bluuebunny/crossref_metadata_2025 huggingface dataset:
P5-13-23: Individualized Treatment Strategies for HER2−Negative Breast Cancer Subtypes.
Authors: Ishikawa T , Shimizu D , Yamada A , Sasaki T , Morita S , Tanabe M , Kawachi K , Nozawa A , Chishima T , Kimura M , Ichikawa Y , Endo I
Published: June 2012

Background Human epidermal growth factor receptor (HER2)-negative breast cancers are more heterogeneous than HER2−positive cancers, and tailored treatment is therefore required for luminal A and triple-negative breast cancer subtypes (LABC and TNBC). We therefore examined predictive factors for the efficacy of standard chemotherapy in LABC and TNBC subtypes.

Methods A total of 109 LABC and 61 TNBC patients were treated with standard neoadjuvant chemotherapy (NAC) consisting of an anthracycline and/or taxane. The pathological treatment response and prognosis were examined for each subtype. Expression levels of the following factors were examined in association with quasi-pathological complete response (QpCR): estrogen- and progesterone-receptor (ER and PgR) status, HER2, nuclear grade, MIB-1, p53, topoisomerase IIα (topoIIα), cytokeratin (CK) 5/6 and epidermal growth factor (EGFR).

Results QpCR rates in LABC and TNBC were 9.1% (10/109) and 54.1% (33/61), respectively. In LABC, the expression of PgR tended to be inversely associated with pathological response (p=0.087), while in TNBC, increased expression of topoIIa (p=0.006) and MIB-1 (p=0.018) were identified as predictors of QpCR. TopoIIα expression was also significantly associated with pathological response in multivariate analysis (p=0.014). The QpCR rate was higher in TNBC lacking CK5/6 and/or EGFR expression, defined as non-basal subtype (p=0.053).

Conclusions Low expression of PgR may be a possible predictor of the efficacy of chemotherapy in LABC, while a high level of proliferative activity, indicated by topoIIa and MIB-1, is associated with chemosensitivity in TNBC. Further subclassification into basaland non basal-subtypes may also be helpful for the development of individualized treatments.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P5-13-23.
P5-14-06: Interaction between Stoma and Tumor Characteristics as a New Prognostic and Predictive Marker in Breast Carcinomas.
Authors: Tagliabue E , Sandri M , Casalini P , Aiello P , Pupa SM , Orlandi R , Balsari A , Triulzi T
Published: June 2012

Background. We recently demonstrated that primary breast tumors can be classified based upon extracellular matrix composition (ECM) (Bergamaschi et al.) suggesting stroma characteristics could influence tumor progression.

Aim of this study is to further investigate the robustness of this classification and its impact in tumor progression and response to therapy.

Methods. The expression profile of ECM-related genes was analyzed by unsupervised hierarchical clustering in 10 independent datasets of breast tumors, counting more than 1000 samples and the prognostic and predictive value of this signature were evaluated in two datasets of not treated patients and in one dataset of neo-adjuvant treated patients, respectively.

Results. Only one of ECM subsets (ECM3) showed a homogeneous gene pattern that consistently allowed the classification of an independent group of tumors in all tested datasets. ECM3 is characterized by highly correlated over expression of 34 ECM genes encoding mainly structural proteins. From 24 to 38% of cases were ECM3-enriched and were mainly estrogen receptor-positive and low grade (p<0.0001). The ECM3 showed inconsistent association with DMFS in the two data sets of not treated patients, whereas the interaction between ECM3 and grade showed strongly and significant association with DMFS (HR=5.35, p=0.0012). In particular, multivariate analysis of covariates available (size, ER, age and ECM) indicated that ECM3 is significantly associated with worse DMFS in grade III patients (HR=2.5, p=0.0386), whereas it is slightly correlated with better DMFS in grade I-II patients (HR=0.6, p=0.088). It is noteworthy that the probability of 10-year DMFS was 90% in ECM3 versus 74% in non-ECM3 patients in differentiated tumors, and 75% in non-ECM3 versus 41 % in ECM3 grade III tumors. The interaction between ECM3 and grade remained significantly associated with DMFS also in ER negative (HR=14.2, p=0.03) and ER positive (HR=3.34, p=0.079) subgroups. ECM3, indeed, was correlated with good and bad prognosis in differentiated and grade III tumors, respectively.

Moreover analysis of a data set of patients treated neoadjuvantly with chemotherapy revealed that, among the grade III tumors, pathological complete response was reached by 9% of ECM3 versus 74% of non-ECM3 tumors (p=0.04). In differentiated tumors no response difference was observed according to ECM.

Conclusions. Our results provide evidence that breast carcinoma progression and response to therapy are influenced by the interaction between tumor and stromal characteristics.

(Partially supported by AIRC).

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P5-14-06.
P5-14-15: Prognostic Impact of Chemotherapy-Induced Amenorrhea (CIA) in Premenopausal Breast Cancer: A Meta-Analysis of Published Literatures.
Authors: Lu J , Zhou Q , Yin W
Published: June 2012

Background: Various studies have reported inconsistent results on the prognostic effect of chemotherapy-induced amenorrhea (CIA) in premenopausal breast cancer. Therefore, we conducted a meta-analysis to assess the effect of CIA on prognosis of premenopausal women with breast cancer.

Methods: The PubMed database was searched for all relevant studies published before March 2011. Relative risks (RRs) were used to estimate the association between CIA on various survival outcomes. Subgroup analyses were also performed by age, nodal status and hormone receptor (HR) status.

Results: With thirteen eligible studies identified, this meta-analysis included 5790 cases and 2159 controls. This meta-analysis demonstrated that CIA was associated with improved DFS (RR= 0.39, 95%CI: 0.32∼0.47, p<0.001) and OS (RR= 0.30, 95%CI: 0.21∼0.44, p<0.001). In the subgroup analyses, the effect of CIA on DFS and OS also existed in HR-positive subjects (for OS, RR= 0.19, 95%CI: 0.07∼0.49, p=0.001; for DFS, RR= 0.47, 95%CI: 0.35∼0.63, p<0.001) while similar results failed to be observed in HR-negative patients (for OS, RR= 0.62, 95%CI: 0.07∼5.75, p=0.671; for DFS, RR= 1.48, 95%CI: 0.50∼4.37, p=0.474). Furthermore, significant difference was achieved between women with and without CIA, irrespective of age and nodal status.*** Conclusion: This meta-analysis clarifies that CIA might contribute to the improvement of prognosis in HR-positive premenopausal breast cancer patients, which is at least partially responsible for the benefit of adjuvant chemotherapy in premenopausal women through chemical castration.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P5-14-15.
P5-14-16: Molecular Classification in Primary Breast Cancer and Corresponding Lymph Node Metastasis Show Impaired Prognostic Profile in the Metastatic Node.
Authors: Falck A-K , Ferno M , Bendahl P-O , Rydén L
Published: June 2012

Introduction The heterogeneity of breast cancer disease has emphasized the need to optimize prediction of outcome by molecular classification of the primary tumor. Clinical management of primary breast cancer is based on biomarkers measured in the primary tumor, whereas biomarker analysis in metastatic lymph nodes is not used for clinical decision making. Recently, microarray-based expression profiling and subsequent immunohistochemical studies have identified biologically distinct subtypes using a combination of biomarkers. The present study aimed to classify primary breast cancer tumors and corresponding lymph node metastases by multiple biomarkers into Luminal A, Luminal B, HER2 type and Triple negative phenotype and compare outcome.

Material and method The study is based on a cohort of patients where biomarker expression (hormone receptor status (ER, PR), human epidermal growth factor receptor 2 (HER2) and Ki67) in primary breast cancer and ipsilateral lymph node metastasis recently were reexamined and individually related to prognosis. In the present study, 85 patients of the original cohort were possible to classify into the four subtypes: Luminal A (ER+, PR+/− and Ki67 ≤ 20%), Luminal B (ER+, PR+/−, HER2+/− and Ki67 > 20%), HER2− type (ER-, HER2+) and Triple negative (ER-, PR-, HER2−). The classifications of primary tumors and corresponding lymph node metastases were compared using the McNemar-Bowker test of symmetry and the molecular subgroups were related to clinical outcome in terms of 5-years distant disease-free survival (DDFS) by Cox analysis with luminal A as the reference group.

In 7 cases (16%) the biomarker classification was discordant between the primary tumor and the lymph node metastasis. All of the discordant cases shifted to a subtype with a worse prognosis according to the lymph node metastasis (p=0.06, Mc Nemar-Bowker test of symmetry). When comparing Luminal A to non-Luminal A in the primary tumor and the lymph node, the deviation from symmetry was significant, p=0.02. DDFS was worse in all subtypes compared to the prognosis for Luminal A using Cox analysis in both primary tumors and lymph node metastasis.

Conclusion Our data shows discordance in classification according to a multiple molecular phenotype between primary tumors and lymph node metastasis in breast cancer patients. The discordant cases were shifted from Luminal A to a subtype where survival analysis showed an impaired prognosis compared to this subgroup. The finding that the molecular phenotype is shifted from a “good” prognostic signature in the primary tumor to a “bad” prognostic signature in the metastatic node can be of clinical importance for the choice of adjuvant systemic therapy in primary breast cancer with lymph node positive disease.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P5-14-16.
P5-14-20: Neoadjuvant Chemotherapy (NCT) in 466 Patients for Operable Breast Cancer: The Prognostic Value of SBR Grade Variation.
Authors: Abrial C , Mouret-Reynier M-A , Wang-Lopez Q , Durando X , Thivat E , Raoelfils I , Gimbergues P , Van Praagh-Doreau I , Nabholtz J-M , Chollet P , Penault-Llorca F
Published: June 2012

Purpose of the study: In a new database harbouring the patients of several prospective phase II neoadjuvant trials (fec 50–100, net, tncf, taxotere/tncf, taxotere alone), the predictive value of tumour factors (SBR grade, RH, Her2, Ki-67 and cycline D1) was studied in 466 women with stage II-III operable breast cancer treated between 1991 and 2006.

Patients and methods: Median age of the patients was 48 years [27-76]. Median diameter of the invasive tumour was 40 mm [10-130]. 382 (82%) patients had a canalar, 57 (12.2%) a lobular, 14 (3%) a mixed or invasive carcinoma, 3 (0.6%) neoplasic cells only and 10 (2.2%) another carcinoma. Before chemotherapy, 33% were grade III SBR. The median number of NCT courses was 6 [1-8] followed by a surgery for 98%, a radiotherapy for 93%, an adjuvant chemotherapy (20%) and/or a hormonotherapy (56%).

Results: Overall response rate was 73% (18% complete). The complete pathological response (pCR) rate was 17.2% according to Chevallier's classification. On 455 patients operated, 324 (71%) had a conservative surgery. On 390 patients with an axillary dissection, 195 (50%) had involved nodes (median number: 2 [1-20]). After a median follow-up of 135 months, DFS and actuarial survival at 120 months were 61.9% and 73%, respectively.

After chemotherapy, we found a significant variation for SBR grade ***: there was five fold more patients for whom we observed a down grading of the SBR grade (grade 2/3 to grade 1) than up grading (grade 1 to grade 2/3), p=6.1.10−6. For patients in objective response, we observed a significant decrease of SBR grade (p=1.7.10−3).

Moreover, the variation of SBR grade was also pronostic. Indeed, the disesase-free survival for patients who presented a SBR down grading after chemotherapy was significantly better (p=0.031) compared to patients for whom SBR grade remained stable or was in up grading. There was a tendancy for overall survival (p=0.15).

Conclusion: Down grading of SBR grade appears linked to response and so may constitute a parameter of better prognostic. At the opposite, up grading of SBR grade is an adverse prognostic factor. To conclude, it is interesting to emphasize that SBR grade variations can be assessed when patients are not in pCR, ie plays a complementary role.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P5-14-20.
P5-14-27: Prognostic Factors of Node-Negative, High Risk and 1–3 Positive Lymph Nodes Breast Cancer by Intrinsic Subtype in Patients with Adjuvant Chemotherapy.
Authors: Kojima Y , Hashimoto K , Harano K , Shimizu C , Yunokawa M , Yonemori K , Tamura K , Katsumata N , Ando M , Kinoshita T , Fujiwara Y
Published: June 2012

Purpose: St. Gallen 2007 categorized high risk, node-negative breast cancer (HNBC) and 1–3 lymph nodes positive BC (LNBC) without HER2 overexpression as intermediate risk. We hypothesized that triple negative BC (TNBC) and hormone-receptor positive without HER2 overexpression (H-BC) in intermediate risk have different prognostic factor in patients with adjuvant chemotherapy (AdC).

Methods: We examined disease-free survival (PFS) and overall survival (OS) of TNBC and H-BC with regard to potential prognostic factors. All the patients included in this study were categorized intermediate risk by St.Gallen 2007 and received AdC.

Results: A total of 470 patients were identified; H-BC (n=360) and TNBC (n=110). Age (<35) was significantly associated with DFS in TNBC while it was not in H-BC (p=0.0.1 and p=0.63, respectively). Tumor size (>2cm) and tumor grade related to DFS in H-BC but not in TNBC. Tumor grade was not associated with DFS in both H-BC and TNBC (p=0.64 and p=0.91, respectively). Lymph node metastasis was a significant factor of DFS only for H-BC (p=0.009 and p=0.19). Conclusions: In TNBC, age was only a significant factor associated with DFS while in H-BC, lymph node status and tumor size rather age related to DFS.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P5-14-27.
P5-18-06: Taxanes and Cyclophosphamide Are Equally Effective in Breast Cancer: A Meta-Analysis of Ten Phase III Trials in Early and Advanced Disease.
Authors: Vriens BE , Lobbezoo DJ , Voogd AC , Veeck J , Tjan-Heijnen VC
Published: June 2012

Background Overall taxanes did not improve survival in metastatic breast cancer trials (n=10 trials), whereas they do so in (neo-)adjuvant breast cancer trials (n=28 trials). We also noticed that in a substantial number of ‘negative’ metastatic trials, taxane-regimens were used without cyclophosphamide. To further explore this, we compared the outcome of studies in early and advanced breast cancer with a similar design, that is all studies substituting taxanes for cyclophosphamide.

Methods We identified 10 phase III taxane-based chemotherapy trials in early and advanced disease, in which taxanes were used instead of cyclophosphamide. They all compared a regimen of an anthracycline combined with a taxane versus an anthracycline combined with cyclophosphamide, i.e., AT versus AC. A pooled analysis was performed using the Review Manager software (RevMan 5) provided by the Cochrane Collaboration.

Results In total, 5 studies in advanced disease, 2 studies in neoadjuvant, and 3 studies in adjuvant disease setting were included and analyzed for their primary endpoint. In metastatic breast cancer studies, the hazard ratio of overall survival was 1.03 (95% CI 0.92 to 1.15) for taxanes as compared to cyclophosphamide. Also, studies on early breast cancer with a similar design showed no improvement for taxanes, resulting in an odds ratio for pCR of 0.91 (95% CI 0.57 to 1.43) in the neo-adjuvant setting and a hazard ratio for 5-year disease-free survival of 0.96 (95% CI 0.84 to 1.09) in the adjuvant setting.

Conclusion Re-assessment of studies of drugs both assessed in metastatic and early breast cancer provides a new tool for improved understanding. This meta-analysis shows that cyclophosphamide in breast cancer patients is equally effective as taxanes, and thus should be considered of pivotal importance in the treatment of metastatic and early breast cancer. Full appreciation of its relevance will prevent replacement of cyclophosphamide in future trials or in daily practice. Funding: Netherlands Organization for Health Research and Development (ZonMw 80–82500-98-10901)

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P5-18-06.
P5-19-01: Final Results of a Three-Arm Randomised Phase II Study of Oral Vinorelbine Plus Capecitabine Versus Oral Vinorelbine and Capecitabine in Sequence Versus Docetaxel Plus Capecitabine in Patients with Metastatic Breast Cancer Previously Treated with Anthracyclines.
Authors: Campone M , Dobrovolskaya N , Tjulandin S , Chen S-C , Fourie S , Mefti F , Konstantinova M , Lefresne F , Meheust N , Jassem J
Published: June 2012

Purpose: Owing to the increasing number of patients treated with anthracycline-based adjuvant chemotherapy, there is a need for new effective and tolerable non-anthracycline based regimens in metastatic breast cancer.

Patients and methods: Patients with HER2−negative metastatic breast cancer previously treated with anthracyclines in (neo) adjuvant setting were randomised to fully oral 3-weekly cycles of the combination of oral vinorelbine with capecitabine (V+C), to the same drugs alternating every 3 cycles (V ⇔ C), or to the combination of docetaxel and capecitabine (D+C). V was given at 80 mg/m2 (after the first cycle at 60 mg/m2) on days 1 and 8 in the V+C arm and weekly in the V ⇔ C arm, C at 1,000 mg/m2 bid from days 1 to 14, and D on day 1 at 75 mg/m2. The primary endpoint was disease control rate (CR+PR+NC ≥ 3 months).

Results: A total of 139 patients were randomly assigned to V+C (44 patients), V ⇔ C (47 patients) and D+C (48 patients). After an independent review, the disease control rate in the intent-to-treat population in the V+C, V ⇔ and D+C arms [95% CI] was 70.5% [54.8−83.2], 37.0% [23.2−52.5] and 70.8% [55.9−83.1]. The response rate was 31.8% [18.6−47.6], 8.7% [2.4−20.8] and 35.4% [22.2−50.5], respectively. The median duration of progression-free survival in the V+C, V ⇔ C and D+C arms [95% CI] was 7.2 months [5.3−8.9], 3.4 months [2.6−5.6] and 8.9 months [7.2−12.0]; the median overall survival [95% CI] was 22.2 [18.8; 29.9], 19.4 [12.5; 35.4] and 24.2 [14.2; 38.5] months, respectively. Lower efficacy observed in the sequential arm could be due to higher prevalence of patients with visceral disease in this arm (91.3% in comparison to 65.9% in V+C and 64.6% in D+C arm). Combinations of V+C or D+C showed similar efficacy and a different toxicity profile; V+C induced less neutropenia, infection, hand-foot syndrome, fatigue/asthenia and alopecia, whereas D+C- less gastrointestinal toxicity.

Conclusions: V+C combination constitutes a valuable fully oral alternative option to D+C in patients with metastatic breast cancer previously treated with anthracyclines in (neo)adjuvant setting, while offering the advantages of an all-oral treatment.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P5-19-01.
P5-19-08: Pooled Analysis of Individual Patient Data from Capecitabine Monotherapy Clinical Trials in Anthracycline-/Taxane-Pretreated Locally Advanced or Metastatic Breast Cancer (LA/MBC).
Authors: Blum JL , Barrios CH , Feldman N , Verma S , McKenna E , Lee S , Scotto N , Gralow J
Published: June 2012

Background

Capecitabine (C) has shown efficacy as monotherapy in anthracycline (A)-/taxane (T)-pretreated LA/MBC in phase II/III trials. Since its approval, the treatment landscape in MBC has evolved, and C has served as the combination partner and/or active comparator in trials of novel agents in pretreated MBC. This pooled analysis of clinical trial data, spanning a 12-yr period, assessed the efficacy and safety of C across treatment lines and the impact of disease characteristics on clinical outcomes with C.

Methods: Individual patient (pt) data were pooled from 7 clinical trials conducted by Roche/GNE from Feb 1996-Jun 2008 where C monotherapy was the test/control regimen for pts with LA/MBC with/without prior A/T therapy. Analyses were performed on an ITT basis. Univariate and multivariate regression analyses assessed the impact of pt/disease characteristics and prior therapy on outcomes.

Results: Data from 805 pts enrolled to intermittent C monotherapy were analyzed: 268 in the 1st-line metastatic setting; 537 in the ≥2nd-line metastatic setting (table).

Median age: 55 yrs (range 23–90). Most pts were Caucasian (81%) with ECOG PS of 0 (55%). Baseline demographics were balanced across treatment lines. Data on A/T treatment history were available for 719 pts; 557 pts received prior A/T therapy, 176 were A-refractory, 234 T-refractory and 154 A- and T-refractory. Pooled overall response rate (ORR) was 21.0% (1.7% complete; 19.3% partial). ORR was 25.0% and 19.0% for pts treated in the 1st- and ≥2nd-line settings (OR 0.70, 95% CI: 0.5−1.0; p=0.0486). Median PFS was 126.9 days (95% CI: 119–132) in the pooled analysis, and significantly longer in pts receiving 1st- vs ≥2nd-line C: 150.0 vs 112.0 days (HR 1.45, 95% CI: 1.23−1.71; p<0.0001). Pooled median OS was 482.1 days (95% CI: 438–516). Significantly longer median OS was seen in pts receiving 1st- vs ≥2nd-line C: 666.0 vs 396.0 days (HR 1.98, 95% CI: 1.62−2.41; p<0.0001). 62 pts (8%) withdrew due to C-related AEs, which were severe in 4% of pts and life threatening in 2%. AEs of special interest occurred in 489 pts (63%); these were severe in 24% of pts and life threatening in 2%. Gastrointestinal disorders were most frequent (417 pts [53%]), and were severe or life threatening in 13% and 1% of pts.

Conclusions: This pooled analysis of pt data (n=805) revealed significantly longer ORR/survival in pts receiving 1st- vs ≥2nd-line C for A-/T-pretreated MBC. Univariate and multivariate analyses are ongoing; results will be presented. Safety data were in-line with findings from phase III trials of C in MBC.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P5-19-08.
P5-19-13: A Randomized Phase II Trial of First-Line Metastatic Breast Cancer (MBC) Patients: Sub-Set Analysis of Albumin-Bound Paclitaxel (ab-pac) Given Weekly at 150 mg/m2.
Authors: Gradishar WJ , Krasnojon D , Cheporov S , Makhson AN , Manikhas GM , Clawson A , Bhar P
Published: June 2012

Background

We previously reported the results of a phase II study evaluating the efficacy and safety of 3 different dosing regimens of ab-pac and docetaxel for the first-line treatment of MBC (Gradishar et al. J ClinOncol. 2009;27:3611). Here we report outcomes for a subset of patients (pts) during treatment with ab-pac at 150 mg/m2 weekly for the first 3 weeks of a 4-week schedule (qw 3/4).

Methods: Patients (N = 300) with previously untreated MBC were randomized to 1 of 4 treatment arms: arm A, ab-pac at 300 mg/m2 q3w; arm B, ab-pac at 100 mg/m2 qw 3/4; arm C, ab-pac at 150 mg/m2 qw 3/4; arm D, docetaxel at 100 mg/m2 q3w. A step-down statistical approach was used for pairwise comparisons of treatment groups. The trial was powered for antitumor activity and safety.

Results: Treatment arm C produced the longest overall survival (OS) (33.8 months) with an 11.6-month longer median OS vs arm B (22.2 months, HR 0.575; P = .008) and a 7.2-month longer median OS vs arm D (26.6 months, HR 0.688; P not statistically significant). Median OS in arm A was 27.7 months. These OS data were consistent with previously published overall response rates (ORR) and progression-free survival (PFS). Forty-seven percent of pts in arm C required dose reduction due to toxicity, including 27% due to neutropenia (np), 15% due to sensory neuropathy (SN), 3% due to allergy/immunology, 1% due to febrile np, and 1% due to ulceration of the skin. The median OS for the subset of pts requiring DRs in arm C was comparable to pts not dose reduced: 35.2 and 31.8, respectively. Pts who were dose reduced in arm C received a median of 2 additional cycles of treatment compared with those without DRs: 10 (range 2 — 27) vs 8 (range 1 — 27). Investigator assessed ORR and PFS were numerically higher in pts dose reduced vs those not reduced. Baseline characteristics were similar between pts requiring DRs vs not.

qw 3/4, first 3 out of 4 weeks; CI, confidence interval; ECOG PS, Eastern Cooperative Oncology Group performance status. aInvestigator assessed. bAll grade 3, no grade 4.

Conclusion: Pts in the ab-pac 150 mg/m2qw 3/4 arm who were dose reduced achieved a similar OS compared to patients who were not dose reduced. No clear trends in baseline characteristics emerged to predict the requirement for dose reduction in the 150 mg/m2 ab-pac arm. The ab-pac 150 mg/m2 qw 3/4 dosing regimen provided a survival advantage in this phase II trial in first-line MBC and dose reductions could be used to manage toxicities and prolong treatment duration without compromising efficacy.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P5-19-13.
P5-21-01: A Renewable Tissue Resource of Phenotypically Stable Human Breast Cancer Xenografts for Preclinical Studies.
Authors: Zhang X , Dobrolecki LE , Lai Q , Landis MD , Wong H , Tsimelzon A , Claerhout S , Contreras A , Gutierrez C , Huang J , Wu M-F , Pavlick AC , Froehlich AM , Hilsenbeck SG , Mills GB , Wiechmann L , Petrovic I , Rimawi MF , Schiff R , Chang JC , Lewis MT
Published: June 2012

Introduction Translational breast cancer research is hampered severely by difficulties in obtaining and studying primary human breast tissue, and by the lack of in vivo preclinical models that accurately reflect patient tumor biology. These limitations are due, in part, to the fact that traditional immunocompromised mouse models are not generally permissive for growth. We sought to circumvent some of these limitations by transplanting and growing human mammary tumors in the mammary fat pad of SCID/Beige immunocompromised mice in the absence of exogenous human fibroblasts.

Aims and Methods To establish a set of stable human breast cancer xenografts for preclinical studies. Human breast cancer biopsies were received, minced into small fragments and then transplanted directly into “cleared” fat pads of recipient SCID/Beige immunocompromised mice. Transplanted fat pads were checked weekly. After initial tumor was palpated and harvested, tumor fragments were transplanted into new SCID/Beige hosts for subsequent transplant generations. Serial immunohistochemical evaluations were performed to confirm human origin and biomarker status. Analytical flow cytometry for evaluating expression of proposed “cancer stem cell” markers, and gene and protein expression analysis were carried out on all stable lines.

Results and Conclusions Xenograft lines were established directly from breast cancer patient samples, without intervening culture in vitro, using the epithelium-free mammary fat pad as the transplantation site. Of the conditions tested, xenograft take rate was highest in the presence of a low-dose estradiol pellet without exogenous human fibroblasts. Thirty six stably transplantable xenograft lines representing 27 patients were established, using pre-treatment, mid-treatment, and/or post-treatment samples. Most patients yielding xenografts were “triple-negative” (ER-PR-HER2−) (n=21), we were able to establish lines from three ER-PR-HER2+ patients, one ER+PR+HER2−, one ER+PR-HER2−and one “triple-positive” (ER+PR+HER2+) patients. Serially passaged xenografts show phenotypic consistency with the tumor of origin at the histopathology level, and remarkable stability across multiple transplant generations at both the genomic, transcriptomic, and proteomic levels. Of 27 lines evaluated fully, thirteen xenografts showed metastasis to the mouse lung. These models thus serve as a renewable, quality-controlled tissue resource, and should prove useful for preclinical evaluation of experimental therapeutics.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P5-21-01.
P5-21-04: Patient Attitudes towards Undergoing Additional Breast Biopsy for Research Purposes.
Authors: Naim FM , Ballinger R , McLauchlan R , Hadjiminas DJ , Hogben K , Palmieri C , Cleator SJ
Published: June 2012

Background

Acquisition of additional breast tissue for research has become integral to breast oncology research but no data exists regarding patient attitudes to additional, non-diagnostic biopsies. Baseline research biopsies may be obtained as part of the diagnostic process as extra passage(s) of the needle through a ‘generic consent’ process, or as an additional biopsy event after carcinoma diagnosis has been confirmed and eligibility for a specific study has been established. This study sought to explore patient perspectives to such biopsies.

Patients and Methods

Patients attending breast clinics were recruited to complete a questionnaire prior to their consultation (and any interaction with researchers or clinicians seeking consent for research biopsies) which examined willingness to undergo an extra fine needle aspirate (FNA) and/or core needle biopsy (CNB) for research either at time of the diagnostic biopsy (B1) or after diagnosis (B2). Descriptions of FNA and CNB procedures were supplied to those with no prior experience. Patient perspectives towards donating surplus tissue left over from a diagnostic procedure and/or surgery for storage for future research was also explored.

Results: 100 patients were recruited, 42% with a prior history of breast carcinoma, 23% with family history of breast carcinoma and 65%/ 42% with previous experience of CNB/FNA respectively for either benign or malignant disease. Willingness to undergo additional biopsy was greater for the B1 than the B2 time point, but equivalent for CNB and FNA (willingness CNB B1 50% vs B2 26%, willingness FNA B1 50% vs B2 29%). A statistically significant increase in willingness to undergo CNB and FNA at B1 and/or B2 was seen in association with prior diagnosis of breast cancer, family history of breast cancer, previous visit to breast clinic and prior experience of breast biopsy. Reasons for willingness or unwillingness were recorded. 83% of patients expressed a willingness to allow surplus tissue to be stored in a biobank for future research.

Conclusions: When asked by questionnaire prior to clinic consultation, additional research biopsy was much more acceptable if undertaken at time of initial diagnostic biopsy rather than at a later time point. Acceptability was equivalent for CNB and FNA. Previous experience of biopsy, previous attendance to breast clinic, a prior history of breast cancer or family history of breast cancer increased acceptability suggesting that willingness is increased with knowledge of the biopsy procedure.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P5-21-04.
PD01-05: Histone Deacetylase Inhibitor LBH589 (Panobinostat) Suppresses the Activated-NFκB Pathway in Acquired Aromatase Inhibitor Resistant Breast Cancer Cells.
Authors: Kubo M , Kanaya N , Liu Z , Chen S
Published: June 2012

Background: Aromatase inhibitors (AIs) are effective in treating postmenopausal patients with estrogen receptor (ER)-positive breast cancer. However, resistance to endocrine therapies remains a major problem in the management of ER-positive breast cancer. Recently, histone deacetylase inhibitors (HDACis) show promise as cancer therapeutics, but the full scope of their utility remains unknown.

Experimental Design: In order to search for critical genes in developing resistance to AI, a microarray study was performed on AI resistant cells derived from MCF-7aro cells that overexpress aromatase and on AI resistant tumors from our mouse experiments. Moreover, using MCF-7aro cells that are resistant to anastrozole (Ana-R), letrozole (Let-R), and exemestane (Exe-R), as well as LTEDaro, we evaluated the significance of NFKB1 and several regulatory genes in AI resistance, and the anti-NFκB1 activity of HDACi LBH589 in vitro and in vivo.

Results: Significant changes (Fold change cutoff > 1.5, false discovery rate < 0.05) in the expression levels of 22 genes among three networks were identified from microarray analysis of our in vitro and in vivo models of acquired AI resistance. NFKB1 is a key player in one of three networks, including 11 genes, and is functionally up-regulated in all AI resistant cells. NFκB1 knockdown suppresses the proliferation of AI resistant cells more than that of MCF-7aro cells. Moreover, AI resistant cell lines are highly sensitive to LBH589 treatment and apoptosis is induced in these cell lines. LBH589 suppressed NFκB1 as demonstrated at both the mRNA and protein levels in LTEDaro and other AI resistant cell lines. Furthermore, the LBH589 treatment abrogated tumor growth in mice, and was associated with significant decreased levels of NFKB1 in tumors.

Conclusions: Our findings provide new insights into how the NFκB1-mediated networks play roles in AI resistance, and strongly support that LBH589 offers a novel therapeutic strategy for patients with AI resistant breast cancer.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr PD01-05.
PD07-04: Lapatinib vs Trastuzumab in Combination with Standard EC-D Chemotherapy in the Neaodjuvant Treatment of HER2+ Patients. Results from the GEICAM 2006–14 Phase II Randomized Trial.
Authors: Alba E , Albanell J , de la Haba J , Barnadas A , Calvo L , Sánchez P , Ramos M , Rojo F , Burgués O , Porras I , Tibau A , Carrasco E , Cámara MC , Lluch A
Published: June 2012

Background: The addition of trastuzumab (T) to neoadjuvant chemotherapy increases pCR rate in patients with HER2 + breast cancer. Lapatinib (L) in combination with chemotherapy is also an active drug in breast cancer patients. This study investigates the efficacy of trastuzumab or lapatinib added to chemotherapy in the neoadjuvant setting.

Methods: Patients (Pt) with tumors greater than 2 cm (or less with positive axilla) and HER2+ (Herceptest 3+ or 2+ with FISH+) that have not received any prior treatment for breast cancer were recruited. Pts were randomized to receive epirubicin 90 mg/m2 plus cyclophosphamide 600 mg/m2 × 4 cycles (cy) followed by docetaxel 100 mg/m2 (with growth colony stimulating factor support) plus either T 8 mg/kg loading dose and then 6 mg/Kg intravenous (EC-DT) or L 1250 mg orally (EC-DL). Patients were stratified according to tumor size (T1-2 vs T3-4), and estrogen receptor status (positive vs negative). The primary end-point was pathological complete response (pCR) in the breast measured by Miller and Payne criteria. Secondary end-points were clinical response (by imaging test), toxicity and correlation between pCR and tumor biomarkers.

Results: From February-09 to October-10, 102 pts were randomized (50 EC-DT, 52 EC-DL). Pts characteristics were well balanced between arms. Median age was 48 years (30-79), 56% of pts were premenopausal, 7% grade I, 45% grade III. 14/59/12/15% were T1/T2/T3/T4 and 69% N+. 58% were estrogen receptor positive and 44% progesterone receptor positive. Three patients were FISH negative after central assessment and were not considered evaluable for efficacy. pCR rate in the breast was 52% (95% CI: 38–66) for EC-DT and 25% (95% CI: 13.5−37.5) for EC-DL (p-value=0.0065). pCR—pN0 was 48% (95% CI: 34–62) for EC-DT and 24% (95% CI: 12 - 35) for EC-DL (p-value=0.01). Clinical response was 77% with EC-DT and 65% with EC-DL (p-value=0.176). Grade III-IV toxicity was similar between arms except for diarrhea that was more frequent in EC-DL 14% than in EC-DT 2% (p-value=0.03); more patients discontinue treatment due to toxicity with EC-DL (1 vs 6; p-value=0.055).

Conclusions: Our study shows that EC-DT was more efficacious and less toxic than EC-DL in the neoadjuvant treatment of HER2+ pts. Biomarker profiling in the tumors, including activation levels of tyrosine-kinase receptors, signaling pathways and surrogate markers (proliferation and apoptosis) is ongoing; their correlation with pCR will be presented at the meeting.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr PD07-04.
PD08-03: Inhibition of MEK/ERK- and JNK-Dependent Expression of Interleukin-6 and Interleukin-8 Targets Basal-Like Breast Cancer Stem Cells.
Authors: Balko JM , Cook RS , Kuba MG , Miller TW , Bhola NE , Sanders ME , Meszoely IM , Dowsett M , Gomez H , Arteaga CL
Published: June 2012

Background: Neoadjuvant chemotherapy (NAC) induces a pathological complete response in approximately 30% of triple-negative or basal-like breast cancers (BLBC). However, patients with residual disease often recur after surgery, presumably due to persistent drug-resistant subpopulations with cancer stem cell (CSC)-like properties. Thus, elimination of this CSC compartment in BLBC has the potential to improve survival by reducing post-surgical metastatic recurrences.

Methods: We sampled 49 post-NAC breast cancer samples including 22 BLBCs and quantified RNA for 355 cancer-related transcripts using Nanostring technology. Transcripts associated with a high Ki67 in the residual disease (a biomarker of early recurrence) were identified and bioinformatically examined for an association with drug resistance and a CSC phenotype. Loss of DUSP4, a negative feedback regulator of ERK1/2 and JNK1/2, was highly associated with a high post-NAC Ki67. We examined the role of loss of DUSP4 in promoting a drug-resistant, CSC phenotype.

Results: Low DUSP4 expression in post-NAC tumors was associated with high ERK1/2 activation and BLBC gene expression. siRNA knockdown of DUSP4 enhanced resistance to anti-cancer chemotherapy (docetaxel, camptothecin). Alternatively, forced DUSP4 expression in breast cancer cell lines abrogated the activation of transcription factors downstream of ERK and JNK and sensitized cells to docetaxel-induced apoptosis. In highly metastastic BLBC cell lines, MEK inhibition with AZD6244 and JNK inhibition with SP600125 significantly reduced mammosphere formation and self-renewal. Inhibition of JNK and MEK reduced IL6 and IL8 expression, two cytokines known to expand the CSC compartment. Reconstitution of exogenous IL6 and IL8 after MEK inhibition restored mammosphere formation potential. Chromatin immunoprecipitation demonstrated that the oncogenic transcription factor ETS-1, an ERK1/2 substrate, binds the IL8 promoter in a MEK1/2-dependent manner. We are currently confirming the association of DUSP4 loss with Ki67, ERK activation, and IL6/IL8 expression in a cohort of 113 post-NAC triple negative breast cancers. Conclusions: Our data demonstrate that loss of DUSP4 in BLBC promotes ERK and JNK activation by impairing negative feedback of these pathways. Activation of ERK and JNK drives expression of IL6 and IL8 expression, possibly through ETS-1 activation. Thus, targeting these signaling pathways may eliminate residual cancer stem cells after neoadjuvant chemotherapy and improve cure rates in BLBC.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr PD08-03.
PD09-03: Phase I/II Study of BKM120 in Combination with Trastuzumab in Patients with HER2 Overexpressing Metastatic Breast Cancer Resistant to Trastuzumab-Containing Therapy.
Authors: Saura C , Bendell J , Jerusalem G , Graña-Suárez B , Su S , Ru Q , De Buck S , Devisme C , Bosch A , Urruticoechea A , Beck JT , DiTomaso E , Rouyrre N , Sternberg DW , Massacesi C , Hirawat S , Dirix L , Baselga J
Published: June 2012

Introduction: HER2 amplification can activate the phosphatidylinositol-3-kinase (PI3K) pathway in breast cancer. Furthermore, trastuzumab (T) resistance can be associated with loss/deregulation of PTEN, which also causes activation of PI3K pathway signaling. BKM120, a potent orally bioavailable pan-class I PI3K inhibitor, could reverse resistance to T in vitro. Furthermore, administration of BKM120 with T demonstrated synergistic activity in preclinical models. The potential for combining BKM120 with T in heavily pretreated HER2+ metastatic breast cancer (MBC) patients (pts) will be determined in this study to substantiate the hypothesis for future development.

Methods: HER2+ MBC pts resistant to T-containing therapy (progression on or within 4 weeks since last T administration) received treatment in the dose-escalation portion of this multicenter trial. Prior anti-HER2−directed therapies were allowed. The primary objective was to determine the maximum tolerated dose (MTD) of BKM120 in combination with the standard dose of weekly T. A Bayesian logistic regression model with overdose control guided the dose escalation.

Results: Seventeen pts with MBC have been enrolled: 5 at 50 mg/day and 12 at 100 mg/day dose of BKM120. Escalation was conducted up to the MTD level of single agent BKM120, 100 mg/day. Patient characteristics: median age 46 years (range 34–70); most pts were heavily pretreated (reported range of prior chemotherapy lines 1–4). In preliminary results (cut-off date 1st June 2011), 7 pts discontinued treatment. BKM120 in combination with T showed a mean peak drug concentration and half-life similar to single-agent BKM120. T trough levels remained above the single agent efficacy threshold. Only one dose-limiting toxicity (G3 asthenia) was reported at the BKM120 dose of 100 mg/day. The MTD was determined to be 100 mg/day. Reported G3 adverse events were asthenia, altered mood, rash, GGT increase, hypokalemia, and hypersensitivity in 1pt each. No G4 toxicity has been observed so far. With regards to activity, 2 partial responses (1 confirmed) and 3 occurrences of disease stabilization have been observed.

Conclusions: BKM120 in combination with T in heavily pretreated HER2+ MBC pts with T-resistance has an acceptable safety profile and has shown encouraging preliminary activity. The Phase II portion of the study is ongoing. An updated analysis of efficacy, the potential correlation with PI3K pathway alteration status overlaying HER2 activation, and pharmacodynamics study will be presented.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr PD09-03.
PL1-1: How To Use Endocrine Therapy for Breast Cancer: Beyond the Basics.
Authors: Winer EP
Published: June 2012

Over two thirds of all breast cancers are hormone receptor positive. The majority of these tumors have some degree of sensitivity to endocrine treatments. The approach to endocrine therapy has become more complex over the past decade, and there are more treatment choices than before. Not only are there choices about which endocrine therapy to use, but also important decisions about how long to continue treatment and how to sequence agents. It has become clear that the benefits and risks of endocrine therapy vary both from tumor to tumor and from person to person. The landscape will soon become even more complicated as targeted therapies are combined with endocrine therapy to prevent or reverse drug resistance. This lecture will focus on a number of key issues: 1) Can we improve upon a 5 year course of tamoxifen in premenopausal women? 2) Given the high risk of late recurrence in many patients with hormone receptor positive disease, which patients will benefit from extended adjuvant therapy and what should this therapy be? 3) Is the use of an aromatase inhibitor alone in postmenopausal women sufficient therapy for all breast cancer patients and how should we select individuals who should be considered for other approaches? 4) In 2011, are there patients with hormone receptor positive disease who should not receive adjuvant endocrine therapy? 5) What approaches can be used to circumvent resistance to hormonal therapy, and how quickly will they be incorporated into clinical practice? The talk will address the dilemmas that are most vexing for clinicians and have immediate relevance for clinical practice.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr PL1-1.
S2-6: ErbB3 Expression Is Required for Maintenance of Normal and Transformed Luminal Breast Epithelial Cells.
Authors: Cook RS , Balko JM , Rinehart C , Miller TW , Polyak K , Prat A , Perou CM , Arteaga CL
Published: June 2012

The ErbB receptor tyrosine kinase family regulates breast development and cancer. ErbB2/HER2 expression correlates with HER2-enriched breast cancers, while epidermal growth factor receptor (EGFR) expression often correlates with triple negative breast cancers. Less is known regarding ErbB3, which harbors only weak kinase activity, but strongly activates phosphatidylinositol-3 kinase (PI3K)/Akt signaling upon heterodimerization with EGFR or ErbB2. We report herein that ERBB3 mRNA expression strongly correlates with Luminal A/B breast cancers, which express lower levels of EGFR and ErbB2. Mammary-specific loss of ErbB3 in mice reduced Akt phosphorylation and caused cell death in the luminal epithelium. A decreased luminal population in ErbB3-deficient epithelium correlated with expansion of the mammary stem/progenitor fraction. In normal breast samples, ERBB3 mRNA expression was highest in mature/progenitor luminal populations, and lowest in the stem/basal population. Loss of ErbB3 shifted gene expression in mammary epithelial cells to resemble a mammary stem cell signature. The genes most greatly impacted by ErbB3 loss produced a signature that correlated with decreased overall survival in tamoxifen-treated patients. Knock-down of ErbB3 in human luminal breast cancer cells caused decreased cell growth under conditions of estrogen deprivation, decreased expression of the gene encoding aromatase, and impaired expression of estrogen receptor-induced genes, including those that encode progesterone receptor and TFF1. Treatment of MCF7 luminal breast cancer xenografts with AMG-888, a humanized monoclonal antibody directed against ErbB3, decreased tumor growth. Combination of AMG-888 with the estrogen receptor inhibitor fulvestrant caused MCF7 tumor regression. Taken together, these results suggest that ErbB3 is required to sustain normal and transformed luminal epithelial cells of the breast, and suggest that targeting ErbB3 may improve the clinical outcome of breast cancers treated with endocrine therapy.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr S2-6.
SR1-2: Breast Cancer and the Environment: A Life Course Approach - Report Release from an IOM Committee.
Authors: Hiatt RA
Published: June 2012

In response to a request from Susan G. Komen for the Cure®, an Institute of Medicine (IOM) committee was convened to review and assess the strength of the science base regarding the relationship between breast cancer and the environment. As part of their charge, the committee members reviewed the evidentiary standards for identifying and measuring cancer risk factors and the methodological challenges involved in conducting research on breast cancer and the environment. They also considered the potential interaction between genetic and environmental risk factors. The committee's report notes evidence-based actions that women could take to reduce their risk of breast cancer and offers recommendations for future research on breast cancer and the environment. The results of this IOM committee's work will be presented.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr SR1-2.
SR2-1: The Genomic Mutation Landscape of Breast Cancer: A TCGA Report.
Authors: Mardis ER
Published: June 2012

An important goal, that of characterizing the breadth and complexity of mutations in breast cancer, is being approached by The Cancer Genome Atlas in its next-generation sequencing and analysis of the exome and whole genome content of breast cancer samples. At present, we have produced, analyzed, and validated mutations by exome capture of matched tumor and normal genomic DNA across all breast cancer subtypes, using over 500 samples as our primary discovery set. The resulting information has been further analyzed by integration with other data types being generated in the project, including RNA-seq, methylation and SNP array data. We have further initiated a study using whole genome sequencing of 21 basal subtype cases. My presentation will provide an overview of the methods of data generation, analysis and integration. I will further provide information about our discoveries to-date and future plans.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr SR2-1.