bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P3-08-04: Impact of CYP3A variation on estrone levels and breast cancer risk
Authors: Ross GM , Johnson N , Orr N , Walker K , Gibson L , Folkerd E , Haynes B , Palles C , Coupland B , Shoemaker M , Jones M , Broderick P , Sawyer E , Kerin M , Tomlinson I , Zvelebil M , Chilcott-Burns S , Tomczyk K , Simpson G , Willianson J , Hillier S , Houlston R , Swerdlow A , Ashworth A , Dowsett M , Peto J , dos Santos I , Fletcher O
Published: August 2014

Background: Epidemiological studies provide strong evidence for a role of endogenous sex steroids in the etiology of breast cancer. Our aim was to identify common variants in genes involved in sex steroid synthesis or metabolism that were associated with premenopausal hormone levels and breast cancer risk.

Methods: We measured urinary estrone glucuronide (E1G) and pregnanediol glucuronide (PG) using a protocol specifically developed to account for cyclic variation in hormone levels during the menstrual cycle, plasma sex hormone-binding globulin (SHBG) and androgenic precursors in up to 763 healthy premenopausal women. We genotyped 642 single nucleotide polymorphisms (SNPs) in these women; a single SNP was further tested for association with breast cancer risk in data from 10,551 breast cancer case patients and 17,535 control subjects. All statistical tests were two-sided.

Results: rs10273424 mapping approximately 50kb centromeric to the cytochrome P450 3A (CYP3A) cluster (7q22.1) was associated with a 21.8% reduction in E1G levels (P = 2.7 × 10−9) and a modest reduction in breast cancer risk in cases diagnosed at or before age 50 (OR = 0.91; P = 0.03) but not older cases (odds ratio (OR) = 1.01; P = 0.82). A rare non-synonymous SHBG SNP was associated with reduced plasma SHBG levels.

Conclusions: Genetic variation in non-coding sequences flanking the CYP3A locus contributes to variance in premenopausal E1G levels and breast cancer risk in younger cases. Since CYP3A4, the most predominantly expressed CYP3A gene, is responsible for metabolism of endogenous and exogenous hormones and hormonal agents such as tamoxifen, used in the treatment of breast cancer this association may have wider implications.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P3-08-04.
Abstract P3-10-02: Modulation of mRNA and miRNAs by the novel histone deacetylase inhibitor, CG-1521 disrupts cytokinesis in SUM149PT inflammatory breast cancer cells
Authors: Chatterjee N , Tenniswood MPR
Published: August 2014

Inflammatory breast cancer (IBC) comprises only 2.0% of all breast cancer cases; however it accounts for 8.0% of breast cancer-specific deaths. IBC is highly aggressive, appears at a younger age, and is more difficult to treat than the common forms of breast cancer. By the time the disease is diagnosed, IBC is usually stage 3 and has already metastasized to neighboring lymphatics. Though many tumors are p53 wild type and/or Her2 positive, the median overall survival among women with IBC is less than 4 years even with aggressive multimodal therapy. To develop IBC-specific therapies, we have compared the effects of two hydroxamic acid histone deacetylase (HDAC) inhibitors, Trichostatin A (TSA) and CG-1521 on the biology of the triple negative SUM149PT IBC cell line. In these cells, CG-1521 and TSA induce dose (0–10 µM) and time dependent (0–96 h) increases in the proportion of cells undergoing cell cycle arrest and cell death, in the presence or absence of 17β-estradiol. To identify the genomic targets of CG-1521, we have performed concurrent microarray analyses of mRNA and miRNA expression followed by qPCR validation. In SUM149PT cells, CG-1521 modulates the expression of approximately 935 transcripts: 341 of these transcripts are up-regulated and 594 are down-regulated after 48h of treatment. Gene ontology analysis demonstrates that many of the down-regulated mRNA transcripts encode proteins that are associated with the spindle assembly checkpoint, chromosome segregation and microtubule based processes including KIF4, PRC1, AURKB and KIF23 (MKLP-1). Strikingly, many genes in these ontologies are potentially targeted by a small number of miRNAs (miR-22, miR-1207-5p & miR-494) which are significantly up-regulated by CG-1521, suggesting that the ability of CG-1521 to down-regulate the transcripts associated with spindle formation and cytokinesis may be significantly modulated by miRNA expression. Immunofluorescence microscopy demonstrates that treatment of SUM149PT cells with CG-1521 results in the appearance of elongated midzone structures visualized by phalloidin staining, similar to the phenotype seen by others after siRNA-mediated knockdown of KIF4 in HeLa cells. These data suggest that CG-1521 affects the central mitotic spindle formation, disrupts furrow formation and abscission, arresting the cells in cytokinesis ultimately leading to cell death. Whether these effects are solely mediated through transcriptional mechanisms or whether CG-1521 also directly affects the activity of these proteins through acetylation remains to be determined. Nevertheless, the data show that histone deacetylase inhibitors induce cell death in IBC cells, and suggest that these small molecules may be useful addition to the armamentarium as adjuvants for the treatment of IBC.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P3-10-02.
Abstract P3-11-01: Matched-pair analysis of patients with male and female breast cancer
Authors: Kim M , Lee SK , Choi M-Y , Kim S , Kim J , Jung SP , Bae SY , Kil WH , Lee JE , Nam SJ
Published: August 2014

Purpose: Male breast cancer (MBC) is extremely rare, accounting for less than 1% of all malignancies in men and only 1% of all breast carcinomas. The treatment and surveillance guidelines on male breast cancers are less recognized. The aim of this study is to evaluate our single institution's experience with MBC over the past 15 years and to contrast differences between female and MBC.

Methods: MBC diagnosed from 1994 to 2010 at the Department of Surgery, Samsung Medical Center (Seoul, Korea) was retrospectively analyzed. Clinical data and tumor characteristics were examined. Each MBC was matched with female counterparts by 1:N varied matching ratio that showed accordance in seven variables (year of diagnosis, age, tumor stage, nodal stage, tumor grade, estrogen receptor(ER), progesterone receptor(PR)).

Results: 39 male/184 female matched-pairs were available for analysis. The median duration of follow-up was 3.8 years. The median age of MBC patients was 50 years and the median size of tumor was 2.0cm. The proportion of positivity of ER and PR status was 97.4% and 84.6%, respectively. Despite of higher positive rate of hormone receptor, the rate of hormone therapy in MBC patients was significant lower than female conterpart (p = 0.002). Men and women with breast cancer had similar disease-free survival (DFS) and disease-specific overall survival (DSS). Five MBC patients had a recurrence during follow up period and 4 of them were expired. The 10-years DFS was 73.1% in men and 80.5% in women (p = 0.348). The 10-years DSS was 74.1% in men and 87.1% in women, respectively (p = 0.207).

Conclusion: This study showed no disease free and overall survival differences between male and female breast cancer patients and revealed that gender is no predictor for survival in breast cancer. Male patients receive obviously less adjuvant treatment compared their female matched patients. It would be better to do more aggressive treatment in MBC to improve the survival outcome.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P3-11-01.
Abstract P3-11-04: Male breast cancer: Overall survival in a single institution.
Authors: Bello MA , Bergmann A , Costa CRA , Pinto RR , Millen EC , Thuler LCS , Bender PFM
Published: August 2014

Background: Male breast cancer is an uncommon disease and the therapy is mainly based on what is know from female breast cancer. Objective: To investigate the clinicopathologic characteristics of male breast cancer and the overall survival in a single institution.

Methods: The clinical data and survival status of 75 male breast cancer treated in a Brazilian public cancer hospital from 2000 to 2009 were collected. The association with clinicopathological characteristics and overall survival was analyzed using Kaplan-Meier curves and the Cox proportional hazards regression (enter method) was used to assess survival differences after adjusting for confounders. The study was approved by National Cancer Institute Research and Ethics Committee (number 128/11).

Results: The median patient age was 64 years (range 33–86). Estrogen receptor (ER) was positive in 58 (77.3%) patients, while progesterone receptor (PR) were positive in 47 (62.7%). Histology type was ductal infiltrant carcinoma for 57 (76.0%) and 51 (68.0%) patients underwent surgery. The median follow-up period was 43,1 months (range 2.7–147.8). The median survival from the diagnosis of breast cancer was 97.0 months (95%CI 53.6 −140.4) with a 61.7% 5-year overall survival (OS). In the final Cox regression model, independent factors associated with increased risk of death were metastasis at diagnosis (HR = 18.1; 95%CI: 5.9–55.2), ≥ 65 years old (HR = 4.3; 95%CI: 1.7–10.5); tumor stages ≥ IIb (HR = 3.5; 95%CI: 1.3–9.7) and smoking (HR = 1.6; 95%CI: 1.04–2.6).

Conclusions: Invasive ductal carcinoma is the main pathologic type. The median survival from the diagnosis of breast cancer was 97.0 months and metastasis at diagnosis, patient age, tumor stage and smoking are independent prognostic factors.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P3-11-04.
Abstract P3-12-05: Clinical features and outcomes of leptomeningeal metastasis in patients with breast cancer: a single center experience
Authors: Jo J-C , Kang MJ , Ahn J-H , Jung KH , Kim JE , Gong G , Kim HH , Ahn SD , Kim SS , Son BH , Ahn SH , Kim S-B
Published: August 2014

Background: Leptomeningeal metastasis (LM) is one of the major problems in managing metastatic breast cancer because of LM typically carries a devastating prognosis and often represents a terminal event. We analyzed the clinical features and outcomes of LM in patients with breast cancer.

Methods: We retrospectively reviewed the medical records of patients who were diagnosed with LM from breast cancer between 2002 and 2012 at Asan Medical Center.

Results: Of the 95 LM patients by cytologically proven (n = 81) or radiologically diagnosed (n = 14), 57 (60%) had an ECOG performance status (PS) ≥ 3, and the median age was 47 years (range, 26–72 years). The patients were diagnosed with LM after a median of 10.3 months (95% CI, 5.5–15.0 months) from the time of diagnosis of metastatic breast cancer. LM was present in 2 patients at the time of initial diagnosis. Twenty-three patients (24.2%) had isolated CNS metastasis, and 6 (6.3%) had only LM without any detectable metastasis sites. At the time of diagnosis of LM, 46 patients (48.4%) presented with coincidental failure of systemic disease control. Seventy-eight patients (82.1%) underwent intrathecal chemotherapy (methotrexate; n=78, thiotepa; n=11), resulting in one-third of cytologic negative conversion (n = 26), and 41 (43.2%) received systemic chemotherapy. The overall median survival time was 3.3 months (95% CI, 2.5–4.2 months) and 7.8% of the patients survived for more than 1 year. Overall survival tended to be better in patients who achieved cytologic negative conversion to intrathecal chemotherapy than those did not (median 4.5 months versus 3.2 months, P = 0.241). Overall survival was not different according to subtypes; hormone receptor (+), HER2 (+), and triple negative (median 3.6 months, 3.3 months, and 3.2 months, P = 0.937). Multivariate analysis demonstrated that ECOG PS ≥ 3 (HR = 2.09, 95% CI 1.21–3.58, P = 0.007), coincidental failure of systemic disease control at LM (HR = 3.01, 95% CI 1.76–5.15, P < 0.001), and systemic chemotherapy after LM (HR = 0.40, 95% CI 0.24–0.68, P = 0.001) were independent factors associated with survival.

Conclusions: The prognosis for patients with LM from breast cancer was still poor. Systemic chemotherapy in addition to intrathecal chemotherapy might confer a survival benefit, even after the detection of LM.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P3-12-05.
Abstract P3-12-06: Clinicopathological Analysis of Breast Cancer Patients with Brain Metastases
Authors: Ushio A , Sawaki M , Fujita T , Hattori M , Kondo N , Horio A , Gondou N , Iwata H
Published: August 2014

Background: Recent advances in chemotherapy have enabled to control the progression of metastases from breast cancer except for the brain, highlighting the symptoms associated with brain metastases. In addition, improvement in our understanding of breast cancer biology changed the treatment strategy, possibly leading to improved outcomes in patients with breast cancer and brain metastases.

Object: The aim of this study was to investigate the correlation between biology of primary tumors and prognosis of patients with breast cancer metastasis to the brain.

Patients and method: Among 3,171 patients with primary breast cancer undergoing surgery at out hospital from 2003 to 2010, 35 patients (1.1%) who developed brain metastases, excluding stage IV at the initial diagnosis. We reviewed 35 patients for a correlation between survival and clinicopathological variables, including stage, hormonal sensitivity, HER2 expression status, the number of brain metastases, and treatment for brain metastases.

Results: The mean age of 35 patients was 52.1 years (range 22–68) and the median follow-up period was 41.0 months. The subtypes of primary tumors included triple-negative (TN) in 14 patients (40%), luminal in 11 patients (31.4%), luminal HER2-positive (luminal HER2) in 2 patients (5.7%), and HER2-positive (HER2) in 2 patients (5.7%). The duration from surgery to recurrence (disease-free survival, DFS, months) was 14.7 in TN, 26.5 in luminal, 29.8 in luminal HER2, and 31.0 in HER2. The duration from the first recurrence to the detection of brain metastases (months) was 7.2 in TN, 14.4 in luminal, 15.8 in luminal HER2, and 4.0 in HER2. The mean survival time after the diagnosis of brain metastases was 8.6 months (range, 0–40) with a survival time of ³ 1 year in 8/35 patients (23%) and that of < 1 year in 21/35 patients (60%). Both DFS and the duration from the first recurrence to the diagnosis of brain metastases were shorter in TN compared with other subtypes. In HER2 subtype, DFS was long but, once recurrence was detected, the duration to the development of brain metastases was short. In multivariate analysis of clinicopathological variables (age, DFS, ER, HG, HER2, the number of brain metastases (solitary versus multiple), the site of initial metastasis (brain versus other organs), and the presence or absence of chemotherapy after brain metastases, revealed solitary brain metastasis (p = 0.002) and the initial metastatic site of brain (p = 0.003) to be independent factors predicting survival.

Conclusion: The clinical outcomes of brain metastases were different among tumor subtypes. Factors predicting survival in patients with breast cancer and brain metastases may be the site of initial metastasis, the number of brain metastasis, and local treatment for brain metastases.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P3-12-06.
Abstract P4-01-09: Screening breast magnetic resonance imaging (MRI) in early-stage breast cancer survivors
Authors: Dowton AA , Meyer ME , Ruddy KJ , Yeh ED , Partridge AH
Published: August 2014

Background: Limited evidence is available regarding the value of screening breast MRI in female early stage breast cancer survivors. We sought to evaluate outcomes of index screening breast MRI in breast cancer survivors in the modern era of breast cancer treatment and imaging.

Methods: Using a large prospective institutional database, we identified all women who underwent breast MRI between 7/2006 and 6/2008, were ≥ 1 year and ≤ 3 years after diagnosis of stage I-IIIa breast cancer, had remaining breast tissue after treatment, and had consented to a record review protocol at our center (95% uptake rate). Sociodemographic, disease, staging and treatment information (see table 1), indications and outcomes for breast MRI, as well as disease status at 2 years after the index MRI were ascertained.

Results: 643 patients were identified as having undergone at least 1 MRI of the breast(s); 248 (38.5%) were diagnostic MRIs (had identified clinical indication or imaging abnormality); 12 (2%) were missing clinical data; 383 (59.5%) were screening MRIs. 338/383 (88%) patients who had screening MRI had stage I-IIIa invasive breast cancer. 220/338 (65%) MRIs performed were index MRIs (index MRI defined as first MRI >1 year post diagnosis). For this preliminary analysis, we include only the 155 patients (70%) who were within 1–3 years of diagnosis at the time of index MRI (see table 1 for patient and disease characteristics).

24/155 (15.5%) screening MRIs were considered not normal. Five women (21%) underwent biopsy based on abnormal MRI finding; 1 resulted in the detection of a local recurrence (4.2%, 95% CI=0.1%–21.1%). 3/131 patients who had a normal index MRI were found to have local/regional recurrence or contralateral breast cancer diagnosed within 2 years of the MRI (2.3%, 95% CI=0.5%–6.6%). A statistically significant difference in the proportions was not observed based on Fisher's test (p = 0.50). 6/155 patients (3.9 %) had a systemic recurrence in the 2 years following index MRI.

Conclusion: The vast majority of first screening breast MRIs obtained 1–3 years following treatment in early stage breast cancer survivors are normal, and very few detect cancer. Planned further analysis in this dataset will evaluate frequency of cancer detection in longer-term survivors. Additional research is necessary to determine the risks and benefits of screening breast MRIs being performed in this population.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-01-09.
Abstract P4-01-13: Practice patterns of MRI utilization for breast cancer treatment within the University of California system as part of the Athena initiative
Authors: Tokin CA , Ojeda H , Mayadev JS , Hylton NM , Fowble BL , Rugo HS , Hwang S , Hurvitz S , Wells C , Blair SL
Published: August 2014

Background: The appropriate utilization of Breast MRI in breast cancer care remains controversial. As part of a quality improvement initiative for breast cancer screening and treatment, we sent out a survey to physicians who treat breast cancer patients. All respondents are participants in the ATHENA initiative, a program which unites physicians, researchers, and patients at the five University of California medical centers.

Objective: To use the ATHENA infrastructure to perform a qualitative analysis of variations in breast cancer care.

Methods: Surveys were sent to 50 physicians in the ATHENA network whose practices are focused on breast cancer. Respondents were presented with clinical scenarios, and asked whether they would recommend MRI always/usually or sometimes/never. Differences were compared by Chi square.

Results: 39 physicians completed the survey (78% response rate). Of these physicians 29% were surgeons, 26% radiation oncologists and 45% medical oncologists. Athena physicians were more likely to order MRI for high risk screening of mutation carriers than not (85% yes vs. 15% no, p < 0.003) but not based on breast density alone or previous history of breast cancer. They were also more likely to order it for monitoring neo-adjuvant chemotherapy (70% yes vs. 30% no, p < 0.03). Although the majority answered that they would order a Breast MRI for new breast cancer the difference between responses was not significantly different (56% yes vs. 44%, p = 0.07).

Conclusion: Athena physicians follow established published guidelines which demonstrate a benefit for Breast MRI screening for BRCA mutation carriers but not based on density or previous history of breast cancer. The Athena network allows a forum for new practice guidelines to be implemented as data becomes available to improve patient outcomes and utilize the best evidence for patient care, where both the patterns of MRI use as well as the outcomes of practice patterns will be evaluated prospectively.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-01-13.
Abstract P4-01-16: Clinicopathological Factors that Predict the Accuracy of MRI in the Preoperative Setting.
Authors: Czechura Tomasz , Kantor Olga , Martz Brigid , Winchester David P , Winchester David J , Yao Katharine
Published: August 2014

Withdrawn by Author

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-01-16.
Abstract P4-02-05: A novel 64Cu-liposomal PET agent (MM-DX-929) predicts response to liposomal chemotherapeutics in preclinical breast cancer models
Authors: Lee H , Zheng J , Gaddy D , Kirpotin D , Dunne M , Drummond D , Allen C , Jaffray D , Hendriks B , Wickham T
Published: August 2014

Background: Liposomal anthracyclines (such as pegylated liposomal doxorubicin (PLD) or HER2-targeted liposomal doxorubicin (MM-302)) are being used and/or evaluated for the clinical management of breast cancer, but responses vary from patient to patient. It is hypothesized that variability in the deposition of liposomal therapeutics within tumors leads to differences in drug exposure, thereby directly influencing tumor response. We have developed MM-DX-929, a novel 64Cu-liposomal PET imaging agent, as a clinically-implementable tool to investigate whether image-based quantification of liposome deposition in tumors can predict treatment response to liposomal chemotherapeutics, including PLD, MM-302 and/or liposomal irinotecan (MM-398).

Objectives: Our primary objective is to demonstrate that the extent of tumor uptake of MM-DX-929 is predictive of tumor response to MM-302 in preclinical breast cancer xenograft models. A secondary objective is to enable clinical translation of MM-DX-929 to enable incorporation into existing therapeutic trials.

Methods: Mice bearing BT474-M3 mammary and subcutaneous tumors were injected intravenously with MM-DX-929 prior to dosing with MM-302. PET/CT imaging was performed at 16h post MM-DX-929 injection, and tumor uptake was determined. Response to treatment was quantified as tumor volume changes measured over a 2-month period by MRI.

Results: Tumor deposition of MM-DX-929 administration correlated well with treatment response to MM-302 (Spearman correlation coefficient of −0.891 and a p-value of 0.0004). MM-DX-929 accumulation in tumors prior to the start of the MM-302 treatment successfully predicted improved tumor growth inhibition following MM-302 treatment.

Conclusion: These findings support trials of MM-DX-929 as a predictive imaging agent to select patients who are most likely to respond to liposomal therapies. The clinical development of MM-DX-929 for identification of breast cancer patients likely to respond to liposomal therapeutics is currently being pursued.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-02-05.
Abstract P4-03-11: SUVmax of FDG-PET/CT is Associated with Chemotherapy Response Assay Test Results and Prognostic Factors in Breast Cancer Patients
Authors: Lee A , Chang J , Bang B , Lee J , Han D , Min S , Yun C , Kim BS , Lim W , Paik N , Moon B-I
Published: August 2014

Backgrounds:18F-fluorodeoxyglucose positron emission tomography with computed tomography (18F-FDG PET/CT) is widely used for cancer evaluation and there are several studies which suggested maximum standard uptake value (SUVmax) may reflect the cancer patients' prognosis. Adenosine triphosphate-based chemotherapy response assay (ATP-CRA) is commonly used as a chemosensitivity assay modality for the treatment of various cancers in clinical settings and there have been several studies on its usefulness in breast cancer. We previously reported chemosensitivity (ATP-CRA) results might reflect patients' prognosis. So we supposed that SUVmax could reflect chemosenitivity (ATP-CRA) results of patients, so performed this study to analyze the relationship between PET/CT SUVmax and chemosenstivity (ATP-CRA) results of these drugs. Furthermore, we evaluated prognostic factors for breast cancer according to SUVmax.

Materials and Methods: 102 breast cancer patients, who underwent PET/CT and chemosensitive (ATP-CRA) test between December 2010 and April 2012, were enrolled in this study. We analyzed, retrospectively, the correlation between SUVmax of PET/CT and chemosenitivity (ATP-CRA) results of doxorubicin/paclitaxel. SUVmax according to prognostic factors were also assessed.

Results: Chemosensitivity (ATP-CRA) results of doxorubicin and paclitaxel have significant positive correlation with SUVmax. Their correlation coefficient were 0.236 (p = 0.020) and 0.216 (p = 0.030), respectively. Patients with larger tumor size, higher histologic and nuclear grade, estrogen receptor negative, progesterone receptor negative, HER2 positive, and Ki67 positive (≥14%) have higher mean SUVmax values (p < 0.05). In molecular subgroup analysis, triple negative group showed higher mean SUVmax values than luminal A group (p < 0.001).

Conclusion: Higher SUVmax values of PET/CT is correlated with better chemosensitive (ATP-CRA) results of doxorubicin and paclitaxel, and poor prognostic factors of breast cancer. However, examination of additional cases might be needed.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-03-11.
Abstract P4-06-10: Epigenetic silencing of glutamine synthetase (Glul) defines glutamine depletion therapy
Authors: Cavicchioli F , Shia A , O'Leary K , Haley V , Crook TR , Thompson AM , Lackner M , Lo Nigro C , Schmid P
Published: August 2014

Background: Methylation-dependent transcriptional silencing of genes involved in amino acid synthesis can provide potential targets for novel synthetic lethality strategies. Glutamine synthetase (Glul) is the key enzyme in the biosynthesis of glutamine. We identified Glul as a novel gene subject to methylation-dependent transcriptional silencing in breast cancer cell lines using a combined functional screen with methylation reversal assays and methylation arrays.

Methods: Methylation reversal assays were performed using 5-aza-2-deoxycytidine and/or trichostatin treatment coupled with whole genome mRNA microarrays (Illumina HT-12 v4 Expression BeadChip Kit). Expression of Glul with and without pharmacological methylation reversal with azacytidine and/or trichostatin was validated using qRT-PCR and Western Blot. Methylation of Glul was analysed using methylation microarrays (Illumina 450K Methylation BeadChip), bisulphite sequencing and pyrosequencing. Sensitivity to glutamine deprivation was assessed using an MTT assay after culturing cells in media with various glutamine concentrations or in complete absence of glutamine. We used a panel of 55 breast cancer cell lines and formalin-fixed paraffin-embedded tissue from a series of 116 stage I-III primary breast cancers with linked mature clinical outcome data that were randomly selected from the Cuneo Tissue Bank. Tissue samples were subject to histopathological review to ensure adequate representation of cancer cells.

Results: Dense methylation of the CpG-island of Glul was detected in 45% of cell lines across all subtypes. Methylation of the CpG island was linked with absent or down-regulated expression of Glul in some but not all cell lines, and Glul expression could be reactivated by azacytidine and trichostatin in these cell lines. Methylation of shore areas was detected in several cell lines but was not associated with transcriptional silencing. Cells with methylation-dependent low or absent Glul expression were highly sensitive to glutamine deprivation, whereas cell lines without Glul methylation were rescued by compensatory up-regulation of Glul. Using pyrosequencing, dense methylation of the CpG island of Glul was found in 32.8% of patients, with an additional 17.2% of patients showing partial methylation. No significant association with a specific breast cancer subtype or outcome was found.

Conclusions: This is the first report of methylation-dependent transcriptional silencing of Glul expression in cancer. Our data demonstrate that a significant proportion of primary breast cancers show methylation of Glul and suggest that glutamine deprivation could be a novel synthetic lethality strategy for these cancers.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-06-10.
Abstract P4-06-14: CD146-suppresses breast tumor invasion via a novel transcription target TIMPv
Authors: Shanmuganathan S , AbdElmageed Z , Fernando A , Gaur R , Ramkumar A , Bhat S , Gupta I , Muzumdar S , Hakkim L , Ouhtit A
Published: August 2014

The function of the cell adhesion receptor CD146, a recently discovered marker of endothelial cells and a tumor promoter of melanoma and other cancers, is controversial in breast cancer (BC). However several lines of evidence supports its role as a tumor suppressor in BC. Further, the molecular mechanisms underpinning this suppression are unknown, neither has the ligand for CD146 been identified. Using a novel validated Enhanced Green Fluorescent Protein (EGFP)-inducible systems of CD146 expression both in vitro and in vivo, we provide here molecular and functional evidence of CD146 and its novel transcriptional target TIMPv (a variant of tissue inhibitor of metallo-proteinases) in underpinning the suppression of BC invasion.

Tetracycline (tet-on) CD146 system was developed in both MCF-7 and MDA-231 BC founder cell lines, and validated using time course RT-PCR and western blot analyses, and fluorescent microscopy. In functional experiments, induction of CD146 inhibited BC cell migration and invasion. TIMPv, the only endogenous protein inhibitor known for metallocarboxypeptidases, was identified by expression profiling as a novel transcriptional target of CD146-signaling, an association validated by quantitative PCR and immunoblotting experiments in a range of breast and melanoma cancer cells. However, siRNA inhibition of CD146 in the SKMel-28 melanoma cell line increased TIMPv expression, suggesting that while TIMPv is a positive transcriptional target of CD146 in BC cells, it is negatively regulated in melanoma cells. Furthermore, using invasion assay, the functional relevance of TIMPv to CD146-suppressed metastasis was demonstrated by selective suppression of TIMPv in CD146-expressing BC inducible cells using RNAi. More interestingly, induction of CD146 expression in vivo, using the tet-on CD146 expression system in BC Xenograft model resulted in suppression of breast tumor growth. Further, Clinical analysis of breast tissue samples by Immunohistochemistry showed that TIMPv expression patterns paralleled those of CD44s during breast tumor progression. Pharmacological and molecular approaches revealed that the activation of NFκB via Akt pathway couples CD146 to the transcription of TIMPv in BC cells.

Our study is the first report to provide a functional molecular link of a novel transcriptional target of CD146, TIMPv, to cancer via a unique axis that underpin CD146-suppressed BC progression; TIMPv is a potential target for guiding the development of novel therapeutic strategies for BC.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-06-14.
Abstract P4-07-01: The Class I Selective PI3K Inhibitor GDC-0941 Enhances the Efficacy of Docetaxel in Human Breast Cancer Models by Increasing the Rate of Apoptosis
Authors: Wallin JJ , Guan J , Prior WW , Lee LB , Ross L , Belmont LD , Koeppen H , Belvin M , Sampath D , Friedman LS
Published: August 2014

Docetaxel (DTX) is commonly used as a front-line treatment option for breast cancer but many patients ultimately relapse and succumb to disease progression. Phosphatidylinositol 3-kinases (PI3K) are lipid kinases that regulate breast tumor cell growth, migration and survival. Given that phosphatidylinositol 3-kinase (PI3K) is frequently activated in breast cancer and induces chemo-resistance, it is an attractive target for combination therapy with standard of care drugs such as DTX. The current study was intended to determine if GDC-0941, an orally bioavailable class I selective PI3K inhibitor, enhances the anti-tumor activity of DTX in human breast cancer models in vitro and in vivo. A panel of 25 breast tumor cell lines representing luminal, HER2+ and basal sub-types were treated with GDC-0941, DTX or the combination of both drugs and assayed for cellular viability, modulation of PI3K pathway markers and apoptosis induction. The combination of GDC-0941 and DTX decreased the cellular viability of breast tumor cell lines in vitro but with variable drug synergy. The addition of both drugs resulted in stronger synergistic effects in a sub-set of tumor cell lines that was not predicted by breast cancer sub-type or mutations within the PI3K pathway. Human xenografts of breast cancer cell lines and patient-derived tumors were utilized to assess efficacy of GDC-0941 and DTX in vivo. In these models, the best combination efficacy was detected when the two drugs were dosed within 1 hour of each other. We also observed that GDC-0941 increased the rate of apoptosis in cells arrested in mitosis upon co-treatment with DTX. Our data provides a preclinical rationale for evaluating GDC-0941 in combination with DTX for breast cancer treatment.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-07-01.
Abstract P4-09-11: Fatty Acid Synthase (FASN) expression in Triple-Negative Breast Cancer
Authors: Viñas G , Oliveras G , Perez-Bueno F , Giro A , Blancafort A , Puig-Vives M , Marcos-Gragera R , Dorca J , Brunet J , Puig T
Published: August 2014

Background: In approximately 15–20% of patients with breast cancer, the tumors do not express estrogen receptor (ER), progesterone receptor (PR) and do not have amplification of HER2. These tumors are called triple-negative breast cancer (TNBC) and patients with these tumors have a poor prognosis. There is no clinically validated, molecularly targeted therapy for TNBC patients and they can be treated only with chemotherapy. Thus, the identification of a novel targeted therapies for TNBC patients would be of great benefit. Fatty acid synthase (FASN) is a multi-enzyme protein that catalyzes fatty acid synthesis. Its main reaction is to synthesize palmitate from acetyl-CoA and malonyl-CoA, in the presence of NADPH, into long-chain saturated fatty acids. Expression levels of FASN are low or undetectable in normal human tissues except for the liver and the adipose tissue. In contrast, high levels of FASN expression have been detected in several human carcinomas. Several reports highlight that FASN overexpression in tumor samples correlates with progression, aggressiveness and metastatic potential of the disease. Previously, our group has shown that FASN activity inhibition induces apoptosis in several human cancer cells. The aim of our study was to evaluate the expression of tumor levels of FASN in triple-negative breast cancer patients.

Methods: FASN tumor expression was retrospectively evaluated in 30 paraffin-embedded core-biopsies of 30 patients with TNBC using the Fatty Acid Synthase polyclonal antibody (Assay design, Enzo Life Sciences, Exeter United Kingdom) and the detection kit EnVision™ (DAKO, Glostrup, Denmark). FASN expression levels were determined by immunohistochemistry (IHC) using the AutostainerPlus Link (DAKO). FASN expression was graded from 0 to 3+, meaning 0–1+ normal amounts of FASN protein compared to non-tumor breast tissue, 2+ moderate amounts and 3+ highest levels of FASN expression. In vitro, we have determined the effect of the FASN-inhibitors, EGCG and G28UCM, on cell viability (measured by the MTT assay) and apoptosis [as assessed by cleavage of poly(ADP-ribose) polymerase (PARP)] of two TNBC cell lines, MDA-MB-231 and MDA-MB-468.

Results: From 30 patients with TNBC, 66.6% (20/30) of the patients had a high expression of FASN (FASN 3+) and 33.3% (10/30) had a moderate expression of FASN (FAS 2+). None of the tumors had lack of FASN expression analysed by immunohistochemistry. In vitro, the FASN-inhibitor G28UCM induced apoptosis and showed low IC50 values of citotoxicity in both, MDA-MB-231 and MDA-MB-468 TNBC cell lines.

Conclusions: FASN is overexpressed in triple-negative breast cancer tumors. The absence of target therapies for this breast cancer subtype and its poor prognosis lead to the exploration of FASN as therapeutic target for TNBC patients.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-09-11.
Abstract P4-12-01: A nomogram based on clinical, imaging and histological data to predict the risk of upgrades to malignancy at surgery in biopsy-diagnosed premalignant lesions of the breast
Authors: Uzan C , Mazouni C , Balleyguier C , Mathieu M-C , Ferchiou M , Delaloge S
Published: August 2014

Background: Population-based mammography screening has resulted in the increased detection of suspicious, non-palpable lesions that require histopathological assessment. When a “pre-malignant” lesion, such as lobular carcinoma in situ (LCIS), atypical ductal or lobular hyperplasia (ADH and ALH) or flat epithelial atypia (FEA), is discovered on the primary biopsy, a surgical excision is recommended due to the risk of underestimation of the lesion. DCIS or invasive carcinoma is finally retrieved in 10–30% of the patients, meaning a large part receive unnecessary surgery. The aim of this study was to define a nomogram integrating clinical, imaging and histological data to predict for the risk of underestimation of the lesion. Intentionally, we decided to include all the premalignant lesions which implied the same management, whereas most of the previous models of the literature are focused in only one or another type of atypia.

Patients and Methods: We collected complete clinical, radiological and double-reading histological data on all patients with a diagnosis of pure atypical lesion (CLIS, ADH, ALH, FEA) on image-guided biopsy performed at the One-Stop Unjt of our breast care center from 2004 to 2011. Univariate and multivariate logistic regression analyses were used to develop a model predicting for the presence of DCIS or invasive carcinoma on the final surgical sample, and build a nomogram. This nomogram was evaluated on a training set of 205 patients treated at IGR.

Results: 205 patients were eligible for the study. Of these 205 patients, 50 cancers (24.4%) were diagnosed at definitive surgery (21 DCIS, 20 ductal and 9 lobular invasive carcinoma). Univariate analysis retrieved age (p = 0.03), number of biopsy cores (p = 0.02) and type of radiological anomalies (p = 0.02) as factors associated with cancer at surgery. The presence of microcalcifications on biopsies of limit significance was included in the multivariate analysis (p = 0.09). The final most informative nomogram included information on patient age (p = 0.03), type of radiological anomalies (p = 0.02) and number of biopsy cores (p = 0.04). The predictive accuracy of the nomogram was 0.71 and 0.68 in the training set before and after bootstrapping, respectively. The calibration of the nomogram was good. The sensitivity, specificity, positive predictive value, and negative predictive values were 68%, 73%, 45%, and 88%, respectively.

Conclusion: This nomogram could help identify a subset of patients with premalignant disease for whom surgery could be spared and who would be eligible for exclusive clinical follow-up. A validation study is currently undergoing in an external dataset.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-12-01.
Abstract P4-12-06: A mammographic density prediction model using environmental factors, endogenous hormones and growth factors in Japanese women
Authors: Yoshimoto N , Nishiyama T , Toyama T , Takahashi S , Shiraki N , Sugiura H , Endo Y , Iwasa M , Asano T , Fujii Y , Yamashita H
Published: August 2014

The incidence of breast cancer in Japanese women has doubled in all age groups over the past two decades, making it important to evaluate breast cancer risk factors in Japanese women. It is well known that mammographic density is positively associated with breast cancer risk in Western countries, and mammographic density is known to be affected by some environmental factors, serum hormones, and growth factors. We performed stepwise variable selection in a multiple regression model with fifteen independent variables as described below, based on the Akaike information criteria (AIC) to build a mammographic density prediction model using a dataset of 1191 women (913 women with breast cancer and 278 disease-free controls). The variables included were: environmental risk factors (body-mass index (BMI), age at menarche, pregnancy, age at first birth, breastfeeding, family history of breast cancer, age at menopause, use of hormone replacement therapy, alcohol intake and smoking), serum hormones and growth factors (estradiol, testosterone, prolactin, insulin-like growth factor 1 (IGF1) and IGF binding protein 3 (IGFBP3)) and mammographic density. The resulting prediction model is:

Mammographic density = + 0.000476 (IGF1) −0.0605 (testosterone) − 0.0508 (IGFBP3) − 0.00683 (age) − 0.0175 (BMI) + 0.00883 (age at menarche) − 0.0153 (breast feeding), (R2 = 0.336).

In this model, IGF1, testosterone, IGFBP3, age, BMI, age at menarche, and breastfeeding were considered to be important factors. IGF1 and age at menarche were positively associated with mammographic density, while on the other hand testosterone, IGFBP3, age, BMI, and breast feeding were negatively associated with mammographic density. Further studies are required to build a modified model incorporating serial measurements of serum hormones and growth factors to take into account time-dependent changes of serum hormones and growth factors, and to assess its accuracy.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-12-06.
Abstract P4-13-08: Diabetes, Related Factors and Breast Cancer Risk
Authors: Boyle P , Boniol M , Koechlin A , Bota M , Robertson C , Leroith D , Rosenstock J , Bolli GB , Autier P
Published: August 2014

Diabetes and breast cancer are both extremely common conditions in women and may share common risk factors. It is natural to investigate any potential common risk factors and to seek biological clarification and improve prospects for prevention.

Therefore, in order to help clarify the potential association between diabetes, related factors and breast cancer risk, a comprehensive literature review and formal meta-analysis was carried out, planned, conducted and reported following PRISMA guidelines regarding meta-analysis of observational studies. Variables studies in relation to breast cancer risk were adiposity, physical activity, glycaemic load, glycaemic index, diabetes, IGF-1, fasting glucose, fasting insulin and C-peptide, adiponectin and metformin and glargine use among patients with diabetes. For all variables except diabetes and breast cancer, only prospective studies were included in meta-analyses. Summary Relative Risks (SRR) and corresponding 95% Confidence Intervals (CI) were calculated from random effect models.

For breast cancer at all ages, the calculated risks were as follows: diabetes (SRR = 1.27 95% CI (1.16, 1.39); physical activity (SRR = 0.88, 95% CI (0.85, 0.92)); glycaemic load (SRR = 1.06, 95% CI (1.00, 1.12)); glycaemic index (SRR = 1.04, 95% CI (0.99, 1.10)); fasting glucose (SRR = 1.12, 95% CI (1.01, 1.24)); serum insulin (SRR = 1.18, 95% CI (0.75, 1.85)); c-peptide (SRR = 1.29, 95% CI (0.91, 1.82)); adiponectin (SRR = 1.16, 95% CI (0.93, 1.46)); metformin (SRR = 1.00, 95% CI (0.69, 1.46)); and glargine (SRR = 1.11, 95% CI (1.00, 1.24)). An increase of 5 units in Body Mass Index (a weight increase if 14.5 kg in a person 1.70 metres tall) was associated in post-menopausal breast cancer (SRR = 1.12, 95% CI (1.08, 1.16)) but not at pre-menopausal ages (SRR = 0.83, 95% CI (0.72, 0.95)). Serum insulin was associated with breast cancer at post-menopausal ages but not at pre-menopausal ages whereas with c-peptide there was a significant association at pre-menopausal ages but not post-menopausal. For IGF-1, Hodge's Standardised Mean Difference (HSMD) was calculated in cohort studies and there was no significant association with breast cancer at all ages (HSMD = 0.003, 95% CI (−0.059, 0.065)), at post-menopausal ages (HSMD = −0.014, 95% CI (−0.106, 0.077)) or at pre-menopausal ages (HSMD = 0.039, 95% CI (−0.038, 0.117)).

The risk of breast cancer is increased among post-menopausal women who have diabetes. Among those factors related to diabetes, key risk factors for breast cancer appear to be adiposity and lack of physical activity which are both related to the risk of developing diabetes. Action on these lifestyle factors should form the basis of a common prevention strategy. There is a need to re-evaluate potential biological mechanisms to explain the increased risk in post-menopausal women with diabetes.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-13-08.
Abstract P4-14-02: National Trends and Indications for Nipple-Sparing Mastectomy: An Analysis Using the Surveillance, Epidemiology, and End Results (SEER) Database
Authors: Agarwal S , Agarwal S , Agarwal J
Published: August 2014

Introduction: Nipple-sparing mastectomy (NSM) is an evolving procedure occasionally used for breast cancer treatment. However, indications for NSM are institution-specific and lack consensus in the literature. This study uses the Surveillance, Epidemiology, and End Results (SEER) database to investigate the oncologic and demographic characteristics of breast cancer patients treated with NSM in the United States from 2005–2009.

Methods: Female breast cancer patients treated from 2005–2009 with nipple-sparing/subcutaneous mastectomy (SEER code 30), defined as removal of breast tissue with preservation of nipple-areolar complex and overlying skin, were included. Variables analyzed included patient age, race, tumor stage, tumor size, lymph node status, and radiotherapy delivery.

Results: A total of 449 female breast cancer patients who underwent NSM were isolated from the SEER database. The number of patients in the SEER database who underwent NSM increased steadily each year, from 66 in 2005 to 133 in 2009. Mean age was 52 years (s.d. 12 years). Breakdown by race showed that 369 (82%) patients were White women, 42 (9%) were Black women, 36 (8%) were of Asian descent. A majority of patients (402) were documented to have ductal or lobular carcinoma. Documented tumor diameter was less than 3 cm in 63% (284) of patients. Invasive cancer was documented in 283 (63%) patients; in situ cancer was documented in 166 (37%) patients. Lymph node involvement was negative in 374 (83%) patients. Radiation therapy was delivered to 91 (20%) patients; however, 64 patients receiving radiation therapy were node-negative. Radiation was delivered to 6% (10) of patients with in situ cancer and 27% (77) of patients with invasive cancer (p < 0.0001).

Conclusions: The number of NSM procedures reported in SEER for breast cancer has increased from 2005–2009. However, there is a lack of consensus in the literature as to which patients may be candidates for NSM. By utilizing a national database, we are able to aggregate and report on the national experience with therapeutic NSM, to date. Patients undergoing NSM for breast cancer were characterized predominantly by tumors less than 3 cm in diameter and with negative lymph node involvement. Patients with both invasive and in situ tumors were considered suitable candidates for NSM. A majority of patients had either ductal or lobular carcinomas. A minority of patients received radiation therapy, although there was a tendency towards providing patients with invasive tumors with radiation therapy. This national data should help guide patient selection for NSM, although future studies are needed to report on the long-term oncologic safety of this procedure.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-14-02.