bluuebunny/crossref_metadata_2025 huggingface dataset:
A Phase I Pilot Study of the Oral mTOR Inhibitor RAD001 in Combination with Capecitabine for Metastatic Breast Cancer.
Authors: Chen C. , Chen M. , Imaoka R. , Svahn T. , Guardino A.
Published: December 2010

Background:RAD001 is an oral inhibitor of mTOR, an intracellular protein kinase downstream of the PI3K/Akt signaling pathway. There is evidence to suggest that Akt promotes breast cancer resistance to cytotoxic treatment and thus targeting this pathway with RAD001 may increase the efficacy of standard chemotherapy. We combined RAD001 in a Phase I dose escalation study with Capecitabine (oral fluorpyrimidine), as treatment of metastatic breast cancer. This study evaluated the oral combination for tolerability and safety. Pharmacokinetics of RAD001 was examined in all dosing cohorts.Methods:This was a single center, Phase I trial. Eligible patients had stage IV breast cancer, have not received endocrine or chemotherapy for 3 weeks prior to initiation and not receiving concurrent endocrine therapy. Patients must not have received prior therapy with capecitabine or an mTOR inhibitor and up to 3 prior chemotherapy regimens were allowable. Pharmacokinetic data are collected weekly for first 2 cycles.All patients received capecitabine (825 mg PO BID) and RAD001 in 3 dose-escalation cohorts: 2.5 mg every other day (Cohort 1), 2.5 mg daily (Cohort 2) and 5 mg daily (cohort 3). The primary endpoint is to determine the DLT of RAD001 + capecitabine, secondary endpoints include tumor response. Treatment continues until disease progression or unacceptable toxicity. Efficacy assessed every 6 weeks for first 4 cycles, then every 9 weeks until end of the 1st year of treatment, then every 12 weeks after.Results: As of April 2009, 12 patients with a median age of 58 years (range 34-75) have been treated at 3 dose levels: 3 in the 1st cohort, 6 in the 2nd cohort and 3 in the 3rd cohort. Toxicities in cohort 1 were generally manageable and expected. They most commonly consisted of nausea, hand-foot syndrome and elevated LFTs. 2 patients in cohort 2 had a grade 3/4 neutropenia and responded to dose reduction. 1 patient had a grade 3/4 elevated lipid panel. Other adverse events (all grade 1/2) included leukopenia, neutropenia, anemia, hypoalbuminemia, elevated LFTs, nausea, hand-foot syndrome, fatigue, sensory neuropathy, and thrombocytopenia. Of the 9 evaluable patients, the clinical benefit rate (CR+PR+SD ≥ 24 weeks) in this heavily pretreated group is 44%. No patient had DLT during cycle 1. 3 patients remain on treatment. Enrollment to the 3rd cohort continues.Conclusion:The all-oral regimen of RAD001 with capecitabine is well tolerated. The maximal tolerated dose (MTD) after 3 dose escalations was not reached. The maximal dose of 5 mg per day of RAD001 was well tolerated. Final toxicity data and preliminary efficacy will be presented at the meeting.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 6112.
3,3'-Diindolylmethane Inhibits Activation of Akt through Inhibition of Hepatocyte Growth Factor Receptor c-Met Signaling.
Authors: Hantz H. , Lee K. , Bjeldanes L.
Published: December 2010

Background: 3,3'-Diindolylmethane (DIM), the major acid condensation product of indole-3-carbinol, is a promising anticancer compound that is bioavailable, non-toxic, and easily obtainable. Although DIM has strong anticancer activity against breast cancer cells in vitro and in vivo, the exact mechanism for this activity is not known. We and others have found that DIM inhibits activation of the Akt pathway in breast cancer cells. Akt is a serine threonine protein kinase involved in many important cellular functions including proliferation and cell survival. Akt is active when phosphorylated at serine-473 and threonine-308. In breast cancer cells, Akt can be activated by various stimuli, including signaling from growth factor receptors like the hepatocyte growth factor (HGF) receptor c-Met, which is often overexpressed in breast cancer. In this project, we show that c-Met is an upstream point of inhibition of the Akt pathway by DIM.Materials and Methods: When MDA-MB-231 breast cancer cells reached 70-80% confluency, we serum-starved them by reducing the fetal bovine serum concentration in the media from 10% to 1% for 20 hours and then treated with DIM or the vehicle control DMSO for 4 hours or the indicated time. When using recombinant growth factors to activate Akt or c-Met, we incubated cells in medium containing no fetal bovine serum and added the growth factors for 10 minutes after incubation with DIM. To analyze protein expression and protein phosphorylation we performed Western blotting. To analyze c-Met kinase activity we used a commercially available kit.Results: DIM does not inhibit activation of Akt downstream of either epidermal growth factor or insulin-like growth factor-1. However, DIM does inhibit Akt activation downstream of HGF and of fetal bovine serum. DIM inhibits tyrosine phosphorylation of c-Met at Y1349 and Y1234/1235. DIM does not affect total c-Met protein or mRNA levels and does not directly inhibit c-Met kinase activity.Discussion: While we have determined that DIM inhibits c-Met phosphorlyation, and therefore HGF signaling, the mechanism of this inhibition requires further study. We hypothesize that DIM induces serine phosphorylation of c-Met, since c-Met can be inhibited by reactive oxygen species (ROS)-induced phosphorylation of serine-985 by protein kinase C (PKC), and DIM increases ROS levels in cells.Components of the Akt pathway are frequent targets for mutations in breast cancer and c-Met and its ligand HGF are associated with lower survival and increased risk of distant metastases in breast cancer. This project represents an important step in determining the mechanism of action for DIM, a promising phytochemical for the treatment of breast cancer.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 6117.
The Role of microRNA Genes in Breast Cancer Progression and Metastasis.
Authors: Johnstone C. , Lucas E. , Kusdra L. , Goga A. , Anderson R.
Published: December 2010

MicroRNAs influence tumor progression and metastasis via action as metastasis promoting genes (for example mir-373 and mir-520) or metastasis suppressor genes (for example mir-335). To identify additional microRNAs capable of driving breast cancer metastasis in vivo, we employed a functional genomics screen using the '4T1' syngeneic (Balb/c) mouse model of breast cancer progression. Approximately 400 microRNA sequences were subcloned into the pMSCV_puro retroviral expression vector and the resultant microRNA expression library transduced into fluorescent non-metastatic 67NR-pMSCV_mCherry and mildy-metastatic 66cl4-pMSCV_mCherry cells and selected with puromycin. The transduced cell lines were then implanted into the mammary fat pads of 20 syngeneic Balb/c mice per cell line. Following establishment of primary tumors, the primary tumors and secondary organs (lung, liver, spine, femurs), including any overt metastatic deposits, were dissected under a fluorescent stereomicroscope and the microRNA inserts amplified from the integrated retroviral genome by PCR.In an alternative approach, we have studied two populations of MDA-MB-231 human breast cancer cells that exhibit different behavior in vitro and in vivo to elucidate processes involved in cellular invasion and metastasis. A late-passage MDA-MB-231 (231variant) population formed invasive clusters when embedded in Matrigel, whereas early-passage (P3) MDA-MB-231 (231) cells did not. Moreover, 231v cells were more motile than 231 cells and showed enhanced tumorigenicity when grown as subcutaneous xenografts in nude mice, whereas proliferation and apoptotic response were indistinguishable between the two lines in vitro. Molecular characterization of the two cell populations cultured in 3D by array-based gene expression profiling revealed elevated expression of NF kappa B target genes in 231v cells (IL1A, IL1B, IL6, PTX3, NR4A2, HMOX1, FLIP). MicroRNA array analysis of 3D cultures also revealed marked induction (approximately 8-fold) of the related NF kappa B target microRNAs, mir-146a and mir-146b (Figure 1). Further evaluation of a panel of human breast cancer cell lines revealed that mir-146a and mir-146b were dramatically elevated in metastatic human breast cancer cell lines that display both epithelial-to-mesenchymal transition and high NF kappa B activity (MDA-MB-231, Hs578T, MDA-MB-436), compared to non-metastatic lines. Experiments to elucidate the function of mir-146a and mir-146b during breast cancer progression are underway. Figure 1: TaqMan qPCR analysis of mir-146a and mir-146b expression in 231 and 231v cells.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 6142.
The Secretory Small GTPase Rab27B Regulates Invasive Tumor Growth and Metastasis through Extracellular Heat Shock Protein 90α.
Authors: Hendrix A. , Maynard D. , Pauwels P. , Braems G. , Denys H. , Van den Broecke R. , Van Belle S. , Cocquyt V. , Bracke M. , Seabra M. , Gahl W. , De Wever O. , Westbroek W.
Published: December 2010

Background: Vesicle exocytosis, controlled by secretory GTPases such as Rab27B, delivers critical pro-invasive factors into the tumor microenvironment. The biological role and expression status of Rab27B in breast cancer was unknown.Methods: Rab27B was studied in human breast cancer cells (MCF-7, T47D, ZR75.1) using GFP-fusion constructs, including Rab27A and Rab27B point mutants defective in GTP-binding or geranylgeranylation. In cell culture, cell-cycle progression was evaluated by flow cytometry and Western blotting, invasion was assessed using Matrigel and collagen type I substrates. Orthotopic tumor growth, local invasion and metastasis were analyzed in mouse xenograft models. Mass spectrometry was performed to identify Rab27B-secreted pro-invasive factors. Rab27B levels in clinical breast cancer were analyzed by quantitative reverse transcription-polymerase chain reaction (n=20) and immunohistochemistry (n=60). Statistical tests were two-sided.Results: Rab27B-upregulation promoted G1/S phase cell cycle transition, F-actin reorganization and invasion in cell culture, and invasive tumor growth and peritoneal metastasis in a xenograft mouse model (at 10 weeks, survival of MCF-7 GFP vs GFP-Rab27B injected mice was 100% vs 62.5%, P=0.0307). Proteomic analysis of purified Rab27B-secretory vesicles and the secretome of Rab27B-expressing breast cancer cells identified heat shock protein (HSP)90α as key pro-invasive factor. HSP90α secretion occurred in a Rab27B-dependent manner and was required for matrix metalloproteinase-2 activation. All Rab27B-mediated functional responses were GTP- and geranylgeranyl-dependent. In clinical samples, upregulation of endogenous Rab27B mRNA and protein correlated with lymph node metastasis (P=0.0002) and differentiation grade (P=0.0014) in estrogen receptor (ER)-positive human breast tumors.Conclusions: Rab27B, a new prognostic marker for ER-positive breast cancer, regulates invasive growth and metastasis.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 6144.
Programmed Cell Death 4 (PDCD4), a Novel Inhibitor of Leptin-Induced Breast Cancer Cell Invasion.
Authors: Gonzalez-Villasana V. , Gonzalez-Villasana V. , Nieves-Alicea R. , McMurtry V. , Tari A. , Gutierrez-Puente Y.
Published: December 2010

Obesity is known to increase the risk of breast tumor development in post menopausal women. Leptin, an adipokine produced in high levels in obese individuals, has been found in breast tumors and been linked with increased incidence of breast cancer metastasis. Breast cancer metastasis is directly associated with breast cancer cell invasion. Previously we demonstrated that leptin increases breast cancer cell invasion. Here we determined whether PDCD4, a novel tumor suppressor, could block leptin-induced breast cancer cell invasion. In MCF-7 breast cancer cells overexpressing PDCD4 (MCF-7/PDCD4), leptin failed to induce invasion, in contrast to the parental MCF-7 cells, indicating that PDCD4 blocks leptin-induced breast cancer cell invasion. MCF-7/PDCD4 cells produced higher levels of the Tissue Inhibitor of Metalloproteinases-2 (TIMP-2) than the parental cells. Pretreatment of MCF-7/PDCD4 cells with TIMP-2 siRNA allowed leptin to induce the invasion of MCF-7/PDCD4 cells, thus indicating that knocking down TIMP-2 expression reversed the anti-invasive effects of PDCD4. The levels of phospho-ERK and phospho-STAT3 were higher in MCF-7/PDCD4 cells treated with leptin and TIMP-2 siRNA than in MCF-7/PDCD4 cells treated with leptin and non-silencing control siRNA. Yet the levels of phospho-JNK were similar in leptin-treated MCF-7/PDCD4 cells transfected with TIMP-2 siRNA or non-silencing control siRNA. Here we report the novel findings that PDCD4 increases TIMP-2 expression to block leptin-induced breast cancer cell invasion by suppressing ERK and STAT3 activities. Metastasis is the primary cause of breast cancer mortality. Novel therapeutic strategies aiming at enhancing PDCD4 expression in breast tumors may be able to stop obesity-related breast tumor progression and prolong patients' lives.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 6153.
Phase III Randomized Trial Comparing Doxorubicin and Cyclophosphamide Followed by Docetaxel (AC→T) with Doxorubicin and Cyclophosphamide Followed by Docetaxel and Trastuzumab (AC→TH) with Docetaxel, Carboplatin and Trastuzumab (TCH) in Her2neu Positive Early Breast Cancer Patients: BCIRG 006 Study.
Authors: Slamon D. , Eiermann W. , Robert N. , Pienkowski T. , Martin M. , Rolski J. , Chan A. , Mackey J. , Liu M. , Pinter T. , Valero V. , Falkson C. , Fornander T. , Shiftan T. , Olsen S. , Buyse M. , Kiskartalyi T. , Landreau V. , Wilson V. , Press M. , Crown J. ,   on Behalf of BCIRG006 Investigators
Published: December 2010

Background: Evaluation of the long-term benefit of biologically-based regimens of trastuzumab in the early breast cancer population, and optimization of trastuzumab integration to maximize efficacy and minimize cardiac toxicity.Material and Methods: We randomized HER2-positive (FISH+) breast cancer patients with axillary lymph node positive or high risk negative, to either standard AC (60/600 mg/m2 q3wk x4) followed by T (100 mg/m2 q3wk x 4) or two trastuzumab-containing regimens; AC followed by T with trastuzumab x 1 year or TCarbo (75 mg/m2/AUC6 q3wk x 6) with trastuzumab x 1 year. Patients were prospectively stratified by number of positive nodes (0, 1-3 vs 4+) and hormone receptor status. Patients with ER and/or PR positive (HR+) tumors received hormone-directed therapy for 5 yrs after chemotherapy. The primary endpoint was disease-free survival (DFS) with 80% power (0.05 significance level) to detect an absolute difference of 7%. Secondary endpoints include overall survival (OS) and safety, including cardiac toxicity (symptomatic events and asymptomatic LVEF decline). The first two protocol-specified analyses for this study were performed at 300 and 450 disease-related events. We now report the results of the third protocol-specified analysis conducted after 650 events, expected by end of June 2009.Results: A total of 3222 patients (1072 in AC-T, 1076 in AC-TH and 1074 in TCH) were recruited between April 2001 and March 2004. Baseline characteristics of the study population will be included. Cox analysis of DFS and OS (unadjusted and adjusted for nodal status) and cardiac toxicity data will be presented for the three treatment arms.Discussion: The results of this trial help define the role of trastuzumab in the breast cancer HER2-positive adjuvant setting, as well as the risks/benefits of adjuvant trastuzumab within the context of overall safety including cardiac toxicity. This latter objective is of particular importance given that many of these women may be cured of their disease in the adjuvant setting.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 62.
Do the ASCO/CAP 2007 HER2 Testing Guidelines Improve Prediction of Benefit to Adjuvant Trastuzumab?: Data from North Central Cancer Treatment Group N9831 Adjuvant Trial.
Authors: Perez E. , Reinholz M. , Dueck A. , Tenner K. , Davidson N. , Gralow J. , Harris L. , Kutteh L. , Jenkins R. , Chen B.
Published: December 2010

Background: In 2007, the American Society of Clinical Oncology and College of American Pathologists (ASCO/CAP) recommended new guidelines to define HER2 positivity by immunohistochemistry (3+ IHC: uniform intense membrane staining of >30% of invasive tumor cells) or fluorescent in situ hybridization (FISH+: HER2/CEP17 ratio >2.2). The original criteria used for enrollment in the pivotal N9831 trial conducted from 2000-2005 were IHC>10% (3+) or FISH ≥2.0.Purpose: To investigate the impact of the ASCO/CAP guidelines on patient (pt) eligibility and disease free survival (DFS) compared to the originally used FDA-approved definitions.Patients/Methods: This analysis included 2268 patients from the HER2+ N9831 adjuvant trastuzumab phase III trial. IHC was centrally performed at the Mayo Clinic using the DAKO HercepTest™ and was re-analyzed to determine the percent of tumor cells with 0, 1+, 2+, and 3+ staining intensities. DFS was compared between pts randomized to standard chemotherapy +/- concurrent trastuzumab (Arms A and C of the trial) within IHC/FISH subgroups using Cox proportional hazards regression stratified by hormone receptor and nodal status.Results: Based on 2268 pts, 83 pts (3.7%) were 3+ by FDA-approved guidelines but not 3+ by the ASCO/CAP guidelines. Among these pts, 77% (64/83) were FISH amplified centrally by ASCO/CAP guidelines. Only 1.5% (34/2268) of pts eligible under FDA-approved definitions from N9831 in this sample set would not meet the ASCO/CAP guidelines. In 1441 patients randomized to standard chemotherapy +/- concurrent trastuzumab, the exploratory DFS hazard ratio (HR) associated with trastuzumab in IHC/FISH subgroups were as follows: ASCO/CAP HER2+ HR=0.59 (95% CI: 0.47-0.75; p<0.001); FDA-approved HER2+ HR=0.60 (95% CI: 0.47-0.75; p<0.001); ASCO/CAP HER2 normal(not positive) HR=0.74 (95% CI: 0.35-1.57; p=0.43); and FDA-approved HER2 normal HR=0.53 (95% CI: 0.22-1.26; p=0.15).Conclusions: A small percentage (1.5%) of N9831 patients did not meet ASCO/CAP 2007 guidelines when applied retrospectively. Trastuzumab effect appeared similar for HER2+ patients regardless of ASCO/CAP or FDA-approved guidelines. Additional DFS analyses by IHC/FISH subgroups will be presented. We gratefully acknowledge support from the Breast Cancer Research Foundation, Genentech, and NIH CA25224 for this study.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 701.
A Population-Based Cross-Over Randomized Controlled Trial of Breast Cancer Screening with Alternate Mammography and Ultrasound for Women Aged 40 to 49 Years in Taiwan.
Authors: Huang C. , Fann C. , Hsu G. , Ho M. , Chang K. , Chen S. , Kon S. , Chen T.
Published: December 2010

Background: As there is paucity of data on population-based screening for breast cancer using mammography and ultrasound for oriental young women aged 40-49 years, the peak of incidence rate and high proportion of dense breast, we aimed to evaluate the relative performance of detecting breast cancer between ultrasound and mammography and also to assess complementary efficacy of ultrasound to mammography screening.Methods: A total of 79,691 female residents aged 40-49 years were invited from community in Taiwan since late 2003. These participants were first randomly assigned to mammography (n=20040), ultrasound (n=20088), and control group (n=39563). The two former groups were further done by a cross-over design with mammography and ultrasound on alternate year until 2008. Detection rate and annual incidence rate of interval cancer as a percentage of the control group (I/E ratio) were compared between mammography and ultrasound.Results: The attendance rate of the first round was 59% (11921/20040) for mammography and 56% (11249/20088) for ultrasound. The repeated attendance rate of both groups was 85% in the second round and 91% in the third round. In the first round of screen, the detection rate of breast cancer for the mammography group (0.34%) was 1.5-fold compared with the ultrasound group (0.22%). The additional detection rate was 0.16% contributed from a subsequent ultrasound screen and 0.36% contributed from a subsequent mammogram screen. The combination of mammography with ultrasound was as three to four times as likely to detect breast cancer compared with the control group (annual incidence rate was 0.17%). The I/E ratio was lower after mammography screening than that after ultrasound screening.Conclusion: The current randomized controlled trial not only demonstrated higher detection rate and better performance using mammography but also indicated the complementary role of ultrasound applied to young Taiwanese women. This further suggests the optimal screening modality for young women in Asian country is to combine mammography with ultrasound.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 73.
Is a Weekly Supervised Upper Limb Exercise Program of Value for Women with Early Breast Cancer?
Authors: Kilbreath S. , Refshauge K. , Beith J. , Simpson J. , Ward L. , Lee M.
Published: December 2010

The aim of this randomised single blinded study was to determine whether an upper limb exercise program, commencing 4-weeks after surgery for early breast cancer, prevents upper limb symptoms, such as pain, swelling, stiffness and weakness.Methods: 156 women, aged 52.5 ± 11.7 y, who had undergone either axillary node dissection (AND; n=94) or sentinel node biopsy (SNB; n=62) for unilateral breast cancer in combination with either mastectomy (n=74) or wide local excision (n=82) within the previous 4 weeks, participated. Women were randomised to either an 8-week exercise program (n=79) or to usual care (n=77); participants were stratified for type of axillary surgery (SNB or AND) and treatment site. The exercise program comprised a home program of daily passive stretching for shoulder muscles and resistance training using Thera-Band® on alternate days for muscles about the shoulder. Women also attended a weekly exercise session. Usual care included routine follow-up with their medical specialists, a breast nurse and a post-operative visit with the physiotherapist and occupational therapist who provided instructions about exercises. The primary outcome was self-reported arm and chest symptoms derived from the EORTC breast cancer specific questionnaire (BR23) which was scored out of 100 with a low score indicative of fewer symptoms. The secondary outcomes included physical measures of shoulder range of motion, strength, and swelling. Presence of lymphoedema (LE) was determined by the inter-limb impedance ratio, using bioimpedance spectroscopy.Results: After the 8-week intervention period, there was no significant difference between groups in either the self-reported or the physical measurements (Table 1).Table 1. Baseline and 8 weeks data (mean ± SD). Usual care group Exercise group  Baseline8 weeksBaseline8 weeksSelf-report symptoms (max 100)    Arm24.5 ± 14.514.6 ± 14.225.9 ± 18.512.5 ± 12.9Breast25.8 ± 16.219.2 ± 17.725.6 ± 19.217.1 ± 16.7Range of motion interlimb difference (deg)    Flexion19.8 ± 19.06.9 ± 7.023.9 ± 19.05.4 ± 7.3Abduction14.5 ± 14.04.7 ± 8.821.1 ± 18.43.8 ± 8.1Inter-limb strength difference (Newtons)    Flexion15.9 ± 21.55.6 ± 19.417.0 ± 21.84.0 ± 15.8Abduction20.0 ± 25.85.7 ± 19.725.2 ± 28.61.0 ± 19.3Swelling (n)    Prevalence of LE (by impedance)7/7714/745/789/74 Conclusion: A weekly, supervised, 8-week upper limb exercise program is not warranted for all women with early breast cancer treated with surgery to the axilla. Usual care in which women are seen post-operatively by a physiotherapist, occupational therapist, and breast nurse with ongoing monitoring by the specialists is sufficient for most women.Acknowledgement: Supported by NSW Cancer Council

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 811.
Contralateral Breast Cancer in BRCA1/2 Mutation Carriers: Risk Assessment by Age at Diagnosis of the First Primary Breast Cancer.
Authors: Hooning M. , Heemskerk-Gerritsen B. , Jager A. , Collee M. , van den Ouweland A. , Menke - Pluymers M. , van Geel B. , van Doorn L. , Tilanus - Linthorst M. , Bartels C. , Seynaeve C.
Published: December 2010

Purpose Age at diagnosis of the first breast cancer is a known predictive factor of the risk of contralateral breast cancer (CBC) in BRCA1/2 mutation carriers. However, accurate risk estimates for different age subgroups are scarce.Methods We studied the risk of CBC in 560 women with a primary breast cancer (PBC) from families with a proven BRCA1 or BRCA2 mutation in relation to the age at diagnosis of the first PBC, taking into account history of oophorectomy. Time at risk was the period between diagnosis of the first PBC and date of contralateral breast cancer, contralateral mastectomy, metastatic disease, death or last follow-up. Oophorectomy was treated as a time-dependent covariate. Patients with bilateral breast cancer diagnosed before the first visit at the family cancer clinic were excluded.Results We observed 96 CBCs in 423 BRCA1 mutation carriers and 21 cases in 137 BRCA2 mutation carriers. Actuarial risk of CBC at 5, 10 and 15 years was 14%, 34% and 39%, respectively, for BRCA1-, and 13%, 22% and 27%, respectively, for BRCA2 mutation carriers. For BRCA1 carriers 10-year actuarial risk of CBC was 54% for women with a PBC ≤ 30 years, 39% for the group with PBC between 31-40 years, 26% for the 41-50 group and 17% for those with PBC > 50 years. In BRCA2 carriers 10-year risk of CBC was 28% for women with a PBC ≤ 40 years, 19% for the 41-50 group and 10% for those with PBC > 50 years. For BRCA1 carriers who had an oophorectomy, 10-year risk of CBC was 33% and 26%, respectively, for those with PBC younger and older than 40 years, against 44% and 24% in women who had no such treatment. In BRCA2 carriers with a history of oophorectomy 10-year risk of CBC was 21% and 14%, respectively, for those with PBC younger and older than 40 years, while the risk was 29% and 17%, respectively, in women without oophorectomy. Effect of adjuvant therapy is being analyzed and will be presented at the meeting.Conclusions The availability of risk estimates of CBC by more detailed age groups at PBC is a helpful tool in the counselling process concerning screening and/or preventive surgery for breast cancer patients with a BRCA1 or BRCA2 mutation.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 903.
Low Risk Ductal Carcinoma In Situ (DCIS) Treated with Breast Conserving Surgery & Accelerated Partial Breast Irradation (APBI): Comparison of the Mammosite Registry Trial with Intergroup Study E5194.
Authors: Goyal S. , Vicini F. , Beitsch P. , Keisch M. , Jeruss J. , Lyden M. , Haffty B.
Published: December 2010

Purpose/Objective(s): Recent data from Intergroup Study E5194 prospectively defined a low risk subset of ductal carcinoma in situ (DCIS) patients where radiation therapy was omitted after local excision alone. At 5 years, they found a 6.8% ipsilateral breast event rate (IBTR) in selected low/intermediate grade (LIG) patients and a 13.7% IBTR in selected high grade (HG) patients. The purpose of our study was to determine the IBTR in DCIS patients treated on the American Society of Breast Surgeons (ASBrS) MammoSite® Breast Brachytherapy registry trial who met the criteria of E5194 treated with local excision and adjuvant APBI.Material/Methods: A total of 194 patients with DCIS were treated between 2002 and 2004 on the Mammosite registry trial; of these, 69 patients met the enrollment criteria for E5194: 1. LIG: low to intermediate grade with a pathological size > 0.3cm but < 2.5cm and margins >3mm (n=40) or 2. HG: high grade, pathological size <1cm and margins >3mm (n=29). All patients were treated with lumpectomy followed by adjuvant APBI (34 Gy in 3.4 Gy fractions). Median follow-up for surviving patients was 50.7 months (range, 0-73.4). Hormonal therapy was given to 51% of these patients (compared to 30% of patients on E5194). The clinical-pathologic data and long term outcomes for each patient were entered into a database for statistical analysis using SAS (v 8.2).Results: The median age of the entire cohort was 62.5 years. In the LIG cohort, the median size was 0.8cm. In the HG cohort, the median size was 0.6cm. In the LIG cohort, the 5-year IBTR was 0%, compared to 6.8% at 5 years in E5194. In the HG cohort, the 5-year IBTR was 5.3% compared to 13.7% at 5years in E5194. The overall the 5-year IBRT was 2% and there were no cases of elsewhere or regional failures in the entire cohort. The 5-year contralateral breast event rate was 0% and 5.6% in LIG and HG patients, respectively (compared to 3.5% and 4.2%, respectively, in E5194). The 5-year disease free survival and overall survival rate was 92.4% and 96.5%, respectively. In all patients with DCIS (n=194) treated with Mammosite on the registry trial, the 5-year actuarial IBTR, DFS, and OS was 3.1%, 93.8% and 98.2%, respectively.Conclusion: Adjuvant accelerated partial breast irradiation using Mammosite is a relatively convenient form of radiotherapy which shortens treatment time to 1 week. This study found that patients who met the criteria of E5194 treated with APBI had extremely low rates of recurrence (0% vs. 6.8% in the low to intermediate grade and 5.3% vs. 13.7% in the high grade). We conclude that all patients with DCIS who were eligible for E5194 have appreciable benefit from adjuvant APBI.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 951.
Our Inner Mouse: Genomic Insights from Mouse Mammary Cancer Models To Guide Pre-clinical Testing
Authors: Green J.
Published: December 2010

Accelerating the pre-clinical testing of novel therapies or combination therapies is an important goal of translational research. This is particularly true for identifying potentially new therapeutic approaches for triple-negative (TN) breast cancer (lacking expression of ER, PR and her2) for which no specifically targeted therapies currently exist. This sub-type of breast cancer, which generally falls into a basal category, has a poorer prognosis than breast cancers expressing ER/PR with a luminal phenotype. Most basal TN breast tumors have lost normal function of the tumor suppressor genes p53 and Rb. Using genetically-engineered mouse tumor models where p53 and Rb have been functionally inactivated through over-expression of SV40 T/t-antigens, we have identified a core gene expression (Tag) signature of about 150 genes involved in cell proliferation, cell replication, DNA repair, and apoptosis that is very relevant to human epithelial cancer. This signature, which is highly represented in the basal TN, C3(1)/SV40 T/t-antigen (Tag) mouse model of breast cancer, segregates human breast cancers with the basal TN phenotype and poor prognosis from luminal type tumors with a better prognosis. Based upon this cross-species analysis, we are determining whether genes within the Tag signature are potential therapeutic targets for basal TN breast cancer using human basal breast cancer cell lines and cell lines derived from the C3(1)/Tag tumors. This is initially being assessed using siRNA, drugs that inhibit specific targets within the signature, drugs in clinical use and combinations of these to find potentially novel therapeutic approaches for basal TN tumors. Additionally, in vivo studies are being performed using both human xenograft and Tag tumor transplant models for preclinical testing of selected therapies. Unlike studies using human xenografts in immunocompromised mice, experiments using Tag tumor transplants can be performed in mice with an intact immune system, adding an important dimension to these pre-clinical studies. Our approach demonstrates the values of cross-species genomic analyses to identify an appropriate model of basal TN breast cancer for pre-clinical testing, potential therapeutic targets, and an accelerated means of performing pre-clinical studies.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr BS1-1.
The Evolving Role of Adjuvant Chemotherapy.
Authors: Winer E.
Published: December 2010

Decision-making for the medical oncologist was once far simpler than it is today. Following the 2000 NIH Consensus Conference on the treatment of early stage breast, almost all women with tumors > 1cm or positive lymph nodes were offered a course of adjuvant chemotherapy. There were some choices to be made in terms of the specific adjuvant regimen, but the oncologist could rely upon the results of large, randomized trials and choose a regimen that had been shown to be beneficial. Indeed, some might argue that a medical oncologist in 2009 could take the same approach. But the breast cancer world has evolved substantially over the past 5-7 years, and such an approach would not take full advantage of our understanding of breast cancer in 2009. We now understand that there is a great degree of intrinsic heterogeneity across breast tumors, and that the benefits associated with adjuvant chemotherapy vary substantially by subtype of disease. At minimum, we should consider three groups of patients separately: 1) individuals with HER2 positive disease; 2) individuals with triple negative disease; and 3) individuals with ER+ and HER2 non-amplified, non-overexpressed disease, who represent the vast majority of the patients with breast cancer in the United States. For patients with HER2+ disease, trastuzumab-based therapy has become the standard of care for all but women who have very small (T1a) tumors in the setting of negative lymph. Even in those women, there are physicians who would recommend a course of adjuvant trastuzumab-based chemotherapy. The major question is what chemotherapy to use in combination with trastuzumab, and this is a matter of considerable debate. In addition, uncertainty exists surrounding the optimal treatment for individuals who have indeterminate HER2 test results. For women with triple negative breast cancer, chemotherapy is also given to almost all patients apart from those with the very smallest tumors in the context of negative lymph nodes. For most patients, one of the standard anthracycline-taxane regimens remains the usual course of therapy. For patients with a low disease burden, first or second generation regimens such as AC, TC, or CMF are reasonable considerations. The most difficult questions arise concerning the use of chemotherapy in women with ER+ and HER2 normal disease. In this setting, the average benefit associated with chemotherapy is far less than in women with HER2+ or triple negative disease. Recent retrospective studies suggest that there are large groups of women who likely derive almost no or almost no benefit from chemotherapy. Increasingly, assays such as the Oncotype Dx Recurrence Score are used to aid clinical decisions, but even these complex genetic assays do not always provide a clear roadmap. Controversies and practical considerations surrounding the use of chemotherapy in women with ER+ and HER2 normal disease will be discussed in detail.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr CS1-2.
What Is the Histone Code, and Why Is It Important?
Authors: Oesterreich S.
Published: December 2010

The field of epigenetics is rapidly expanding, and has changed and revised traditional paradigms of inheritance. Epigenetics, literally “beyond genetics” or “in addition to genetics”, is defined as the study of heritable changes in gene expression that occur without a change in DNA sequence. Within this presentation, I will focus on modifications of histones, an important mechanism to convey epigenetic information. Histones, small proteins which serve as spools for DNA wrapped in nucleosomes, are critical regulators of the dynamic state of chromatin. They are subject to numerous posttranslational modifications, including acetylation, methylation, ubiquitylation, and phosphorylation. The best characterized modifications are acetylations and methylations. Increased acetylation of histones by histone acetyltransferases is associated with gene activation due to weakened charge attraction between DNA and histone; histone deacetylases in contrast remove acetyl groups leading to gene inactivation. Histone methylation mediated by histone methyltransferases has either positive effects or negative effects on gene expression depending on location and association of other protein complexes. In general, it is difficult to predict gene regulation based on the study of a single histone modification, since it is the combination of these modifications, also called the histone code, which ultimately controls gene expression. I will discuss details of the histone modification, how they regulate gene expression, and also their relevance in disease, especially breast cancer.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr ES4-2.
Overview of the Randomised Trials of Radiotherapy in Early Breast Cancer.
Authors: Darby S. ,   on Behalf of the Early Breast Cancer Trialists' Collaborative Group
Published: December 2010

The Early Breast Cancer Trialists' Collaborative Group (EBCTCG) was set up in the mid-1980s to co-ordinate 5-yearly meta-analyses of centrally collated individual patient data from women in all randomised trials of the treatment of early breast cancer. The present cycle considers trials that began by 2000 and includes follow-up to 2006 where available. The latest results from the overview of trials of radiotherapy versus the same management but without radiotherapy will be presented.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr MS3-1.
How Will the '4:1 Ratio' Change with Increasingly Effective Systemic Therapy.
Authors: Harris J.
Published: December 2010

The 4:1 ratio between a reduction in local recurrence at 5 years and a reduction in mortality at 15 years has been well established in clinical practice. The Ratio is particularly useful to estimate the survival benefit of adding RT, particularly in the postmastectomy setting where the use of RT is not routine. It is worth noting, however, that the 4:1 ratio is an average effect that is largely based on trials without systemic therapy and does not account for the interrelated issues of (1) competing risks of local recurrence and distant metastases, (2) biologic heterogeneity, and (3) the impact of (improving) systemic therapy. If the ratio becomes less than 4:1, it means that local treatment is even more important for long-term survival. In this talk, emerging data will be presented illustrating how the '4:1 ratio' is influenced by competing risks of local recurrence and distant metastases, biologic heterogeneity, and the impact of improving systemic therapy. The current data suggest that over the short to intermediate time frame, with increasingly effective systemic therapy, the risk of LR will be less, but the ratio will also be less. Current systemic therapy is most effective in the setting of microscopic residual disease or micrometastases. As systemic therapies are better able to sterilize distant disease, improvements in local-regional control have become more important. As systemic therapy continues to improve such that it can control both distant and local-regional disease, the eventual goal is that local therapy could be completely eliminated; however, that is likely to be sometime into the future.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr MS3-2.
An Epidemiological Perspective on the Causes and Prevention of Breast Cancer.
Authors: Beral V.
Published: December 2010

Breast cancer is the most common cancer in women worldwide, with over 1 million new cases occurring annually. Incidence rates used to be 6-7 times more common in developed than in less developed countries, but the gap is narrowing, as breast cancer rates in less developed countries are now increasing rapidly. The contribution to the total burden of breast cancer from the well-established and potentially modifiable causes will be described. Most of the difference between breast cancer rates in developed and less developed countries is because of the small average family size in developed countries and the relatively short duration of lactation. However, it is neither desirable nor feasible for women to revert to childbearing patterns of the past to prevent breast cancer. If we seriously want an acceptable way of achieving a sizeable reduction in breast cancer incidence we need work out the appropriate way of mimicking the effects of childbearing on the breast. The epidemiological evidence points to persistent, lifelong, protection from short-term exposures to hormones of late pregnancy and lactation (probably not from oestrogens and progesterone alone). It should therefore be possible to develop a short-term 'hormonal vaccine' to provide long-term protection against breast cancer.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr P1-1.
Non-image guided fine needle aspiration of palpable axillary lymph nodes in breast cancer patients.
Authors: Marti JL , Ayo D , Levine P , Hernandez O , Rescigno J , Axelrod DM
Published: December 2010

Abstract #1013

Introduction  While the utility of ultrasound-guided axillary lymph node FNA is well established, there is a paucity of data on non-image guided office-based FNA of palpable axillary lymphadenopathy. We investigate the sensitivity and specificity of non-image guided FNA of axillary lymphadenopathy in patients presenting with breast cancer, and report factors correlating with a positive FNA result.  Methods  Retrospective review of 80 consecutive patients who underwent office-based FNA of palpable axillary lymph nodes between 2004-2008, with cytology results compared with histology during axillary sentinel node or lymph node dissection. Pearson correlation, chi-square and Fisher exact tests were used to determine correlation with other clinical and pathologic data.  Results  Mean age was 56 with a median tumor size of 2.2 cm and 39% have pNO disease. Non-image guided axillary FNA was 82% sensitive, 100% specific, and 89% accurate. Positive predictive value was 100% and negative predictive value was 76%. Therefore only 24% of patients who had a negative FNA had axillary involvement compared to 61% for the entire group.  A significant correlation or association was identified between positive FNA cytology and breast tumor size (p=0.02), number of pathologic positive lymph nodes (p<0.0001), presentation with a palpable breast mass (p=0.003), radiographic lymphadenopathy (p=0.0005), lymphovascular invasion (p=0.003), stage of disease (P<0.0001), and N stage (p<0.0001). There was a borderline significant association with her2neu receptor status (p=0.08).  Age, lymph node size, ER/PR status, Ki-67 index, tumor histology, multicentric disease, degree of differentiation and nuclear grade were all not significantly correlated or associated with positive FNA.  Conclusions  Non-image guided FNA of palpable axillary lymphadenopathy in breast cancer patients is a very sensitive and specific test. Prompt determination of lymph node positivity benefits select patients, permitting avoidance of ultrasound, sentinel lymph node biopsy, or delay in receiving neoadjuvant therapy. This results in time and cost savings for the patient and the health care system, and expedites definitive management of the patient.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 1013.
Identification of BRCA1 and BRCA2 genetic modifiers.
Authors: Weyandt JD , Snyder C , Lynch HT , Gillanders E , Holmes TN , Bailey-Wilson J , Ellsworth RE
Published: December 2010

Abstract #1040

Background: Although mutations in BRCA1 and BRCA2 represent the primary lesions in hereditary breast and ovarian cancer (HBOC) families, these mutations are marked by incomplete penetrance, as 15-20% of carriers never develop either cancer. In addition, the expressivity of these gene mutations varies with manifestation in breast, ovary, or both breast and ovary, suggesting other genes may modify the effects of BRCA1 and BRCA2 mutations.  Methods: DNA was isolated from nine large HBOC families with known BRCA1 and BRCA2 mutations. SNP data was generated using the GeneChip Mapping 10K Array (Affymetrix, Santa Clara, CA). The data were checked for deviation from Hardy-Weinberg equilibrium and Mendelian errors. SNPs with low completion rates were not used in the analyses. Two-point parametric linkage analysis was performed under dominant and recessive models using VITESSE and GAS.  Results: Genotypes were generated for 301 individuals from nine families and included mutation carriers, and non-carriers with and without cancer. When all families were considered together using a dominant model, LOD scores >3.0 were detected for SNPs on chromosomes 6q25.3 and 18q21.3 and >2.0 for chromosomes 2q24, 5p13 and 5q33. When families were segregated by mutation type, linkage was detected at chromosome 22q12 (LOD =2.65) in BRCA1 families, and at 2q24 (LOD=2.17) in BRCA2 families.  Conclusions: Chromosomal regions harboring putative genetic modifiers have been identified that may contribute to pathogenesis in both BRCA1 and BRCA2 families. For example, frequent gain of 2q24 has been detected in BRCA1 carriers with ovarian cancer, loss of 5q in breast tumors from BRCA1 patients, and SNP rs720321 is intragenic to the apoptosis gene BCL2 on chromosome 18q21, expression of which has been associated with BRCA2 tumors. Linkage to chromosome 22q12 was specific to BRCA1 families, of note, the loss of the CHEK2 gene, located on chromosome 22q12, may promote tumor progression in BRCA1 tumors. Further elucidation of the underlying genetic lesion from each of these chromosomal regions may improve the risk assessment and treatment regimens of women with BRCA1 and BRCA2 mutations.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 1040.
Breast cancer instructs dendritic cells to express OX40 ligand and to prime pro-inflammatory type 2 CD4+ T cells that facilitate tumor development.
Authors: Palucka AK , Pedroza A , Aspord C , Gallegos M , Burton E , Banchereau J
Published: December 2010

Abstract #1050

We found that breast cancer tumors are infiltrated with mature dendritic cells (DCs), which cluster with CD4+ T cells. CD4+ T cells infiltrating breast cancer tumors secrete type 1 (IFN-g), pro-inflammatory (TNF) as well as high levels of type 2 (IL-4 and IL-13) cytokines. Immunofluorescence staining of tissue sections revealed intense IL-13 staining on breast cancer cells. The expression of phosphorylated STAT6 in breast cancer cells suggests that IL-13 actually delivers signals to cancer cells.  To determine the link between breast cancer, DCs and CD4+ T cells, we implanted human breast cancer cell lines in NOD/SCID b2m-/- mice engrafted with human CD34+ hematopoietic progenitor cells (HPCs) and autologous T cells. There, CD4+ T cells promote early tumor development. This can be prevented by administration of IL-13 antagonists and is dependent on DCs. Furthermore, these humanized mice demonstrate a signature of IL-13.  Similarly to patient samples, in humanized mice breast cancer but not melanoma tumors are infiltrated with large numbers of human myeloid DCs. DC isolated from breast tumor and its draining lymph nodes induce the production of high level of IL4, IL13 and TNF by naive CD4 T cells further demonstrating that breast tumor modulates DC to induce an inflammatory Th2 response.  Accordingly, immunofluorescence staining of breast cancer tissue sections from patients shows high expression of OX40 ligand on DCs. This can be ascribed to activity of breast cancer cell-derived soluble factors. Finally, blocking OX40 ligand prevents generation of IL-13 secreting CD4+ T cells.  Thus, breast cancer targets DCs to generate pro-inflammatory type 2 immunity that promotes its development.  Supported by U19 AIO57234, RO-1 CA89440, CA78846, and CA85540.

Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 1050.