bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P1-08-13: Predictive value of intrinsic luminal subtypes in premenopausal women with endocrine-responsive early breast cancer: Results from Austrian breast and colorectal cancer study group trial 5
Authors: Bago-Horvath Z , Rudas M , Jakesz R , Greil R , Dietze O , Suppan C , Lax S , Singer CF , Filipits M , Gnant M
Published: August 2014

Purpose: The aim of the present study was to assess the predictive value of intrinsic luminal subtypes in premenopausal hormone receptor-positive early breast cancer patients who received adjuvant endocrine treatment or chemotherapy.

Patients and Methods: Intrinsic luminal breast cancer subtypes were centrally assessed by immunohistochemistry on whole tissue sections of breast cancer patients who had been enrolled in the Austrian Breast and Colorectal Cancer Study Group (ABCSG) Trial 5 and received either 3 years of goserelin plus 5 years of tamoxifen or six cycles of cyclophosphamide, methotrexate, and fluorouracil (CMF). Luminal A disease was defined as Ki67 <20%, Luminal B/HER2-negative subtype as Ki67≥20%. Luminal B/HER2-positive subtype displayed HER2 overexpression by IHC or amplification by ISH. Recurrence-free survival (RFS) and overall survival (OS) were analyzed using Cox models adjusted for clinical and pathological factors.

Results: 155 (36%), 217 (50%) and 59 (14%) of 431 tumors were classified as Luminal A, Luminal B/HER2-negative and Luminal B/HER2-positive, respectively. Luminal B subtypes were associated with poor outcome. Patients with Luminal B tumors had a significantly shorter RFS (adjusted hazard ratio [HR] for recurrence: 2.29, 95% confidence interval [CI] 1.44-3.64, P < 0.001) and OS (adjusted HR for death: 3.97, 95% CI 1.94-8.08, P < 0.001) as compared to patients with Luminal A disease. No interaction between intrinsic luminal subtypes and treatment was observed (test for interaction: P = 0.42 for RFS; P = 0.32 for OS). Combination endocrine treatment with goserelin/tamoxifen tended to be more effective than CMF chemotherapy in both luminal A and luminal B subtypes.

Conclusion: Intrinsic luminal subtype is an independent prognostic factor for recurrence and death in premenopausal women with early-stage, hormone receptor positive breast cancer but is not predictive for outcome of adjuvant treatment with either goserelin/tamoxifen or chemotherapy with CMF.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P1-08-13.
Abstract P1-08-31: Putative role of genetic variants of eNOS in survival and toxicity of patients given antiangiogenic therapy
Authors: Del Re M , Fontana A , Del Re I , Bona E , Salvadori B , Ferrarini I , Landucci E , Fancelli S , Bertolini I , Michelotti A , Falcone A , Danesi R
Published: August 2014

Background: The development of tumor angiogensis is mainly driven by vascular endothelial growth factor (VEGF), which is strongly overexpressed in many cancers. VEGF induces the expression of the endothelial nitric oxide (NO) synthase (eNOS) and the resultant overproduction of NO may be associated with recruitment of inflammatory cells, disruption of endothelial barrier, edema and impaired drug delivery within tumors (1). Functional polymorphisms in the eNOS gene, including -786C>T and 894G>T, have been associated with impaired production of NO and higher incidence of hypertension (HT) (2), diabetic nephropathy (3) and glaucoma (4). Since suppression of VEGF-eNOS signal transduction by antiangiogenic drugs may normalize tumor vasculature by restoring interstitial fluid pressure and drug distribution in tumors, but may induce HT in patients, the purpose of this study was to examine the association between the major eNOS variants -786C>T and 894G>T with treatment outcome and risk of HT in metastatic breast cancer (MBC) patients given bevacizumab. Methods: Sixty-five MBC patients given bevacizumab as per approved indication were enrolled. Main characteristics were: mean age 49.5 years (range 29-73) at first diagnosis, 53 years (range 34-74) at metastatic progression and PS 0-1. Four subjects with HT and 1 patient with compensated cardiovascular disease were also included. First-line chemotherapy for metastatic disease was taxol plus bevacizumab. Germline DNA was extracted from peripheral blood with the Qiamp Mini Kit (Qiagen, Milano, Italy) and examined for eNOS -786C>T and 894G>T variants by Real Time PCR (Life Sciences 7900HT platform) and automatic sequencing (Life Sciences 3100 Avant). The study was approved by the local Ethics Committee. Results: Genotype frequencies are reported below (Table 1). The presence of -786TT genotype was associated with longer PFS compared with the other genotypes (median PFS 95%CI, CC/CT = 9 vs TT = 12 months, Log-rank [Mantel Cox] test p = 0.0066), but not with any grade of HT. Also the 894GT/TT was associated with longer PFS compaed with the GG homozygous wild-type genotype (median PFS 95%CI, GG = 7,5 vs GT/TT = 10 months, Log-rank [Mantel-Cox] test p = 0,0497). The incidence and severity of HT did not vary among genotypes. Conclusion: Patients bearing deficient eNOS variant did not experience higher risk or severity of HT with respect to the wild-type allele but enjoied a longer PFS.

Table 1. Genotype frequenciesSNPFrequencies%Frequencies%Frequencies%-786C>TCCCTTT 37%33.8%26.2%894G>TGGGTTT 41.5%53.8%4.6%

References

1. Goel S et al. Physiol Rev 2011;91:1071

2. Niu W, Qi Y. PLoS One 2011;6:e24266

3. Rahimi Z et al. Dis Markers 2013;34:437

4. Awadalla MS et al. Invest Ophtalmol Vis Sci 2013;54:2108.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P1-08-31.
Abstract P1-08-35: Androgen receptor expression and changes in serum dehydroepiandrosterone sulfate levels in locally advanced breast cancer patients who received neoadjuvant chemotherapy
Authors: Park S , Lee JS , Park JM , Park HS , Kim SI , Park B-W
Published: August 2014

Purpose: The associations between androgen receptor (AR) expression, serum dehydroepiandrosterone-sulfate (DHEA-S) level and response to neoadjuvant chemotherapy (NCT) were investigated in locally advanced breast cancer patients who received NCT.

Methods: Between May 2008 and April 2013, a total of 346 patients underwent NCT mainly based on anthracycline with or without taxane regimens. Biomarkers including AR were immunohistochemically determined using biopsy specimens before NCT and tumors with ≥ 1% nuclear staining were considered positive for AR. Changes in serum DHEA-S levels before and after NCT were examined by chemiluminescent immunoassay (Access DHEA-S, Beckman Coulter, Inc., Brea, CA) in 205 (59.2%) patients. Pathologic complete response (pCR) was defined as the absence of residual invasive carcinomas in the surgical specimens of breast and lymph nodes. Breast cancer subtypes were categorized by immunohistochemistry of estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2), and Ki-67.

Results: Mean age at diagnosis was 49.7 years (range, 26–76). Clinical tumor size was ≥ 2cm in 243 (70.2%) patients and 318 (91.9%) showed node-positive disease at initial presentation. AR-positive tumor was determined in 195 (56.4%) patients and 95 (27.5%) achieved pCR after NCT. AR-negative tumors were significantly associated with grade III, ER-negative, PR-negative, HER2-negative, high Ki-67 index, and triple-negative subtype tumors. By univariate logistic regression analysis, AR-negative tumor showed significantly higher odds ratio of 1.650 (95% CI, 1.026–2.654; p-value, 0.039) for achievement of pCR. Nevertheless, not AR status but breast cancer subtype, longer duration of NCT and use of targeted agents remained to be significant for pCR by multivariate model. In the analysis of subgroup by breast cancer subtypes, AR-negative tumor was associated with pCR in 75 patients with the low proliferative luminal A subtype, while AR-positive tumor achieved higher pCR in 59 patients with the HER2-positive luminal B subtype. In 205 patients available for serum DHEA-S levels, there was no difference in DHEA-S between pCR and non-pCR groups, although a trend of decrease in DHEA-S after NCT was noted among postmenopausal AR-positive or luminal A subtype women who achieved pCR.

Conclusions: AR expression might predict pCR in locally advanced breast cancer patients who undergo NCT, but this implication could be different according to breast cancer subtypes. Clinical significance of changes in serum DHEA-S levels during NCT remains to be elucidated.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P1-08-35.
Abstract P1-08-36: Neoadjuvant trial of bevacizumab (B) and trastuzumab (T) in combination with weekly paclitaxel (P) as neoadjuvant treatment in HER2-positive breast cancer: A phase II trial (AVANTHER). Analysis of biomarkers
Authors: Fernández M , Martinez N , Calvo I , Garcia E , Ugidos L , Herrero M , Quijano Y , Durán H , Garcia M , Suarez A , López-Rios F , Perez D , Perea S , Hidalgo M , Garcia-Estevez L
Published: August 2014

Background: AVANTHER is a phase II trial of preoperative systemic therapy combining B with T and P in a weekly regimen in HER2 positive breast cancer to assess safety and efficacy of the combination. The primary endpoint was pathological complete response (pCR), defined as the absence of invasive disease at the time of histological study. Tissue samples were collected at baseline and the relationship between the expression of PTEN, PI3KCA, TOPO 2a, VEGF, activation of insulin growth factor 1-receptor (IGF1R) and pCR was assessed.

Methods: Patients (pts) with centrally-confirmed HER2–positive (IHC 3+ or FISH amplified) breast cancer (stage II or III including locally advanced) received neoadjuvant chemotherapy (NC) with weekly P (80mg/m2/week) for 12 weeks in combination with weekly T (4mg/kg loading dose and 2 mg/kg maintenance) and B (15mg/kg every 3 weeks) for 4 cycles. After surgery all pts received T (1 year) and liposomal doxorubicin plus cyclophosphamide every 3 weeks (4 cycles). Tumor samples were evaluated for the expression of PTEN and TOPO 2α by fluorescent in situ hybridization (FISH), IGF-1R, VEGF, and EGFR using standard immunohistochemistry (IHC). Status of PI3KCA was determined using Fluidigm-based real-time PCR.

Results: A total of 44 pts were enrolled. Median tumor size: 3.9 cm. Nine (21%) pts had stage IIA; 19 (45%) stage IIB; 10 (24%) stage IIIA and 4 (10%) stage IIIB. Twenty-nine (69%) pts had estrogen positive receptors. PCR achieved was 42.9%. PTEN expression was normal in all pCR group and 1 patient (pt) (3.8%) had a PTEN deletion in group with no pCR. PI3KCA was mutated in 3pts (16.7%) (1pt E542k and 2 pts H1047r) in the pCR group and 4 pts (15.4%) (1pt E545k, 1pt G1049r and 2 pts H1047r) in the no pCR. Topo 2α was amplified in 8 pts in both groups. The expression of IGFR 1 was negative in 44.4% in the pCR group and 15.4% in patients with no pCR (p = 0.04). VEGF was positive in 7pts (39%) in pCR group and 10pts (38.5%) in no pCR.

Conclusions: Our results are in agreement with some prior studies that show that expression of IGFR could be correlated with trastuzumab resistance. Therefore, these findings raise a hypothesis to be validated in prospective studies.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P1-08-36.
Abstract P1-08-38: HER2 heterogeneity affects trastuzumab responses and survival in patients with HER2-postive metastatic breast cancer
Authors: Lee HJ , Seo AN , Kim EJ , Jang MH , Suh KJ , Ryu HS , Kim YJ , Kim JH , Im S-A , Gong G , Park IA , Jung KH , Park SY
Published: August 2014

Purpose: Heterogeneity of HER2 gene amplification is found in a subset of breast cancers. However, it is not known whether breast cancers with heterogeneous HER2 amplification respond differently to HER2-targeted therapy than those with homogeneous amplification. In this study, we investigated the relationship between HER2 heterogeneity and trastuzumab resistance and clinical outcomes in patients with HER2-positive metastatic breast cancer.

Patients and methods: We studied tumor tissues from 127 patients with HER2-positive metastatic breast cancers who had received trastuzumab-based chemotherapy. Regional and genetic heterogeneity of HER2 amplification was determined in three different areas of each tumor by immunohistochemistry (IHC) and silver in situ hybridization. We also assessed the overall levels of HER2 amplification, and the proportion of tumor cells with a HER2/CEP17 ratio >2.2 or HER2 overexpression (IHC score of 3+). HER2status including HER2 heterogeneity was correlated with trastuzumab responses and survival of the patients.

Results: HER2regional and genetic heterogeneity was confirmed in 7.8% and 3.6% of cases, respectively. Poor response to trastuzumab was associated with overall low-level amplification, HER2regional heterogeneity, HER2/CEP17 ratio >2.2 in <80% of tumor cells, and HER2 IHC score of 3+ in <75% of tumor cells. In survival analyses, low-level HER2 amplification, HER2regional heterogeneity, and HER2/CEP17 ratio >2.2 in <80% of tumor cells were associated with shorter time to progression and lower overall survival in univariate and multivariate analyses.

Conclusion: Accurate assessment of HER2 status including HER2 heterogeneity is important in predicting trastuzumab responses and outcomes in patients with HER2-positive metastatic breast cancer.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P1-08-38.
Abstract P1-09-06: Breast cancer in the Pan-American region: Inequities in incidence and mortality rates according to the human development index
Authors: Barrios CH , Werutsky G , Martínez-Mesa J
Published: August 2014

Background: Breast cancer (BC) is considered a public health problem in countries of the Pan-American Health Organization (PAHO). Data from GLOBOCAN 2008 shows BC age-adjusted incidence and mortality rates of 57.1 and 13.7 per 100,000 inhabitants in the region. The aim of the present study is to evaluate the association between BC incidence and mortality rates with the human development index (HDI) in PAHO countries.

Methods: This is an ecological analysis including 29 countries (PAHO) with reported data both in GLOBOCAN 2008 and in the 2013 United Nation Development Reports (UNDP). In alphabetical order the participant countries were Argentina, Bahamas, Barbados, Belize, Bolivia, Brazil, Canada, Chile, Colombia, Costa Rica, Cuba, Dominican Republic, Ecuador, El Salvador, Guatemala, Guyana, Haiti, Honduras, Jamaica, Mexico, Nicaragua, Panama, Paraguay, Peru, Suriname, Trinidad and Tobago, United States of America, Uruguay and Venezuela. HDI is a composite statistic of life expectancy, education, and income and was analyzed as a continuous score. Age-adjusted BC incidence and mortality rates were taken from GLOBOCAN 2008 and log-transformed due to skewness. Pearson correlation and simple linear regression were performed using Stata 12 (Stata Corp., College Station, USA).

Results: A positive correlation was found between HDI and log-transformed age-adjusted BC incidence and mortality rates. The correlation coefficient between HDI and BC incidence rate was 0.68 (p-value<0.001). The correlation with BC mortality rate was 0.49 (p-value = 0.007). Linear regression showed that an increase in one HDI unit lead to a gain of 3.51 points (se = 0.72; p-value<0.001) in the incidence rate and 2.14 points (se = 0.73; p<0.007) in the mortality rate.

Conclusion: HDI inequities are important and should be considered in the analysis of the difference in BC incidence and mortality rates seen in PAHO countries.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P1-09-06.
Abstract P1-09-10: Disparities in presentation of breast cancer in a geriatric population
Authors: Ursem C , Camacho F , Anderson R , Kimmick G
Published: August 2014

Background: Although there are known to be disparities by socioeconomic status (SES) in breast cancer presentation, this has not been as well studied in the elderly. We examined older women in North Carolina (NC) using insurance status as an indicator of SES. Dual Medicaid/Medicare (dMM) status was used as a surrogate for low SES and Medicare only (M) as a surrogate for higher SES.

Methods: From the 1999-2002 NC Central Cancer Registry, we identified women age ≥65 years presenting with nonmetastatic breast cancer, having surgery within 60 days of diagnosis, no neoadjuvant therapy, and insured by Medicare only or dual Medicaid/Medicare. We used Chi-square tests to compare demographic and tumor characteristics, including: age, race, ACE comorbidity index, tumor size, lymph node status (LN), ER/PR status and HER2 status.

Results: We identified n = 3088 women with mean age 75 (SD 6.69) years, including 560 dMM and 2528 M insured women. We found that the dMM patients were older than M patients, with 57.7% ≥75 years, vs. 42.6% (p<0.001) and mean age 76.9 vs. 74.5 years (p<0.001), as well as composed of more African Americans, 34.6% vs. 6.6% (p<0.001). The dMM group also had significantly more comorbidity, with ACE comorbidity index: none 21.4% vs. 40.0%, minimal 45.2% vs. 43.2%, moderate 15.5% vs. 9.8% and severe 17.9% vs. 6.9% (p<0.001). The dMM patients were diagnosed more often with regional as compared to local disease, 26.3% vs. 19.7% (p<0.001) and had larger tumors at diagnosis, mean 23.46mm vs. 18.50mm (p<0.001). Additionally, the dMM patients were found to have more LN involvement: 0 LN in 73.4% vs. 80.0%, 1-3 LN in 19.6% vs. 15.5%, 4-9 LN in 5.7% vs. 3.5% and ≥ 10 LN in 1.3% vs. 1.1% (p = 0.004). We did not find significant differences in tumor histology, grade, ER/PR subtype or HER2 status.

Conclusions: We found that in a population of elderly breast cancer patients, lower SES as defined by Medicaid insurance status was associated with older age, African American race and more advanced comorbidity. These same patients presented with move advanced disease, characterized by larger tumors and more LN involvement.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P1-09-10.
Abstract P1-09-18: Doctors with breast cancer are more likely to opt for mastectomy: Fact or fiction?
Authors: O'Connell RL , Gui G , Rusby JE
Published: August 2014

Introduction

Doctors who are diagnosed with breast cancer make up a small but unique subset of women owing to their medical knowledge. Anecdote suggests that doctors with breast cancer are more likely to opt for mastectomy than non-medically qualified patients.

The primary aim of this study was to compare mastectomy rates in medically-qualified doctors with degree-educated controls with breast cancer.

Methods

Research ethics approval was obtained. Patients diagnosed with breast cancer between 1/1/2006 and 31/12/2011 and who had signed a generic research consent form were included in this observational study. Every patient was screened by occupation independently by 2 investigators to identify medically-qualified doctors and suitable (degree-educated) controls such as teachers, lawyers etc. Those with a medical background (e.g.nurse, physiotherapist etc) but not a medical doctor were excluded. Further exclusions were necessary if patients who were reviewed for a second opinion, reconstruction only or radiotherapy only or were male, surgery was not performed, BRCA positivity (might opt for bilateral mastectomy), unusual histology (e.g. lymphoma, sarcoma), recurrence or had a past history of contralateral breast cancer. Doctors and controls were compared by age, tumour characteristics and treatments using Student's t test for continuous variables and Fisher's exact test for categorical variables.

Results:

In total 8623 patients were reviewed and/or treated for breast cancer of whom 5215 had signed the generic research consent form. Overall 607 were either doctors or appropriate occupation controls. After exclusions 473 patients were eligible to be included in the analysis. Of these: 51 were medically-qualified doctors and 422 were degree-educated (controls).

Comparison of tumour characteristics and treatment between doctors and controls DoctorsControlspNumber51422 Mean age (years)53.350.80.11Mean total tumour size (mm)32.231.60.89     Number (%)Number (%)pDCIS alone10(20)53 (13)0.18Node positive16 (31)157 (37)0.44ER positive45 (88)337 (80)0.18HER2 positive12 (24)113 (27)0.73    Neoadjuvant chemotherapy6 (12)58 (14)0.83Adjuvant chemotherapy14 (27)154 (36)0.21    mastectomy17 (33)145 (34)1.00Immediate reconstruction9 (53)65 (45)0.61Post mastectomy radiotherapy5 (29)68 (47)0.20Comparison of age, tumour characteristics, mastectomy, chemotherapy and post mastectomy radiotherapy rates between doctors and the control group

There was no statistical difference in age and tumour characteristics between the doctors with breast cancer and the control group. There was no statistical difference in chemotherapy rate, mastectomy rate, or uptake of immediate breast reconstruction or post-mastectomy radiotherapy in mastectomy patients between the doctors with breast cancer and the control group.

Conclusion

Doctors in this study were not statistically more likely to undergo mastectomy than controls.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P1-09-18.
Abstract P1-10-01: Patient advocate involvement shapes UK’s first national breast cancer tissue bank - The breast cancer campaign tissue bank
Authors: Morgan A , Gath J , MacKenzie M , Matthews A , Stobart H , Turner L , Wilcox M
Published: August 2014

The Breast Cancer Campaign Tissue Bank (BCCTB) was initiated in 2010 after the charity Breast Cancer Campaign heard from breast cancer experts that insufficient high quality breast cancer tissue was preventing research which would impact people with breast cancer1. The charity decided to involve patient advocates in the development of the BCCTB from the beginning.

Two advocates reviewed the initial applications to form the BCCTB. These included both single and multi-centre applications so they suggested that, rather than choosing one centre to form the Bank, the charity consider a collaboration of centres. Throughout the development of the BCCTB, the patient advocates asked the ‘elephant in the room’ questions and brought a different perspective. They also helped to design the Bank's processes, including the consent process and asking that all tissue applicants produce a lay summary of their research.

There are now two advocates on the Board which provides strategic direction to the BCCTB. The advocates have equal status to other Board members. There are also three advocates on the Tissue Access Committee which controls the use of tissue in the Bank. Lay summaries of projects have to be approved by all advocates before an application can proceed. This means that no precious tissue is released without the approval of a lay person, who can ensure maximum patient benefit will be achieved from any research. The Bank's data return policy was also driven by the patient advocates who were keen that the tissues donated were used to their maximum benefit. This has resulted in the first publication from BCCTB2

Patient advocates have been involved in promoting the use of tissue from the bank presenting at conferences and at BCCTB road shows. Patient advocates have also been involved in asking potential donors to consent to donate to the tissue bank.

The involvement of patient advocates in the development and oversight of the BCCTB has shown researchers that patient and public involvement works. The advocates can also help promote the Bank and the benefits of tissue donation to both professionals and lay groups.

The patient advocates involved in the Breast Cancer Campaign Tissue Bank are members of the charity Independent Cancer Patients’ Voice

1. Thompson P, Brennan K, Cox A, Gee J, Harcourt D, Harris A, Harvie M, Holen I, Howell A, Nicholson R, Steel M, Streuli C. Evaluation of the current knowledge limitations in breast cancer research: a gap analysis. Breast Cancer Res 2008: 10(2); R26.

2. Speirs V, Morgan A. Investment biobanking – increased returns on tissue samples. Nat Rev Clin Onc 2013; 10:128-129.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P1-10-01.
Abstract P1-13-09: Sensitive liquid chromatography-tandem mass spectrometry method for serum estradiol and estrone assessment without derivatisation, overcoming cross reactivity with exemestane
Authors: Lintermans A , Pauwels S , Vermeersch P , Jans I , Billen J , Van Asten K , Vanderschueren D , Neven P
Published: August 2014

Objective Measuring serum estrogen levels in postmenopausal women on oral aromatase inhibitors is important but challenging. Direct immunoassays are the most widely used techniques for measuring serum estradiol (E2) in research and clinical laboratories. Unfortunately, commercially available immunoassays for E2 lack precision and accuracy in the low range (<20 ng/l) and are susceptible to interference. In addition, particularly in exemestane users biochemical monitoring of E2 is problematic because exemestane metabolites cross react in most immunoassays for E2. We developed a sensitive, routinely applicable liquid chromatography-tandem mass spectrometry (LC-MSMS) method for E2 and estrone (E1) quantification in human serum without the need for derivatization or an extended extraction protocol.

Materials and methods Serum E2 levels, assessed with LC-MSMS and electrochemiluminescence estradiol assay (ECLIA) on Modular E170 from Roche Diagnostics, of 10 breast cancer patients treated with exemestane were compared. Limit of quantification (LOQ) was 1.3 ng/l for LC-MSMS (coefficient of variation <20% and signal-to-noise ratio > 10, own validation) and 5 pg/ml for ECLIA (Roche, according to manufacturer).

Results Patients treated with exemestane showed much higher E2 levels when measured by ECLIA compared to E2 levels determined by LC-MSMS (Table 1). All patients, but one, had E2 values below LOQ when assessed with LC-MSMS whereas ECLIA measurements showed higher levels with a large variety (mean: 16.5 ng/l; range: 7-33 ng/l).

Results for E2 via ECLIA and LC-MSMS for patients treated with exemestanePatientE2 LC-MSMS ng/lE2 ECLIA ng/l1<1.3212<1.373<1.3124<1.3335<1.3156<1.3227<1.398<1.32692.51010<1.310

Conclusion E2 levels in exemestane users measured with ECLIA from Roche are overestimated, probably due to cross reactivity of its metabolites. Therefore, this assay can not be used in these patients. Here, we demonstrate that this hurdle can be overcome by assessment of estrogen values with LC-MSMS. Our group was able to develop such a sensitive method, without derivatization.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P1-13-09.
Abstract P1-13-13: Association between the serum concentrations of tamoxifen and its metabolites and age during steady state breast cancer treatment
Authors: Lien E , Søiland H , Lundgren S , Ås T , Steen VM , Mellgren G , Gjerde J
Published: August 2014

It has been suggested that the concentrations of tamoxifen and its demethylated metabolites increase with age. Here, we measured the serum concentrations of the active tamoxifen metabolites, 4OHtamoxifen (4OHtam), 4OHNdesmethyltamoxifen (4OHNDtam, Endoxifen), tamoxifen and its demethylated metabolites. Their relations to age were examined. 151 breast estrogen receptor and/or progesterone receptor positive cancer patients were included. Tamoxifen and its metabolites were measured by liquid chromatography-tandem mass spectrometry. Their serum concentrations were related to the age of the patients. The concentrations of 4OHNDtam, tamoxifen, NDtam (N-desmethyltamoxifen), and NDDtam (N-desdimethyltamoxifen) were positively correlated to age (n = 151, P = 0.017, 0.045, 0.011, and 0.001 respectively). Up to tenfold inter-patient variation in the serum concentrations was observed. The median (inter-patient range) of the concentrations 4OHNDtam in the age groups 30-49, 50-69 and >69 years were 44 (65) ng/ml, 51 (116) ng/ml, and 54 (159) ng/ml, respectively. We conclude that the serum concentrations of 4OHNDtam (endoxifen), tamoxifen and its demethylated metabolites increase with age during steady state tamoxifen treatment. The observed high inter-patient range in serum concentrations of tamoxifen and its metabolites, especially in the highest age group, suggest that use of therapeutic monitoring of tamoxifen and its metabolites is warranted.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P1-13-13.
Abstract P1-13-16: Women commencing with anastrozole, letrozole or tamoxifen: The impact of comorbidity and demographics on initial choice
Authors: Boyle F , Kemp A , Preen DB , Saunders C , Bulsara M , Malacova E , Roughead EE
Published: August 2014

Background: Australian clinical guidelines recommend post-surgical endocrine therapy for all women with early, hormone-dependent breast cancer. Guidelines specify tamoxifen as first-line therapy for pre-menopausal women, and either tamoxifen or an aromatase inhibitor for post-menopausal women depending on their risk of recurrence which is based on tumour size and other prognostic indicators. Tamoxifen and the aromatase inhibitors have different side effect profiles and therefore comorbidity may also influence choice of therapy. We examined comorbidities, and the clinical and demographic characteristics of women initiated on different endocrine therapies for breast cancer in Australian practice.

Method: We identified the first dispensing of tamoxifen, anastrozole or letrozole for women diagnosed with invasive breast cancer in the 45 and Up Study between January 2003 and December 2009 (n = 1383). Unit-level pharmacy and medical service claims, hospital separations, and Cancer Registry records were linked to self-reported data to determine menopause status at diagnosis, tumour size, age, change in subsidy restrictions, and specified comorbidities which may affect choice of therapy (i.e. arthritis, endometrial cancer, osteoporosis, thromboembolism). Chi-square tests and multivariate generalised linear models were used to compare the characteristics of post-menopausal women initiated on different therapies.

Results: The majority of pre-menopausal women commenced therapy with tamoxifen (93%). Tamoxifen was the predominant therapy for post-menopausal women with tumours ≤1cm (54%), and anastrozole most common for those with larger tumours (49%). Post-menopausal women with osteoporosis were less likely to commence an aromatase inhibitor compared with tamoxifen (anastrozole RR = 0.7, 95%CI = 0.5-0.9, P = 0.019; letrozole RR = 0.6, 95%CI = 0.3-0.9, P = 0.033) and those with arthritis were 1.6-times more likely to commence with letrozole than anastrozole (95%CI = 1.6-2.3, P = 0.004). Tamoxifen was more likely to be initiated in women with large tumours who also had comorbid arthritis or were aged ≥75 years. Changes in subsidy restrictions were associated with significant increases in the proportion of post-menopausal women commencing an aromatase inhibitor rather than tamoxifen (anastrozole RR = 7.0; letrozole RR = 18.5).

Conclusions: The findings show clinical guidelines regarding endocrine therapies are being followed in Australian practice, particularly in relation to menopause status and tumour size as a recurrence risk factor. The findings also indicate there is interplay of comorbidity and choice of therapy for older women and those with arthritis or osteoporosis.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P1-13-16.
Abstract P1-15-03: Preoperative endocrine treatment with letrozole ± triptorelin in patients with ER (estrogen receptor) and PgR (progesterone receptor) positive locally advanced breast cancer
Authors: Mazza M , Luini A , Veronesi P , Intra M , Bagnardi V , Sangalli F , Iorfida M , Munzone E , Colleoni M
Published: August 2014

Background: Neoadjuvant endocrine therapy demonstrated activity in endocrine responsive locally advanced breast cancer. Proper duration of neoadjuvant therapy has not yet established. We aimed to assess the efficacy and safety of longer duration of letrozole alone or combined with triptorelin in pre-or postmenopausal breast cancer patients.

Methods: Premenopausal and postmenopausal patients (pts) with ER and PgR-positive > 50% of the cells, HER2-negative, T2-T4b breast cancer were considered eligible. Patients received letrozole 2,5 mg per day (plus triptorelin 3,75 mg/month in premenopausal pts) for 3-9 months. Tumor response was measured by caliper, ultrasound (US) and mammography. The primary endpoint was overall tumour response (ORR) (complete response plus partial response), during the neoadjuvant treatment period for the intention-to-treat population.

Results: Between 2009 and 2013, 54 pts were enrolled and 46 (34 pre- and 12 post-menopausal) pts were evaluable for ORR. Median age was 44 and 55 years, respectively. 44 patients are evaluable. The ORR was 77% in the premenopausal group and 67% in the postmenopausal group. One premenopausal patient had a pathological complete response (pCR). The mean time to complete/partial response was 4.9 months (95% CI: 3.8-6.0) in the premenopausal group and 3.6 months (95% CI: 0.8-6.4) in the postmenopausal group. Overall, 56% of premenopausal and 58% of postmenopausal pts underwent breast conservative surgery. Ki67-LI after surgery had a mean decrease of 33% (95% CI: 16%-50%, p-value = 0.0005) and 42% (95% CI: 18%-65%, p-value = 0.0030) in pre-and postmenopausal pts, respectively. Therapy was well tolerated in both groups with no grade 3/4 toxicity. The most common adverse events in both groups were hot flashes, fatigue, arthralgias/stiffness, and myalgias.

Conclusions: The results of this preliminary analysis support neoadjuvant endocrine therapy for a duration of up to 9 months. The combination of letrozole plus triptorelin might represent an alternative neoadjuvant treatment option for premenopausal women with early stage endocrine-responsive breast cancer.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P1-15-03.
Abstract P1-16-01: Skeletal-related events (SRE) prevented in breast cancer patients with bone metastases in the United States (US)
Authors: Arellano J , Chen K , Halperin M , Danese M
Published: August 2014

Background: SREs are potentially debilitating consequences of bone metastases in advanced stage cancers. Bone-targeting agents (BTAs), including denosumab and intravenous zoledronic acid (ZA) are approved to prevent SREs. The purpose of this study was to estimate the annual number of SREs that might occur in US women with breast cancer without intervention, and to estimate the SREs that could be prevented with denosumab or with ZA.

Methods: A model was developed using US prevalence estimates (Li, et al. 2011), combined with 2012 Census projections to estimate the number of SREs in women with bone metastases from breast cancer in 2012. SREs were defined as fracture, radiation to bone, surgery to bone, or spinal cord compression. The SRE rate in untreated patients was estimated by pooling rates from 8 treatment groups in 4 trials (Lipton 2000; Rosen 2003; Kohno 2005; Stopeck 2010), applying the appropriate relative hazard (RH) of first and subsequent SRE from a network meta-analysis (NMA;Ford 2012), and weighting by the sample size. The denosumab and ZA SRE rates were derived from the pooled rate and the RH estimates from the NMA. Ineligibility was defined as one of the following: all uninsured patients (US Census) and patients discontinuing due to adverse events or non-compliance (ZA 4.6%, denosumab 3.7%; Stopeck 2010). Patients with kidney clearance < 30 ml/min (1.3%; Launay-Vacher 2007) were considered ineligible to receive ZA. Using Monte Carlo sampling, we derived mean estimates and 95% simulation intervals (SI) based on 1,000 samples.

Results: Overall, we estimated 96,598 women with breast cancer had bone metastases at any given time in 2012. We estimated 164,673 (95% SI 110,153 to 220,770) SREs would have occurred in 2012 if no women were treated. Assuming all women eligible to receive ZA were treated with ZA (n = 83,870), the number of SREs would have been 108,233 (decline of 56,440, 95% SI 13,744 to 100,098); assuming all women eligible to received denosumab were treated with denosumab (n = 85,780), the number of SREs would have been 86,992 (decline of 77,681, 95% SI 38,327 to 120,521). The incremental difference between denosumab and ZA was 21,242 (95% SI 9,312 to 36,674).

Conclusions: The use of BTAs to reduce SRE risk, including denosumab, could result in reductions in the number of adverse skeletal complications in the US.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P1-16-01.
Abstract P1-17-02: Male breast carcinoma: A clinicopathological and immunohistochemical characterization study
Authors: Zhou R , Yu L , Yu B , Zhou S , Bi R , Shui R , Lu H , Cai X , Yang W
Published: August 2014

AIM Male breast carcinoma is a relatively rare disease. This study retrospectively investigated the clinicopathological features of male breast carcinoma in Chinese population, and classified the molecular subtype based on surrogate immunohistochemical definitions. The expression of GCDFP15, MGB, AR and FOXP1 were also evaluated.

METHODS Clinical and pathological data on 73 cases of male breast carcinoma were collected and microscopic pathological features were examined. Immunostaining for ER, PR, HER2, Ki-67, CK5/6, EGFR, GCDFP15, MGB, AR and FOXP1 was performed and the molecular subtype was classified.

RESULTS Invasive carcinoma of no special type was the major histological type in our study group (71.2%, 52/73). Except for 4 cases of carcinoma in situ, forty-two male invasive carcinomas (42/69, 60.9%) were classified as luminal A subtype while 24 cases (24/69, 34.8%) were classified as luminal B subtype. Only 2 cases of basal-like subtype and 1 case of HER2 positive subtype were identified in this group. Nuclear immunostaining of AR and FOXP1 could be observed in 82.4% (48/57) and 71.9% (41/57) of the cases respectively. Cases of luminal A subtype expressed GCDFP15 (73.5%, 25/34) and MGB (58.8%, 20/34) more frequently than those of luminal B subtype (34.8%, 8/23 and 43.5%, 10/23, respectively).

CONCLUSIONS Invasive carcinoma of no special type was the most common histological type in male breast carcinoma. Our study revealed that luminal A and B subtypes were the major types of male breast carcinoma. AR and FOXP1 were highly expressed in male breast cancer. Luminal A subtype tends to express GCDFP15 and MGB more frequently than luminal B subtype.

KEYWORDS: breast carcinoma, male, molecular subtype, immunohistochemistry.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P1-17-02.
Abstract P2-01-05: Mammographic microcalcifications and breast cancer tumorigenesis: A radiologic-pathologic analysis
Authors: Naseem M , Murray J , Hilton JF , Manoharan AS , Polenz CK , Han D , Hogeveen S , Heersink RL , Bookwala A , Muradali D , Karamchandani J , Bell D , Brezden-Masley CB
Published: August 2014

Background: Microcalcifications (MCs) are tiny deposits of calcium in breast soft tissue. They serve as key diagnostic radiological features for localization of malignancy. Approximately 30% of early invasive breast cancers have fine, granular MCs detectable on mammography; however, their role in breast cancer tumorigenesis is currently unknown. The purpose of this study was to investigate the relationship between mammographic MCs and breast cancer pathology.

Methods: A retrospective chart review was performed for 1015 women treated for breast cancer between 2000-2012 at St. Michael's Hospital. Demographic information (age and menopausal status), tumor pathology (size, histology, grade, nodal status and lymphovascular invasion), hormonal status (ER and PR), HER-2 overexpression and presence of MCs were collected for breast cancer patients. Chi-square tests were performed for categorical variables and t-tests were performed for continuous variables. All tests were two-sided and p-values less than 0.05 were considered statistically significant.

Results: A total of 1015 patient charts were included; 78 (7.7%) patients had metastatic carcinoma and were excluded from analysis. About 38.3% (287/1015) of the patients presented with mammographic MCs. Patients were more likely to have MCs if they were HER-2 positive (52.9%) as opposed to being HER-2 negative (33.8%) (p<0.001). There was a significant association between MCs and having heterogeneous breast density (p = 0.031) and having multifocal disease (p = 0.044). Patients with invasive ductal carcinomas (40.9%) were more likely to present with MCs than were patients with other tumor histology (p = 0.001). There was a positive correlation between MCs and tumor grade (p = 0.057), with grade III tumors presenting with the most MCs (41.3%), followed by grade II (39.8%) and grade I (30.7%). There was no significant association between mean age, mean tumor size, ER and PR status with the presence of MCs.

Conclusion: This is the largest study analyzing data over a 12 year period, suggesting that the appearance of MCs on mammograms is strongly associated with HER-2 overexpression, invasive ductal carcinoma, heterogeneous breast density and multifocal breast cancers. Since HER-2 is implicated in mediating aggressive tumor growth and metastasis, future studies should investigate the molecular pathways connecting HER-2 overexpression and MC development. This would help better understand the role of MCs in breast cancer tumorigenesis.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P2-01-05.
Abstract P2-01-06: How much agreement can we expect on BI-RADS mammographic findings? Observer agreement among 10 expert mammographers
Authors: Wisner DJ , Feig S , Joe B , Hargreaves J , Ojeda-Fournier H , Bassett L , Aminololama-Shakeri S , DeGuzman JQ , Flowers CI , Campbell JE , Elson S , Retallack H , Wells C
Published: August 2014

Purpose: To determine the agreement between expert readers on mammographic findings and calcification patterns.

Materials and Methods: Ten academic radiologists from 5 centers reviewed 250 de-identified mammographic cases without prior exams which were previously assessed as BI-RADS 4 or 5 with subsequent pathologic diagnosis by percutaneous or surgical biopsy. For benign cases diagnosed by percutaneous biopsy, 1 year of benign or negative imaging follow-up was required. Using standardized forms, each radiologist assessed the presence of any suspicious mammographic findings (microcalcifications, asymmetry (1-vew), focal asymmetry (2-view), architectural distortion), and the morphology (none, round/punctate, amorphous, coarse heterogeneous, fine pleomorphic, fine linear branching) and distribution (none, diffuse, regional, grouped, linear, segmental) of any identified microcalcifications. Agreement between radiologists for presence/absence of findings, morphology, and distribution of calcifications was determined by calculating the Kappa (k) coefficient with 95% confidence interval (95% CI). The kappa coefficient proposed strength of agreement is ≤0 = poor, .01-.20 = slight, .21-.40 = fair, .41-.60 = moderate, .61-.80 = substantial, and .81-1 = almost perfect, as established by Landis and Koch.1

Results: Of the 250 lesions, 156 (62%) were benign and 94 (38%) malignant. Agreement among the 10 expert readers was strongest for recognizing the presence/absence of calcifications (k = 0.82, 95% CI: 0.80-84), “almost perfect”). There was substantial agreement among the readers for the identification of a mass (k = 0.67, 95% CI: 0.66-69), whereas agreement was fair for the presence of a focal (2-view) asymmetry (k = 0.21, 95% CI: 0.1900.23) or architectural distortion (k = 0.28, 95%CI: 0.26-0.30). Agreement for asymmetries (1-view) was slight (k = 0.09, 95%CI: 0.08-0.11). Among the 6 categories of microcalcification distribution and morphology, reader agreement was moderate (distribution k = 0.60, 95%CI:0.59-0.61; morphology k = 0.51, 95%CI: 0.50-0.52).

Conclusion: When asked to characterize suspicious mammographic findings, this sampling of 10 expert academic breast imagers across 5 centers revealed varying strength of agreement for different findings, ranging from slight to almost perfect. Strongest agreement (“almost perfect”) was found for identifying the presence or absence of microcalcifications, although agreement drops to moderate when readers are asked to specify microcalcification morphology and distribution.

1 Landis JR, Koch GG. The measurement of observer agreement for categorical data. Biometrics.1977;33:159-174.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P2-01-06.
Abstract P2-01-07: Analysis of the relationship between breast cancer risk-related gene polymorphisms, ESR1/6q25.1-rs2046210 and mammographic breast density
Authors: Nogami T , Taira N , Mizoo T , Nishiyama K , Iwamoto T , Motoki T , Shien T , Matsuoka J , Doihara H , Ishihara S , Kawai H , Kawasaki K , Ogasawara Y , Ishibe Y
Published: August 2014

Background: Based on the results of a case-control study performed in Japan (Okayama and Kagawa), it was revealed that gene polymorphisms of the estrogen receptor (single nucleotide polymorphisms, SNPs), ESR1/6q25.1-rs2046210 and rs3757318, were closely related to breast cancer risk in Japanese women, and that the odds ratios per allele of individual SNPs were 1.37 [95% CI: 1.1 - 1.7] and 1.33 [1.04 - 1.7], respectively (San Antonio Breast Cancer Symposium 2012, Mizoo et al.). To examine the effects of these SNPs on the pathogenic mechanism of breast cancer, we analyzed the relationship of SNPs with reproductive/physiological factors (age of menarche and number of births), physical factors (height and Body Mass Index [BMI]), family history and mammographic breast density.

Methods: Among the patients enrolled in the case-control study, the patients who could be evaluated regarding their lifestyle, SNPs and mammographic breast density were divided into the case group (N = 394) and the control group (N = 511) as the dataset for the study. SNPs were analyzed using the TaqMan genotyping assay® for blood samples. In the analysis of the relationship between SNPs and breast cancer risk, the odds ratios (OR) and 95% confidence intervals (95%CI) were calculated using logistic regression analysis. For the analysis of the relationship between SNPs and the factors, t-test and chi-square test were used.

Results: In our dataset, the age-adjusted OR [95% CI] of ESR1/6q25.1-rs2046210 and rs3757318 were 2.22 [1.41 - 3.55] (AA vs. GG) and 2.12 [1.2 - 3.8] (AA vs. GG), respectively, suggesting that they would be significant risk factors. As a result of analyzing the relationship between SNPs and other factors, a significantly lower BMI was observed for the risk allele AA/AG of rs2046210, compared to GG (p = 0.014, t-test). Regarding the mammographic breast density, a significantly higher proportion of mammary glands with high density was confirmed for the risk allele AA/AG of rs2046210, compared to GG (p = 0.0389, chi-square test). The tendency towards a high mammographic breast density in the rs2046210 risk allele AA/AG was also confirmed based upon a stratified analysis of the case and control groups before and after menopause, and a significant correlation was observed, especially before menopause (p = 0.026, chi-square test).

Conclusion: Mammographic high breast density is generally considered as one of the breast cancer risks, and it was suggested that gene polymorphisms of ESR1/6q25.1-rs2046210 might affect mammographic breast density.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P2-01-07.