods

Our retrospective study included 135 patients with stage IIIB or IIIC breast cancer seen at a large county hospital in Dallas, TX between 2008 and 2013. Patients were stratified into two groups based on whether they had chest wall invasion (T4) or not. Cases of inflammatory breast cancer (T4d) were excluded. Kaplan-Meier survival curves, the Log-Rank test, univariate and multivariate methods were used for statistical analysis. We evaluated the effects of chest wall involvement, ethnicity, and hormone receptor/Her2 status on overall survival.

Results

In patients with stage III breast cancer, those with chest wall invasion (T4) have a significantly decreased 5-year overall survival (OS) compared to patients with significant regional nodal spread (N3) (62% vs. 81%, p = 0.022, HR 2.3, 95% CI: 1.1-4.9). Univariate analysis of hormone receptor, Her2 status, African-American or Hispanic ethnicity, the presence of chest wall involvement, and significant regional lymph node involvement (N3 disease or not) showed that ER receptor status and the presence of chest wall involvement were the only significant prognostic factors (p = 0.006 and p = 0.022, respectively). Notably, the combination of ER-negativity and the presence of chest wall invasion predicts for worse prognosis than either alone (p = 0.0011, HR 1.6, 95% CI: 1.2-2.2).

Conclusion and Discussion

Chest wall invasion was an independent predictor for decreased overall survival in stage III breast cancer. This could explain in part why patients with stage IIIB breast cancer have worse overall survival than patients with stage IIIC. There may be an intrinsic biological difference between breast cancer cells that directly invade surrounding chest wall and those that have nodal spread as the main route of metastasis. An inflammatory component in those with chest wall invasion may allow these tumors to be more aggressive and additional research is needed to target potential genes for therapy.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P6-06-39.
Abstract P6-06-40: Analysis of outcome according to risk factors in the randomized phase III TURANDOT trial evaluating first-line bevacizumab-containing therapy for HER2-negative locally recurrent/metastatic breast cancer (LR/mBC)
Authors: Brodowicz T , Pienkowski T , Beslija S , Melichar B , Lang I , Inbar MJ , Anghel R , Spanik S , Ahlers S , Zielinski C
Published: August 2014

Background: The randomized phase III TURANDOT trial compared first-line bevacizumab (BEV) + paclitaxel (PAC) vs BEV + capecitabine (CAP) in HER2-negative LR/mBC [Lang, Lancet Oncol 2013]. At the prespecified interim analysis there was no detectable difference in overall survival (OS; primary endpoint) between treatment groups, but the secondary endpoints of progression-free survival (PFS) and objective response rate (ORR) favored BEV-PAC. We sought to identify patient populations defined by risk factors that may be most appropriately treated with one or other of the regimens.

Methods: Patients with HER2-negative LR/mBC who had received no prior chemotherapy for LR/mBC were randomized to either BEV-PAC (BEV 10 mg/kg d1 & 15 + PAC 90 mg/m2 d1, 8, & 15 q4w) or BEV-CAP (BEV 15 mg/kg d1 + CAP 1000 mg/m2 bid d1-14 q3w). The study population was categorized into three cohorts: triple-negative (TNBC), high-risk hormone receptor-positive (HR+) and low-risk HR+. High-risk and low-risk HR+ were defined, respectively, as having ≥2 vs ≤1 of the following four risk factors: disease-free interval ≤24 months; visceral metastases; prior (neo)adjuvant anthracycline and/or taxane; ≥3 metastatic sites.

Results: Baseline characteristics, efficacy, and safety by treatment arm are summarized below for the three cohorts. Although PFS results in all cohorts favored BEV-PAC, interim OS results showed a trend in favor of BEV-PAC in TNBC patients and in favor of BEV-CAP in low-risk HR+ patients. Grade ≥3 adverse events were less common with BEV-CAP than BEV-PAC in all three cohorts.

 TNBCHigh-risk HR+Low-risk HR+ BEV-PAC (n = 63)BEV-CAP (n = 67)BEV-PAC (n = 146)BEV-CAP (n = 162)BEV-PAC (n = 75)BEV-CAP (n = 50)Median age, years545658576161ECOG PS 0,%756068666366PFS      Events, n (%)50 (79)54 (81)93 (64)125 (77)33 (44)35 (70)Median, months (95% CI)9.0 (7.8-10.7)5.6 (4.9-8.0)11.1 (10.4-13.4)8.3 (7.1-10.7)14.4 (10.4-20.5)11.5 (8.1-16.3)HR (95% CI)a1.37 (0.93-2.02)1.29 (0.98-1.69)1.39 (0.86-2.25)OS      Events, n (%)28 (44)34 (51)50 (34)52 (32)18 (24)11 (22)1-year OS rate,%786380828590HR (95% CI)a1.33 (0.80-2.19)0.97 (0.66-1.43)0.80 (0.38-1.69)Grade ≥3 AEs,%634261516148aBEV-CAP vs BEV-PAC

Conclusion: The simple risk factor index is prognostic for both PFS and OS and may be used to guide treatment choice when selecting BEV-containing therapy, balancing outcome with safety profile and patient preference. Final analysis of OS is expected in 2014.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P6-06-40.