bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract P5-01-13: Flat Epithelial Atypia: Management and outcome in three Dutch teaching hospitals
Authors: Ghuijs PM , de Vries B , Strobbe LJA , van Deurzen CHM , Heuts EM , Keymeulen KBMI , Lobbes MBI , Wauters CAP , Van de Vijver KKBT , Smidt ML 
Published: August 2014

Abstract
Background: Flat Epithelial Atypia (FEA) is a presumably neoplastic alteration of terminal duct-lobular units, characterized by the replacement of native luminal epithelium by ductal cells demonstrating low-grade cytologic atypia. The architecture shows stratification of epithelial cells. FEA is often accompanied by microcalcifications and therefore discovered in biopsies following screening mammography. FEA is frequently seen in association with ADH (atypical ductal hyperplasia), DCIS (ductal carcinoma in situ), lobular neoplasia and invasive tubular carcinomas. There is emerging evidence suggesting FEA may represent a precursor to DCIS. The risk of subsequent breast carcinoma remains to be defined. The aim of this study is therefore to inventorise the management and outcome of solitary FEA in histological biopsies in three Dutch teaching hospitals.
Materials and Methods: Data of this retrospective multicentre study were collected in a database. Local pathology databases were screened with the terms: ‘FEA’, ‘Flat Epithelial Atypia’, ‘columnar atypia’ and Dutch equivalents. Results were manually screened, only including solitary FEA.
Patient files were viewed for information on presentation, mammography, ultrasound and management: surgery vs follow-up. In case of excision, definitive pathology was added.
Results: We included 103 patients showing only solitary FEA in the primary biopsy. Management of these patients consisted of follow-up for 60 patients (58,3%) and surgery for 43 patients (41,7%, 49 excisions): lumpectomy (42) or mastectomy (7). Reason for choosing mastectomy was preventive in case of contralateral breast cancer or increased familial or genetic risk.
Definitive pathology of lumpectomy or mastectomy showed no abnormalities or solitary FEA in 31 patients; other findings were ADH in 7, LCIS in 4 and DCIS in 7 patients. Some patients showed more than one finding. Invasive breast cancer (IBC) was detected in 3 patients. Only one mastectomy showed invasive disease, located in a different lobe, however.
No incidents occurred in the follow-up group.
Conclusions: No consistent management exists concerning solitary FEA in these three hospitals. Also, one hospital used the diagnosis of FEA inconsistently and interchangingly with other terms. Lack of this study is the retrospective gathering of data, making it difficult to detect the reasons for the chosen management. DCIS or IBC was discovered in 20,4% of all surgical specimens. It was concluded that FEA should be seen as a red flag, indicating the possible presence of a more malignant lesion. Additional research is warranted concerning long term follow-up for this patient group.
Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P5-01-13.
Abstract P5-01-14: Discrepancy between routine and expert pathologists in histological assessment of early stage non palpable breast cancer and its impact on loco regional and systemic treatment
Authors: Postma EL , Verkooijen HM , van Diest PJ , Willems SM , van den Bosch MA , van Hillegersberg R 
Published: August 2014

Abstract
Background: Histopathological parameters are essential for deciding on further adjuvant treatment in patients with breast cancer after surgery. Here, we assessed the impact of inter observer variability on further treatment strategy in patients with clinically node negative, non palpable breast carcinomas.
Methods: Clinical and histopathological data of 314 patients with clinically node negative non palpable invasive breast cancer were analysed. Histological assessment of the primary tumour and sentinel nodes was first performed in a routine setting, subsequently central review took place. In cases of discordance between local en central assessment, we determined the impact on locoregional and systemic treatment strategy.
Results: Discordance between local and central review was seen in 13% of the patients (kappa 0.60 95% CI 0.50–0.71) for type, in 12% (k = 0.796 95% CI 0.73–0.86) for grade, in 1% for ER status (k = 0.898 95% CI 0.80–1.0), in 2% for PR status(0.940 95% CI 0.89–0.99). Discrepancy in the assessment of the sentinel node(s) was seen in 2% of the patients. (k = 0.954 95% CI 0.92–0.98). Practising current Dutch Guidelines, loco regional treatment would be affected in 2% (7/310), systemic treatment in 5% (16/310) and both in 1% (2/310) of the patients.
For patients in whom central review would have led to additional systemic treatment (3%), the 10 years mortality and recurrence rate would decrease with a median of 4.6% and 15% respectively.
Conclusion: Inter observer variation in the histopathological assessment of non-palpable breast carcinoma specimens and sentinel nodes results in a different loco regional or systemic treatment advice in 8% of the patients.
Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P5-01-14.
Abstract P5-02-03: Large-scale genomic instability consistently identifies BRCA1/2 inactivation in breast cancers
Authors: Stern M-H , Popova T , Manié E , Dubois T , Sigal-Zafrani B , Bollet M , Sastre-Garau X , Vincent-Salomon A , Houdayer C , Stoppa-Lyonnet D 
Published: August 2014

Abstract
The goal of this study was to identify genomic markers predicting actual BRCA1/2 inactivation in breast cancers. By comparing genomic profiles of BRCA1/2 mutated and wildtype basal-like carcinomas (BLC) we developed a signature of BRCAness. The signature accounts for the level of the large scale genomic instability and was demonstrated to predict BRCA1/2 impairment with 100% sensitivity and 90% specificity in the large series of BLCs. We introduced a bioinformatics procedure evaluating the number of large scale rearrangements (denoted as Large Scale Transition or LST) in the genome and showed that the number of LSTs discriminate homologous recombination deficient and proficient tumors (Popova et al, Cancer Research 2012 Aug).
The efficiency of the signature of BRCAness represented by the number of LSTs was then evaluated for the luminal breast tumors and ovarian carcinomas. Luminal breast tumors and ovarian carcinomas could be stratified into two groups according to the number of LSTs. The performance of the LST signature in all types of breast cancers will be presented.
Measurement of LST is a highly reliable, simple and cost effective method to identify patients who should benefit of a BRCA1/2 testing, and to select tumors which may respond to treatment targeting DNA repair deficiencies (platinium salts, PARP inhibitors).
Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P5-02-03.
Abstract P5-03-08: A fluorescence STAT3 reporter preferentially expressed in human breast cancer tumor-initiating cells
Authors: Wei W , Tweardy D , Zhang M , Roarty K , Rosen J , Lewis M 
Published: August 2014

Abstract
Cancer initiating/stem cells are thought to be responsible for treatment-resistance, disease-recurrence and metastasis. Current breast cancer stem cell markers lack direct correlation with stem cell function, which makes the targeted therapeutic design difficult. The STAT3 pathway is a critical regulator of the function of normal stem cells and evidence is accumulating for its important role in breast cancer stem cells, as well as associated “niche” cells. However, due to lack of method for pathway-activity-based live cell separation, whether STAT3 functions in the cancer stem cells themselves or in surrounding niche cells, or both remains unknown. To answer this question, we have constructed lentiviral fluorescent reporters, which enables FACS-sorting of cells differ in STAT3 activity as well as in vivo/in situ localization of STAT3 responsive cells. Our study for the first time separated the STAT3 positive and STAT3 negative cells from the same breast tumor in live, and for the first time provided functional evidences that the STAT3 positive cells are enriched for breast cancer stem cells compared to STAT3 negative cells, as tumor cells positive for reporter activity perform better in both mammosphere formation and dilution transplantation assays compared to the negatives.
Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P5-03-08.
Abstract P5-03-09: Receptor Activator of Nuclear Factor Kappa B (RANK) as a potential therapeutic target in triple-negative breast cancer
Authors: Reyes ME , Masuda H , Zhang D , Reuben JM , Woodward W , Darnay BG , Hortobagyi GN , Ueno NT 
Published: August 2014

Abstract
Background: Triple-negative breast cancer (TNBC) has a poorer prognosis in comparison to other breast cancer subtypes because of its high metastatic rate. There is evidence that cancer stem cells (CSCs), known to have an epithelial-to-mesenchymal (EMT) phenotype, may contribute to the metastasis of breast cancer. Denosumab (an anti-RANK ligand antibody) is currently being explored as a treatment for breast cancer in phase II and III clinical trials. RANK is thought to regulate mammary stem cells. Therefore, we hypothesized that RANK promotes tumorigenicity and metastasis of TNBC via the regulation of EMT and enrichment of CSCs. We sought to determine the effects of RANK expression on the tumorigenic and metastatic activities of TNBC cells and whether RANK is a potential therapeutic target in TNBC.
Methods: Primary breast tumor specimens from the MD Anderson Cancer Center Tumor Bank were stratified by ER/HER2 status. We analyzed RANK mRNA expression levels, obtained by Affymetrix gene chip array, in ER+/HER2-negative breast tumors (n = 22) and ER−/HER2-negative breast tumors including TNBC tumors (n = 18). To study the effects of RANK suppression in TNBC cells, we generated two stable RANK shRNA MDA-MB-231 TNBC cell lines and a negative shRNA cell line as a control and examined whether knockdown of RANK expression correlated with a decrease in mesenchymal markers, vimentin and snail, and stem cell marker, ALDH1, invasion, migration, and tumorigenicity in TNBC cells.
Results: An analysis of RANK mRNA expression levels among human primary breast cancer specimens revealed increased RANK expression in ER−/HER2-negative primary breast tumors including TNBC tumors, compared with ER+/HER2-negative primary breast tumors (p = 0.034). Knocking down RANK in MDA-MB-231 cells significantly inhibited migration (p = 0.0005) and invasion (p = 0.0006) and reduced the stem cell subpopulation, as shown by a decrease in mammospheres (p = 0.0188) and ALDH1 activity. Further, in a three-dimensional (3D) cell culture model, we observed significant inhibition of projections in MDA-MB-231 cells with knockdown of RANK (p &lt; 0.05), which correlated with downregulation of the mesenchymal markers vimentin and snail shown by qRT PCR analysis.
Conclusion: Our data demonstrate that RANK may promote the tumorigenesis of TNBC by enriching cancer stem cells. In future studies, we will investigate the effect of RANK inhibition on metastasis in vivo using a TNBC xenograft model. Our long-term goal is to develop RANK-targeted therapy for TNBC.
Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P5-03-09.
Abstract P5-03-12: Targeting breast cancer stem cells using the autopahgy inhibitor N-Acetyl cysteine
Authors: Dave B , Granados S , Mitra S , Chang JC 
Published: August 2014

Abstract
Introduction: One in eight women is diagnosed with breast cancer in the United States. The most aggressive form of breast cancer is the “triple negative breast cancer” (TNBCs), where there is a lack of expression of all three receptors, namely ER, PR and HER2 and a lack of targeted therapies lead to the highest relapse rate in breast cancer. This makes it imperative to identify and target the mechanism of relapse of this cancer. We have recently identified autophagy as a mechanism of tumor resistant cell survival in breast cancer. We have demonstrated an increase in autophagy in chemotherapy treated patients. Further, we have shown that addition of a stem cell inhibitor against NOTCH reduces autophagy and stem cells population. In order to eliminate these tumor resistant cells from surviving chemotherapy, we plan to target the cell survival pathway of autophagy using N-acetyl csyteine as a novel inhibitor.
Materials and Methods: We treated three triple negative cell lines (SUM159, BT549 and MDA-MB231) with varying concentrations of N-acetyl cysteine and determined its impact on tumor initiating cells via mammosphere formation and FACS sorting of CD44hi/CD24low cells. N-acetyl cysteine affects mitochondrial metabolism so we tested its impact on mitochondrial DNA mass.
Results: N-acetyl cysteine significantly decreased TIC population as evidenced by the remarkable reduction in mammosphere formation efficiency and levels of CD44hi/24low cells at 1 and 10uM in all three cell lines. In two cells lines we have demonstrated that there is a significant increase in mitochondrial mass upon treatment of NAC.
Conclusion: We have currently determined that N-acetyl cysteine works via autophagy and eliminates tumor initiating cell population. We have also demonstrated that this involves changing the mitochondrial mass and overall changes in the metabolism of these cells. This novel interlink between mitochondrial metabolism and autophagy provided a new insight into the role of tumor initiating cells in breast cancer and possible new approaches to treat therapy resistance in triple negative breast cancer.
Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P5-03-12.
Abstract P5-03-13: Detection of tumor initiating cells (TIC) among the peripherally circulating epithelial tumor cells from patients with breast cancer
Authors: Pachmann K , Zimon D , Pizon M , Carl S , Rabenstein C , Camara O , Pachmann U 
Published: August 2014

Abstract
Background: The detection of tumor cells circulating in the peripheral blood of patients with breast cancer is a sign that cells have been able to leave the primary tumor and survive in the circulation. However, in order to form metastases they require additional properties such as the ability to adhere, self renew and grow. Here we present data that a variable fraction among the circulating tumor cells detected by the maintrac® approach express mRNA of the stem cell gene nanog and of the adhesion molecule vimentin and are capable of forming tumor spheres a property ascribed to TIC.
Patients and methods: Between 10 to 50 circulating epithelial antigen positive cells detected by the maintrac® approach were selected randomly from each of 20 patients with breast cancer before and after surgery and isolated using automated capillary aspiration and deposited individually onto slides for expression profiling. In addition, the circulating tumor cells were cultured without isolation among the white blood cells from 5 patients with breast cancer in different stages of disease using culture methods favoring growth of epithelial cells.
Results: Although no EpCAM mRNA positive cells expressing stem cell genes or the adhesion molecule vimentin were detected before surgery, 10 to 20% of the cells were found to be positive for mRNA of these genes after surgery. Most surprisingly, it was possible to grow tumor spheres (Fig 1) from these circulating cells without previous isolation in all patients in a fraction comprising between 1% of the epithelial antigen positive cells in patients with newly diagnosed tumor up to 10% in a patient with advanced breast cancer.
Conclusion: We here show, that among the peripherally circulating tumor cells a variable fraction is able to express stem cell and adhesion properties and can be grown into tumor spheres, a property ascribed to cells capable of initiating tumors and metastases.
Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P5-03-13.
Abstract P5-03-14: Expression of ALDH1 in metastasizing axillary lymphnodes in breast cancer
Authors: Shi A , Dong Y , Bi L , Xu N , Fan Z , Li S , Yang H , Li Y 
Published: August 2014

Abstract
Background: There is increasing evidences that a wide variety of malignancies, including breast cancer, may be driven by a small subset of ‘tumor-initiating cells’ or ‘cancer stem cells’ (CSC) which are able to form tumors in immunocompromised mice as well as to generate the phenotypic heterogeneity of the initial tumor. Enzyme aldehyde dehydrogenase (ALDH1) has been reported as a possible marker for mammary CSC. These cells are a source of tumor recurrence and metastasis, and are resistant to chemotherapy, radiotherapy and hormone therapy.
Objective: Assuming that the detection of CSC in axillary lymph nodes is more effective to predicting cancer outcome than the widely used detection of cancer cells in axillary lymph nodes, we measure ALDH1 levels to predict their presence into axillary lymph nodes on development of cancer and anticipate outcomes.
Methods: ALDH1 protein was detected by an immunohistochemical technique in 229 cases of breast cancer diagnosed from 2002 to 2011 Follow-up ranged from 11.5 months to 96.9 months, with a mean of 73.9 months. A survival assay was used to determine the relationship between distant metastatic rate and survival rate.
Results: ALDHl expression was detected in 79cases and the Positive rate in metastatic axillary lymph nodes was 34.5%. Negative ER, PR status were related to the ALDH1 positive cases(P = 0.012). See Table 1. Mortality rate between ALDH1 positive cases (50.8%) and negative cases (28.8%) were significantly different (P = 0.001). See Table 2. Further, survival analysis of recurrence-free survivals (RFS) and survival rate decreased significantly between ALDHl positive and negative cases (P = 0.001) (see table 2) and COX analysis shows that ALDH1 expression is an independent predictor of poor outcome in breast cancer(P = 0.011).
Discussion: What cancer stem cells migrate to the axillary nodes have more important prediction than that the matastesis of normal cancar cells in axillary node. It might be a role resulting in dying in breast cancer.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P5-03-14.
Abstract P5-04-01: Expression of epithelial-mesenchymal transition (EMT)-related markers in primary tumors and matched lymph node metastases in breast cancer patients
Authors: Zaczek AJ , Ahrends T , Markiewicz A , Seroczynska B , Szade J , Welnicka-Jaskiewicz M , Jassem J 
Published: August 2014

Abstract
Background. Malignant tumors may include multiple cancer cell populations with various metastatic potential. More aggressive subpopulations might easier be captured in lymph nodes metastases (LNM) than in primary tumors (PT). We evaluated mRNA and protein levels of master EMT regulators: TWIST1, SNAIL and SLUG, protein levels of EMT-related markers: E-cadherin, vimentin, and expression of classical breast cancer receptors: HER2, ER and PR in PT and corresponding LNM. The results were correlated with clinicopathological data and patient outcomes.
Methods. Formalin-fixed paraffin-embedded (FFPE) samples from PT and matched LNM from 42 stage II-III breast cancer patients were examined. Expression of TWIST1, SNAIL and SLUG was measured by RT-qPCR, with median as cut off for positive result. Protein expression was examined by immunohistochemistry on tissue microarrays. For TWIST1, SNAIL, SLUG, E-cadherin and vimentin &gt;10% of positively stained cells defined positive result. ER and PR were scored according to Allred system, and HER2 - according to HercepTest criteria. Results were considered concordant if PT and LNM were both positive or both negative. Concordance was measured by estimating Cohen's kappa coefficient (κ), with κ value equal 1 indicating perfect agreement. Disease-free survival (DFS) and overall survival (OS) were compared using F-Cox test. Hazard ratios (HRs) with 95% confidence intervals (95% CI) were computed using Cox regression analysis.
Results. On average, expression of TWIST1, SNAIL and SLUG was significantly higher in LNM compared to PT (P &lt; 0.00001 for all). Gene and protein levels of TWIST1, SNAIL and SLUG were highly discordant between PT and matched LNM (Tab 1). Receptor conversion, particularly loss of ER and PgR, occurred in 18% and 29% of cases, respectively. Increased expression of TWIST1 and SNAIL in LNM was associated with shorter OS (p = 0.045 and p = 0.022, respectively) and DFS (p = 0.02 and p = 0.01, respectively), whereas their expression in PT had no prognostic impact. Negative-to-positive switch of TWIST, SNAIL and vimentin correlated with shorter OS and DFS. Conversion of ER and PR had no significant impact on survival.
Conclusions. LNM are enriched in cells with more aggressive phenotype, marked by elevated levels of EMT regulators. High expression of TWIST1 and SNAIL in LNM, as well as negative-to-positive conversion of both TWIST1 and SNAIL confer worse prognosis, confirming the correlation of EMT with aggressive disease behavior.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P5-04-01.
Abstract P5-04-02: DYRK2 regulates breast cancer invasion via Snail/E-cadherin pathway
Authors: Mimoto R , Imawari Y , Kamio M , Kato K , Nogi H , Toriumi Y , Takeyama H , Yoshida K , Uchida K 
Published: August 2014

Abstract
Introduction: Epithelial-Mesenchymal Transition (EMT) is characterized by the loss of cellular adhesion molecule E-cadherin, and plays a fundamental role during early steps of breast cancer invasion. An important regulator of E-cadherin expression is Snail, a zinc finger transcriptional repressor. Snail is posphorylated by GSK3β and then degraded by βTrCP mediated ubiquitination. Here we found another kinase, DYRK2, regulates the stability of Snail. Knockdown of DYRK2 promoted the EMT and cancer invasion both in vitro and in vivo experiments.
Methods: Cell lines: MCF-7 cells were transfected with pSuper vector (pSuper control) and pSuper shRNA DYRK2 (shRNA-DYRK2) with G418 to isolate stable cell lines.
Immunoblotting: Expression of pertinent molecular markers was determined by western blotting.
RT-PCR: RNA was extracted using TRIsure and 500 ng of total RNA was amplified using Primer Script One Step RT-PCR Kit Ver.2.
Invasion assay: Various cancer cell lines were seeded on the top of the upper chamber with serum free medium while the bottom chambers were filled with medium containing 10% FBS.
In vivo metastasis assay: Cells in PBS were injected into the left ventricle of 7-week-old female nude mice. Metastases to distant organs were confirmed by IVIS2000.
Immunohistochemistry: We acquired paraffin-embedded tissue sections from primary breast tumor cores and from Jikei University of Medicine. Immunohistochemistry for DYRK2 was performed.
Results: In MCF-7 cells, knockdown of DYRK2 increased Snail in the protein level but not in mRNA level. Upon DYRK2 knockdown, MG132 treatment had no effect on additional increase in Snail. DYRK2 knockdown decreased Snail ubiquitination. Stable DYRK2 depletion led to Snail accumulation and E-cadherin abrogation. Fibroblast marker, Vimentin, emerged in DYRK2 depleted cells. For further analysis, we carried out invasion assays. The invasion potential in DYRK2-depleted cells was substantially higher than that in control cells. In xenograft model, we used nude mice received intracardial injections of pSuper control or shRNA-DYRK2 cells. 6 weeks after injections, a significant increase in bone and lung metastasis was observed in shRNA-DYRK2 group. The patients with tumors expressed low DYRK2 showed significantly poor prognosis.
Conclusion: In breast cancer cells, Snail is phosphorylated by DYRK2 and then acquires the ability to be ubiquitinated. In the absence of DYRK2, Snail is unable to be degraded by ubiquitin-proteasome machinery. Accumulation of Snail promotes EMT and cancer invasion, so low expression of DYRK2 leads to poor prognosis in breast cancer.
Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P5-04-02.