bluuebunny/crossref_metadata_2025_split huggingface dataset:
P3-01-12: Prognostic Impact of RANK, RANKL and OPG Gene Expression in ER Positive Primary Breast Cancer.
Authors: Ruckhaeberle E , Mueller V , Schmidt M , Saenger N , Hanker L , Gaetje R , Ahr A , Holtrich U , Karn T , Rody A , Kaufmann M 
Published: June 2012

Abstract
Background
The cell surface receptor RANK (receptor activator of NFκB), its ligand (RANKL) and the decoy receptor of RANKL osteoprotegerin (OPG) play an important functional role in bone physiology and in bone metastasis by regulating osteoclasts. Just recently it was shown that tumor-infiltrating lymphocytes can stimulate breast cancer metastases through RANK-RANKL signalling.
Material and methods: We analyzed gene expression of RANK, RANKL and OPG in a combined Affymetrix dataset of 307 ER positive breast cancers from our institutions which were either untreated of treated with chemotherapy. Kaplan Meier analysis of disease free survival and Cox regression analysis was applied to examine the prognostic value of the different markers.
Results: We observed no significant difference in survival when samples were analyzed according to either RANK or RANKL mRNA expression. In contrast when samples were stratified in quartiles of OPG expression a positive linear relationship of survival with the expression of OPG was observed. Moreover since OPG demonstrated a bimodal type of expression a cutoff value can be derived from the expression data. Using this cutoff value a hazard ration of 2.14 (95% CI 1.27−3.61; P=0.004 for low OPG expression was detected. OPG expression correlated with lower proportion of grade 3 tumors (15.7% vs 27%; P=0.022) and a higher proportion of PgR positive samples (86.2% vs 71.4%; P=0.002). No significant differences were observed for lymph node status, age, tumor size and HER2 status. In multivariate analysis only lymph node status remained significant while OPG, Ki67, age, grade, and PgR only displayed a trend towards significance.
Conclusion: Expression of osteoprotegerin seems to correlate with good prognosis in ER postive breast cancer. These data are in line with in vitro studies demonstration that OPG inhibits RANKL induced migration of tumor cells.
Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-01-12.
P3-02-01: A Novel Bispecific, Hexavalent, Antibody (HexAb) Inhibits Anchorage-Independent Growth and Reduces Invasiveness of Triple-Negative Breast Cancer Cell Lines In Vitro.
Authors: Goldenberg DM , Wang Y , Trisal P , Cardillo TM , Rossi EA , Chang C-H 
Published: June 2012

Abstract
Background
Combination therapy using two different monoclonal antibodies to achieve improved efficacy without increased toxicity is being pursued in various preclinical and clinical studies but, in principle, could be accomplished with a single bispecific antibody (bsAb). We note the elevated expression of the type-I insulin-like growth factor receptor (IGF-1R) and the trophoblast cell-surface marker (Trop-2) in diverse epithelial tumors, including breast cancers, and the increasing evidence of their correlation with poor survival. Therefore, we chose to explore the potential of 1R-(E1)-(E1), the bispecific HexAb constructed from hR1 (humanized anti-IGF-1R) and hRS7 (humanized anti-Trop-2) for treating breast cancers. Methods: 1R-(E1)-(E1) was generated by the Dock-and-Lock (DNL) method to comprise a full IgG of hR1 linked to two stabilized dimers of hRS7 Fab, and evaluated in three HER2−negative breast cancer cell lines: the non-evasive MCF-7, the moderately invasive MDA-MB-468, and the highly invasive MDA-MB-231. The in vitro characterizations of 1R-(E1)-(E1) included flow cytometry to determine cell binding, soft agar assay to evaluate the effect on anchorage-independent growth, and BD matrigel chambers to assess invasion properties. Statistical differences (P values) between two populations were determined by Student's t-test.
Results: All three cell lines were found to express both IGF-1R and Trop-2, with MCF-7 and MDA-MB-468 notably higher in IGF-1R and Trop-2, respectively. As expected, 1R-(E1)-(E1) bound to all three cell lines. At 50 μg/mL, 1R-(E1)-(E1) reduced the invasion of MDA-MB-468 to less than 10% of the untreated control, whereas under the same conditions, MDA-MB-231 appeared to be resistant, while the parental antibodies showed no effects. The ability of 1R-(E1)-(E1) to inhibit anchorage-independent growth was demonstrated at 200 nM in MDA-MB-231, with a statistically significant difference (P&lt;0.041) when compared with samples treated with parental antibodies at the same concentrations. Cells treated with 1R-(E1)-(E1) produced few and much smaller colonies, the largest size of which was less than 1/10 of the untreated cells. The parental hR1 alone, but not hRS7, had some effect on inhibiting the growth of MDA-MB-231 in soft agar, presumably resulting from the down-regulation of IGF-1R.
Conclusions: These promising results warrant further evaluation of 1R-(E1)-(E1) in other breast cancer cell lines, as well as the exploration of new bispecific HexAbs that comprise different antibodies capable of targeting other solid cancers.
Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-02-01.
P3-02-02: Novel Ranpirnase-Based ImmunoRNases Display Potent Cytotoxicity in Diverse Human Breast Cancer Cell Lines.
Authors: Goldenberg DM , Liu D , Wang Y , Rossi EA , Chang C-H 
Published: June 2012

Abstract
Ranpirnase (Rap) is an amphibian ribonuclease originally isolated from the oocytes of Rana pipiens. Rap shows anti-tumor activity in diverse cancers and its potency can be enhanced by chemically linking or recombinantly fusing Rap to a tumor-targeting antibody, as demonstrated for CD22- or CD74-expresing hematological malignancies, as well as a variety of Trop-2-expressing cell lines derived from breast, cervical, lung, pancreatic, and prostate cancers. The Dock-and-Lock (DNL) platform enables the design and generation of targetable therapeutics that are multivalent, multispecific, and multifunctional. Here, we report the successful application of the DNL method to generate a novel class of Rap-based immunoRNases, each of which features a pair of dimeric Rap molecules covalently tethered to a select monoclonal antibody at the carboxyl termini of the heavy chains. Two such constructs, designated E1-Rap and 22-Rap, were developed with hRS7 (humanized anti-Trop-2) and epratuzumab (humanized anti-CD22), respectively, purified to near homogeneity, and evaluated in a panel of human breast cancer lines, including the basal-like, triple-negative subtype (MDA-MB-468, MDA-MB-231, BT20, HCC1806, and HCC1395), the luminal B, HER2−negative subtype (MCF-7), and the HER2−positive subtype (SKBR3), all except HCC1395 expressing high to moderate levels of Trop-2, and none expressing CD22. As demonstrated by flow cytometry, E1-Rap and hRS7 bound equivalently to MDA-MB-468, indicating the affinity of E1-Rap for Trop-2 is not compromised. Surprisingly, 22-Rap, but not epratuzumab, also bound substantially to MDA-MB-468, albeit to a lesser extent than E1-Rap. We thus postulate that the four highly basic Rap molecules in the DNL conjugate may confer a spatial configuration to largely enhance their interaction with negatively-charged cell surface proteins, such as heparan sulfate proteoglycans. Whereas the individual DNL component (IgG or Rap) alone or in combination showed negligible in vitro cytotoxicity in all seven breast cancer cell lines examined, E1-Rap exhibited EC50 values of 1 nM or less in MDA-MB-468 (0.03 nM), MCF-7 (0.1 nM), BT20 (0.18 nM), HCC1806 (0.19 nM), and SKBR3 (1.29 nM). In comparison, the potency of 22-Rap was at least 10-fold lower than E1-Rap in MDA-MB-468, BT20 and HCC1806, with an EC50 of ∼2 nM. Neither E1-Rap nor 22-Rap was very effective in inhibiting the proliferation of the more aggressive MDA-MB-231, with an EC50 above 50 nM. In the Trop-2-negative HCC1395, the dose-response curves obtained for E1-Rap and 22-Rap were nearly identical (EC50 ∼100 nM), as they should be if the cytotoxicity was mediated mainly through the Rap component. The results of Immunofluorescence microscopy showed E1-Rap was effectively internalized and localized in the cytosol. Thus, these ImmunoRNases are potentially new cancer therapeutics for breast and other solid tumors.
Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-02-02.
P3-03-01: Low Expression of microRNA-210 Is an Independent Good Prognostic Factor in Japanese Triple-Negative Breast Cancer Patients.
Authors: Toyama T , Kondo N , Endo Y , Sugiura H , Yoshimoto N , Iwasa M , Takahashi S , Iwase H , Fujii Y , Yamashita H 
Published: June 2012

Abstract
Background: microRNAs (miRNAs) have emerged as a new class of non-coding genes involved in regulating cell proliferation, differentiation, and viability. Recent studies have identified miR-210 among a set of hypoxia-regulated miRNAs and demonstrated a direct regulatory role of hypoxia-inducible factor-1 alpha (HIF-1 a) in its transcription. High expression of miR-210 has been reported to be a poor prognostic factor in several types of cancers including breast.
Materials and Methods: TaqMan MicroRNA assays for miR-210 expression were performed in 219 breast cancers (58 triple-negative (TNBCs), and 161 ER-positive and HER2−negative). Correlations between miR-210 expression and clinicopathological factors were analyzed. The effects of several variables on survival were tested by Cox proportional hazards regression analysis.
Results: miR-210 expression in TNBCs was significantly higher than in ER-positive and HER2−negative breast cancers (p&lt;0.001). Patients whose TNBCs had low miR-210 expression experienced significantly better disease-free and overall survival compared with high miR-210 expressors (p=0.02 and p=0.05, respectively). Notably, among 40 node-negative TNBCs, 5-year disease-free survival was approximately 60% in patients whose tumors had high or intermediate miR-210 expression (n=26), while no patients with low miR-210 expression (n=14) suffered recurrent disease. Cox univariate and multivariate analyses demonstrated that low expression of miR-210 was an independent good prognostic factor in TNBCs.

Discussion: Although prognosis of patients with TNBCs is poor, those whose tumors expressed low levels of miR-210 had a more favorable prognosis. Thus, the degree of miR-210 expression might be a clinically useful prognostic factor for decision-making regarding treatment in the adjuvant setting, especially in node-negative TNBC patients.
Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-03-01.
P3-03-03: Congruence between Patterns of microRNA Expression and Histologic Grading of Invasive Breast Carcinomas.
Authors: Ellsworth DL , Croft DT , Field LA , Deyarmin B , Kane J , Ellsworth RE , Hooke JA , Shriver CD 
Published: June 2012

Abstract
Background
Histologic grading may be used as an indicator of prognosis in breast cancer; patients with low-grade carcinomas have ∼85% ten-year survival compared to just 45% survival in patients with high-grade disease. Although useful for risk stratification, assigning nuclear grade is subjective, and a large proportion of carcinomas are classified as intermediate-grade with uncertain prognosis, thus limiting clinical utility. MicroRNAs (miRNAs) regulate gene expression and serve an important role in breast cancer development. In this study we examined miRNA expression profiles in low-grade and high-grade breast carcinomas to determine if miRNA expression is associated with pathological classifications of tumor grade.
Methods: Breast tumors were obtained from 69 patients enrolled in the Clinical Breast Care Project. Samples were partitioned into low-grade (n=30) or high-grade (n=39) categories using the Nottingham Histologic Score. Following laser microdissection of frozen tissue sections, miRNA was isolated from pure populations of breast tumor cells and hybridized to Affymetrix GeneChip® miRNA arrays containing over 800 human miRNA probes. Expression profiles were analyzed with Partek Genomics Suite using the miRNA Expression Module.
Results: We identified 30 unique miRNAs that showed differential expression at a False Discovery Rate (FDR) p&lt;0.05 between low-grade and high-grade breast carcinomas. Gene targets for these miRNAs function primarily in metabolism and cell communication. Expression of hsa-miR-18a and hsa-miR-572 was significantly different between histologic grades at an FDR p&lt;1×10−8 and hierarchical clustering based on these miRNAs correctly classified 97% (29/30) of low-grade and 90% (35/39) of high-grade tumors. miR-18a has been shown to inhibit ER signaling and promote cellular differentiation, while the role of miR-572 in breast carcinogenesis is not well known.
Conclusions: Dysregulation of miRNAs may accompany changes in cellular morphology typically used in histologic classification of breast carcinomas. Patterns of miRNA expression may improve reproducibility and clinical utility of tumor grading and may prove useful for prediction of recurrence and survival for patients with intermediate-grade carcinomas.
Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-03-03.
P3-03-07: A Novel Approach Integrating microRNA and mRNA Signatures of HMAPK Signaling Is Highly Predictive of ER- Status and Outcome in Breast Cancer – Role of HMAPK microRNAs in Repression of ER and p27.
Authors: Miller P , Clarke J , Koru-Sengul T , Brinkman J , El-Ashry D 
Published: June 2012

Abstract
Deregulation of the MAPK signaling pathway in breast cancer is known to facilitate the down-regulation of the estrogen receptor and to contribute to the aggressive nature of ER negative and triple negative breast cancers. We have identified a microRNA signature indicative of hyperactive MAPK (hMAPK) signaling, which complements a previously established hyperactive MAPK gene expression signature. We have shown that hMAPK signaling also alters the regulatory activity of many microRNAs, including particular microRNAs with established roles in the biology of breast cancer, miR-221/222 and miR-22. Expression correlation with both the hMAPK microRNA signature and hMAPK mRNA signature is significantly associated with ER-negative status, increased tumor grade, high proliferation rate, and, importantly, poor disease specific survival among breast cancer patients, regardless of ER status. The hMAPK microRNA signature contains 127 microRNAs, 47 up-regulated and 70 down-regulated. Of note, hMAPK up-regulated microRNAs include miR-221/222 and 22, both of which have been demonstrated to target ER while miR-221/222 targets the cell cycle regulatory protein p27. Down-regulated microRNAs include miR-375, which positively regulates ER expression by down-regulating expression of an ER repressor. miR-221/222 and miR-22 exhibit both enhanced expression and enhanced regulatory activity in the context of hMAPK signaling, indicating an important role for these microRNAs in the biology of hMAPK signaling in breast cancer. In addition to these microRNAs, an unbiased approach of determining MAPK regulated microRNAs targeting the 3’ UTRs of both ER and p27 will identify novel microRNAs involved in the MAPK regulated repression of ER and p27. These data not only suggest a regulatory role for microRNAs whose expression and biological activity are altered under conditions of hyperactivation of MAPK signaling in establishing and maintaining ER negativity and tumor aggression, but also indicate that hMAPK signaling may represent a novel aggressive tumor biology that is indicative of poor disease outcome in breast cancer.
Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-03-07.
P3-04-04: Patterns of Distant Metastasis According to the Molecular Subtypes of Breast Cancer; Results of 529 Breast Cancer Patients.
Authors: Hwang MJ , Seo YJ , Lee JH , Son GT , Choi JE , Bae YK , Kang SH , Lee SJ 
Published: June 2012

Abstract
Backgrounds; Distant metastasis from breast cancer arises from various sites. But few studies concerning factors that can predict metastasis patterns in breast cancer has been reported. In this study, we analyzed the effect of breast cancer molecular subtypes on distant metastasis patterns and tried to determine factors that predict metastasis sites.
Patients and methods; From January 1995 to January 2004 at Yeungnam university hospital, patients diagnosed with the primary invasive breast cancer and received treatments were included in this study. Patients with bilateral breast cancer or distant metastasis at diagnosis were excluded. After analyzing estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor 2 (HER2), epidermal growth factor (EGFR) and cytokeratin (CK) 5/6 status, we classified patients into 5 categories, luminal A, luminal B, HER2−enriched, basal-like, and normal (triple negative nonbasal) breast cancers. Distant metastatic patterns of each category were analyzed.
Results; 529 patients were eligible for tissue microassay analysis and median follow-up period was 7.7 years. In this period, total 82 patients (15.5%) had locoregional relapse or distant metastasis and distant metastasis were identified in 54 patients (10.2%). Each distant metastatic rate was 10.5% (33/313) in luminal A, 8.7% (4/46) in luminal B, 10.2% (6/59) in HER2−enriched, 7.9% (7/89) in basal-like, and 18.2% (4/22) in normal subtype. Most frequent site of distant metastasis in all patients was bone and such result was consistent with Luminal A subtype. Liver metastasis was most frequent in Luminal B subtype, lung metastasis in HER2 enriched and normal subtype and brain metastasis in basal-like subtype.

Conclusions; There was a definite association between breast cancer molecular subtype and distant metastatic pattern. If more patients and prolonged follow up periods are analyzed, we would be able to determine the best follow up intervals, methods and treatment directions concerning subtypes of breast cancer.
Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-04-04.
P3-04-06: Comparison of MammaPrint, BluePrint, and TargetPrint with Clinical Parameters in Patients with Breast Cancer: Findings from a Prospective United States Cohort.
Authors: Nguyen B , Sinha R , Kerlin D , Barone J , Garcia A , Yao K , Rivera E , Stork-Sloots L , Deck K 
Published: June 2012

Abstract
Background: MammaPrint (MP) is a powerful predictor of disease outcome in early stage breast cancer. In addition, TargetPrint (TP), a quality approved microarray-based test that measures the mRNA expression level of ER, PR and HER2 and an 80 gene expression Molecular Subtyping profile BluePrint (BP) were developed. In the present study, MP, BP and TP were measured in a prospective U.S. breast cancer patient cohort.
Methods: MP results were evaluated in fresh tumor samples from 141 breast cancer patients (T1-4N0-2; median age 62 [35-97 yr]) collected by core needle biopsy or from a surgical specimen between July 2008 and February 2011. We compared treatment advice as recommended by NCCN guidelines and classification according to MP. In addition, we compared IHC/FISHER, PR and HER2 assessments with TP. The MP and BP results were used to subtype the patients into molecular subgroups.
Results: For the group of patients (n=69) for which NCCN recommends the use of a multi-gene signature for determining chemotherapy treatment recommendations, 50 patients were classified as High Risk and 19 as Low Risk by MP. Comparison of TP with IHC/FISH indicated a concordance of 98% for ER, 91% for PR, and 94% for HER2. For a subgroup of 63 patients combined MP and BP results were available; 22 patients were Luminal-type/MP Low Risk, 31 patients were Luminal-type/MP High risk, 1 patient was Her2-type/MP Low Risk, 2 patients were Her2-type/MP High Risk and 7 patients were Basal-type/MP High Risk.
Conclusion:The multi-gene signature MammaPrint, as well as BluePrint and TargetPrint provides additional information for treatment guidance. By combining MammaPrint with the BluePrint molecular subtyping profile, specific groups of patients can be recognized that are at high risk of recurrence and that would possibly benefit from specific treatment. This study shows that TargetPrint provides high quality second opinion for local IHC/FISH assessment.
Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-04-06.
P3-04-07: Physiological Concentrations of Genistein and 17b-Estradiol Inhibit MDA-MB-231 Breast Cancer Cell Proliferation by Increasing Bax/Bcl2 Ratio and Decreasing pERK1/2 Expression.
Authors: Rajah TT , Peine KJ , Du N , Serret CA 
Published: June 2012

Abstract
Background: Our previous results were the first to show that physiological concentrations (1 μM) of genistein, a soy component, in the presence of17β-estradiol (1 nM) inhibited the cell proliferation of MDA-MB-231 (Estrogen receptor (ER) β positive) breast cancer cells. These results are relevant in premenopausal women with breast cancer of the ERα-negative and ERβ-positive type, especially given the increasing trends of soy intake among the US population in the past decade. The aim of the present study was to identify the mechanism by which genistein plus 17β-estradiol inhibits the cell proliferation of ERα-negative and ERb-positive breast cancer cells. Our hypothesis is that the balance of signaling actions by genistein plus 17β-estradiol from different signaling pathways is likely to lead to the cell's choice to either proliferate or enter the apoptotic pathway. For this purpose, the effect of low and high concentrations of genistein (1 μM and 100 μM) in the presence or absence of 17β-estradiol (1 nM) was studied on the expression of cell signaling proteins involved in cell proliferation, survival and apoptosis (pERK1/2, pAkt, Bax and Bcl2) and correlated to cell proliferation and apoptosis in MDA-MB-231 (ERβ positive and ERα negative) breast cancer cells.
Methods: Cell proliferation was determined by the MTT assay, apoptosis determined microscopically by the use of acridine orange and ethidium bromide dyes and the expression of cell signaling proteins by western blotting.
Results: Our results show that 1μM genistein plus 17β-estradiol significantly increased apoptosis (p&lt;0.05) as compared to the control (12.47% vs 5.87% respectively). Increased expression of Bax/Bcl2 (2.5 fold) along with a decreased expression of phosphorylated ERK1/2 (2 fold) was observed in cells treated with 1μM genistein plus 17β-estradiol as compared to the control cells. Phosphorylated Akt did not show any differences in the treatment condition as compared to the control. High concentrations of genistein (100 μM) in the presence or absence of 17β-estradiol also increased apoptosis; however these changes could not be correlated to the expression of Bax/Bcl2, or pERK1/2.
Conclusion: In conclusion, our results show that physiological concentrations of genistein in the presence of17β-estradiol inhibit cell growth through apoptosis via increased Bax/Bcl2 and a concomitant decrease in pERK1/2 expression. Our results also suggest that different concentrations of genistein elicit cell responses through different signaling mechanisms. These results are especially relevant to the cohort of premenopausal women with breast cancer of the ERβ positive and ERα negative type.
Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-04-07.