bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P5-10-11: Clinical significance of microRNA expression as a prognostic factor in early N+ breast cancer (BC)
Authors: Ciruelos EM , de Velasco GA , Castañeda C , Rodriguez-Peralto JL , Gamez A , Sepúlveda JM , Cortés-Funes H , Castellano DE , Fresno JA
Published: August 2014

Background: microRNAs (miRNAs) are small noncoding RNA molecules that post-transcriptionally regulate gene expression. In the present study, we describe miRNA expression in early node-positive BC and identify a 8-miRNA score that could contribute to prognosis assessment.

Methods: First, microarray analysis was performed using RNA samples extracted from primary breast cancer. The expression of miRNAs was analyzed by RT-qPCR using the TaqMan Arrays from Applied Biosystems. After a suitable normalization of the 677 miRNAs analyzed, 123 presented a good correlation between their levels of expression in frozen tissue and paraffin tissue. A supervised analysis was done in order to identify MIR that could predict relapse.

Results: 172 patients with node-positive chemotherapy-treated early BC were included in the present study. The median age at diagnosis was (53.7) years, 128 cases (74.4%) had positive estrogen receptor, and 163 patients (94,7%) received adjuvant chemotherapy. After a median follow up of 8,1 years, 71p (41.2%) relapsed. Supervised analyses identified 8 microRNAs (has miR-30e, miR-30a, miR-21, miR-210, miR-93, miR-150, miR-99b, miR-572) associated with higher risk of relapse (5 year PFS 50% vs 90%, HR 3.75, 95% CI 2.27–6.19) and with estimated overall survival (median not reached, HR 4.51, 95% CI 2.36–8.61). Interestingly, miRNAs defined 2 prognostic groups (high and low risk) regardless of type of adjuvant chemotherapy (with or without anthracyclines) and histological subtype (luminal A, B or triple negative).

Conclusions: This study suggests that miRNA expression could contribute to assess molecular prognosis of breast cancer and identifies clusters of miRNAs that could improve outcome prediction. A validation study in node-negative and triple negative disease is planned.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P5-10-11.
Abstract P5-14-02: Women's responses to changes in US Preventive Services Task Force mammography screening guidelines: results from focus groups among ethnically diverse women
Authors: Bluethmann SM , Allen JD , Hernandez C , Opdyke KM , Gates-Ferris K , Hurlbert M , Harden E
Published: August 2014

Introduction: In 2009, the US Preventive Services Task Force (USPSTF) changed its recommendations on the age and frequency for routine mammography. To this point, responses to this change among ethnically diverse women have been not been well examined. The objective of this qualitative study is to describe women's awareness of the change in mammography screening guidelines by the U.S. Preventive Services Task Force and to describe their attitudes toward this change.

Methods: White, Black and Hispanic women ages 40–49 years were recruited from a variety of community settings in the Greater Boston, MA area to participate in focus groups (k = 10; N=73). Groups were segmented by race/ethnicity (Black = 29%; White=29%; Hispanic=15%). Women were asked if they were aware of the change in USPSTF guidelines, and if so, their understanding about reasons for this change and intention to comply. Focus groups were audio-taped and transcribed verbatim. Thematic content analysis was used to cull recurring discussion themes.

Results: Most women in this study were not aware of changes in the USPSTF mammography screening guidelines. Those who were aware of the guideline change were highly suspicious that it was motivated by a desire for cost savings on the part of insurance companies and/or providers. Concerns regarding the accuracy of mammography, pain associated with screening, and fear of receiving positive test results were prevalent. Nevertheless, most said that they did not intend to comply with changes in guidelines; many believed that regular (yearly) mammography should start at a young age (40 or before) and continue indefinitely.

Conclusion: Most women in this sample were unaware about changes in mammography screening guidelines and lacked understanding regarding underlying reasons for the change. Communication about the rationale for changes in mammography screening guidelines has left many women unconvinced about the potential downsides of screening and has generated a high degree of mistrust of insurance companies and medical providers.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P5-14-02.
Abstract P5-14-03: Breast cancer screening and follow-up of abnormal mammogram results: A population-based study comparing results from an urban university cancer center to a national database.
Authors: Mitchell EP , Avery TP , Jaslow RC , Berger AC , Lee SY , Vaughan-Briggs C , Bonat J
Published: August 2014

Background: To improve access to screening, the National Breast and Cervical Cancer Early Detection Program (NBCCEDP) was developed through the Centers for Disease Control and Prevention (CDC) to provide low-income, and uninsured underserved women access to timely breast and cervical cancer screening and diagnostic services. This population-based study investigates the demographics and diagnostic outcomes of women who underwent breast cancer screening through this program established at an urban university cancer center compared to data obtained from the national program database.

Methods: The Kimmel Cancer Center at Jefferson (KCC) launched its screening program with resources from the CDC, the State of Pennsylvania, and the Susan G. Komen Foundation in 2008. The core of the program is staff lead by a Social Worker/Navigator who connects patients to education and screening services, institutional information and guidance, and follow-up to minimize barriers to access, lessen dropout, and ensure follow-through for timely diagnosis and treatment. Working with our Community-Based partner organizations, uninsured and underinsured women in Philadelphia are able to seamlessly travel from education and screening through to treatment and support. All KCC patients evaluated through this program from 2008–2011 were included and records of the NBCCDP database 2006–2010 for this study and analyses.

Results: KCC has a substantially larger African American (54.44% vs. 13.8%), smaller Hispanic (8.59% vs. 27.6%), larger percentage of abnormal mammograms (25.96% vs. 14%), higher breast cancer diagnosed per mammogram (2.13 vs 1.0) and a much younger population than the national cohort. Fewer than 1% of KCC patients have been lost to follow-up.

Conclusions: The KCC Breast and Cervical Screening and Treatment Program Services reaches a highly vulnerable and at-risk population, has a higher abnormal mammogram and breast cancer detection rate, and a higher continued participation rate than the national cohort. The Social/Worker/Navigator has a distinct role in providing follow-up for abnormal findings to minimize no-shows by providing creative problem-solving, support, counseling, finding resources to minimize barriers, and contributes significantly to the ease of operation and the continued participation of patients in the program.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P5-14-03.
Abstract P5-17-04: Management of Antiangiogenics' Renovascular Safety in breast cancer. Subgroup and intermediate results of the MARS Study.
Authors: Launay-Vacher V , Janus N , Daniel C , Rey J-B , Ray-Coquard I , Gligorov J , Spano J-P , Thery J-C , Jouannaud C , Goldwasser F , Mir O , Morere J-F , Oudard S , Azizi M , Dorent R , Deray G , Beuzeboc P
Published: August 2014

Background: Hypertension (HTN) and proteinuria (Pu) are class-side-effects of anti-VEGF drugs (AVD), related to the inhibition of the VEGF pathway. The MARS study has been conducted to assess the renovascular tolerance of these drugs in the clinical setting.

Methods: The MARS study is a multicentric prospective observational study of the renovascular safety of AVD in AVD-naive patients. in 7 centres from 2009 to 2011. There was no intervention on the choice of the AVD. Data collected included: gender, age, serum creatinine (SCr), diabetes, HTN, hematuria (Hu) and dipstick Pu. This sub-group analysis presents the intermediate results for the first 184 patients with breast cancer (BC) receiving bevacizumab who completed the 1-year follow-up (f/u) (out of 337 BC patients in total).

Results: All these 184 patients received bevacizumab (mean and median durations of treatment: 7.8 and 8 months, respectively). Median age at inclusion was 60 years. Bone, visceral and cerebral metastasis frequencies were 75.0, 52.2 and 7.1%, respectively. Diabetes and HTN prevalences were 4.3% and 10.3%, respectively. Baseline renal assessment retrieved: Pu 13.0%, Hu 8.2%, mean aMDRD 98.8 ml/min/1.73m2 and 1.1% had aMDRD<60. The incidence of de novo Pu during f/u was 16.3% (Table). 58.3% of patients with Pu at inclusion improved. Among patients with de novo Pu, 42.3% afterwards improved/normalized. No grade 3/4 Pu has been reported and no hematuria. 12.7% developed HTN. In addition, renal function decreased by −3.4 ml/min/1.73m2/year and 4 patients had aMDRD<60 at the end of f/u. 33.2% increased their SCr: 27.7% grade 1, 4.9% grade 2, and 0.5% grade 3. All patients with grade 2–3 returned to normal or grade 1 during f/u. No thrombotic micro-angiopathy has been reported.

Conclusion: These results show that 1) No thrombotic micro-angiopathy remains rare, 2) Pu developed in 16.3% of patients, with no grade 3/4, 3) less than 13% developed HTN, and 4) renal function was only slightly impaired with transient elevations in SCr. Furthermore, in case of a renovascular effect, investigators followed the recommendations from the French Society of Nephrology (Halimi JM et Al. Nephrol Ther 2008) and no treatment withdrawal for unmanageable renovascular toxicity occurred.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P5-17-04.
Abstract P5-18-10: Chemotherapy can enhance trastuzumab-mediated ADCC
Authors: Tagliabue E , Sfondrini L , Regondi V , Casalini P , Pupa SM , Sommariva M , Balsari A , Triulzi T
Published: August 2014

Trastuzumab, a recombinant humanized monoclonal antibody directed to the HER2 protein, has shown survival benefits in women with HER2-positive breast cancer, and treatment is now FDA-approved in combination with chemotherapy. However, some patients do not respond clinically to trastuzumab, pointing to the need for further definition of trastuzumab killing activity on tumor cells to optimize this therapy. Recent clinical data indicate a synergistic therapeutic effect between trastuzumab and taxanes in HER2-positive breast cancer patients, but the mechanism(s) underlying this synergy remains unclear. Because drug-stressed cells can dynamically regulate factors that favor activity of immune effector cells, and because trastuzumab-mediated ADCC is crucially dependent on NK cell activity, we hypothesize that this synergy reflects enhanced trastuzumab-mediated ADCC on tumor cells due to improved rather than impaired participation of immune effectors resulting from drug-induced stress. To test this hypothesis, we investigated the effect of chemotherapy in HER2-positive breast carcinoma models on NK receptor ligands and correlated the changes in these molecules with the ability of trastuzumab to mediate ADCC.

Flow cytometry revealed a 4-fold upmodulated surface expression of NKG2D (MICA, MICB, ULBP1, ULBP2) and DNAM-1 (CD112 and CD155) ligands in HER2-positive breast carcinoma cell lines BT474 and MDA-MB-361 treated for 6 hr with taxotere at a 72-hr IC50 dose, consistent with results of Western blot analysis using soluble lysates obtained from chemotherapy-treated HER2-positive breast carcinoma xenografts. The enhanced expression of NKG2D and DNAM-1 ligands in breast carcinoma cells was accompanied by about a 50% increase in trastuzumab-mediated in vitro ADCC in chemotherapy-treated versus untreated cells. Antibodies blocking NKG2D and DNAM-1, but not control monoclonal antibody, abolished the chemotherapy-induced increase of in vitro trastuzumab-dependent ADCC. Increased expression of NK ligands in taxotere-treated BT474 cells was associated with cytoskeletal damage assessed by confocal microscopy; at 72 hr after chemotherapy, NK ligands returned to basal levels, pointing to the importance of duration of NK ligand enhancement for the chemotherapy-induced trastuzumab-mediated ADCC.

Together, our results indicate that chemotherapy can modulate expression levels of NK-activating ligands on human breast carcinoma cells in correlation with trastuzumab-mediated ADCC, raising the possibility of identifying optimal chemotherapy administration schedules to maximize activity of trastuzumab-mediated ADCC effectors. (Partially supported by AIRC)

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P5-18-10.
Abstract P6-04-15: The involvement of LMTK3 in endocrine resistance is mediated via multiple signaling pathways
Authors: Giamas G , Xu Y , Filipovic A , Grothey A , Coombes CR , Stebbing J
Published: August 2014

We have previously identified new kinases that are able to phosphorylate and/or modulate ERα activity via a high-throughput RNA interference screening. Amongst the most potent novel regulators was LMTK3, a previously uncharacterized molecule. We found that LMTK3 confers its anti-tumor effects predominantly via ERα-regulated signaling. Evolutionary analyses revealed LMTK3 was the only kinase that has adaptively evolved and subjected to Darwinian positive selection, an intriguing and suggestive result given the unique susceptibility of humans to ERα+ breast cancer compared to the great apes (especially for chimpanzees [Pan troglodytes], our closest living relatives). Furthermore, in a large cohort of human breast cancer patients (n = 613), LMTK3 protein levels and intronic polymorphisms were significantly associated with disease-free and overall survival and predicted response to endocrine therapy. Finally, using an established orthotopic mouse model we demonstrated that inhibition of LMTK3 significantly reduces tumor volume and bioluminescence.

Now, by performing: i) mass spectrometry (MS), ii) SILAC-based quantitative proteomics and iii) genome microarray experiments of ERα+ breast cancer cell lines (Tam-sensitive and Tam-resistant) in the presence or absence of various treatments (Tam, E2, ICI), we reveal certain genes, including well-defined oncogenes and tumor-suppressors that are statistically up- or down-regulated.

Interestingly, amongst the newly identified LMTK3-regulated genes/proteins were targets belonging to already established signalling pathways implicated in cell survival, cytoskeletal rearrangement, transformation, and autophagy.

In addition, inhibition of LMTK3 re-sensitised a Tam treatment breast cancer mouse model, to Tam treatment, leading to decreased tumour growth. Moreover, in patients' plasma samples, acquired LMTK3 gene amplification (copy number variation) was associated with relapse whilst receiving tamoxifen. In aggregate, these data support a role for LMTK3 in both innate (intrinsic) and acquired (adaptive) endocrine resistance in breast cancer.

Further results and mechanistic data concerning LMTK3 actions will be presented, placing for the first time this new kinase in the maps of signal transduction.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P6-04-15.
Abstract P6-04-19: Neutralizing antibody to human GP88 (progranulin) restores sensitivity to tamoxifen and inhibits breast tumor growth in mouse xenografts
Authors: Serrero G , Dong J , Hayashi J
Published: August 2014

The 88 kDa glycoprotein GP88 (Progranulin, PCDGF, acrogranin) is the largest member of the granulin/epithelin family of growth modulators characterized by 7.5 cysteine-rich granulin/epithelin repeats. Our laboratory has demonstrated that GP88 represents an ideal therapeutic and diagnostic target in breast cancer. Our published studies have demonstrated that: 1) GP88 expression increases with tumorigenesis; 2) inhibition of GP88 expression by antisense transfection in MDA-MB-468 cells inhibited by 90% tumor formation in nude mice; 3) in ER+ breast cancer cells, GP88 stimulates proliferation and its overexpression confers estrogen independence and resistance to several anti-estrogens and aromatase inhibitor; 4) In Her-2 overexpressing breast tumors, GP88 stimulated Her-2 phosphorylation and conferred trastuzumab resistance; 5) GP88 is expressed in 80% invasive ductal carcinoma and 60% of ductal carcinoma whereas it is negative in lobular carcinoma, benign lesions and normal mammary epithelial cells; 6) pathological studies with 530 cases of ER+ invasive ductal carcinoma showed that GP88 tumor expression measured by immunohistochemistry (IHC) is a prognostic indicator of recurrence in early stage breast cancer patients; High GP88 tissue expression was associated with a 4-fold increase in risk of recurrence at 5 years; 7) GP88 is secreted and can be detected in the serum of breast cancer patients at an increased level when compared to healthy subjects. Based on these results, we developed a neutralizing anti-GP88 antibody AG1. AG1 was expressed in a high yield CHO cell line and formulated. AG1 inhibits GP88 biological effect (proliferation and migration) in a dose-dependent fashion in vitro. Here we will report the results of studies investigating the effect of AG1 on the tumor development of tamoxifen sensitive (MCF-7) and resistant (TamR) estrogen receptor positive breast cancer cells lines injected in nude mice. We show that AG1 alone (5mg/kg) inhibited tumor growth of MCF-7 cells injected into nude mice in a similar efficacy as tamoxifen. Interestingly, in tamoxifen resistant cells derived from MCF-7, the anti-GP88 antibody AG1 inhibited by more than 50% tumor formation of TamR cells in combination with tamoxifen, whereas tamoxifen only had no effect.

These data suggest that inhibiting GP88 provides a novel and alternative therapeutic pathway for the restoration of tamoxifen sensitivity in ER+ breast cancer.

This works is supported by grant 2R44CA124179 from the National Cancer Institute and 02–2010-010 from the Avon Foundation for Women.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P6-04-19.
Abstract P6-04-30: COUP-TFII suppresses NFκB activation in endocrine-resistant breast cancer cells
Authors: Litchfield LM , Klinge CM
Published: August 2014

COUP-TFII is an orphan nuclear receptor that functions as either a transcriptional activator or repressor. We previously reported that COUP-TFII expression is reduced in endocrine-resistant LCC9 breast cancer cells and that overexpression of COUP-TFII restored the ability of the antiestrogen tamoxifen to inhibit cell proliferation. An increase in NFκB activity and p65/RELA expression has been reported in endocrine-resistant breast cancer cells. Because NFκB activation results in increased expression of anti-apoptotic and pro-proliferative genes, increased NFκB signaling promotes the survival of cancer cells. The goal of the current study was to further elucidate the mechanism by which COUP-TFII enables cells to maintain endocrine sensitivity and to determine if COUP-TFII can modulate NFκB activity in tamoxifen-resistant breast cancer cells. To examine whether COUP-TFII can suppress NFκB activity, MCF-7 (tamoxifen-sensitive, TAM-S) and LCC9 (tamoxifen-resistant, TAM-R) breast cancer cells were transfected with a NFκB luciferase reporter and treated with TNFα to activate NFκB. NFκB activity was elevated in LCC9 TAM-R cells and co-transfection with COUP-TFII reduced NFκB activity. COUP-TFII overexpression led to a statistically significant reduction in the expression of NFκB subunits RELA, RELB, NFKB1, and REL in LCC9 cells. Significantly reduced expression of NFκB target genes IL6, ICAM1, TNFAIP3, and CCL2 was also evident upon COUP-TFII overexpression, reflecting the suppression of the NFκB pathway by COUP-TFII. Inhibition of NFκB activation by COUP-TFII also led to a reduction in cell viability in TAM-R cells in response to tamoxifen treatment. Together these data indicate a novel role for COUP-TFII in suppression of NFκB activity which may explain the ability of COUP-TFII to promote tamoxifen-sensitivity. COUP-TFII may be both a useful biomarker to predict tamoxifen-sensitivity as well as a target to restore endocrine sensitivity to resistant cells.

Supported by NIEHS T32 ES011564 predoctoral fellowship to L.M.L. and Susan G. Komen for the Cure grant KG080365 to C.M.K.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P6-04-30.
Abstract P6-05-09: Unravelling the global effect of estrogen on breast cancer cell proteome using quantitative proteomics.
Authors: Pavlou MP , Drabovich AP , Dimitromanolakis A , Diamandis EP
Published: August 2014

Estrogens exert their function through genomic and non-genomic pathways mediated mainly by estrogen receptors. Estrogen signalling is highly complex and once activated initiates cancer cell proliferation and survival, playing a pivotal role in breast cancer development and progression. The identification of estrogen regulated genes is the first step towards elucidating the mechanisms of estrogen function. Although numerous studies have investigated the effect of estradiol at the mRNA level, assessment of global changes at the protein level has been limited mainly due to technological limitations.

Here, we present the most extensive proteomic study in the quest of proteins whose expression is regulated or associated with estradiol action. To facilitate accurate quantification of proteins, we first developed and optimized a proteomic protocol for cell lysis and sample preparation, based on a mass spectrometry-compatible detergent. Following our protocol coupled to stable isotope labelling with amino acids in cell culture (SILAC) of MCF-7 breast cancer cells, we quantified approximately 4,000 proteins and identified 153 proteins differentially expressed upon estradiol stimulation. Notably, known estrogen regulated proteins such as trefoil 1(TFF1) and progesterone receptor (PGR) showed a four- and three-fold increase respectively, at 48 hours after estradiol stimulation. Forty eight of 153 differentially expressed proteins have been found to contain estrogen receptor elements (EREs) indicating direct regulation by estradiol. Protein-protein network analysis (STRING) revealed an extensive protein network related to cell proliferation. Differential expression of proteins was verified in three estrogen receptor positive breast cancer cell lines (MCF-7, HCC-1428, BT-483) using a targeted mass spectrometry-based quantitative approach; selected reaction monitoring (SRM). A multiplex SRM assay was developed for the simultaneous quantification of 56 proteins. The same assay was utilized to study the kinetics of these proteins in time and in presence of inhibitors of estrogen receptor facilitating the distinction between direct and indirect protein targets.

To our knowledge, such an extensive network of estrogen-regulated genes has never been previously studied at the protein level. Interestingly, numerous differentially expressed proteins identified in the present study have not been connected to estrogen signalling or breast cancer before. The role of these proteins in breast carcinogenesis and their potential as therapeutic targets warrants further investigation.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P6-05-09.
Abstract P6-07-05: Prognosis of 368 women with primary breast cancer during pregnancy: results from an international collaborative trial
Authors: Amant F , von Minckwitz G , Han SN , Bontenbal M , Ring A , Giermek J , Fehm T , Wildiers H , Linn SC , Schlehe B , Neven P , Westenend PJ , Müller V , Van Calsteren K , Rack B , Nekljudova V , Harbeck N , Lenhard M , Witteveen PO , Kaufmann M , Van Calster B , Loibl S
Published: August 2014

Objective: We aimed to add to the limited outcome data on breast cancer during pregnancy (BCP), and compared disease-free survival (DFS), and overall survival (OS) between patients with BCP and breast cancer not diagnosed during pregnancy, adjusted for known prognostic factors.

Methods: We analyzed a pro- and retrospectively compiled multi-centric registry of BCP patients (www.cancerinpregnancy.org and GBG/BIG 0203). Outcome was compared to a control group of patients who did not have associated pregnancies. The main analysis is a Cox proportional hazards regression of disease free survival (DFS) and overall survival (OS) on exposure (pregnant or not). The analysis was weighted by propensity score matching to correct for any bias that may have been associated with differences in baseline characteristics, with the following set of covariates: age, stage, grade, hormone receptor status, HER2 status, histology, type of chemotherapy and trastuzumab. Sensitivity analysis was also performed on age only, and also on chemotherapy only.

Results: From April 2003-December 2011, 447 women with BCP were registered, 368 women with BCP were eligible for this analysis. The control group consisited of 2739 women (ratio 1:7.44).

Median age was 33 years (interquartile range 30–36) for the cases and 41 years (interquartile range 37–43) for the controls. Median follow-up was 57 months. There were no statistically significant differences noted for DFS and OS studied after propensity-score matching, and sensitivity analysis.

Conclusion: Patients diagnosed with breast cancer during pregnancy do not have a worse prognosis compared to patients who are not pregnant after matching for prognostic factors and treatment. This information is important when BCP patients are counseled regarding prognosis and adds to the possibility of treatment during pregnancy.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P6-07-05.
Abstract P6-07-07: Influence of comorbidity on survival after early breast cancer
Authors: Ewertz Marianne , Jensen Maj-Britt , Land Lotte H , Dalton Susanne O
Published: August 2014

Withdrawn by Author

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P6-07-07.
Abstract P6-07-13: Local relapse and survival
Authors: Harland R , Prathap P , Lionaki A , Mahmood N
Published: August 2014

Aim: Local recurrence may precede distant relapse and death. Many factors which predict local recurrence also predict death. Few studies have investigated the contribution of local recurrence to mortality independent of prior prognostic factors such as nodal status. We wished to estimate the extent to which overall survival is affected specifically by local recurrence.

Methods: 114 of of 2945 patients aged 19 to 94 years (mean 58.3) who had either mastectomy or breast conservation for breast cancer between 1991 and 2011 subsequently suffered an ipsilateral local recurrence. We matched, with one exception (due to extreme young age), each patient who had a local recurrence with 2 controls matched for calendar year of presentation, age, nodal status, grade, and tumour size. A Cox regression analysis of survival from the first therapeutic operation was done on the case matched subset with local recurrence coded dichotomously and age coded continuously as the absolute deviation from the population mean (as both young and old age affect survival adversely) as predictors. To confirm independence of the effect of local recurrence on the hazard of death a further Cox regression analysis of survival as above was done for the whole series with local recurrence, Grade 3, >3 nodes involved, tumour diameter >40mm, coded dichotomously, and age coded as above as independent variables.

Results: The results of the analyses are given in the table below.

The estimate of hazard of death associated with local recurrence was similar in both analyses suggesting minimal confounding effect. As expected large tumour size, high grade and heavy nodal involvement predicted survival independently.

Conclusion: Local recurrence was associated with only a modest increase of about 50% in the hazard of death independent of tumour stage, size and grade which remained the most significant predictors within this model. Extremes of age were associated with increasing hazard of death and should be similarly coded when analysing unadjusted survival.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P6-07-13.
Abstract P6-07-19: Prognostic relevance of PR and detection mode in Luminal Her-2 negative breast cancer
Authors: Brouckaert O , Van Calster B , Paridaens R , Wildiers H , Van Limbergen E , Weltens C , Christiaens M-R , Neven P ,   Multidisciplinary Breast Centre
Published: August 2014

Introduction: Although known to hold prognostic value, PR status is not used to discriminate luminal A from luminal B HER2 negative breast cancers. We will here assess whether PR (and also detection mode) is an independent prognostic variable in luminal HER2 negative breast cancer and whether the prognostic value of PR is grade dependent.

Patients and methods: Primary operable breast cancer patients diagnosed between 1/1/2000 and 31/12/2009 treated in UZ Leuven were retrieved from the prospectively managed database (n = 4318). Her-2 status was missing in 93 and ER/PR in 2 patients. Of the remaining patients, 3330 were luminal (ER and/or PR positive) HER2 negative (full cohort details: Brouckaert O., et al. Ann Oncol. 2012 Apr 6). Missing values were observed in 137 patients (4.1%), and these were imputed using ‘multivariate imputation by chained equations’. A competing risks proportional hazards model was used for multivariable analysis (includes age, size, grade, screening, palpability, nodal status, histology, type of surgery, radiotherapy, endocrine and chemotherapy) of distant metastasis free interval (DMFI) and breast cancer specific survival (BCSS). We used a predefined set of predictors, investigated non-linearity of the predictor effects, checked the proportional hazards assumption, and tested for interactions with grade.

Results: Median follow-up was 76 months and 5-year survival probability 93%. PR is an independent prognostic variable but this is grade dependent for DMFI, but not for BCSS (interaction p = 0.15), although PR positivity was mainly protective for grade 3 breast cancers again (table 1). Palpability is a strong prognostic variable for DMFI and BCSS and was strongly correlated with screen detection (yes/no) and here, we found no evidence of grade dependency.

Conclusion: PR status and palpability may further refine prognosis of patients with luminal HER2 negative breast cancers. For PR, prognostic value is grade dependent, but for palpability, this is grade independent.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P6-07-19.
Abstract P6-07-24: Prognostic Tools in Early Breast Cancer: Predicting benefit of adjuvant chemotherapy
Authors: Parkes EE , Davidson C , James CR , Hanna GG
Published: August 2014

Introduction: Adjuvant chemotherapy (CT) in early breast cancer reduces the risk of mortality. However, absolute reductions in mortality can be small. For patient with low risk disease, prognostic tools such as ‘Adjuvant! Online’ and ‘Predict’ can be used to estimate the benefit of adjuvant chemotherapy. We compare the survival gains estimated using ‘Adjuvant! Online’ and ‘Predict’ in routine clinical practice, assessing the characteristics of patients in which ‘Adjuvant! Online’ and ‘Predict’ disagree.

Methods: In a retrospective study using the hospital electronic database, the clinical and pathological details of all patients with early breast cancer referred for adjuvant therapy at the Northern Ireland Cancer Centre in a 3 month period in 2011 were collected and were entered in to ‘Adjuvant! Online’ and ‘Predict’ to assess percentage benefit (absolute reduction in mortality at 10 years) from CT. We categorised patients into three prognostic groups: those where risk from CT outweighs benefit (<2% predicted benefit), marginal benefit (2 to 5%) and significant benefit from CT (>5%) We excluded patients with metastatic disease at presentation, DCIS, a second primary breast cancer or receiving neo-adjuvant treatment.

Results: Of the 250 patients identified, 61 (24.4%) fell in to different prognostic groups depending on whether ‘Adjuvant! Online’ or ‘Predict’ was used to calculate benefit from CT. In those in whom there was disagreement, ‘Adjuvant! Online’ suggested marginal or significant benefit in 75.4% of patients, compared to 55.7% using ‘Predict’. Nine patients had “major” comorbidities, which is weighted only in ‘Adjuvant! Online’, and were excluded in subsequent analysis. Of those without major comorbidities, ‘Adjuvant! Online’ offered at least 2% benefit in 85% of cases, and ‘Predict’ in only 58.3%. The majority (93.3%) of cases were ER positive, and node negative (85.0%). This difference was notable in women aged 65 or less, with 86.2% with >2% benefit using ‘Adjuvant! Online’, compared to 55.1% using ‘Predict’. ‘Adjuvant! Online’ estimates of benefit were on average 3.7% higher for this age group. HER2 status had little impact, with similar recommendations using either ‘Adjuvant! Online’ or ‘Predict’.

Conclusions: This study highlights lack of concordance between two available online prognostic tools, notably in ER positive, node negative patients. For patients with a marginal benefit from adjuvant chemotherapy, care must be used when making adjuvant treatment decisions as other prognostic tools may be of help in this group of patients.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P6-07-24.
Abstract P6-07-28: Assessment of T stage in the multiple breast carcinomas
Authors: Gong G , Jo J-H , Lee HJ , Kang J
Published: August 2014

Background: Tumor size is an important predictor of nodal metastasis, recurrence, distant metastasis in invasive breast carcinomas. T stage assignment of the multiple simultaneous ipsilateral breast carcinomas is based on the largest tumor with the suffix “(m)” to indicate multiplicity in the AJCC staging, and sum of the sizes should not be used. The aim of this study is to compare current AJCC staging and two test models using the aggregate of all tumor foci and the aggregate of all tumor volume in the single and multiple breast carcinomas.

Methods: We reviewed original pathologic reports of 1145 consecutive patients with invasive ductal carcinoma, 2003–2004. 131 (11.4%) patients had multiple masses. Three factors were used to compare T stages; (1) the greatest dimension of the largest invasive component (T), (2) the aggregate of all tumor foci (TA), and (3) the aggregate of all tumor volume (TV). T stage follows AJCC (7th ed, 2010) system. TA and TV were categorized as followings: TA1 < 2cm, 2cm ≤ TA2 < 5cm and TA3 ≥ 5cm; TV1 < 8cm³ (average diameter < 2cm), 8 cm³ ≤ TV2 < 125cm³ and TV3 ≥ 125cm³ (average diameter ≥ 5cm). Clinicopathologic parameters were analyzed according to T stage.

Results: Axillary lymph node metastasis was more common in the cases with multiple masses (64/131, 48.9%) than single mass (397/1014, 39.2%) and the number of the metastatic lymph node was also higher in multiple cases. Multiple breast carcinomas tended to show frequent recurrence, and disease specific death compared with the single cases in the same stage. (p < 0.05) Among 606 cases of T1, 34 cases were reclassified to TA2 and in 487 cases of the T2, 23 cases were reclassified to TA3. TV also showed reclassification of the cases. Multiplicity, T stage, TA stage, and TV stage were significant factors for nodal positivity in the univariate analysis (p < 0.05). Nodal positivity was remained in the multivariate analysis for recurrence and disease specific death, but the other factors were not significant for nodal involvement, recurrence and disease specific death.

Conclusion: Multiple breast carcinomas has adverse patients' outcome than single carcinoma with same T stage of AJCC system and a better staging method is required to cover multiple carcinomas.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P6-07-28.
Abstract P6-07-30: The clinical significance of pathologic complete response using different definitions after neoadjuvant chemotherapy in HER2 positive breast cancer patients according to hormonal receptor status.
Authors: Tanioka M , Hirokaga K , Kitao A , Matsumoto K , Yoshida S , Miki M , Maekawa Y , Takao S , Negoro S
Published: August 2014

Background: For patients (pts) with HER2 positive (HER2+) breast cancer, the achievement of pathological complete response (pCR) after neoadjuvant chemotherapy is regarded as a surrogate endpoint of prognosis, however, the exact definition of pCR and its prognostic impact on survival in HER2+ breast cancer subtypes when stratified by hormonal receptor (HR) status is uncertain.

Methods: We retrospectively investigated 163 HER2+ pts who received neoadjuvant chemotherapy in Hyogo Cancer Center between 2003 and 2010. HR+ was defined as estrogen or progesterone receptor positive. pCR at surgery was defined as no evidence of invasive and noninvasive residual in the breast or axillary nodes (ypT0 ypN0), no evidence of invasive residual in the breast or axillary nodes; noninvasive breast residuals allowed (ypT0/is ypN0), or no evidence of invasive residual in the breast; noninvasive breast residuals and infiltrated lymph nodes allowed (ypT0/is ypN0/+). Logistic regression analysis and logrank test were performed to identify HR status as a predictor of pCR and its prognostic significance on recurrence-free survival (RFS).

Results: The median age at diagnosis was 55 years (range, 25–84) and the median follow-up period was 61 months (range, 15–104). Stage I, IIA, IIB, III were 4, 57, 56, 46 pts, HER2+HR+ and HER2+HR- were 89 and 74 pts, respectively. The rates of pts receiving anthracycline, taxane and trastuzumab were 66, 82 and 51%, respectively. After surgery, 98% of HER2+HR+ pts received hormonal therapy. Five-year RFS rates were significantly higher in HER2+HR- pts with pCR compared to non-pCR, however, which were similar in HER2+HR+ pts with pCR compared to non-pCR regardless of the pCR definitions.

In logistic regression analysis, significantly fewer HER2+HR+ pts achieved pCR compared to HER2+HR- pts regardless of the pCR definitions.

Conclusions: HER2+HR+ breast cancer pts had lower achievement rate of pCR, which was less prognostic compared to that in HER2+HR- pts regardless of pCR definitions. pCR is a suitable surrogate end point for HER2+HR- pts but not for HER2+HR+ pts in clinical trial settings.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P6-07-30.
Abstract P6-11-01: Intermittent High Dose Proton Pump Inhibitor Improves Progression Free Survival as Compared to Standard Chemotherapy in the First Line Treatment of Patients with Metastatic Breast Cancer
Authors: Hu X , Wang B , Sun S , Chiesi A , Wang J , Zhang J , Fais S
Published: August 2014

Purpose: High dose proton pump inhibitor (PPI) has proved to be potentially effective when combined with chemotherapy in preclinical data. This study (NCT01069081) was designed to investigate whether the efficacy of chemotherapy could be improved with the addition of proton pump inhibitor (PPI) in patients with metastatic breast cancer.

Patients and Methods: Females elder than 18 years old with histologically confirmed metastatic breast cancer were eligible for participation. Patients enrolled were randomly assigned to three arms: Arm A, docetaxol 75 mg/m2 on d4, followed by cisplatin 75 mg/m2 on d4, repeated every 21 days until maxinum 6 cycles or presence of disease progression or intolerable toxicity; Arm B, the same chemotherapy plus esomeprazole 80 mg p.o. bid three days on and the subsequent four days off, beginning on d1 repeated weekly up to disease progression, intolerable toxicity, patient's withdrawal, or a maximum of 66 weeks; Arm C, the same as Arm B with the only difference being dose of esomeprazole at 100 mg p.o. bid. The primary endpoint was progression free survival (PFS), and the secondary endpoints were time to progression (TTP), objective response rate (ORR), safety profile, and overall survival (OS).

Results: From Aug. 2009 to Dec. 2011, 100 women signed informed consent form (ICF) and 94 patients have undergone at least one injection of chemotherapy. Three patients had severe hypersensitivity reactions, two occurring after the first injection of docetaxol and one in the second cycle. After a median follow up of 17 months, 68 (72.3%) patients got disease progression and 23 (24.5%) patients died. Median PFS for the whole group (n = 94), arm A (n = 33), arm B (n = 30), arm C (n = 31) were 8.9, 7.5, 10.9, and 9.5 months, respectively (p = 0.082). A significant difference was observed between patients who had taken PPI and who not with median PFS of 9.5 and 7.5 months, respectively (p = 0.030). Among 17 patients with triple negative breast cancer, this difference was bigger with median PFS of 9.5 and 3.3 months, respectively (p = 0.014). The overall response rates for the whole group, arm A, arm B, arm C were 58.5%, 51.5%, 63.3%, 61.3%, respectively.

Conclusion: This trial is the first randomized study to demonstrate antitumor effects of intermittent high dose PPI in patients with metastatic breast cancer. Docetaxel and cisplatin doublet is comparable to the other first-line chemotherapeutical regimens and the addition of proton pump inhibitor to the doublet improves efficacy with no adding toxicity, especially in patients with triple negative breast cancer.

Key Words Proton Pump Inhibitor (PPI), Triple Negative Breast Cancer (TNBC), Phase II Study, Metastatic Breast Cancer, Chemotherapy

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P6-11-01.
Abstract P6-11-07: The Cancer Stem Cell-Targeting Wnt Inhibitor VS-507 Reduces Breast Cancer Growth and Metastasis
Authors: Ring JE , Kolev VN , Padval MV , Keegan M , Vidal CM , Neill AA , Shapiro IM , Pachter JA , Xu Q
Published: August 2014

Cancer stem cells (CSCs) support tumor viability and growth through their ability to self-renew and differentiate into heterogeneous tumor tissue. CSCs are typically resistant to standard cytotoxic agents, leading to their enrichment and the consequent regrowth of refractory tumors. While this population has been challenging to directly target therapeutically, we are actively developing novel agents, such as VS-507, that selectively target the cancer stem cell subpopulation in vivo.

Previously, we and others have shown that VS-507, a cancer stem-cell specific agent, inhibits Wnt signaling with corresponding reduction of the LRP6 protein, a Frizzled co-receptor upregulated in breast cancer cell lines. In the current study, we have continued to examine the effect of VS-507 on the Wnt/Beta-catenin signaling pathway to further elucidate its mechanism of action. We determined that VS-507 also decreases expression of the second Frizzled co-receptor LRP5 in MDA-MB-231 breast cancer cells stimulated by Wnt3A. The combined inhibition of the co-receptors LRP5 and LRP6 may contribute to the observed inhibition of Beta-catenin-mediated transcription by VS-507 in a TOP-Flash assay. Accordingly, VS-507 reduced expression of Axin2, a transcriptional target of Beta-catenin.

Because human breast cancer cell lines represent a mixed population of CSC and non-CSC cells, we evaluated VS-507 across a panel of cell lines by monitoring changes in viability and in the percentage of cancer stem cells. In SUM159 triple negative breast cancer cells, VS-507 had micromolar potency against the bulk population with preferential nanomolar potency against the ALDEFLUOR+ CSC population with similar effects observed in the Hoechst side population (SP) CSC assay. In contrast, cytotoxic anticancer drugs such as paclitaxel and cisplatin increased the percentage of ALDEFLUOR+ cancer stem cells under similar conditions. In vivo, oral administration of VS-507 (15 mg/kg, QD daily) as a single agent partially inhibited MDA-MB-231 tumor growth, consistent with its observed inhibition of the cancer stem cell subpopulation. Furthermore, VS-507 inhibited lung metastasis in a 4T1.2 murine breast cancer model. In summary, these results further elucidate the mechanism by which VS-507 inhibits Wnt/Beta-catenin signaling. VS-507 reduces tumor cell proliferation in vitro with preferential effects on cancer stem cells which translate to inhibition of tumor growth, enhancement of efficacy of cytotoxic agents such as docetaxel, and inhibition of metastasis in mouse models. These data provide additional support for the development of VS-507 as a novel anti-cancer stem cell agent.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P6-11-07.
Abstract P6-11-13: In vitro antineoplastic evaluation of rationally designed naphtoquinone-derived drugs in triple-negative breast cancer cell line
Authors: Madeira KP , Daltoé RD , Herlinger AL , Guimarães IS , Allochio Filho JF , Teixeira SF , Valadão IC , Greco S , Rangel LBA
Published: August 2014

Background: Breast cancer (BC) comprises multiple diseases harboring different genetic alterations, which subtypes respond differently to treatment, and are associated to distinct clinical outcomes. Likewise, triple negative breast cancer (TNBCs) are a heterogeneous subgroup of BC, immunophenotypically negative for estrogen and progesterone receptor, and HER2, that account for 10–15% of all invasive BC, affecting mostly African-American and Hispanic pre-menopausal women. Considering that TNBC have poor prognosis, and cannot be effectively treated with current targeted therapies, the discovery of new treatment options is imperative.

Methods: Novel naphtoquinone-derived drugs were rationally designed to act through multiple pathways aiming the avoidance of drug-resistant phenotype acquisition by tumor cells, and were synthesized following high efficiency and low cost method. Drugs antineoplastic efficacy (AE) was accessed in claudin-low TNBC cell line, MDA-MB-231, through the evaluation of cellular metabolic viability (CMV) (MTT method). Drugs structures are protected by patent. Cells were cultured in RPMI media supplemented with 10% (v/v) FBS and antibiotics until subconfluence; then, 1.5×105 cells/well were subcultured for 72h prior to treatment with drugs (10−4, 10−5, 10−6, 10−7, and 10−8 M). After 24h, CMV was assessed. Experiments in which the lineage was treated with cisplatin, doxorubicin or paclitaxel were run in parallel. The mean and standard-deviation of the absorbancies were used to calculate CMV and drugs IC50 (PrismaGraphPad version 5.1).

Findings: We screened the AE of 43 novel naftoquinones-derived drugs in MDA-MB-231 lineage; seven have decreased its CMV by, at least, 50%, as: PIC1 (IC50 1.15×10−4M; CMV decrease of 50%); PIC6 (IC50 4.24×10−5M; CMV decrease of 70%); PIC10 (IC50 5.07×10−5M; CMV decrease of 70%); PIC 20 (IC50 1.38×10−5M; CMV decrease of 90%); PIC21 (IC50 5.00×10−5M; CMV decrease of 70%); S5 (IC50 7.26×10−5M; CMV decrease of 60%); M20 (IC50 7.94×10−5M; CMV decrease of 60%). Their AE was significantly higher than that of cisplatin (IC50 1.56×10−4M; CMV decrease < 10%), doxorubicin (IC50 1.76×10−4M; CMV decrease of 38%), and paclitaxel (IC50 5.05×10−7M; CMV decrease of 20%).

Interpretation: Altogether, our results present potential novel antineoplastic drugs to treat TNBC from a panel of in house rationally designed naftoquinones-derived compounds, developing an innovative and economically viable project.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P6-11-13.