bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract S5-2: HERA TRIAL: 2 years versus 1 year of trastuzumab after adjuvant chemotherapy in women with HER2-positive early breast cancer at 8 years of median follow up
Authors: Goldhirsch A , Piccart-Gebhart MJ , Procter M , de Azambuja E , Weber HA , Untch M , Smith I , Gianni L , Jackisch C , Cameron D , Bell R , Dowsett M , Gelber RD , Leyland-Jones B , Baselga J ,   the HERA Study Team
Published: August 2014

Background: One year (yr) of trastuzumab (T) significantly improves disease-free (DFS) and overall survival (OS) in patients with HER2-positive early breast cancer (EBC) and is considered the standard of care. HERA is the only randomized trial investigating whether longer duration of T can further improve efficacy outcome.

Materials and Methods: HERA (BIG 01–01) is an international, multicenter, phase III randomized trial involving 5102 women with HER2-positive EBC. Pts were randomized, after completion of primary therapy [surgery, chemotherapy and radiotherapy as indicated], to T every 3 weeks for 1 yr, 2 years (yrs), or observation. This landmark efficacy analysis compares the outcome of pts randomized to either 2 yrs or 1 yr of T who were disease-free at 1 yr after randomization (N = 1553 for 2 yrs, and N=1552 for 1 yr). The primary endpoint is DFS and secondary endpoints are OS and time to distant recurrence (TTDR). Updated efficacy analyses of the T arms vs. observation at 8-yrs of median follow-up (FU) are also presented.

Results: On 12 April 2012 HERA reached the target number of 725 DFS events needed for 80% power to detect a true hazard ratio (HR) of 0.80 for the comparison of 2 yrs vs. 1 yr of T. The unadjusted HR for an event in the 2-yr vs. 1-yr T arms was 0.99 (95% CI 0.85–1.14; p = 0.86). OS in the two arms was comparable [HR=1.05 (95% CI 0.86–1.28; p = 0.63)]. TTDR results were similar. The primary cardiac endpoint* was comparable (1.0% vs. 0.8% for 2-yr and 1-yr arms, respectively), but the secondary cardiac endpoint** was higher in the 2-yr arm (7.2% vs. 4.1%). Importantly, the durable benefit in DFS and OS for both 1 yr and 2 yrs of T compared with observation remains stable at 8 yrs of median FU. Subgroup analyses including by hormone receptor status will be available for the meeting.

Conclusions: These results confirm that 1 yr of adjuvant T remains the standard of care for HER2-positive EBC pts. It is also reassuring that the significant improvement in DFS and OS persists over time and that the incidence of cardiac endpoints remains low at a median FU of 8 yrs.

* NYHA class III or IV, confirmed by a cardiologist, and LVEF < 50% and ≥ 10% below baseline, OR cardiac death.

** LVEF < 50% and ≥ 10% below baseline confirmed by repeat assessment, excluding patients with a primary cardiac endpoint.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr S5-2.
Abstract S5-5: Trastuzumab plus adjuvant chemotherapy for HER2-positive breast cancer: Final planned joint analysis of overall survival (OS) from NSABP B-31 and NCCTG N9831
Authors: Romond E , Suman VJ , Jeong J-H , Sledge GW , Geyer CE , Martino S , Rastogi P , Gralow J , Swain SM , Winer E , Colon-Otero G , Hudis C , Paik S , Davidson N , Mamounas EP , Zujewski JA , Wolmark N , Perez EA ,   National Surgical Adjuvant Breast and Bowel Project (NSABP) Operations and Biostatistical Centers
Published: August 2014

Background: Preplanned combined analysis of the National Surgical Adjuvant Breast and Bowel Project (NSABP) B-31 trial and North Central Cancer Treatment Group (NCCTG) (Alliance) N9831 Trial compared adjuvant chemotherapy with or without trastuzumab in women with HER2-positive breast cancer. The initial results were reported in 2005 and updated in early 2011

Methods: 4045 patients with HER2-positive operable breast cancer were enrolled to receive doxorubicin plus cyclophosphamide followed by paclitaxel with or without trastuzumab in both trials. The definitive analysis for OS will occur when 710 events have been reported.

Results: Median time on study is 8 years. The required number of events have occurred for the definitive analysis for OS to be conducted by the end of September 2012. Updated analyses of disease free survival and related subgroups will also be presented.

Supported by NCI grants U10-CA-12027, -69651, -37377, -69974, -44066, U24-CA-114732, CA25224; Breast Cancer Research Foundation; Genentech 35–03

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr S5-5.
Abstract BS01-1: Genetic and non-genetic mechanisms contribute to longterm clonal growth dynamics and therapy resistance
Authors: Dick JE
Published: August 2014

The cellular and molecular basis for intra-tumoral heterogeneity is poorly understood. Tumor cells can be genetically diverse due to mutations and clonal evolution resulting in intra-tumoral functional heterogeneity. Often proposed as mutually exclusive, cancer stem cell (CSC) models postulate that tumors are cellular hierarchies sustained by CSC heterogeneity due to epigenetic differences (i.e. long term tumor propagation only derives from CSC). Two lines of evidence support the CSC model in leukemia. We have recently developed gene signatures specific to either AML LSC or normal HSC and found they share a set of genes that define a common stemness program. Only these stem cell related gene signatures were significant independent predictors of patient survival in large clinical databases. Thus, determinants of stemness influence clinical outcome of AML across a spectrum of mutations indicating that many genetic abnormalities coalesce around stem cell properties. Second, we have carried out a series of combined genetic and functional studies of the LSC from either B-ALL or AML that point to commonalities between clonal evolution and CSC models of cancer. LSC from diagnostic patient samples were genetically diverse and reconstruction of their genetic ancestry showed that multiple LSC subclones were related through a complex branching evolutionary process. The discovery that specific genetic events influence L-IC frequency and that genetically distinct L-IC evolve through a complex evolutionary process indicates that genetic and functional heterogeneity are highly inter-related. Finally, we have also begun to study paired diagnostic (Dx) and relapse (Rx) samples and found that rare Dx subclones are chemotherapy resistant and become enriched at Rx. Collectively, our study points to the need to develop therapies that effective target all genetic subclones present in a tumor and also ensure that cells possessing stemness properties are eliminated to prevent further evolution and recurrence.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr BS01-1.
Abstract OT2-6-02: Denosumab versus placebo as adjuvant treatment for women with early-stage breast cancer at high risk of disease recurrence (D-CARE): A global, placebo-controlled, randomized, double-blind, phase 3 clinical trial
Authors: Goss PE , Barrios CH , Chan A , Finkelstein DM , Iwata H , Martin M , Braun A , Zhou Y , Maniar T , Coleman RE
Published: August 2014

Background: In women with early-stage breast cancer, the skeleton represents one of the most common sites for metastases and represents approximately 40% of all first recurrences. Preclinical studies demonstrated that inhibition of RANKL significantly delays skeletal tumor formation, reduces skeletal tumor burden, and prolongs survival of tumor-bearing mice. Denosumab, a human IgG2 monoclonal antibody, binds RANKL with high specificity and affinity thus inhibiting RANKL activity. Denosumab has been approved for the prevention of skeletal-related events (SREs) in patients with established bone metastases from solid tumors.

Purpose: The D-CARE trial is designed to assess if denosumab treatment prolongs bone metastasis-free survival (BMFS; primary endpoint) in the adjuvant breast cancer setting. Secondary endpoints include disease-free survival (DFS) and overall survival. Additional endpoints include safety, patient reported outcomes (pain, health utilities), as well as exploratory including breast density, time to first on-study SRE following the development of bone metastasis and biomarkers.

Methods: In this global, randomized, double-blind, and placebo-controlled phase 3 trial, approximately 4500 women with stage II or III breast cancer at high risk for recurrence were randomized. High risk is defined as biopsy evidence of breast cancer in regional lymph nodes, tumor size > 5 cm (T3), or locally advanced disease (T4). Patients must be receiving or planned to receive standard-of-care adjuvant/neoadjuvant therapy (chemo-, endocrine, or HER-2 targeted), alone or in combination. Patients with a prior history of breast cancer (except DCIS or LCIS) or distant metastasis, oral bisphosphonate (BP) use within 1 year of randomization, or any intravenous BP use, were not eligible. Patients were randomized 1:1 to receive denosumab 120 mg or placebo subcutaneously monthly for 6 months, then every 3 months for a total of 5 years of treatment. Supplemental vitamin D (≥ 400 IU) and calcium (≥ 500 mg) are required. The primary efficacy and two interim efficacy analyses are planned and will be event-driven. The trial, sponsored by Amgen Inc. and registered with the ClinicalTrials.gov identifier NCT01077154, began enrolling patients in June 2010 and completed enrollment in late 2012.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr OT2-6-02.
Abstract P1-01-14: Extracorporeal shock wave therapy for lymphedema after axillary lymphadenectomy
Authors: Imboden S , Herzig D , Rabaglio M , Hohermuth R , Mueller MD , Günthert AR , Eser P , Schmid J-P
Published: August 2014

Introduction: Lymphedema is a frequent complication of local treatment for breast cancer. Substantial differences in the incidence of lymphedema have been reported depending on the extent of surgery, presence of lymph node metastasis and postoperative radiotherapy. The classical treatment of lymphedema includes manual lymph drainage and compressive bandages. No pharmacological treatment has been shown to be effective. In animal experiments extracorporeal shock wave therapy (ESWT) has been shown to stimulate angiogenesis and lymphatic vessel regeneration. However, whether ESWT reduces lymphedema after axillary lymphadenectomy is unknown.

Patients and Method: We randomized patients with lymphedema after axillary lymphadenectomy (2004 to 2011) into a treatment and a placebo group. Informed consent was obtained from all patients. All but one patient had at least one positive lymph node. A mean of 21.9 (5-43) lymph nodes were removed. 8/10 had an adjuvant radiotherapy of the axilla. ESWT of the whole edematous limb was performed once a week for 10 weeks (2000 shots, energy density 0,25-0,69 mJ/mm2). The placebo group underwent similar treatment, but the shock waves were absorbed in the probe. At baseline and at the end of treatment a clinical and oncological control was performed, including tolerability of the therapy (pain, local skin changes or infections) and oncological safety (recurrence rate). The response to ESWT was measured by volumetry (water displacement of the whole arm) and peripheral quantitative computed tomography (pQCT, change in total cross sectional area of the most swollen part of the arm). Also questionnaires on subjective body feeling, activity score and body image were applied.

Results: We report the preliminary results of the first 10 patients (placebo n = 5, verum n = 5) after 10 weeks. The median change water displacement volume in the verum group corresponded to -192.5 ml (25% percentile: -366.3 ml, 75% percentile -148.8 ml) vs. -12.5 ml (25% percentile: -192.8 ml, 75% percentile: 147.5 ml) in the placebo group (p-value for Mann-Whitney U test: 0.093). Cross sectional area decreased by -3.05% (25% percentile: -8.73%, 75% percentile: 1.49%) in the verum group vs. -1.36% (25% percentile -3.47%, 75% percentile 4.20%) in the placebo group (p-value: 0.394). No treatment had to be interrupted due to local complications. Conclusion: ESWT is well tolerated in patients treated for lymphedema after axillary lymphadenectomy. Our preliminary results show a promising trend towards both objective and subjective improvement of the lymphedema. The statistical significance of these improvements has to be confirmed after inclusion of all patients (N = 30).

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P1-01-14.
Abstract P2-09-02: BMN 673 is a PARP inhibitor in clinical development for the treatment of breast cancer patients with deleterious germline BRCA 1 and 2 mutations
Authors: Mina LA , Ramanathan RK , Wainberg ZA , Byers LA , Chugh R , Sachdev JC , Matei D , Zhang C , Henshaw JW , Dorr A , Kaye SB , de Bono JS
Published: August 2014

Background: BMN 673 is the most potent and specific inhibitor of poly-ADP ribose polymerase (PARP) 1 and 2 in clinical development (IC50<1nM). In BRCA-related tumors, which are genetically dependent on PARP-mediated DNA repair by base excision repair, PARP inhibition induces synthetic lethality. Initial phase 1 results have been presented (de Bono et al JCO 31:5s, 2013 suppl; abstr 2580), showing good tolerability and anti-tumor activity with an MTD on a daily, oral dosing schedule of 1000 μ/day.

Methods: In this 2-stage study, patients with solid tumors including BRCA-related cancers, were enrolled during dose-escalation followed by an expansion phase at the MTD in breast, ovarian, and pancreatic cancer patients with deleterious germline BRCA mutations and in small cell lung cancer and Ewing sarcoma patients to further characterize safety and efficacy. This abstract summarizes demographics and safety for all patients and efficacy for patients with BRCA-related cancers.

Results: As of 17May2013, a total of 1 pancreatic, 18 breast (17F/1M), and 28 ovarian cancer pts with germline BRCA mutations were enrolled at doses from 100-1100 μg/day. All breast cancer patients were treated from 900-1100 μg/day. The median (range) age for all 70 patients is 51.5 (18-81), PS 0 (0-1) and # of prior therapies 4 (1-13), with 47 patients having deleterious BRCA mutations. An MTD of 1000 μg/day was established with thrombocytopenia being dose-limiting. Related adverse events occurring in > 10% of all 70 patients included fatigue, nausea, alopecia, anemia, thrombocytopenia and neutropenia. One patient has had related grade 4 thrombocytopenia. Grade 3 related AE's included fatigue in 1 patient (1%), anemia and thrombocytopenia in 9 each (13%) and neutropenia in 4 (6%). Dose reductions occurred in 11 pts due to myelosuppression. No patients discontinued for adverse events.

Response in Germline BRCA PatientsBRCA Tumor TypeNDose Range (μ/day)ResponseBreast18900-11001CR/6PR/5SD≥12 weeksOvarian28100-11001CR/10PR/19GCIG/4SD≥24 weeksPancreas110001 SD ≥ 12weeksTwo of 2 BRCA breast responders had responded to prior platinum while 0 of 4 non-responders to prior platinum responded to BMN 673. Five of 12 breast cancer pts with no prior platinum have responded.

Conclusions: BMN 673 is well tolerated with impressive anti-tumor activity in pts with deleterious germ line BRCA mutations. Myelosuppression and fatigue are the primary side effects associated with need for dose reduction. A phase 3 trial in metastatic breast cancer patients with deleterious germ line BRCA mutations is planned with single-agent, once-daily oral dosing of 1000 μg (1 mg) per day.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P2-09-02.
Abstract P2-11-17: L-dex ratio in detecting and diagnosing breast cancer-related lymphedema: Reliability, sensitivity, and specificity
Authors: Fu MR , Cleland CM , Guth AA , Kayal M , Haber J , Cartwright F , Kleinman R , Kang Y , Scagliola J , Axelrod D
Published: August 2014

Background: It remains a great challenge to accurately diagnose breast cancer-related lymphedema. Advances in bioelectrical impedance analysis (BIA) permit the assessment of lymphedema by directly measuring lymph fluid changes. Despite its value in assessing lymphedema, the use of BIA in clinical settings is still very limited. In part, this may be due to anecdotal complaints from clinicians about BIA's ability to identify true case of lymphedema in the clinical settings using L-Dex ratio >+10 as the cutoff point for lymphedema diagnosis. This may also be due to lack of large clinical data to support the reliability, sensitivity, and specificity of using L-Dex ratio. The objective of the study was to examine the reliability, sensitivity, and specificity of BIA in a clinical setting.

Methods: BIA was used to measure lymph fluid changes. Data were collected from 250 women, including healthy female adults, breast cancer survivors with lymphedema, and those at risk for lymphedema. Reliability, sensitivity, specificity and area under the ROC curve were estimated.

Results: BIA ratio, as indicated by L-Dex ratio, was highly reliable among healthy women (ICC = 0.99; 95% CI = 0.99 - 0.99), survivors at-risk for lymphedema (ICC = 0.99; 95% CI = 0.99 - 0.99), and all women (ICC = 0.85; 95% CI = 0.81 - 0.87); reliability was acceptable for survivors with lymphedema (ICC = 0.69; 95% CI = 0.54 to 0.80). The L-Dex ratio with a diagnostic cutoff of >+7.1 discriminated between at-risk breast cancer survivors and those with lymphedema with 80% sensitivity and 90% specificity (AUC = 0.86).

Conclusion: The L-Dex ratio may have a role in clinical practice despite its cost to add confidence in the diagnosis of arm lymphedema among breast cancer survivors. Lymphedema is a progressive and debilitating condition and early treatment usually leads to better clinical outcomes, given that BIA using a diagnostic cutoff of L-Dex ratio > +7.1 still misses 20% of true lymphedema cases, it is important for clinicians to integrate other assessment methods (such as self-report, clinical observation, or perometry) to ensure the accurate diagnosis of lymphedema.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P2-11-17.
Abstract P2-14-01: Breast cancer patients with HER2 low-expression: An under-recognized group at significant risk for recurrence
Authors: Vreeland TJ , John BS , Trappey AF , Schneble EJ , Hale DF , Clifton GT , Shumway NM , Perez SA , Papamichail M , Ponniah S , Peoples GE , Mittendorf EA
Published: August 2014

Background:

HER2 over-expression is associated with more aggressive malignant disease. The introduction of trastuzumab and other HER2-directed therapies, however, has led to improved prognosis for patients (pts) with HER2 over-expressing (OE) tumors. Currently, no HER2-targeted therapies are available for patients with HER2 low-expressing (LE) (1+, 2+ by IHC) tumors. We are conducting a randomized, controlled Phase II trial of multiple peptide vaccines enrolling patients with any level of HER2 expression (1+, 2+ and 3+). Here, we report survival data based on levels of HER2 expression in our unvaccinated, control pts.

Methods:

After standard of care therapy, disease-free, high-risk BCa pts were randomized to receive either peptide+GM-CSF (Vaccine Group, VG) or GM-CSF alone (Control Group, CG) in six, monthly doses followed by four boosters every six months. Pts were prospectively followed for recurrence. Demographic information was available for all pts and was compared between groups using chi square or fisher exact tests. Disease-Free Survival (DFS) was compared using log rank.

Results:

To date, we have enrolled 196 pts in the CG. 96 pts had HER2 OE tumors, 100 had LE tumors. The only significant demographic difference between the CG OE and LE groups was more ER/PR positive patients in LE (LE 72% vs OE 51%, p = 0.008). 83% of CG OE pts received trastuzumab, 3% of CG LE pts received trastuzumab. At a median f/u of 30 mo, DFS was significantly higher for CG OE vs CG LE (92.5% v 65.5%, p = 0.001).

Conclusions:

In the cohort of control pts from our ongoing vaccine trial, conducted in an era when Tz has been standard of care therapy for patients with HER2 OE tumors, we have shown that HER2 LE pts are at higher risk of recurrence than OE pts, despite having more ER/PR positive. This calls for increased efforts to develop novel therapies for patients with HER2 LE disease. We have previously shown a trend towards increased DFS with the HER2 vaccines, AE37 (p = 0.13, median f/u 22 mo) and E75 (p = 0.16, median f/u 60mo) in HER2 LE pts, suggesting that these vaccines may represent one such novel therapeutic approach.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P2-14-01.
Abstract P2-16-01: A randomized, phase II, multicenter, double-blind, placebo-controlled trial evaluating onartuzumab with or without bevacizumab in combination with weekly paclitaxel in locally recurrent or metastatic triple-negative breast cancer (TNBC)
Authors: Dieras V , Yardley D , Romieu G , Valero V , Isakoff S , Koeppen H , Thurm H , Teng M , Campone M , Mocci S
Published: August 2014

Background: Dysregulation of the hepatocyte growth factor (HGF)/MET pathway has been observed in many malignancies. In patients with TNBC, overexpression of MET has been shown to be associated with poor prognosis. Onartuzumab (MetMAb) is a recombinant, humanized, monovalent, monoclonal antibody directed against MET, which selectively blocks ligand binding and subsequent HGF activation. A recent phase 2 trial demonstrated improved progression-free survival (PFS) and overall survival (OS) when onartuzumab was added to erlotinib for patients with MET 2+/3+ non-small-cell lung cancer (NSCLC).

Methods: Patients with locally recurrent or metastatic TNBC who had received 0 or 1 prior therapies for metastatic disease were enrolled, provided tumor sample, and randomized (1:1:1) to receive paclitaxel (P) + bevacizumab (B) + onartuzumab (O); P+O; or P+B. Dosing regimens were given IV q4w: P 90 mg/m2 d1, 8 & 15; B 10 mg/kg d1 & 15; O 10 mg/kg d1 & 15. Treatment continued until disease progression or unacceptable toxicity or death; crossover was not permitted. Stratification was by number of metastatic sites (<3, ≥3), number of prior MBC regimens (1 vs 2), and disease-free interval for first-line patients (≤6 months, >6 months). The primary endpoint was PFS; secondary endpoints included OS, objective response rate (ORR), and safety. MET immunohistochemistry (IHC) was performed centrally for exploratory analyses.

Results: 185 patients were enrolled from 30 US and 37 EU sites: median age was 53 years (range 30-79); 77% and 23% were treated in the first- and second-line settings, respectively. Baseline characteristics were balanced. Efficacy data for the ITT population are shown in Table 1. The distribution of MET expression using the NSCLC diagnostic algorithm is shown in Table 2. The MET 2+/3+ group was too small for efficacy analyses. Frequently observed AEs (all grades) included peripheral edema (58% vs 59% vs 18%), fatigue (47% vs 41% vs 48%) and diarrhea (53% vs 22% vs 50%), in the P+B+O vs P+O vs P+B arms, respectively. No new safety signals were observed.

Table 1Table 1ITT populationP+B+O n = 63P+O n = 60P+B (control) n = 62Median PFS, months7.35.47.2HR vs control (95% CI)1.08 (0.69 - 1.70)1.74 (1.13 - 2.78)-Median OS, months14.713.417.4HR vs control (95% CI)1.35 (0.74 - 2.44)1.88 (1.00 - 3.54)-ORR,% (95% CI)42 (29 - 57)28 (16 - 41)55 (41 - 68)

Conclusions: In the ITT population, the addition of O to either P+B or to P alone did not improve PFS and other clinical outcomes compared with P+B alone. Further exploratory subgroup analyses are ongoing.

Table 2Table 2MET IHC score01+2+3+Not availablen (%)81 (44)75 (41)21 (11)1 (0.5)7 (4)

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P2-16-01.
Abstract P2-17-01: The PI3K inhibitor GDC-0032 is selectively potent against PIK3CA mutant breast cancer cell lines and tumors
Authors: Wallin JJ , Edgar KA , Guan J , Sampath D , Nannini M , Belvin M , Friedman LS
Published: August 2014

Mutations in the phosphoinositide-3 kinase alpha isoform (PIK3CA) are frequent in breast cancer and activate the PI3K signaling pathway. We discovered GDC-0032, a selective, potent, orally bioavailable inhibitor of PI3Ka with a Ki = 0.2nM, and with reduced inhibitory activity against PI3Kβ. This selectivity profile, and excellent pharmacokinetic and pharmaceutical properties, allowed for greater efficacy in vivo at the maximum tolerated dose relative to a pan Class I PI3K inhibitor in PIK3CA mutant xenografts. Notably, GDC-0032 preferentially inhibited PIK3CA mutant cells relative to cells with wild-type PI3K. GDC-0032 potently inhibits signal transduction downstream of PI3K and induces apoptosis at low concentrations in breast cancer cell lines and xenograft models that harbor PIK3CA mutations. The mutant-bias of GDC-0032 is linked to unique properties of GDC-0032, including cellular potency against the mutant isoform and reduction of receptor tyrosine kinase (RTK) signaling.

Endocrine therapies such as letrozole are commonly used treatment options for metastatic Hormone Receptor positive (HR+) breast cancer but many patients ultimately relapse. Due to the importance of PI3K in breast cancer, PI3K inhibitors such as GDC-0032 are attractive for combination with endocrine therapies. GDC-0032 was evaluated in breast cancer lines and models in combination with letrozole, and assayed for cellular viability, modulation of PI3K pathway, modulation of ER pathway markers, and apoptosis induction. The combination of GDC-0032 and letrozole decreased cellular viability and increased apoptosis relative to either single agent. We observed cross-talk between the PI3K and ER pathways that suggest a mechanism of action for the combination. In a secreted factor screen we found that multiple soluble factors render breast cancer cells non-responsive to letrozole. It was discovered that many of these factors signal through the PI3K pathway and GDC-0032 in combination with letrozole was able to overcome the growth inhibition caused by the soluble factor. We also established letrozole resistant cell lines that grow independently of any estrogen source. These letrozole resistant lines have elevated PI3K pathway signaling and are still sensitive to GDC-0032. Taken together, these data provide rationale for evaluating GDC-0032 in combination with endocrine therapies for ER+ breast cancer treatment in the clinic.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P2-17-01.
Abstract P3-13-06: A prospective phase II trial of vinorelbine plus oxaliplatin in second- or third-line metastatic triple-negative breast cancer
Authors: Wang Z , Zhang J , Wang L , Hu X , Wang B , Cao J , Lv F , Zhen C , Zhang S , Shao Z
Published: August 2014

Purpose: Patients with metastatic triple-negative breast cancer (mTNBC) typically have a poor prognosis and limited treatment options. Previous study showed biweekly combination of vinorelbine and oxaliplatin (NVBOX) at doses of 30 mg/m2 and 90 mg/m2, respectively, is highly active and well tolerated as first-line treatment for patients with metastatic breast cancer. The purpose of this study (NCT 01528826) was to prospectively evaluate the efficacy and toxicity of NVBOX in second- or third-line mTNBC.

Methods: Eligible patients were female with 18–70 years old and an Eastern Cooperative Oncology Group (ECOG) performance status of 0 to 1, and had mTNBC that had progressed after 1or 2 prior chemotherapy regimens in the metastatic setting. A period of 4 weeks (NVBOX twice) was considered as one treatment cycle and the maximum cycles were 6. The primary endpoint was progression-free survival (PFS). Secondary endpoints included overall survival (OS), objective response rate (ORR), and safety.

Results: Between Dec 2011 and Nov 2012, forty-four patients were recruited [median age 47; 77.3% with viceral metastasis, 100% had been exposed to anthracyclines and/or taxanes; 56.8% cis/carbo-platin pretreated for MBC; 38.6% with time to progression (TTP) of 1-2 previous regimens before recruitment ≤ 3 months]. The overall response rate was 31.6% (1 complete response, 11 partial responses) and 10 achieved stable diseases (7 lasting more than 6 months) in 38 evaluable patients. After a median follow-up of 12.8 months, the median PFS and OS were 4.3 (95% CI, 3.6–5.0) months and 12.6 (95% CI, 8.1–17.0) months, respectively. PFS was significantly shorter in patients with disease free interval (DFI) ≤ 1year (HR = 2.10; 95% CI, 1.05–4.21; P = 0.037) and TTP of 1-2 previous regimens before recruitment ≤ 3 months (HR = 3.39; 95% CI, 1.66–6.89; P < 0.001). Also, these two factors were two independent predictors for the poor OS (HR = 5.45; 95% CI, 2.08–14.32; P < 0.001 and HR = 4.09; 95% CI, 1.73–9.68; P < 0.001). For 34 second line patients, prior platinum in the first line was a factor significantly compromise the PFS (HR = 2.29; 95% CI, 1.03–5.10; P = 0.043). Dose adjustment happened in 14 patients (31.8%) due to adverse events (AEs). Grade 3/4 hematologic toxicities were neutropenia (70.5%), thrombocytopenia (27.3%) and anemia (15.9%). Four patients experienced febrile neutropenia. The most frequent grade 3/4 non-hematologic toxicities were constipation (20.5%) and vomiting (11.4%). Two patients developed grade 3 NVBOX-related peripheral neurotoxicity. There were no treatment-related deaths.

Conclusion: We conclude that biweekly NVBOX regimen is effective with a good safety profile in the second- or third-line mTNBC, which warrants further investigation in a phase III study.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P3-13-06.
Abstract P4-01-18: A discrete population of p63+/K5/14+ cells implicated in the pathogenesis of salivary gland-like tumors of the breast
Authors: Boecker W , Stenman G , Loening T , Andersson MK , Berg T , Buchwalow I
Published: August 2014

Background: Salivary gland-like tumors are currently subdivided into those with and those without myoepithelial differentiation. Furthermore, they show a wide range of differentiation patterns including squamous differentiation. Many authors believe that the myoepithelial cell plays a major role in this context. However, currently little is known about the evolution and cellular differentiation of these tumors.

Material and Methods: For that reason, we performed an in situ Triple Immunofluorescence Lineage/Differentiation Tracing (isTILT) and qRT-PCR study of basal (K5/K14), glandular (K7/K8/18), and epidermal-specific squamous (K10) keratins, p63, and smooth muscle actin (SMA; myoepithelial marker) with the aim to trace different cell lineages and define their cellular hierarchy in tumors. We investigated 50 cases of salivary gland-like tumors of the breast (11 Adenoid-cystic carcinomas, 9 (adeno-)myoepithelial tumors, 8 adeno-squamous carcinomas including 12 syringomas of the nipple, 10 squamous cell carcinomas), and their corresponding counterparts. In 4 cases, the corresponding cell cultures were also analysed.

Results: All tumor types contained p63+/K5/14+ progenitor cells in varying frequencies from a few percent up to 15%. These tumor cells were found to differentiate to glandular (K8/18-positive) or to myoepithelial (SMA-positive) lineage specific cells; they were also shown to generate various heterologeous cell differentiations such as squamous and mesenchymal progenies. p63 was partly retained in the myoepithelial and squamous cells lineages but not in the glandular cells lineage. These data were corroborated by RNA-expression results. The corresponding counterpart tumors of the salivary/lacrimal glands were found to have identical cellular compositions.

Conclusions: Our findings clearly demonstrate that p63+/K5/14+ tumor cells in salivary gland-like tumors of the breast and in their counterparts are key players in tumor development and differentiation. They undergo a transition from their original p63+/K5/14+ precursor state to new p63+/K5/14+/SMA+ myoepithelial and/or p63+/K10+ squamous lineages, and/or to a K7+/K8/18+ glandular state, which can be pictured by isTILT-experiments and RNA-expression studies. Given the phenotypic similarity of p63+/K5/14+ tumor cells to physiological counterparts in the normal breast and salivary duct epithelium and considering the fact that these cells may give rise to a number of different progenies with specific differentiation lineages, we suggest that this cell and its physiological differentiation potential provides an important ontogenetic key to a better understanding of these tumors.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-01-18.
Abstract P5-08-05: The influence of doxorubicin on PTEN and PI3K-Akt-mTOR signaling in human breast cancer
Authors: Austreid E , Knappskog S , Lønning PE , Eikesdal HP
Published: August 2014

Background:

De novo as well as acquired resistance are common features of anthracycline therapy in breast cancer. The PTEN tumor suppressor gene is frequently inactivated in human breast cancer, leading to induction of PI3K-Akt-mTOR signaling, which facilitates cell growth, survival, invasion and ultimately tumor progression. Upregulation of the PI3K-Akt-mTOR pathway has been suggested as a possible mechanism of anthracycline resistance, but there is limited data in the clinical setting, including its influence on PTEN expression.

Methods:

The early PTEN response to doxorubicin was assessed in vitro in MCF7 (estrogen receptor positive) and MDA-MB231 (triple negative) human breast cancer cell lines 24 hours after the initiation of chemotherapy. Tumor samples from thirty patients with locally advanced breast cancer were analyzed for PTEN expression levels and PI3K-Akt-mTOR signaling before and after neoadjuvant anthracycline therapy. The early effect of anthracyclines on PTEN expression is currently being evaluated in human breast cancer samples 24 hours after the initiation of chemotherapy. Mice bearing MCF7 and MDA-MB231 xenografts will be used to assess the potential of PI3K-Akt-mTOR signal transduction inhibitors in preventing anthracycline resistance.

Results:

PTEN expression was found to be profoundly downregulated in both MCF7 and MDA-MB231 cell lines, 24 hours after exposure to doxorubicin at their respective IC50. An increase in Akt1 protein levels in both cell lines was observed in the same setting. In 19 out of 30 human breast cancers PTEN levels increased after a median of 16 weeks neoadjuvant doxorubicin treatment. No difference between estrogen receptor positive and negative breast cancers was observed. The increase in PTEN was accompanied by an increase in fibrotic tissue in the tumor interior. There was no correlation between PTEN level or treatment-induced alterations in PTEN expression and response rates or recurrence-free survival (RFS). The results from our currently ongoing experiments will be presented.

Conclusion:

PTEN expression increases in the majority of human breast cancers during long-term exposure to doxorubicin. However, PTEN is greatly downregulated, with corresponding Akt1 increase, 24 hours after exposure to high doses of doxorubicin in vitro. These results indicate different effects of short- and long-term exposure to doxorubicin, which may be of importance to the development of anthracycline resistance, and to understand at what time a combinational approach with inhibitors of the PI3K-Akt-mTOR pathway will be beneficial. Current efforts are aimed at evaluating the potential of such inhibitors to counteract doxorubicin resistance in this setting.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P5-08-05.
Abstract S4-02: Integrated genomic analyses of members of protein kinase C family identifies subtype specific alterations as novel therapeutic targets
Authors: Natrajan RC , Leonidou A , Brough R , Frankum J , Wai PT , Ng CK , Reis-Filho JS , Lord CJ , Ashworth A
Published: August 2014

Introduction

Members of the protein kinase C family are serine/threonine kinases that are involved in proliferation, apoptosis, cell survival and migration, and have been implicated in tumorigenesis. Recently PRKCE has been found to be up-regulated in triple-negative (TN) breast cancers and has been proposed as a target for therapeutic intervention. The aims of this study were to determine i) whether different members of the PKC family are dysregulated and/ or mutated in specific subtypes of breast cancer, ii) to investigate the impact of silencing or overexpression of members of the PKC family in cell line models representative of the different breast cancer subtypes.

Material and methods:

We obtained expression and mutational data from the cancer genome atlas (TCGA) project from 567 and 640 samples subjected to microarray-based gene expression profiling and whole exome sequencing, respectively. Pair-wise SAM analysis of TCGA gene expression data was performed to identify differential expression between subgroups (ER+, HER2+ and TN). Potential driver mutations were identified through the algorithm CHASM. Subtype specific dependencies were identified from the re-analysis of publicly available siRNA kinome-wide screens in a panel of 20 breast cancer cell lines. In vitro assessment of gene overexpression was assessed in MCF10A cells by wound healing scratch assays and 3D growth in Matrigel.

Results

PRKCA, B, I and Q were expressed at significantly higher levels in TN breast cancers. Higher expression of PRKCE was significantly associated with ER-negativity, whereas high PRKCD expression was associated with ER-positivity. Analysis of siRNA kinome-wide screen data resulted in a significant reduction in survival associated with PRKCI and PRKCE in ER-negative cells, PRKCQ in triple-negative cells, and PRKCD in ER-positive cells. Furthermore meta-analysis of published exome and whole genome sequencing data identified potentially activating recurrent kinase domain mutations in PRKCB (0.93%), Q (1.25%) and Z (0.93%), with mutations in PRKCZ being associated with higher gene expression. Forced expression of wild-type PRKCQ and PRKCZ in MCF10A cells resulted in the formation of irregular acini in 3D cell culture and expression of wild-type PRKCZ resulted in increased migration.

Conclusions

Differential expression of members of the protein kinase C family, are associated with different molecular subtypes of breast cancer. Furthermore, we have shown that breast cancer cells are dependent upon expression of these family members in vitro, which are associated with different cellular phenotypes.

Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr S4-02.
Abstract A28: Latino farm workers and non-farm workers and their experiences in providing biospecimens for research.
Authors: Thompson Beti , Carosso Elizabeth , Islas Ilda , Ibarra Genoveva
Published: August 2014

Objectives: The goal of this project was to re-contact 40 of the 100 adult participants (Hispanic farm worker and non-farm workers), who participated in a pesticides exposure study, to learn about their experience providing biospecimens for research. The 40 randomly selected participants are members of a larger cohort that has been participating in pesticides exposure research for over 10 years. During their participation in the pesticides exposure study these participants were asked to provide two 10mL tubes of blood, two buccal cell samples and four urine samples during the seven-day collection period. Two dust samples were also collected from the home and a 7-day dust sample was collected from the vehicle. This protocol was repeated a total of three times (during two different pesticide spray seasons and one non-spray season) over the course of one year. Twelve of the 40 participants interviewed also provided 20 saliva samples, 2 hair samples and one blood sample over 10 weeks for a stress and cortisol study.

Methods: Two months after completing the pesticides exposure study, we re-contacted a random selection of 40 participants to take part in a one-on-one interview. All participants are Hispanic farm workers and non-farm workers and all live in the Yakima Valley of Washington. Interview questions were selected to learn about the participant's feelings toward research, the barriers and facilitators to participating in research and providing biological samples, expectations surrounding their biological samples and thoughts about providing tumor samples.

Results: A total of 42 participants were re-contacted; 40 agreed to participate and completed an interview, 1 refused and 1 was lost to follow-up. Each participant who completed an interview was provided a $20 gift card. Qualitative analysis is summarizing the beliefs and opinions of Hispanics about donating biospecimens and their understanding of what such donation means. Their attitudes to research are also being explored. Results will be available in mid-July, 2012.

Conclusion: Speculation about Latinos giving biospecimens suggests that Hispanics are more reluctant to participate than non-Hispanic Whites. This was not our experience. The personal meaning of donation and the meaning of research is important to better understand Latinos participation in biobanking.

Citation Format: Beti Thompson, Elizabeth Carosso, Ilda Islas, Genoveva Ibarra. Latino farm workers and non-farm workers and their experiences in providing biospecimens for research. [abstract]. In: Proceedings of the Fifth AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2012 Oct 27-30; San Diego, CA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2012;21(10 Suppl):Abstract nr A28.
Abstract A30: Informed consent for cancer biobank research: Case series of patients with cancer disparities in rural Maine.
Authors: Hock Janet M. , Morrison Deborah G. , Farah Christopher C.
Published: August 2014

One critical issue in cancer research biobanking is improving representation of America's minority and medically underserved populations. Rural populations often suffer disparities related to poverty and healthcare access, limiting their participation in cancer research. We report a case series of newly diagnosed cancer patients at Eastern Maine Medical Center (EMMC), a rural comprehensive medical center. Of 85 eligible cases, 83 (98%) consented to participate in biobanking research; 2 years later, survivors (87%) consented to 2 additional requests.

We used NCI/ISBER Best Practices to create biobank infrastructure after approval by EMMC IRB and OHP HRPO. Informed consent forms were at Flesch-Kincaid 8th Grade reading level, and supplemented by NCI educational brochures. Cases lived 50-150 miles from EMMC. Of 108 candidates, 83 eligible cases (49 lung cancer; 21 breast cancer; 13 other cancers) consented to questionnaires and donation of blood and/or tissue. Of the remaining 23 cases, 2 declined; cancer was not confirmed in 12; 4 died, and 7 were ineligible. Within 2 years, 14 cases died. Of those alive, 60/69 (87%) were re-contacted and consented to additional data collection, and transfer of biospecimens to NCI's biorepository. Most cases exhibited co-morbidities with 1-3 chronic diseases (mostly cardiovascular), prior cancer history and long-term tobacco and/or alcohol consumption. Nearly 90% reported 1+ relative with cancer, while 69%, reported 1+ prior cancers. Anecdotally, willingness to consent was based on altruistic hopes that research would generate knowledge to reduce cancer incidence. In summary, cancer patients from disadvantaged rural communities will consent to participate in biobank research.

Citation Format: Janet M. Hock, Deborah G. Morrison, Christopher C. Farah. Informed consent for cancer biobank research: Case series of patients with cancer disparities in rural Maine. [abstract]. In: Proceedings of the Fifth AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2012 Oct 27-30; San Diego, CA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2012;21(10 Suppl):Abstract nr A30.
Abstract A38: Nanoreformulation of platinum-based drugs to address disparate cancer treatment outcomes.
Authors: Lillard James W. , Singh S. , Singh R.
Published: August 2014

Modest responses and systemic toxicities during treatment of advanced cancer limits cancer drug efficacy, especially in African Americans. We determined the utility of cisplatin-loaded biodegradable XPclad nanoparticles coated with folate-conjugated polycaprolactone-polyethylene glycol. XPclad nanoparticles under in vitro conditions exhibited sustained release of the encapsulated cisplatin. Prostate cancer (PCa) cell lines (PC3, LNCaP, PTEN-CaP2, etc.), but not normal prostatic epithelial cells (PrEC and RWPE-1), expressed high levels of folate receptor α (FRα/FR1). Similar to cancer cells, Treg cells but not T helper cells express high affinity folate receptor (FRδ/FR4). Treatment using 20 μM of cisplatin solution resulted in > 90% cell death of PCa cells, PrEC, and RWPE-1 cells as well as purified CD4+CD25HiFoxP3+ T cells (Tregs) and CD4+CD25LoFoxP3- T cells (T helper). However, XPclad nanoparticles, containing cisplatin (equivalent to a 200 nM dose), were taken up and induced the same level of apoptosis in PCa and Treg cells as cisplatin solution, but did not significantly affect PrEC or RWPE-1 cells or T helper cells. Finally, we show that PC3 tumor-bearing mice that received either cisplatin solution (8 mg/kg/week) or cisplatin-encapsulated folate-coated XPclad nanoparticles (equivalent cisplatin dose = 0.08 mg/kg/week) resulted in significant tumor regression. Mice receiving cisplatin solution had significantly higher kidney and liver toxicities than naïve or XPclad nanoparticle-treated mice. Our studies show XPclad nanoparticles enhance tumor therapy efficacy with significantly lower to no toxicity and suggest that FR-targeted nanotherapy modulates the destruction of both tumor cells and Treg cells for increased efficacy. GRANT SUPPORT: G12 RR03034, U54 CA118638 and P60MD00525

Citation Format: James W. Lillard, S. Singh, R. Singh. Nanoreformulation of platinum-based drugs to address disparate cancer treatment outcomes. [abstract]. In: Proceedings of the Fifth AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2012 Oct 27-30; San Diego, CA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2012;21(10 Suppl):Abstract nr A38.