bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P5-10-10: The miR-34a is down-regulated in breast cancer and breast stem cells and a potential to eradicating breast cancer via a systemic delivery of a VISA –miR-34a nanoparticle system
Authors: Xie X , Li L , Xie X , Wei W , Kong Y , Wu M , Yang L , Gao J , Xiao X , Tang J , Xie Z , Wang X , Liu P , Li X , Guo J
Published: August 2014

Breast cancer is the most common disease in women around the world and the current treatment strategies are not potent enough for the patients, especially those who are the triple-negative type of molecular classifications. Therefore, novel and more effective treatments are pressingly needed. Of the current methods, target therapy, which not only retains cancer-specific expression but also limits toxicity, is a new strategy for treatment of cancers. In this study, it was to investigate miR-34a expression in breast caner specimen and its relationship with patient's clinical status, and develop targeted miR-34a delivery system as a potential method for breast cancer therapy. miR-34a expression was investigated by qRT-PCR and related to clinicopathologic significance and found to be down-regulated in breast cancer as compared with normal adjacent tissues and involved in breast cancer stem cell through down-regulation of CD44, ZEB1, and Bmi1. We designed targeted miR-34a expression using T-VISA system (hTERT promoter driven VP16-Gal4-WPRE Integrated Systemic Amplifier) liposomed-based nanoparticles and evaluated the antitumor effect in breast cancer cells in vitro and in orthotopic animal model as well as its systemic toxicity. The T-VISA-miR-34a system robust targeted to breast cancer cells and breast cancer stem cells and prolonged duration expression of miR-34a. Furthermore, a systemic delivery of a VISA –miR-34a nanoparticle system targeted efficiently expression of miR-34a to tumors and could significantly inhibit tumor growth and prolong mouse survival in multiple living imaging xenograft and syngeneic models of orthotopic breast cancer and without toxicity in intact mice. Our study demonstrated that miR-34a expression was down-regulated in breast cancer and breast cancer stem cells through down-regulation of CD44, ZEB1, and Bmi1. The T-VISA-miR-34a nanoparticle system showed robust antitumor effects in breast cancer and could be a potential therapeutic approach to eradicating breast cancer.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P5-10-10.
Abstract P5-10-15: Genetic variants located in beta2 adrenergic receptor gene (ADRB2) and miRNA let-7 binding site alter breast cancer susceptibility: a case control analysis
Authors: Du Y , Lu J
Published: August 2014

Previous studies have shown that a potential association may exist between risk of breast cancer and polymorphisms in the beta-adrenergic receptor gene. ADRB2 is a target for miRNA let-7 and it is reported that low let-7 expression probably increases risk of breast cancer. In this study, we investigated the relationship between genetic variants located in alpha2 and beta2 adrenergic receptor genes and miRNA let-7 binding sites with breast cancer risk in benign mammary disease and sporadic breast cancer. We assessed frequency distributions of these variants in 741 patients with histologically confirmed breast cancer and 315 controls with benign mammary disease. We initially demonstrated that rs1042713, located in ADRB2 and rs11292, located in miRNA let-7 binding site were associated with susceptibility to breast cancer. A significant association was observed between GG, AG+GG genotype compared to AA genotype of SNP rs1042713 and a decreased breast cancer risk (adjusted OR = 0.79, 95%CI: 0.64–0.97, p = 0.025; adjusted OR = 0.72, 95%CI: 0.54–0.96, p = 0.028). We also found a significant association between TC, TC+CC genotype compared to TT genotype of SNP rs11292 and an increased breast cancer risk (adjusted OR = 1.44, 95%CI: 1.01–2.06, p = 0.044; adjusted OR = 1.43, 95%CI: 1.01–2.02, p = 0.044). The allele of G of SNP rs1042713 was associated with a decreased risk of breast cancer (OR = 0.79, 95%CI: 0.66–0.96, p = 0.018). Taken together, our study demonstrated that genetic variants located in ADRB2 and miRNA let-7 binding site could alter breast cancer risk.

Keywords: ADRB2 gene, miRNA let-7, polymorphism, breast cancer

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P5-10-15.
Abstract P5-10-16: miRNAs as novel therapeutic adjuvants for the prognosis and treatment of triple negative breast cancers
Authors: Zoghi B , Chang Y-F , Subbarayalu P , Plyler JR , Rao M
Published: August 2014

Approximately 15–20% of all breast cancers account for triple negative breast cancers (TNBCs) that exhibit an aggressive, distinct metastatic pattern and poor prognosis resulting in a disproportionate number of breast cancer deaths. More than 50% of patients with TNBC develop chemoresistance and do not respond to chemotherapeutic drugs, leading to early relapse and shorter survival. Understanding the mechanisms underlying such resistance is therefore crucial for the development of new, efficacious cancer drugs. In spite of extensive inquiry in this field, little is known about the key molecules/signaling pathways that regulate this phenomenon. Recently, we have discovered that small non-coding RNAs-miRNAs play critical roles in mediating drug sensitivity/resistance in TNBCs. We have identified miRNAs that are differentially expressed between chemo-resistant and sensitive breast cancer cells. In particular, through high-throughput miRNA inhibitor library screens, we have identified miRNA inhibitors that sensitize resistant TNBC cells to paclitaxel, a drug commonly used to treat triple negative breast cancers. Interestingly, our studies revealed that specific miRNAs including miR-185 (patent pending) that sensitize resistant TNBCs to paclitaxel are expressed at significantly lower levels in relapsed metastatic TNBC patient sera compared to sera from their healthy siblings. Furthermore, using liposome- or biocompatible PLGA nanoparticle-based approaches, we show that systemic delivery of one of the sensitizer miRNAs suppresses breast cancer lung metastasis without any hepatotoxicity in preclinical mouse tumor models. Importantly, we show that one of the mechanisms by which sensitizer miRNAs regulate paclitaxel sensitivity is by targeting Stathmin, a microtubule destabilizer protein that can bind to tubulin dimers and stimulate microtubule destabilization. These findings suggest that tumor-specific miRNAs render selective cell cytotoxicity in a drug-specific manner, that these miRNAs may serve as detection markers for identifying patients who might benefit most from specific drug treatment, and that these miRNAs may represent novel therapeutic tools for the treatment of the TNBCs. Since miRNAs are endogenously expressed and can be easily manipulated using synthetic oligoribonucleotides, we believe that they represent more attractive targets than the single gene or gene product targeted by conventional cancer treatments that are typically prone to drug resistance. Since paclitaxel is used for the treatment of many cancers, the long-term implications of this study are likely not limited to breast cancer alone but may apply to other tumor types.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P5-10-16.
Abstract P5-12-03: Developing the ePromotora: Increasing Promotora's Access to Breast Cancer Electronic Resources
Authors: Lopez AM , Ryan J , Valencia A , El-Khayat Y , Nunez A
Published: August 2014

Background: Disparities in breast cancer screening and survival have been attributed to multiple factors including limited access to breast health education. Innovative information technologies facilitate health education; however, access to information technology is not uniform. The resultant systematic gap in health knowledge has led to a new health disparity where higher socioeconomic individuals acquire health information electronically more easily than the disadvantaged. Promotoras or Community Health Workers (CHWs) facilitate access to culturally appropriate cancer health information, screening and care. Hypothesis: An electronic asynchronous breast cancer educational intervention developed by the Arizona Telemedicine Program and the Arizona Health Sciences Library will improve CHW computer literacy and will enhance health education communication.

Specific Aims: to develop, implement and assess a 6-session e-promotora training curriculum with a focus on breast health.

Methods: The first step in developing a training curriculum was a needs assessment and a computer literacy assessment. This assessment served as the foundation for the computer literacy curriculum which focused on breast health content. The educational intervention was delivered as a combination of on-line, webinar, and face-to-face seminars. To meet the needs of the participants all sessions were delivered bilingually, Spanish and English. Program evaluation included pre and post knowledge assessments, session assignments, program satisfaction and 6-month follow up focus groups or structured interviews to evaluate the utility, implementation and dissemination of knowledge gained.

Results: 26 CHWs have completed the training. All participants identified as Latinas, average age – 51 years of age and educational attainment – 58% (15/26) reported high school or less. Pre and post assessment data reveal high program satisfaction scores, increased participant knowledge and confidence in knowledge, consistent and ongoing use of information learned and computer skills gained and effective dissemination of health information gained.

Conclusions: The e-promotora training curriculum effectively improves CHW computer literacy, facilitates dissemination of electronic health information and helps address the disparities in electronic health information.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P5-12-03.
Abstract P5-14-05: Compliance with radiation therapy in breast cancer patients in Southeastern Kentucky
Authors: Elsoueidi R , Adane E , Dignan M
Published: August 2014

Background: Nearly a quarter of Americans live in rural areas, which consistently report higher cancer mortality rates than urban and suburban areas. The worse outcome for cancer patients in these areas has been attributed in part to compliance with treatment recommendations. Southeastern Kentucky is a rural area with a low income population and barriers such as distance and lack of public transportation to access health care and compliance with treatment is a major concern.

Methods: Data on patients with breast cancer treated with radiation therapy at Appalachian Regional Healthcare cancer center which serves the area of Southeastern Kentucky were collected. Age, economic status (measured by availability of medical insurance), distance of patient's residence from the hospital, were evaluated as potential predictors of compliance with recommendations to obtain radiation therapy. Data were analyzed with Fisher's exact test.

Results: A total of 80 patients with breast cancer received adjuvant radiation treatment from 2008 to 2011. Of these 80 patients, 24 (30%) had Medicaid, 31 (39%) had Medicare, and 25 (31%) had commercial health insurance. Only two (2.5%) patients did not complete treatment due to transportation. An additional 10 patients (13%) had treatment interruption due to weather, transportation or other financial reasons. In patients who completed treatment, a total of 2640 days of treatment were provided. A total of 55 (2.1%) days of treatment were missed. Of the 10 patients who had interruption, 5 of them had Medicaid and the number of missed days by the Medicaid patients was (37/55), which is 67% of the total days missed. No statistically significant difference was detected in the number of patients missing treatment between Medicaid and other insurance types (p = 0.16). None of the factors evaluated predicted patient compliance, although a trend toward lower compliance was noted in patients who had Medicaid.

Conclusion: Despite being a rural area with multiple barriers to access healthcare, the rate of full compliance (defined as completion of the entire course of radiation therapy) with adjuvant radiation therapy in breast cancer patients in Southeastern Kentucky was 97.5%, which is similar to the rates in clinical trials that generally exceed 90%. A trend toward lower compliance was noted in patients who had Medicaid although it was not statistically significant.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P5-14-05.
Abstract P5-15-04: CTX and CTX-related direct medication costs saved by testing biomarkers uPA and PAI-1 in primary breast cancer: Results of a prospective multi-center study at Certified Breast Centers in Germany
Authors: Jacobs VR , Augustin D , Wischnik A , Kiechle M , Hoess C , Steinkohl O , Rack B , Kapitza T , Krase P
Published: August 2014

Background: Biomarkers uPA and PAI-1 are guideline recommended by ASCO, USA, and AGO, Germany, to be used in primary breast cancer to avoid unnecessary CTX in medium risk recurrence patients [G2, N−, HR+, HER2neu−, >35 years]. This study was performed to verify in normal clinical settings how many CTX cycles and how much CTX-related direct medication costs can be avoided by uPA/PAI-1 testing.

Methods: Prospective, non-interventional, multi-center study over two years among six Certified Breast Centers in Germany to analyze application of uPA/PAI-1 and consecutive decision making in the clinical setting for AOK Bayern-insured patients. CTX regimen and cycles avoided were identified for each case and direct costs for CTX and CTX-related medication costs for concomitant medication calculated for each patient individually according to body weight and body surface as well as potential FN prophylaxis according to physicians' decision. All medication costs were taken from the pharmaceutical price list for Germany Rote Liste of 2012. EURO [€] to US Dollar conversion rate as of June 12 2012: € 1.00 = US$ 1.25.

Results: In n=93 breast cancers n=35 CTX (37.6%; FEC n=25, FEC+DOC n=8 and TC n=2) were avoided according to uPA/PAI-1 test result. Consecutively 210 CTX cycles or 12.1 years of CTX application were saved improving patients' quality of life. uPA/PAI-1 testing saved direct medication costs for avoided CTX of US$ 221.816, CTX-related concomitant medication of US$ 34.353 and G-CSF prophylaxis of US$ 25.749, overall US$ 281.918. At process costs for a single uPA/PAI-1 test calculated at US$ 359, uPA/PAI-1 testing resulted in additional costs of US$ 33.387 for all breast cancer cases. Overall, testing of uPA/PAI-1 has been proven to be cost-effective regarding direct medication costs alone at a return-on-investment ratio of 8.4:1. Indirect cost savings further increase this ROI.

Conclusions: Innovative and individual cancer diagnostics like biomarkers uPA/PAI-1 can decrease need of CTX and increase patients' quality of life and thereby reduce costs for health care systems. Since application of this test is inadequate at present time measures are suggested to fully implement such cost-effective diagnostics.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P5-15-04.
Abstract P5-15-06: Societal economics of the 21-gene Recurrence Score® in estrogen-receptor-positive early-stage breast cancer in Japan
Authors: Yamauchi H , Nakagawa C , Yamashige S , Takei H , Yagata H , Yoshida A , Hayashi N , Hornberger J , Yu T , Chien R , Chao C , Yoshizawa C , Nakamura S
Published: August 2014

Background: Breast cancer incidence and breast-cancer mortality have been increasing in Japan. In estrogen-receptor-positive breast cancer, adding chemotherapy as adjuvant treatment is not definitive benefit for all women. Primary data on the clinical validity and decision impact of the 21-gene Recurrence Score for guiding the use of adjuvant chemotherapy has been studied among a Japanese population.

Objective: The cost-effectiveness of the clinically validated 21-gene Recurrence Score for estrogen-receptor-positive, lymph-node-negative, early-stage breast cancer (ESBC) was assessed from a Japanese societal perspective.

Method: The proportion of patients with low, intermediate, and high risk of recurrence by the 21-gene Recurrence Score and the influence of the Recurrence Score on use of adjuvant chemotherapy were obtained in a study of 104 patients. A Markov model was used to track time until distant recurrence and time from distant recurrence to death. Effect of adjuvant chemotherapy based on 21-gene Recurrence Score was based on published clinical validation studies. Direct and indirect medical costs were obtained from the referral center. Utilities associated with progression and adverse events associated with chemotherapy were extracted from the literature. Sensitivity analyses were performed to assess the key drivers of cost-effectiveness and the robustness of the findings to variations in the input estimates.

Results: Forty-eight percent of patients were identified by the 21-gene Recurrence Score as low-risk, 36% as intermediate-risk, and 16% as high-risk. In treatment decision by using the 21-gene Recurrence Score, chemotherapy use overall decreased by 19%, resulting in an average increase of 0.235 QALYs. Testing with the 21-gene Recurrence Score cost ¥350,000 per patient with ¥153,490 lower acute costs because fewer women were treated with adjuvant chemotherapy. Eighteen percent more women who were identified as high risk were recommended adjuvant chemotherapy, which results in longer progression-free survival. On average across all risk groups, cost of monitoring until recurrence per patient increased by ¥3,572 and costs associated with recurrence declined by ¥43,687. Use of the 21-gene Recurrence Score resulted in a net cost of ¥665,455 ($8,386) per QALY gained.

Conclusions: The change in decisions resulting from use of the 21-gene Recurrence Score in women with estrogen-receptor-positive, lymph-node-negative, ESBC is projected to be societally cost-effective in Japan.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P5-15-06.
Abstract P5-16-01: Survival advantage in patients with metastatic breast cancer receiving endocrine therapy plus Sialyl Tn-KLH vaccine: post hoc analysis of a large randomized trial
Authors: Ibrahim NK , Murray JL , Zhou D , Mittendorf EA , Sample D , Tautchin M , Miles D
Published: August 2014

Background: A multicenter, double blinded, randomized phase III trial of the therapeutic cancer vaccine STn-KLH was completed in an international cohort of 1,028 women with metastatic breast cancer who had either no evidence of disease or nonprogressive disease following first-line chemotherapy. This trial is registered with ClinicalTrials.gov (No. NCT00003638). The outcomes showed that STn-KLH was safe and relatively well tolerated but had neither a positive nor negative effect on time to progression (TTP) or overall survival (OS) duration in the intent-to-treat population when compared with KLH control alone. The purpose of this post hoc analysis is to explore the potential benefit of combining an antiestrogen with MUC1 vaccines in metastatic breast cancer patients.

Methods: The data were further explored to determine if a retrospective, reassigned endocrine subset patient stratification produces subgroups that may have experienced benefit in TTP or survival compared with the phase III trial ITT analysis.

Results: Women treated with concomitant endocrine therapy, a pre stratified subset comprising approximately one third of the original study population, achieved a clinical benefit both in terms of TTP and survival compared with women who did not receive endocrine therapy. Moreover, women in the endocrine-treatment subset who mounted a median or greater antibody response (titer >1:320 toward bovine submaxillary mucin) to the STn-KLH vaccine experienced significantly longer median survival than their trial counterparts who mounted a below-median antibody response.

Conclusion: Unlike maintenance chemotherapy, with its associated cumulative toxicity, the combination of endocrine and STn-KLH therapy may offer clinical benefit with few adverse effects for women with metastatic breast cancer.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P5-16-01.
Abstract P5-17-02: Molecular response in breast cancer tumors treated with neoadjuvant chemotherapy with and without bevacizumab: Results from NeoAva - a randomized phase II study.
Authors: Engebraaten Olav , Silwal-Pandit Laxmi , Fleischer Thomas , Borgen Elin , Garred Øystein , Fangberget Anne , Holmen Marit M , Schlichting Ellen , Skjerven Helle , Lundgren Steinar , Gribbestad Ingrid S , Krohn Marit , Edvardsen Hege , Kristensen Vessela N , Mills Gordon , Wist Erik , Børresen-Dale Anne-Lise
Published: August 2014

Withdrawn by Author

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P5-17-02.
Abstract P5-18-18: Lapatinib versus trastuzumab, or both, added to preoperative chemotherapy for breast cancer: A meta-analysis of randomized evidence
Authors: Valachis A , Nearchou A , Lind P
Published: August 2014

Purpose: We compared the efficacy and safety of the addition of lapatinib versus trastuzumab, or the combination, to neoadjuvant chemotherapy in HER2-positive breast cancer.

Methods: Potentially eligible trials were identified through PubMed and Cochrane Library searches and abstracts of major international conferences. The endpoints that we assessed were pathologic complete response (pCR) rate and toxicity. Pooled relative risks (RR) were estimated for each endpoint with fixed or random effects models, depending on between-studies heterogeneity.

Results: Six trials were identified with 1494 eligible patients. The probability to achieve pCR was higher for the trastuzumab plus chemotherapy arm than lapatinib plus chemotherapy (RR 1.25, 95% Confidence Interval (CI): 1.08–1.43; p-value = 0.003) (6 trials; 1494 patients). Furthermore, the probability for pCR was significantly higher in the group given both lapatinib and trastuzumab than in the group given trastuzumab alone (RR 1.39, 95% CI: 1.20–1.63; p-value < 0.001) (4 trials; 779 patients). Diarrhea and dermatologic toxicities were statistically more frequent in patients receiving lapatinib than trastuzumab. The combination was associated with a higher rate of diarrhea than trastuzumab alone. No differences were observed regarding cardiac adverse events among patients receiving trastuzumab, lapatinib, or the combination.

Conclusion: These data support the superiority of a dual-HER2 inhibition for the treatment of HER2-positive breast cancer in the neoadjuvant setting. The direct comparison of trastuzumab and lapatinib demonstrated that lapatinib is inferior in terms of pCR and toxicity.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P5-18-18.
Abstract P5-18-21: A Phase II randomized trial of lapatinib with either vinorelbine or capecitabine as first- and second-line therapy for ErbB2-overexpressing metastatic breast cancer (MBC)
Authors: Janni W , Sarosiek T , Pikiel J , Karaszewska B , Staroslawska E , Salat C , Caglevic C , Potemski P , Brain E , Briggs K , de Silvio M , Sapunar F , Papadimitriou C
Published: August 2014

Background: Lapatinib (L), a dual kinase inhibitor of epidermal growth factor receptor and ErbB2, is effective in the treatment of ErbB2+ MBC in combination with capecitabine (C) following progression after trastuzumab, anthracyclines, and taxanes. Vinorelbine (V) is an important chemotherapy option in MBC, and multiple Phase II trials have been conducted in combination with trastuzumab. This randomized, open-label, multicenter, Phase II study (LAP112620, VITAL) evaluated the efficacy and safety of L with either V or C in women with ErbB2+ MBC.

Methods: Patients with MBC who had received ≤1 chemotherapy regimen in the metastatic setting were randomized 2:1 to either L 1250 mg orally once daily (QD) continuously plus V 20 mg/m2 intravenously on Days 1 and 8, every third week, or L 1250 mg orally QD continuously plus C 2000 mg/m2/day orally in 2 doses 12 hours apart on Days 1–14 every third week. Patients were stratified by prior receipt of therapy for MBC (Y/N) and site of metastatic disease (visceral/soft tissue or bone-only). The primary endpoint of progression-free survival (PFS) was assessed once all subjects had been followed for a minimum of 6 months or had otherwise progressed, died or withdrawn, if sooner. The primary focus was to evaluate PFS in the L plus V arm with a descriptive intent only. Other endpoints included overall response rate, overall survival, and safety. Patients progressing on one treatment arm were given the option of crossing over to the other arm.

Results: 112 patients were randomized. The results and conclusions sections will be updated once the primary analysis has been completed in September 2012.

ClinicalTrials.gov - NCT01013740

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P5-18-21.
Abstract P5-18-23: The prognosis of T1a, T1b N0 M0, HER2+ patients in Korea.
Authors: Lee JW , Moon H-G , Han W , Noh D-Y
Published: August 2014

Backgrounds: Increased expression of HER-2 or amplification of the HER-2/neu gene has been associated with a worse prognosis than with other breast cancer phenotypes. Especially in cases of size of 0.5∼1cm and node-negative status, adjuvant chemotherapy with or without Trastuzumab treatment is suggested by recent National Comprehensive Cancer Network(NCCN) guidelines. In Korea, the breast cancer patient pool is more younger than western data, and there could be some ethnic differences.

Methods: The Korean Breast Cancer Society (KBCS) has been collaborating with the Korean Central Cancer Registry (KCCR) to improve the completeness and validity of the breast cancer registry and the mortality data. Data of T1a, bN0M0 breast cancer patients from 1999 to 2006 from KBCS on-line registry was obtained, and a total of 2,962 patients were included for survival analysis.

Results: 711 patients (24.0%) was HER-2 positive. Hormone receptor and HER2 status was used to classify 4 molecular subtypes; Hormone receptor(HR)+/HER2−, HR+/HER2+, HR−/HER2+, HR−/HER2−. The median follow-up was for 55.82 month (±24.196). Breast cancer specific survival rate was lowest at HR−/HER2+ subtype(97.9%). It's 1.0∼1.8% lower than other subtypes. Between all T1N0M0 cases, especially T1b HR−/HER2+ cases showed lowest survival of 96.4%.

Conclusion: This study calculated the nationwide survival rates of Korean breast cancer patients with T1a, bN0M0 disease for the first time. HER2 gene positivity was associated with worse prognosis and it's concordant with western studies. But prognosis of T1N0M0 lesion is much better than that of other nations. So, the usage of chemotherapy and Trastuzumab treatment should be considered more carefully in Korea. Further analysis of the long-term survival and a nationwide collaborative study should be performed to estimate the survival trend of Korean breast cancer patients.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P5-18-23.
Abstract P5-18-25: Bispecific Fynomer-antibody fusion proteins targeting two epitopes on HER2
Authors: Grabulovski D , Brack S , Toller I , Mourlane F , Bertschinger J
Published: August 2014

Background: Fynomers are small binding proteins (7 kDa) derived from the human SH3 domain of Fyn kinase. They can be engineered to yield specific and high-affinity binding domains to target proteins of interest. In the past, we have isolated Fynomers with excellent physico-chemical properties binding to 16 different antigens. One important application of the Fynomer technology represents the genetic fusion of a Fynomer to an antibody to provide bispecific fusion proteins with enhanced activity compared to the unmodified antibody.

Method: Using phage display technology we have isolated Fynomers binding to an epitope on HER2 which is different from the epitopes recognized by trastuzumab and pertuzumab. After genetic fusion of the HER2 binding Fynomers to pertuzumab the resulting bispecific fusion proteins were evaluated in vitro and in vivo for their antitumoral activity.

Results: The fusion of Fynomers to pertuzumab did not alter the favorable biophysical properties of pertuzumab: First, the Fynomer-antibody fusion proteins could be purified with the same high yields from the supernatant of transiently transfected CHO cells as the unmodified antibody pertuzumab (in the range of 100 mg/L). Second, the purified fusion proteins were monomeric and showed no signs of aggregation even after four months of storage at 4 °C or −20 °C in PBS as determined by size exclusion chromatography. Third, the fusion of Fynomers to antibodies did not alter their Fc-mediated effector functions. Antibody-dependent cellular cytotoxicity (ADCC) was maintained, and the affinity to the neonatal Fc-receptor (FcRn) was similar as observed for pertuzumab. In addition, the bispecific Fynomer-antibody fusion proteins demonstrated excellent growth inhibition of tumor cells in vitro compared to pertuzumab and trastuzumab. In particular, the most potent molecule COVA208 exhibited impressive efficacy in vivo in tumor xenograft mouse models.

Conclusion: These encouraging preclinical results indicate that the bispecific Fynomer-antibody fusions have highly promising properties with a great potential for further preclinical and clinical development, both alone and in combination with trastuzumab.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P5-18-25.
Abstract P5-19-01: Targeting the HER3-phosphatidyl inositol-3 kinase pathway in breast cancers
Authors: Cook RS , Morrison MM , Arteaga CL , Perou CM
Published: August 2014

Background: Using a mouse model of HER3 ablation in the mammary epithelium, we previously showed that HER3 is required for the genesis of transgenic HER2-overexpressing breast cancers. However, increasing evidence suggests that transgenic models of HER2 overexpression result in luminal B-like mammary tumors, which resemble human breast tumors that express estrogen receptor (ER) but are often not responsive to endocrine therapy. Furthermore, we found that HER3 is required in the untransformed mouse mammary epithelium for differentiation and survival of luminal cells, but not basal cells. Herein we report that HER3 drives growth and survival of luminal breast cancers in a manner that parallels its role in maintaining the untransformed luminal epithelium in the breast.

Results: Expression profiling of a large human breast cancer dataset demonstrated that HER3 expression was highest in Luminal breast cancers as compared to different molecular subtypes of breast cancers, including HER2-enriched, claudin-low, and basal-like breast cancers. In luminal breast cancer cell lines MCF7, ZR75-1, and MDA-MB-361, HER3 targeting using siRNA or a monoclonal antibody against HER3 decreased Akt, S6, and Erk1/2 phosphorylation, cell growth and cell survival under conditions of estrogen deprivation. Luminal breast tumor xenografts treated with U3-1287, a humanized monoclonal anti-ErbB3 antibody, or EZN-3920, an anti-sense oligonucleotide against ErbB3, decreased tumor growth and phosphatidyl inositol-3 kinase (PI3K)/mTOR signaling in vivo. Interestingly, luminal breast tumor xenografts responded to the ER inhibitor fulvestrant with increased HER3 expression and phosphorylation, and with increased PI3K/mTOR signaling as shown by enhanced S6 phosphorylation. Fulvestrant-mediated upregulation of HER3 was also seen in matched clinical breast tumor specimens harvested before treatment and after 21 days of fulvestrant treatment. AMG-888 blocked fulvestrant-induced HER3 and phospho-S6 upregulation in vivo, resulting in regression of luminal breast tumor xenografts.

Conclusions: These data suggest that HER3 targeting may improve the outcome of luminal breast cancers treated with endocrine therapy, and are consistent with our findings that a HER3-dependent expression signature positively correlates with relapse in tamoxifen-treated patients. Taken together, these results define a novel role for HER3 in luminal breast cancer cells, and support future investigations into therapeutic strategies to target HER3 in luminal breast cancers.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P5-19-01.
Abstract P5-19-02: Selective PI3K and dual PI3K/mTOR inhibitors enhance the efficacy of endocrine therapies in breast cancer models
Authors: Friedman L , Ross LB , Wallin J , Guan J , Prior WW , Wu E , Nannini M , Sampath D
Published: August 2014

Purpose: Phosphoinositide 3-kinases (PI3K) are lipid kinases that can regulate breast tumor cell growth, migration and survival. Standard of care drugs such as estrogen receptor (ER) antagonists including fulvestrant and tamoxifen, and aromatase inhibitors such as letrozole are indicated for the treatment of hormone receptor positive breast cancer. The current study is focused on investigating preclinical activity in breast cancer models, for GDC-0941, a class I PI3K inhibitor, GDC-0032, a PI3K inhibitor, and GDC-0980, a dual mTOR kinase and class I PI3K inhibitor. Investigation into PI3K inhibitor efficacy in combination with endocrine therapies is also explored.

Experimental Design: A panel of ER+ breast cancer cell lines were treated with GDC-0941, GDC-0032 and GDC-0980 either as single agents or in combination with fulvestrant or tamoxifen and assayed for cellular effects. MCF-7 cells ectopically expressing aromatase were utilized to test the efficacy of aromatase inhibitors in combination with PI3K inhibitors in vitro. In addition, human xenografts of breast cancer cell lines were employed to assess combination efficacy of PI3K inhibitors with fulvestrant and tamoxifen in vivo.

Results: Combination of GDC-0941, GDC-0032 or GDC-0980 with endocrine therapies resulted in a decrease in cellular viability and an increase in cell death. Synergy of PI3K inhibitor combinations with fulvestrant or tamoxifen was assessed using Combination Index (C.I.), and C.I. values as low as 0.1 indicated strong synergy in some contexts. Combination activity of PI3K inhibitors and letrozole was also observed in MCF7 cells expressing aromatase. In MCF-7 xenografts, the combination of GDC-0980, GDC-0032 and GDC-0941 enhanced activity of fulvestrant resulting in tumor regressions and tumor growth delay (116% tumor growth inhibition (TGI) for GDC-0980 and 91% TGI for GDC-0941 and GDC-0032). In addition, the combination of GDC-0941 or GDC-0032 with tamoxifen enhanced the efficacy of tamoxifen in vivo (83%TGI for GDC-0941 and 102%TGI for GDC-0032). Mechanism of action and biomarker studies are underway.

Conclusion: Collectively, the non-clinical efficacy data provide a strong rationale to evaluate the combination of PI3K inhibitors with anti-estrogen therapy in hormone receptor positive breast cancer.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P5-19-02.
Abstract P5-20-11: Bevacizumab in metastatic breast cancer: a retrospective matched-pair analysis.
Authors: Gampenrieder SP , Romeder F , Muß C , Pircher M , Ressler S , Rinnerthaler G , Bartsch R , Sattlberger C , Mlineritsch B , Greil R
Published: August 2014

Background: In November 2011 the FDA withdrew accelerated approval of the breast cancer indication for bevacizumab, because “the drug has not been shown to be safe and effective for this use” (FDA Commissioner M. A. Hamburg, MD). The objective of this analysis was to evaluate efficacy and safety of bevacizumab in non-selected patients with stage IV breast cancer.

Patients and methods: All patients with metastatic breast cancer treated with bevacizumab in combination with chemotherapy in our institution between 2005 and 2011 were retrospectively analysed. At least 1 cycle of bevacizumab was required for safety analysis, 3 applications for efficacy evaluation. A control group was matched according to the following criteria: receptor status, line of treatment, chemotherapy backbone, visceral or non-visceral disease and age.

Results: A total of 213 patients fulfilled the inclusion criteria. All of them were evaluable for toxicity, 199 for response. 503 potential controls allowed a complete matching for 103 patients. Fifty-eight percent of patients received first-line, 15% second-line and 27% of patients beyond second-line treatment. Visceral metastases were present in 69% of patients in both groups. The most common chemotherapy backbones were paclitaxel, capecitabine and docetaxel with 44%, 32% and 19%, respectively.

Most common grade 3/4 toxicities related to bevacizumab were hypertension (27%), proteinuria (1%), thromboembolism (9%) and bleeding events (1%). Interestingly, patients developing hypertension during bevacizumab treatment had a more favourable outcome (PSF 11.7 vs. 5.7 months, p < .001; OS 27.4 vs. 14.5 months, p < .001). Since the median time until apparent blood pressure elevation was only 1.4 months, this provides a simple biologic biomarker for bevacizumab efficacy.

Conclusion: Bevacizumab in combination with chemotherapy appears to be an effective, well-tolerated regimen in the treatment of advanced breast cancer even in a non-selected, partly intensively pre-treated population. Bevacizumab induced hypertension predicted a better PFS and OS.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P5-20-11.
Abstract P5-21-04: Real-world use and effectiveness of adjuvant trastuzumab in 2665 consecutive breast cancer patients
Authors: Tjan-Heijnen VCG , Seferina SC , Lobbezoo DJA , Voogd AC , Dercksen MW , van den Berkmortel F , van Kampen RJW , van de Wouw AJ , Joore MA , Borm GF
Published: August 2014

Background The impact of drug prescriptions in the real world as opposed to strict trial prescription is only rarely assessed even though it is well recognized that incorrect use may harm a patient and may have a huge impact on health care resources. We aimed to assess how and to whom trastuzumab was given since its introduction in September 2005 as adjuvant treatment, and to assess its efficacy in daily practice.

Patient and methods We set up a registry of all patients diagnosed with stage I-III invasive breast cancer in 5 Dutch hospitals during the years 2005–2007. Patients were included irrespective of HER2 status and of treatment received. We present the main patient and tumor characteristics, treatment delivered and 5-year treatment outcomes.

Results Of 2665 patients included, n=479 (18%) had a HER2 positive tumor. As of September 2005, adjuvant trastuzumab was given to 96% of patients with a HER2 positive tumor receiving adjuvant chemotherapy. The median age of these patients was 51 (27–72) years. Their median tumor size was 23 (5–65) mm, 55% had node-positive, 57% ER-positive and 47% PR-positive disease. In 83% of patients, trastuzumab was delivered in sequence after (taxane-based) chemotherapy, mainly in the first years after introduction, and in 91% in a 3-weekly regimen. Trastuzumab was delivered for a median of 49 (IQR 45–52) weeks. For patients with a HER2 positive tumor treated with adjuvant trastuzumab (n = 232), the five-year disease-free survival was 81%. For patients with a HER2 positive tumor who were not treated with trastuzumab (n = 247) the five-year disease-free survival was 75% (p = 0.13). Corresponding 5-year overall survival rates were 91% and 78%, respectively (p = 0.0002).

Conclusion In the Netherlands, trastuzumab was rapidly implemented since its introduction in September 2005. Characteristics of patients treated with trastuzumab resembled those included in the HERA study. Similarly, outcomes were in agreement with reported phase III studies. More detailed and multivariate analyses will be shown at the meeting.

Funding Netherlands Organization for Health Research and Development (ZonMw: 80–82500-98-9056) and Roche Netherlands.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P5-21-04.
Abstract P6-01-03: Decreasing the metastatic potential in Triple Negative Breast Cancer through the miR-17 cluster
Authors: Jin L , Simone B , Sano Y , Lim M , Zhao S , Savage JE , Baserga R , Camphausen K , Simone NL
Published: August 2014

Background: In animal models of breast cancer, caloric restriction (CR), or a 25–30% reduction of calories, has decreased tumor initiation and progression. We sought to determine if CR could be used as a novel therapeutic intervention to enhance cytotoxic therapies such as radiation (RT) and alter the molecular profile of triple negative breast cancer (TNBC), which displays a poor prognosis.

Methods/Results: In two murine models of TNBC (4T1 which is highly metastatic and 67NR which is locally aggressive), mice with palpable tumors were treated with radiation, caloric restriction (30% intake reduction) or both. 3 times per week measurements revealed significant tumor regression with RT or diet modification, and an even greater regression is observed combining diet modification with RT. Concurrently, many reports have shown that TNBC is most distinct from other breast tumor subtypes due to up-regulation of the miR-17-92 cluster which we have shown by RT-PCR is downregulated with both CR and radiation (RT) and even further decreased with the combination. cDNA arrays done on all conditions revealed that the top 5 statistically significant GO terms with high fold changes were all associated with extracellular matrix and metastases. In silico analysis demonstrated 5 ECM related genes that were targets of the miR-17 cluster: collagen 4 alpha 1 (COL4A1), collagen 4 alpha 3 (COL4A3), laminin alpha 3 (LAMA3), metallopeptidase inhibitor 2 and 3 (TIMP2 and TIMP3) and these were inversely correlated with the miRs.

Conclusions: CR, or altering the metabolism of breast cancer, can be combined with RT to decrease progression of TNBC tumors and that the miR-17-92 cluster is also decreased with these conditions. cDNA arrays suggest that extracellular matrix (ECM) proteins play a large role in the mechanism of action of CR and RT combined. Combination therapy with CR and RT can decrease the metastatic potential in TNBC and in part does this through the miR-17-92 cluster by way of controlling ECM.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P6-01-03.
Abstract P6-01-04: DDB2 a new regulator of metabolism and cell death in human breast tumor cells
Authors: Klotz R , Besancenot V , Brunner E , Becuwe P , Grandemange S
Published: August 2014

The protein Damaged DNA Binding-2 (DDB2) is well known for its role in DNA repair by nucleotide excision repair. Interestingly, DDB2 is differentially expressed in breast cancer expressing the estrogen receptor alpha or not. Recent works performed in our laboratory showed a new role of DDB2 in the control of proliferation and invasive abilities in different breast tumor cells through its involvement in the transcriptional regulation of target genes. Two genes involved in tumorigenic processes, MnSOD (manganese superoxide dismutase) and IκBα (inhibitor alpha of Nuclear Factor-kappa B), have been found to be regulated by DDB2. In addition, transcriptomic analyses on cells that differentially express DDB2 showed that several genes involved in the regulation of cellular metabolism are modulated. Our aim is now to focus on the effects of DDB2 expression on cellular metabolism and glycolysis. The results indicated that the overexpression of DDB2 leads to a respiratory chain dysfunction and an increase of the glycolytic pathway. Moreover, we observed an increased production of reactive oxygen species in these cells, compared to parental cells. As mitochondria are involved in cell death, we performed different experiments to evaluate the impact of DDB2 in the response to anticancer agents (Doxorubicin, and 5-fluorouracile (5-FU)) commonly used in the treatment of breast cancer. Interestingly, the cells exhibit a greater sensitivity to anticancer drugs when DDB2 is overexpressed. As these two agents are related to DNA damaged, we have also used other molecules, the apoptotic inducer, TNFα (Tumor Necrosis Factor alpha), and the Paclitaxel (an antimicrotubule agent) to precise the role of DDB2 on cell death. Similar results were obtained, thus demonstrating the influence of DDB2 overexpression in the response to cell death. The identification of molecular mechanisms responsible for these cellular modifications could place DDB2 and these target genes as predictive markers of sensitivity to anticancer drugs in breast cancer.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P6-01-04.
Abstract P6-01-05: Enhancement of18F-FDG uptake and glycolysis by epidermal growth factor via PI3K activation in T47D breast cancer cells
Authors: Lee EJ , Park JW , Cung Q , Jung K-H , Lee JH , Paik J-Y , Lee K-H
Published: August 2014

It is well known that the binding of epidermal growth factor (EGF) to EGF receptors (EGFR) stimulates proliferation of breast cancer cells via activation of mitogen-activated protein kinase (MAPK) and phosphatidylinositol 3-kinase (PI3K)-protein kinase B (Akt) signaling pathways. There was a report that glucose consumption and lactate production were increased in MDA-MB-468 breast cancer cells following EGF exposure. However, the effect of EGF in enhancing glucose uptake and signaling pathways that are involved have not been clearly revealed. We thus investigated the role of EGF in stimulation of18F-FDG uptake in T47D breast cancer cells, and further elucidated the molecular mechanisms that are involved.

Exposure of T47D cells to 100 ng/mL EGF for 24 h caused a substantial augmentation of18F-FDG uptake to even greater extents to 310.8% ± 30.1% of control cells (P < 0.001), and this effect of EGF showed dose- and time-dependency. Increased glucose uptake by EGF occurred through enhanced membrane GLUT-1 expression as well as hexokinase activity. Inhibition experiments with cycloheximide showed that the metabolic effect by EGF required new protein biosynthesis. Stimulation of glucose uptake by EGF was dependent on sufficient EGFR expression because18F-FDG uptake in MCF-7 cells weakly expressing EGFR was not affected by EGF treatment. Although EGF was also a stimulator of T47D cell proliferation (137.3 ± 7.3% of basal levels at 24 h, P < 0.001), the metabolic effect occurred in a manner that clearly preceded and surpassed the proliferative effect. EGF-stimulated proliferation was significantly inhibited by the EGFR inhibitor BIBX1382, the PI3K inhibitor LY294002 and the specific MAPK inhibitor PD98059. Unlike proliferation, the ability of EGF to augment18F-FDG uptake was found to be dependent on PI3K but not on MAPK activity.

These findings yield the insight into our understanding of the role of EGF and the molecular basis that are involved in glucose metabolism of breast cancer cells.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P6-01-05.