bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P5-21-02: Hormone receptor status and endocrine therapy in a prospective observation study on trastuzumab (Herceptin®) in the adjuvant treatment of breast cancer
Authors: Dall P , Friedrichs K , Petersen V , Hinke A , Brucker C , Schmidt P , von der Assen A , Jungberg P , Bohnsteen B
Published: August 2014

Background: Based on the findings from large randomized studies, Trastuzumab (T) was registered in 2006 for the treatment of early stage, HER2+ breast cancer (BC), after surgery, (neo)adjuvant chemotherapy (CT) and/or radiotherapy. However, limited systematic evidence is available on preceding, concomitant or sequential endocrine treatment (ET) based on the hormone receptor (HR) status. This large prospective observation study, reflecting the unrestricted routine practice of HER2 antibody treatment in Germany, allows to examine this topic.

Methods: At present, more than 4000 patients (pts) have been enrolled in this ongoing study; 3446 pts from more than 316 centres were already sufficiently documented to be analysed. Details on concomitant treatment was collected at baseline and for 12 months after start of T.

Results: With respect to an overall HR positivity (HR+), i.e. estrogen and/or progesterone receptor (ER, PR) being positive, of 2181/3446 (63%), these HER2+ pts do not differ from the general BC population. Both hormone receptor types are positive in 47%, while ER+ only or PR+ only was detected in 13% and 3%, respectively. Adjuvant ET of any type, starting before or during T treatment, was given in 87% of HR+ and in 3% of HR- pts. 6% of HR+ pts received T and ET without concomitant CT. Details on the adjuvant treatment options are provided in the table:

After a median follow-up of 26.4 months and the observation of a total of 296 relapse events (9%; HR-: 12%; HR+: 7%) relapse-free survival (RFS) was significantly shorter in the HR- compared to the HR+ group (at 3 years: 86 vs 92%, hazard ratio = 0.56, p < 0.0001).

Conclusion: The vast majority of HER2+ BC pts receives concomitant endocrine therapy in case of a positive HR status. Regardless of the regulatory status in Germany, a small proportion of patients with co-positive early BC received T and ET without concomitant CT. No differences between receptor subgroups could be detected with respect to type of CT and its onset relative to T. HR status remains a paramount prognostic factor in this HER2+ sub-population of BC pts, with risk of early relapse (after a median follow-up of 2 years) almost twice as high in HR- tumors.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P5-21-02.
Abstract P6-02-07: In vitro 3D Model of Breast Tumor Stroma
Authors: Jaganathan H , Mitra S , Dave B , Godin B
Published: August 2014

One in eight women is diagnosed with breast cancer in the United States every year. Despite advancements for early diagnosis and initial treatment of breast cancer, the complexity and multiplicity of biological factors and barriers in tumor microenvironment prevent potent active agents from reaching their targets in cancer cells. There has been a growing body of evidence about the importance of three-dimensional (3D) culture systems in discovery and modeling of biological processes with in vivo relevance. A few examples that have been reported in 3D monocultures include different growth patterns of normal and malignant breast cancer cells in 3D cultures of laminin-rich gels and effect on cell polarity pathways. These effects are not observed in conventional two-dimensional (2D) cultures, showing that cells organized in a 3D matrix can shed light on novel mechanisms in breast cancer biology. Moreover, neither 2D nor 3D monoculture models available today mimic the complexity of clinically observed tumor microenvironment (tumor stroma), which also contains fibroblasts, macrophages and endothelial cells and represents an important physiological barrier for efficient delivery of therapeutics to breast tumors. In this work we propose and investigate an in vitro system mimicking the tumor microenvironment in which relevant cell populations in tumor microenvironment are co-cultured with breast cancer cells*. The system uses the Nano3D® platform based on magnetic levitation. 3D tumor models for breast cancer were designed and optimized by co-culturing SUM159 and MDA-MB-231, triple negative breast cancer cells, with 293T fibroblasts. The cells were co-cultured at different ratios and for a various time periods. Further, to assess the similarity of the in vitro system to the in vivo disease, we tested an an orthotopic model of breast cancer in SCID beige mice. Histological analysis of frozen tissues from in vitro and in vivo studies confirmed that the in vitro 3D system based on the mixture of fibroblasts and cancer cells has structural similarities to the in vivo observed tumor stroma. Engineering an in vitro 3D tumor model that accurately represents the in vivo tumor will further benefit research efforts in breast cancer diagnosis and therapy.

*Patent pending – inventors: B. Godin, G. Souza, and B. Dave

Acknowledgments: The authors acknowledge a financial support from the Susan G. Komen Postdoctoral Fellowship #PDF12229449.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P6-02-07.
Abstract P6-04-12: Novel selective estrogen receptors degraders regress tumors in pre-clinical models of endocrine-resistant breast cancer
Authors: Hager JH , Darimont B , Joseph J , Govek S , Grillot K , Aparicio A , Bischoff E , Kahraman M , Kaufman J , Lai A , Lee K-J , Lu N , Nagasawa J , Prudente R , Qian J , Sensintaffar J , Shao G , Heyman R , Rix P , Smith ND
Published: August 2014

80% of all breast cancers express the estrogen receptor alpha (ERα) and thus are treated with anti-hormonal therapies that directly block ER function (e.g. Tamoxifen) or hormone synthesis (Aromatase Inhibitors). While these therapies are initially effective, acquired resistance invariably emerges. Importantly, the majority of these tumors continue to depend on ERα for growth and survival. The emerging evidence that ERα can signal in both a ligand-dependent and ligand-independent manner supports the development of agents that are not only competitive ERα antagonists but also reduce steady state levels of the receptor and thus limit both modes of signaling.

We have identified novel, non-steroidal ERα antagonists that induce degradation of ERα in breast cancer cell lines at picomolar concentrations resulting in significant reduction in steady state ERα protein levels. Using peptide-based conformational profiling, we show that these Selective Estrogen Receptor Degraders (SERDs) induce estrogen receptor conformations that are distinct from both fulvestrant and tamoxifen indicating a unique mechanism of action. This unique biological profile coupled with good oral pharmacokinetics produces tumor regressions in both tamoxifen-sensitive and -resistant models of breast cancer in vivo. Recent pre-clinical and clinical data indicate that PI3K pathway signaling can contribute to the endocrine resistant state. In a preclinical model of tamoxifen resistant breast cancer in which SERD monotherapy only produces tumor growth inhibition, SERD therapy in combination with torc1/2 inhibition results in frank tumor regressions. These orally bioavailable SERDs hold promise as a next generation therapy for the treatment of ER+ breast cancer as monotherapy, as well as in combination with agents that target other pathways involved in both intrinsic and acquired endocrine resistance.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P6-04-12.
Abstract P6-04-17: The androgen metabolite-dependent growth in hormone receptor positive breast cancer as a novel aromatase inhibitor-resistance mechanism
Authors: Hanamura T , Niwa T , Nishikawa S , Konno H , Ghono T , Kobayashi Y , Kurosumi M , Takei H , Yamaguchi Y , Ito K-I , Hayashi S-I
Published: August 2014

Introduction: Aromatase inhibitors (AIs) have been reported to exert their anti-proliferative effects not only by reducing the production of estrogens but also by unmasking the inhibitory effect of androgens such as testosterone (TS) or dihydrotestosterone (DHT) in postmenopausal women with hormone receptor-positive breast cancer. The behavior of androgens in AI-resistance mechanisms is not sufficiently understood. 5α-androstane-3β,17β-diol (3β-diol) generated from DHT by 3β-hydroxysteroid dehydrogenase type 1 (3β-HSD type 1: HSD3B1) has androgenic activity and substantial estrogenic activity, representing a potential mechanism of AI resistance.

Methods: To investigate these issues, ERE-GFP-transfected MCF-7-E10 cells were cultured for 3 months under steroid-depleted, TS-supplemented conditions which is the similar as the AI treatment. Among the surviving cells, two stable variants that show ER activity depending on androgen metabolites were selected as AD-EDR (androgen metabolite-dependent and estrogen depletion-resistant) by monitoring GFP expression. Using these cell lines, we investigated the process of adaptation to androgen-abundant conditions and the role of androgens in AI-resistance mechanisms.

Results: AD-EDR cell lines showed increased growth and induction of estrogen-responsive genes rather than androgen-responsive genes by androgens or 3β-diol. Further analysis revealed increased expressions of HSD3B1 and reduced expression of androgen receptor (AR) in these cell lines. In parental MCF-7-E10 cells, ectopic expression of HSD3B1 or inhibition of AR resulted in adaptation to estrogen-deprived and androgen-abundant conditions. In coculture with stromal cells replicating the local estrogen production from androgen, AD-EDR cell lines showed AI resistance compared with parental MCF-7-E10 cells. Immunohistochemistry and real-time PCR analyses on 9 pairs of primary and recurrent tissue samples from AI-resistant breast cancer revealed the decrease of AR protein expression in all cases and increase of HSD3B1 mRNA expression in 5.

Conclusion: In the present study, we successfully cloned two stable variants that show ER activity depending on androgen metabolites. Investigation of these cell lines suggested that the increased function of 3β-HSD type 1 and reduced function of AR contribute to AI resistance by enhancing the androgen metabolite-dependent growth and reducing the inhibitory effect of androgens. Our data of clinical samples suggest that this mechanism also acts as an AI-resistance in clinical breast cancer in some cases.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P6-04-17.
Abstract P6-04-26: Tamoxifen may block estrogen induced secretion of certain cytokines to interrupt tumor associated macrophage infiltration in breast cancer
Authors: Ding J , Chen CM , Jin W , Shao Z , Wu J
Published: August 2014

The role of tumor microenvironment during the initiation and progression of breast cancer is now realized to be of critical importance, both for understanding of the fundamental cancer biology and exploiting relatively new mechanism for breast cancer metastasis. Macrophage is a one of the most common components in the breast cancer microenvironment. Normally, the macrophage in tumor microenvironment is referred to tumor associated macrophage (TAM), which shares some attributes of alternatively activated macrophage. Many pre-clinical and clinical studies demonstrate an inverse correlation between TAM infiltration and patients' prognosis indicating a macrophage supporting role for tumor progression. It is well documented that selective patients with breast cancer can benefit from anti-estrogen therapy. However, the mechanisms involved are still not fully elucidated. Our previous study indicates that highly tumorigenic breast cancer cell line MDA-MB-231 can educate monocyte differentiation into alternatively activated macrophage, while weakly tumorigenic cell line MCF-7 (without estrogen supplementation) can not. In the present study, we explored the effects of estrogen and tamoxifen on the secreting of certain cytokines, which are required for monocyte chemotaxis and differentiation. Breast cancer cell lines with different estrogen receptor status were applied to estimate the level of cytokines (IL4, IL6, M-CSF, GM-CSF, and MCP-1) before and after cultured with 17β-estradiol, tamoxifen or both. We found that weakly invasive breast cancer cell lines (MCF-7, T47D, MDA-MB-468, and SKBR3) generally expressed lower levels of such cytokines compared with highly invasive breast cancer cell lines (MDA-MB-231, MDA-MB-231-BO, MDA-MB-231 HM, Bcap 37, BT549, and Hs578T) at baseline. However, the discrepancies could be compensated partially by exposure to 10nM 17β-estradiol in estrogen receptor (ER) positive cell lines (MCF-7 and T47D). Moreover, the compensation was substantially blocked by 2μM tamoxifen. On the contrary, tamoxifen, alone, didn't affect the secretion of such cytokines mentioned above in vitro. Based on these findings, we tentatively concluded that some patients with ER positive breast cancers benefiting from anti-estrogen therapy may partially attributes to blocking monocyte differentiation into TAM, which destroyed the tumor microenvironment. These findings suggest the future possibility of using TAM as a novel therapeutic target in patients with anti-estrogen resistance and primary triple-negative breast cancers (TNBC) with no effectively therapeutic measures nowadays, which are defined by lack of estrogen receptor, progesterone receptor and ERBB2 gene amplification, representing approximately 16% of all breast cancers.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P6-04-26.
Abstract P6-04-29: Vitamin D induces expression of estrogen receptor and restores endocrine therapy response in estrogen receptor-negative breast cancer
Authors: Santos N , Diaz L , Ordaz D , Garcia J , Barrera D , Avila E , Halhali A , Medina H , Camacho J , Larrea F , Garcia R
Published: August 2014

Background: Approximately 30% of all breast tumors do not express estrogen receptor (ER) and patients with these tumors present poor prognosis and respond poorly to hormone therapy. Calcitriol through its vitamin D receptor (VDR) exerts antiproliferative, apoptotic and pro-differentiating effects in cancer. Calcitriols effects upon ER expression in breast cancer cells is controversial. Therefore, in order to clarify this issue, the aim of the present study was to determine if calcitriol induces ERα expression in ERα-negative breast cancer cells and could restore antiestrogen responses. The evaluation of calcitriol effects was performed in terms of proliferation and regulation of the following genes: Cyclin D1, involved in cell cycle, and Ether-à-go-go 1 (Eag1), related to cell proliferation and tumor progression.

Methods: Cultured cells derived from ERα-negative breast tumors and an established ERα-negative breast cancer cell line (SUM 229) were used in this study. These cells were treated with calcitriol and reverse transcription-PCR or western blotting analyses were performed to assess ERα expression. Growth assays with XTT were used to evaluate the antiproliferative response to the antiestrogens fulvestran and tamoxifen. Gene expression analysis for Cyclin D1 and Eag1 was evaluated by real time PCR in cells treated simultaneously with calcitriol plus estradiol or fulvestran.

Results: The treatment with calcitriol in ER-negative breast cancer cells resulted in the induction of ERα. This effect was specifically mediated through the vitamin D3 receptor (VDR), since the VDR antagonist TEI-9647 effectively inhibited the ability of calcitriol to stimulate ERα gene expression. Consequently, the induction of ERα by calcitriol restores the response to antiestrogens in breast cancer cells by inhibiting cell proliferation. Co-treatment of calcitriol and antiestrogens down-regulated Cyclin D1 and Eag1 gene expression.

Conclusion: Calcitriol induced the expression of ERα and restored antiestrogenic responses in ERα-negative breast cancer cells. Moreover, fulvestran down regulated mRNA expression of Cyclin D1 and Eag1 when ERα-negative cells were pre-treated with calcitriol. These results suggest that the combined treatment with calcitriol and antiestrogens could be a new therapeutic strategy for ERα-negative breast cancer patients.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P6-04-29.
Abstract P6-05-07: Amplified in breast cancer 1 and ankyrin repeat containing cofactor 1 mediate estrogen induced repression of the ErbB2 oncogene
Authors: Garee JP , Riegel AT
Published: August 2014

The oncogene amplified in breast cancer 1 (AIB1/SRC3) is a nuclear receptor coactivator and plays an important role in the regulation of gene transcription. Additionally, AIB1 is important in malignant progression as AIB1 has been shown to be a gene amplified in breast cancer and is also over-expressed in a variety of cancer types. Previous work from our laboratory has identified a splice variant of AIB1 termed AIB1-Δ4. This splice variant arises from the splicing of exon 4 from the mature mRNA. This leads to the formation of an N terminally truncated form of the AIB1 protein, which is a more potent coactivator of steroid dependent transcription compared to the full length AIB1. This suggests that the N-terminal fragment of AIB1, which is not present in AIB1-Δ4, may act to dampen AIB1 mediated transcriptional activation. The presence of this N terminal region suggests this region may be a site of interaction with a repressive protein, which could interact with AIB1 and not AIB1-Δ4. We have identified ANCO1 as an N terminal AIB1 binding protein that acts to repress AIB1 mediated transcriptional activity. Co-immunoprecipitation experiments have indicated interaction between AIB1 and ANCO1 but not between AIB1-Δ4 and ANCO1 indicating ANCO1 binds in the N terminal region of AIB1. AIB1 has a clinical implications and association with the oncogene ErbB2, which is an estrogen repressed gene with hormone effect acting through an estrogen response element (ERE) in intron 1 of the ErbB2 gene. We wanted to determine if the AIB1/ANCO1 complex played a role in hormone regulation of ErbB2. Silencing of AIB1 and ANCO1 by shRNA led to a reversal of estrogen-induced repression of ErbB2 indicating this protein complex may play a role in mediating this repression. Examining binding of these proteins to the intronic ERE of ErbB2 we determined that AIB1 and ANCO1 are recruited in the presence of estrogen. Additionally, repressive proteins such as HDAC3, HDAC4 and N-CoR are recruited along with the AIB1/ANCO1 complex. While AIB1-Δ4 can still be recruited to the intronic ERE no additional repressive proteins are recruited. This suggests a mechanism by which the AIB1/ANCO1 complex can recruit repressive proteins to repress ErbB2 gene transcription, while AIB1-Δ4 can escape this repressive mechanism. This may potentially have interesting clinical implications, as high levels of AIB1-Δ4 may be responsible for increasing the expression levels of the ErbB2 oncogene.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P6-05-07.
Abstract P6-05-14: Estrogen-induced genes in ductal carcinoma in situ(DCIS): their comparison with invasive ductal carcinoma.
Authors: Ebata A , Suzuki T , Takagi K , Miki Y , Onodera Y , Nakamura Y , Fujishima F , Ishida K , Watanabe M , Tamaki K , Ishida T , Ohuchi N , Sasano H
Published: August 2014

It is well known that estrogens play important roles in both the pathogenesis and development of invasive ductal carcinoma (IDC) of human breast. However, molecular features of estrogen actions have remained largely unclear in pure ductal carcinoma in situ (pDCIS), regarded as a precursor lesion of many IDCs. This is partly due to the fact that gene expression profiles of estrogen-responsive genes have not been examined in pDCIS. Therefore, we first examined the profiles of estrogen-induced genes in estrogen receptor (ER)-positive pDCIS and DCIS (DCIS-c) and IDC (IDC-c) components of IDC cases (n = 4, respectively) by microarray analysis. Estrogen-induced genes identified in this study were tentatively classified into three different groups in the hierarchical clustering analysis, and 33% of the genes were predominantly expressed in pDCIS rather than DCIS-c or IDC-c cases. Among these genes, the status of MYB (c-MYB), RBBP7 (RbAp46) and BIRC5 (survivin) expression in carcinoma cells was significantly higher in ER-positive pDCIS(n = 53) than that in ER-positive DCIS-c (n = 27) or IDC-c (n = 27) by subsequent immunohistochemical analysis of the corresponding genes (P < 0.0001, P = 0.03 and P = 0.0003, respectively). In particular, the status of c-MYB immunoreactivity was inversely (P = 0.006) correlated with Ki-67 in the pDCIS cases. These results suggest that expression profiles of estrogen-induced genes in pDCIS may be different from those in IDC, and c-MYB, RbAp46 and survivin may play particularly important roles among estrogen induced genes in ER-positive pDCIS.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P6-05-14.
Abstract P6-06-02: Characterization of an exosome-associated apoptosis-inducing activity produced by triple negative breast cancer cells.
Authors: Georgoulia NE , Iliopoulos D , Mitchison TJ
Published: August 2014

Purpose: We sought to investigate whether breast cancer cells can suppress the proliferation of other breast cancer cells in order to understand the biological significance of cell heterogeneity in breast cancer.

Materials and Methods: 20 breast cancer cell lines, corresponding to all molecular profiles of breast cancer, were cultured in a 20×20 conditioned medium protocol. Cell proliferation and apoptosis were evaluated by EdU (5-ethynyl-2′-deoxyuridine) and PARP (Poly ADP ribose) cleaved labeling respectively, and documented by fluorescence microscopy. Serum free conditioned media (CM) were prepared from all the cell lines, 24hrs of incubation, and screened for their anti-proliferative effect against the different breast cancer tumor cells. Active media were ultra-centrifuged and both the supernatants as well as the pellets were assessed for the anti-proliferative activity.

Results: Using this 20×20 matrix, CM obtained from two out of six triple negative breast cancer cells displayed anti-proliferative activity against the majority of cell lines, irrespectively of their molecular phenotype including triple negative cells. No significant anti-proliferative activity was observed in CMs from luminal or HER2(+) cell lines. However the two triple negative CMs induced a significant decrease in the cell lines' proliferation index, which was associated with a concomitant increase in cells' apoptotic index. Ultracentrifugation revealed that the apoptosis-inducing activity was retained in the pellet but not the supernatant of the CM. Negative staining electron microscopy (EM) revealed the presence of exosomes in the CM's ultra-centrifuged pellet, which stained positive for cd63 surface protein marker using an immunogold assay.

Conclusions: These data strongly suggest that some triple negative breast cancer cell lines posses exosome-mediated apoptosis-inducing activity. This activity may have biological and clinical relevance since it could be a factor participating in the selection of tumor cell subpopulations of different degrees of aggressiveness. Further proteomic and microRNA profiling of the exosomes is ongoing in order to characterize this exosome-mediated apoptosis-inducing activity.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P6-06-02.
Abstract P6-07-04: Evaluation of Adjuvant! Online for primary operable grade 2 breast cancers with 10-year follow-up
Authors: Van Calster B , Brouckaert O , Wildiers H , Christiaens M-R , Weltens C , Paridaens R , Van Limbergen E , Neven P ,   Multidisciplinary Breast Centre, UZ Leuven
Published: August 2014

Background: In primary operable breast cancers, Adjuvant!Online (AOL) uses demographics and clinic-pathological markers to predict the 10-year breast cancer specific survival (BCSS) with and without adjuvant systemic therapy. Grade 2 BC is a difficult group for adjuvant treatment decisions. We compared the observed with the AOL-estimated BCSS rate in our centre in consecutive women primary operated for a grade 2 BC.

Patients and Methods: All patients with grade 2 primary operable breast cancer from UZL diagnosed between 1/1/2000 and 31/3/2002. After 10 years of follow-up, patients were evaluated for BCSS and dead from other cause. For each patient, the adjuvant! Online algorithm (version 8) was applied to predict BCSS taking the patients' adjuvant therapy into account. Adjuvant! Online was validated by assessing the calibration and discrimination. Discrimination was assessed by the area under the ROC curve. The expected BCSS rate was obtained by averaging the predicted probabilities of BCSS. Next, logistic calibration was performed by predicting observed BCSS within 10 years of follow-up with the odds of the predicted probability of BCSS (odds BCSS). In this analysis, the intercept should be 0 and the coefficient for odds BCSS 1 for perfect calibration of the predicted probabilities. A likelihood ratio test is used to test the null hypothesis of perfect calibration.

Results: Six patients were lost to follow-up. The median age at diagnosis of the remaining 445 patients was 56 years (IQR 48 to 67 years, range 26 to 89 years). In 433/445 cases (97%), the prediction from AOL was available. The median AOL predicted probability for BCSS was 89.1% (IQR 83.2 to 94.6%, range 20.3 to 98.9%). The observed BCSS rate was 93.3% whereas the AOL-expected rate was 86.7%. The likelihood ratio test for perfect calibration was statistically significant (p < 0.0001), with an intercept of −1.55 (95% CI −2.33 to −0.84) and a coefficient for odds BCSS of 0.55 (95% CI 0.19 to 0.92). AOL strongly underestimated the probability of BCSS. The AUC was 0.65 (95% CI 0.53 to 0.76). For patients that did not die from BC within 10 years, the median estimated BCSS probability by AOL was 89.6% (IQR 83.5 to 94.6 %), whereas for patients that died from BC the median estimated probability was 83.9 % (IQR 77.9 to 94.5%).

Conclusion: Although Adjuvant! Online achieved moderate discrimination between those that did and those that did not die from BC within 10 years FU, it clearly underestimated the probability of BCSS in our sample.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P6-07-04.
Abstract P6-07-21: Signs and symptoms at central nervous system (CNS) relapse in breast cancer patients with Leptomeningeal carcinomatosis related with shorter survival after recurrence
Authors: Gomez HL , Schwarz LJ , Vásquez J , Neciosup SP , Pinto JA , Vidaurre T , Ferreyros G , Vallejos CS
Published: August 2014

Background: There are signs and symptoms detected at CNS relapse in breast cancer patients that could be identified as a poor prognosis feature of the disease.

Objectives: To describe the commonest signs and symptoms at CNS relapse and its relation with post SNC - recurrence survival in patients with leptomeningeal carcinomatosis (LMC).

Methods: We evaluated a retrospective cohort of 2597 women with breast cancer treated at the Instituto Nacional de Enfermedades Neoplásicas (Lima-Perú) in the period 2000–2005. Clinical data was retrieved from medical records. Breast cancer phenotypes were determined by IHC expression of hormonal receptors (HR) and HER2 and categorized in [HR+, HER2−], triple-negative (TN) [HR−, HER2−] and Her-2 [HR+/−, HER2 +]. The main endpoint was Post-CNS recurrence survival.

Results: A total of 157 cases had CNS relapse from which 124 had only brain metastases, 19 only leptomeningeal carcinomatosis (LMC), and 14 both. Forty three cases were [HR+, HER2−] (5 LMC), 68 were TN (13 LMC) and 51 were [HR+/−, HER2 +] (15 LMC). 152 cases were in clinical stages I-III at diagnosis (96.8% of all SNC relapses). There was no association between IHC phenotypes and age at CNS relapse, ECOG Performance Status (p = 0.837), extracraneal metastases (p = 0.304), control of primary tumor (p = 0.164), number or volume of brain metastases (p = 0.706). Significant association was found between phenotype and time from diagnosis to development of CNS metastases (≤8 months: 16.3% in TN, 7.3% in Her-2 and 0% for [HR+, HER2−]; p = 0.017) and histological grade (grade III: 74.4% in TN, 55.% in Her-2 and 30% for [HR+, HER2−]; p = 0.006). Symptoms/signs commonest in LMC patients were cephalea (7 cases, 72%), meningism (6, 24%), nausea (6, 24%), vomiting (5, 20%), ataxia (5, 20%), facial palsy (4, 16%), somnolence (4, 16%), paraparesia (3, 12%), hiporeflexia (3, 12%), apraxia (3, 12%), arreflexia (3, 12%), poor sphincter control (3, 12%), hemiparesia (2, 8%), seizures (2, 8%), bradypsichia (2, 8%), neuropathic pain (1, 4%). Meningism was most frequent in TN cases (50%). Post recurrence survival was shorter in LMC TN patients (3.61mo vs 4.89mo to [HR+, HER2−] vs 5.95mo to [HR+/−, HER2 +; p = 0.044] and in LMC patients with meningism (1.38mo vs 5.26mo; p = 0.02). Multivariate analysis identify meningism as the worse risk factor for survival in patients relapsed with LMC (HR: 4.33 compared with patients without LMC; p = 0.066).

Conclusions: In LMC patients, meningism was identified as a sign related to shorter survival after the SNC relapse.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P6-07-21.
Abstract P6-07-32: Prognosis of metastatic breast cancer subtypes: the hormone receptor/HER2 positive subtype is associated with the most favorable outcome
Authors: Tjan-Heijnen VCG , Lobbezoo DJA , van Kampen RJW , Voogd AC , Dercksen MW , van den Berkmortel F , Smilde TJ , van de Wouw AJ , Peters FPJ , van Riel JMGH , Peters NAJB , Borm GF
Published: August 2014

Background: In early breast cancer the different subtypes and their prognostic relevance are well known, contrary to the knowledge on prognostic relevance of subtypes in metastatic breast cancer. This study presents survival estimates after diagnosis of distant metastases of breast cancer subtypes in a recent cohort in which the treatment is representative of current clinical practice.

Patients and methods: All patients diagnosed with metastatic breast cancer in one of eight participating hospitals in the South-East part of the Netherlands between 2007–2009 were included and all medical charts were reviewed. Patients were categorized in 4 subtypes based on the hormone receptor (HR) and human epidermal growth factor receptor 2 (HER2) status of the primary tumor; HR positive/HER2 negative, HR positive/HER2 positive, HR negative/HER2 positive and triple negative (TN). Metastatic survival was estimated using the Kaplan-Meier product-limit method. Cox proportional hazards model was used to determine the prognostic impact of breast cancer subtype, adjusted for possible confounders.

Results: A total of 815 patients were included; the HR+/HER2− subtype comprising 66%, the HR−/HER2+ subtype 8%, the TN subtype 15% and the HR+/HER2+ subtype 11% of patients. The four subtypes were associated with different metastatic survival times; the HR+/HER2+ subtype was associated with the best metastatic survival (median, 32.9 months), compared to 22.1 months for the HR+/HER2− subtype, 19.5 months for the HR−/HER2+ subtype and 7.7 months for the TN subtype. Aside from subtype, other prognostic factors of metastatic survival were site of metastases and metastatic-free interval.

Conclusion: Of the four subtypes, the HR+/HER2+ subtype was associated with the most favorable outcome, in terms of metastatic survival.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P6-07-32.
Abstract P6-07-35: Dragonfly Effect or Ironic Paradox: Prognostic Implication of Postsurgical Drain in Breast Cancer Patients
Authors: Yin W , Lin Y , Shen Z , Shao Z-M , Lu J
Published: August 2014

Background: Surgical removal of primary breast cancer may result in subsequent alterations of host milieu. Massive clinical data have indicated that surgical extirpation favorably modifies the natural history for some patients, but it may also precipitate the metastatic outgrowth for others. Drainage fluid, harvested from a post-surgical wound, appears to be a physiological response to surgical trauma. However, few clinicians have recognized that the volume of wound drain might mirror the postoperative recurrence kinetics.

Methods: A total of 1439 patients were selected retrospectively from a large database of patients who underwent surgery between Jan 1, 2000 and Dec 31, 2002 in Fudan University Shanghai Cancer Center, Shanghai, China. The Spearman rank correlation analysis was conducted to evaluate the association between drainage volume of postoperative day 1 (POD1) and clinicopathological parameters, which included age at diagnosis, body weight at diagnosis, total number of excised axillary lymph nodes (ALNs), time to drainage volume less than 30ml (TTV30), total protein and albumin concentrations in serum. Survival curves were performed with Kaplan–Meier method and annual recurrence hazard was estimated by hazard function.

Results: Drainage volume of POD1 was positively correlated with age at diagnosis (Spearman's P < 0.001), weight at diagnosis (P <0 .001) and number of excised ALNs (P <0 .001) and TTV30 (P <0 .001), whereas it was inversely correlated with total protein concentration in serum (p = 0.028). We also observed that drainage volume of POD1 was significantly higher in patients with larger tumors (P < 0.001) and ALNs involvement (P = 0.030).

In the univariate survival analysis, there was less recurrence and death in patients with lower drainage volume of POD1 (both p = 0.006). Cox proportional hazards regression analysis revealed that drainage volume of POD1 was an independent predictor of recurrence-free survival (RFS; p = 0.035) and overall survival (OS; p = 0.047). Additionally, triple negative patients with larger volume of POD1 seemed to have poorer RFS (log-rank p = 0.022; Cox model P < 0.05) and OS (log-rank p = 0.015; Cox model P < 0.05).

Conclusions: Our study suggests that postsurgical drainage volume was strongly correlated with prognosis in breast cancer patients. The aim of this study was not only to enlighten novel prognostic factors, but also to infer that some host related characteristics such as extent of reaction to surgery play very important roles in the growth of micro-metastases. Therefore, integration of both host and tumor related factors be more precise to predict relapse risk than current modalities such as the St Gallen criteria, Oncotype-21 and Mammaprint etc. Further research on the underlying mechanism of this phenomena may lead the way to new adjuvant treatment targets and strategies.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P6-07-35.
Abstract P6-07-42: Patients with Nipple-areola Paget's Disease and Underlying Invasive Breast Carcinoma had a Very Poor Survival: a Matched Cohort Study
Authors: Ling H , Xu X-L , Liu Z-B , Shao Z-M
Published: August 2014

Purpose: Nipple-areola Paget's disease together with invasive breast cancer is a rare kind of disease. This study is to investigate the characteristic and survival of it.

Patients: From 2001 to 2005, all patients diagnosed as nipple-areola Paget's disease with underlying invasive breast carcinoma in Fudan University Shanghai Cancer Center (FUSCC) were recruited in this study (PD group).

Patients diagnosed as invasive breast cancer without Paget's disease were recruited in control group for clinical and pathologic features comparison.

Since tumors in PD group expressed more high risk factors (see results), a matched study was done to investigate whether the poor survival of PD group was by nature or due to the overexpression of high risk factors. The matched group was derived from all the patients who received mastectomy for invasive breast cancer (with no Paget's disease) during 2001 to 2005 in FUSCC. The match was conducted according to four variables: dimension of tumor, lymph node status, hormone receptor status and Her2 status. After matching, patients were randomly selected in a 3:1 ratio to patients in PD group.

Results: 52 patients were recruited in PD group. 22 (42.3%) patients' primary symptom were nipple change, 25 (48.1%) patients' primary symptom were mass in breast. Physical examination found no nipple-areola change in 24 patients at diagnosis, their Paget's disease were found unexpectedly by routine nipple-areola section examination after mastectomy.

More than 8000 invasive breast cancers were diagnosed during 2001 to 2005. 700 patients (140 consecutive patients per year, average monthly patient amount) were recruited in control group. Compared with control group, tumors in PD group had larger size, more axillary lymph node involvement, lower hormone receptor expression, higher HER2 expression and worse survival.

156 patients were recruited in the matched group. The 5-year relapse-free survival and overall survival were both lower for patients in PD group than those in the matched group (table1).

Conclusions: Patients with invasive breast cancer and Paget's disease tended to have larger size, more lymph nodes involvement, lower hormone receptor expression, higher HER2 expression and worse survival than those without PD. The matched study confirmed the survival disadvantage of patients with Paget's disease and underlying invasive breast cancer. Thus, oncologists should regard such patients as having high risk for relapse. Pathologists should exam the nipple-areola section carefully after mastectomy although the patient has no macroscopic nipple-areola change.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P6-07-42.
Abstract P6-08-10: Cancer as self: a novel assessment of patient identity as it relates to a cancer diagnosis
Authors: Horst KC , Fero KE , Haimovitz K , Dweck CS
Published: August 2014

Introduction: Previous studies have demonstrated that an individual's attitude towards self may impact how one faces challenges. Those who view themselves as malleable (growth mindset) approach challenges and problems as a learning opportunity and an opportunity to improve and change. In contrast, those who are more fixed in their approach (fixed mindset) have difficulty with challenges or setbacks. In the present study, we applied this conceptual framework to a cohort of breast cancer patients to evaluate their attitudes towards the cancer diagnosis. The goal of this study is to test the internal validity of this questionnaire and to correlate its findings with patient-reported measures of depression, anxiety and insomnia.

Methods: Twenty consecutive breast cancer patients undergoing radiation therapy completed an electronic survey consisting of 4 scales: the Hospital Anxiety and Depression Scale (HADS; 14 items), the Insomnia Severity Index (ISI; 7 items), a validated psychological questionnaire assessing fixed versus growth mindset (8 items), and an experimental questionnaire evaluating whether the patient views their disease as self-defining. Demographic and clinical variables were also collected. A principal components factor (PCF) analysis was used to examine the internal structure of the new questionnaire. Hierarchical linear regression analyses were performed to assess its relationship to other variables.

Results: Twenty patients completed the survey during the first week of radiotherapy. Median age was 57 (range, 31–81). Five patients (25%) had Stage 0 disease, 9 (45%) Stage I, 0 (0%) Stage II, 5 (25%) Stage III, and 1 (5%) Stage IV. Seven patients (35%) received chemotherapy while 13 (65%) did not. Eleven patients (55%) had significant others while undergoing treatment and 9 (45%) were single. The PCF analysis confirmed that the items of the experimental questionnaire were internally consistent (α = .88), such that all were retained in the final scale for subsequent analyses. The extent to which patients saw cancer as self defining was statistically significantly associated with their reported levels of depression (r = .60, p < .005) and anxiety (r = .60, p < .005). There was a similar trend with reported levels of insomnia which was not statistically significant (p = 0.2). Of the demographic and clinical variables, only stage of cancer was associated with depression and anxiety.

Conclusions: Patients undergoing treatment for breast cancer who view the cancer diagnosis as self-defining are more likely to self-report depression, anxiety, and insomnia. In this cohort, the experimental questionnaire demonstrated strong internal validity. An intervention that alters patients' perception of identity as it relates to the cancer diagnosis could be a promising compliment to the standard treatment of depression, anxiety, and insomnia.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P6-08-10.
Abstract P6-10-03: The PKC inhibitor PKC412 antagonizes breast cancer cell growth and enhances tamoxifen sensitivity
Authors: Shou J , Chew SA , Mitsiades N , Kumar V , Fu X , Chamness G , Osborne K , Schiff R
Published: August 2014

Background: AIB1 (SRC-3, NCoA3), a member of the p160/steroid receptor coactivators family, plays a critical role in cell growth and proliferation. In estrogen receptor-alpha positive (ER+) breast cancer (BC) cells, it coactivates estrogen- and additional transcription factors-dependent gene transcription, reducing the antagonistic activity of tamoxifen and resulting in tamoxifen resistance (TR). We have previously shown that BC patients whose tumors expressed high levels of both AIB1 and HER-2 had worse outcomes with tamoxifen therapy, suggesting that AIB1 may be an important diagnostic and therapeutic target. Our findings that knocking down AIB1 attenuates ER signaling and inhibits breast cancer cell growth further indicate that the manipulation of AIB1 level could be an approach to treating BC and overcoming TR. Recently, it has been shown that protein kinase C (PKC) isoforms phosphorylate AIB1 and prevent its proteasome-mediated degradation. The present study was carried out to test if the multi-targeted kinase and PKC inhibitor PKC412 (midostaurin) is capable of promoting degradation of AIB1, inhibiting BC cell growth, and promoting tamoxifen antagonistic activity.

Methods: The ER+ MCF7, T47D, and ZR75-B BC cells and their tamoxifen-resistant derivatives (TR) were used. The Methylene Blue assay was employed to measure cell viability of BC cell lines after treatment with PKC412 or the combination of PKC412 with tamoxifen. To determine the impact of PKC412 on AIB1 and ER proteins and mRNA levels, we used immunoblotting and RT-qPCR, respectively.

Results: PKC412 successfully inhibited the growth of MCF7, T47D, and ZR75-B cells, and their tamoxifen-resistant derivatives. Treatment with PKC412 depleted AIB1 protein and reduced the level of ER protein without significant alteration in AIB1 mRNA level. Consequently, ER signaling was disrupted, as reflected by decreased expression of ER target genes such as GREB1. PKC412 also enhanced tamoxifen's antagonistic activity in the parental cell lines and sensitized tamoxifen-resistant MCF7 and ZR75-B cells to tamoxifen.

Conclusions: The results of this study suggest that the multi-targeted kinase and PKC inhibitor PKC412 can post-translationally destabilize AIB1 protein and ER, inhibiting BC cell growth and viability. PKC412 enhances tamoxifen's antagonistic activity on BC growth; furthermore, it sensitizes tamoxifen-resistant cells to tamoxifen. Thus, PKC412 is a promising agent to treat BC and, in combination with tamoxifen, to delay or overcome TR.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P6-10-03.
Abstract P6-11-02: Inhibition of mTOR and Fatty Acid Synthase (FASN) overcome acquired resistance to Trastuzumab, Lapatinib and both in HER2+ breast cancer
Authors: Puig T , Blancafort A , Giro A , Viqueira A , Viñas G , Massaguer A , Carrion-Salip D , Bolos MV , Urruticoechea A , Oliveras G
Published: August 2014

Introduction: Resistance to dual biological blockade of HER2 positive breast cancer arises as a novel therapeutic challenge with high clinical relevance. We have shown that inhibition of fatty acid synthase (FASN) has anticancer activity and enhances the effect of chemotherapy and anti-HER2 drugs in pre-clinical breast cancer models. Furthermore, our group has recently observed that inhibition of FASN can reverse resistance to anti-HER2 therapies.

The development of FASN inhibitors has consequently appeared as a novel anti-target modality for treating cancer. However, the clinical use of FASN inhibitors, such as Cerulenin, C75 and Epigallocatechin 3-gallate (EGCG) is limited by anorexia and induced body weight loss or by its low in vivo potency and stability. We synthesized novel EGCG-related inhibitors to improve their use as anti-tumor agents. Within these, G28UCM was selected for its inhibitory effect of FASN activity and selective cytotoxicity in tumor cells.

Recently, it has been reported that mTOR blockade acts synergistically with HER2 inhibition to induce cell death and tumor regression in resistant breast cancer models.

Materials and Methods: We have developed long term HER2+/ FASN+ breast cancer cell lines (SKBr3) resistant to the HER2-monoclonal antibody Trastuzumab (SKTR), the EGFR/HER2-tyrosine kinase inhibitor Lapatinib (SKLR) or both (SKLTR). Once established, we have characterized these cells by studying a panel of EGF receptors signaling proteins with western blot analysis, changes in adherence to extracellular matrix proteins and invasion capacity with colorimetric assays. Using MTT assay, we have assessed the effect of the mTOR-inhibitor, Temsirolimus, and G28UCM on viability of parental and resistant cells. The isobologram method has been used to estimate the possible synergistic effect between both treatments.

Results: Resistant cells maintained downstream HER2 pathway activation by stimulating the expression/activation of alternative EGF family receptors and/or those specific ligands. Moreover, these cells increased adherence to extracellular matrix proteins and invasion capacity.

Different combination regiment of Temsirolimus with G28UCM displayed a strong synergistic effect in inducing cell death of both, parental and resistant HER2+ breast cancer cells.

Conclusions: The inhibition of mTOR and FASN is a potential novel therapeutic strategy in dual resistant HER2 positive breast cancer.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P6-11-02.