bluuebunny/crossref_metadata_2025 huggingface dataset:
P3-13-08: Survival Analysis and Recurrence Patterns in Locally Advanced Breast Cancer Following Neoadjuvant Chemotherapy and Preoperative Concurrent Chemo Radiotherapy.
Authors: Shaw RJ , Lara F , Robles CD , Vilar D
Published: June 2012

Introduction: Although not widely used, preoperative concurrent chemo radiotherapy (CCRth) following poor response to neo adjuvant chemotherapy (CT) has been recently associated with an improved prognosis in patients with locally advanced breast cancer (LABC) and high recurrence risk, showing acceptable toxicity profiles, acceptable post surgical complication rates and increases in both clinical and pathologic response rates and local control rates. The objective of this study was to assess disease free survival (DFS), overall survival (OS) and patterns of recurrence in a cohort of patients with LABC treated with pre operative CCRth following poor response to neo adjuvant CT.

Methods: This longitudinal study included a total of 287 patients with LABC treated with pre operative CCRth following poor response to neo adjuvant CT between February 2000 and December 2002. Demographic, clinical/pathologic, therapeutic and follow up data were collected from patient charts to determine DFS, OS and recurrence patterns.

Results: Stages IIB, IIIA and IIIB accounted for 22.3, 37.6 and 40% of the population, respectively. Clinical response rates compared to pathologic response rates following anthracyline based CT and CCRth were minimal or stable in 16% vs 62.4%, partial in 53 vs 76.3% and complete in 30 vs 23.3%. No significant treatment related toxicities were identified. Post surgical complications included flap ischemia in 16%, partial flap necrosis in 18%, both ischemia and partial flap necrosis in 13.2% and partial wound dehiscence in 7.7%. With an average follow up of 64.1 ± 37.2 months, (0-126.3 months), 36.9% of the population relapsed and 55.1% remained disease free. With regard to recurrence patterns, 22.6% presented loco regional relapse, 67.7% recurred at a distant site and 9.8% showed presented both. Median OS was 64.2 months (0-126 months). Five and ten year survival probability was shown to be 75% and 60%, respectively.

Conclusions Despite improvements in local control rates and overall outcomes with current therapeutic regimens combining chemotherapy, surgery, and radiotherapy, 30 to 40% of patients with locally advanced breast cancer develop post treatment loco regional recurrence and 5 year overall survival remains low (50%). Multimodal treatment integrating pre operative CCRth is therefore a valid alternative in the LABC setting, showing promising results in regard to OS, DFS and recurrence rates. Prospective clinical studies to evaluate its use are warranted.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-13-08.
P3-13-11: Survival Benefit Associated with Post-Mastectomy Radiation for Node-Positive, Intermediate-Risk Breast Cancer Patients: Results of a Study from the SEER Registry Data.
Authors: Horowitz DP , Ng J , Burri RJ
Published: June 2012

Background

The role of post-mastectomy radiation therapy (PMRT) for breast cancer patients with intermediate-risk (1-3 positive nodes) disease remains controversial. This Surveillance, Epidemiology, and End Results (SEER) study investigated the impact of PMRT in the intermediate-risk breast cancer patient population on cause-specific survival (CSS) and overall survival (OS). The study also investigated the impact of estrogen receptor (ER) status, which has been reported in the SEER database since 2004, on the impact of PMRT in this patient population on CSS and OS.

Methods: The SEER database was used to identify all breast cancer patients who were treated between 2004–2007, who had undergone mastectomy, and who had T1-2 tumors and 1–3 pathologically positive lymph nodes. The status of radiation therapy, ethnicity, tumor grade, TNM stage, year of diagnosis, number of LNs sampled, number of LNs positive, and estrogen/progesterone receptor status was recorded. Patients missing pathologic or treatment-related information were excluded. Statistical analysis for CSS and OS was performed using the Kaplan-Meier method and analyzed using the log-rank test. Multivariable analysis was performed using the Cox proportional hazards regression model. Statistical analyses were performed using PASW, version 18.

Results: 10,517 patients were identified, with a median follow-up of 21.8 months (range 0–47 months). 2339 patients (22.2%) received PMRT. Younger patients, increasing tumor size, increasing tumor grade, and ER negative tumors were more likely to receive PMRT. PMRT was associated with improved OS, 92.8% (95% CI 91.294.4%) versus 88.7% (95% CI 87.7−89.7%), p < 0.001. However, there was no overall CSS benefit to PMRT, p = 0.197. In the subgroup with ER positive tumors, PMRT was associated with both improved OS, 95.5% (95% CI 93.9−97.1%) versus 91.0% (95% CI 90.0−92.0%), p < 0.001, and improved CSS, 97.6% (95% CI 96.6−98.8%) versus 96.1% (95% CI 95.3−96.9%), p = 0.009. Multivariate analysis showed that PMRT was significantly associated with improved survival (HR 0.744, p = 0.011); increasing age, tumor grade, ER negative tumors, increasing tumor stage, and increasing positive nodal ratio were associated with increased risk of death.

Conclusion: Patients with intermediate risk breast cancer who received PMRT have improved OS compared to those who did not receive PMRT. For patients with ER positive tumors, PMRT also appears to be associated with improved CSS. Greater consideration for post-mastectomy radiation therapy use may be warranted for this patient population.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-13-11.
P3-14-02: Sequential Versus Upfront Intensified Neoadjuvant Chemotherapy in Patients with Large Resectable or Locally Advanced Breast Cancer (INTENS), Toxicity Results from a Phase III Study of the Dutch Breast Cancer Trialists' Group (BOOG).
Authors: Vriens BE , Van de Vijver KK , Boetes C , van Gastel SM , Wals J , Smilde TJ , van Warmerdam LJ , van Laarhoven HW , van Spronsen DJ , Borm GF , Tjan-Heijnen VC
Published: June 2012

Background Taxanes have an established role as (neo-)adjuvant treatment of breast cancer. In the present study, we compared 4 AC - 4 T with 6 cycles of TAC in the neo-adjuvant setting (A=adriamycine, C=cyclophosphamide, T=docetaxel). Previously, we reported that AC-T resulted in a trend for improved outcome (odds ratio pCR of the breast 1.61; 95% CI 0.79−3.33). Now we report the safety data.

Methods Women presenting with breast cancer, cT2≥3cm, cT3, cT4 and/or cN+, with measurable disease and no prior treatment, age ≥18 and ≤70 years and Karnofsky Score ≥70% were eligible. Patients were randomized to AC (60/600 mg/m2 q3wk x 4 cycles) followed by T (100 mg/m2 q3wk x 4 cycles) without primary G-CSF prophylaxis, or to TAC (75/50/500 mg/m2 q3wk x 6 cycles) with primary G-CSF prophylaxis. If indicated, trastuzumab and/or endocrine therapy were given as adjuvant treatment. This present analysis focuses on the toxicity profile of the two treatment arms.

Results In total, 201 patients (n=100 AC-T, n=101 TAC) were included between February 2006 and April 2009. Baseline characteristics (AC-T/TAC) were well balanced. Patients in the AC-T arm had more frequently grade 3 / 4 toxicities as compared to the TAC arm, respectively in 57% and 28% (p=0.001). Grade 3 / 4 neutropenia without fever was more frequently reported with AC-T (35% vs. 4%; p=0.001). Grade 3 / 4 febrile neutropenia was also more frequent with AC-T (17% versus 5%; p=0.0062) and significantly increased during docetaxel treatment after AC. Notably, diarrhea was also more frequently seen in the AC-T arm (4% versus 0%, p=0.0423). Other grade 3 / 4 toxicities more frequently reported in the AC-T arm were neuropathy - sensory (5% vs. 0%; p=0.229) and pain other than muscle or bone pain (4% vs. 0%; p=0.0423). There were no grade 3 / 4 toxicities more frequently observed in the TAC arm.

Conclusion In the comparison of two taxane-anthracycline-cyclophosphamide regimens in the neo-adjuvant setting, it is observed that the sequential approach with a lower cumulative dose tends to have a slightly better efficacy outcome, but at the cost of increased grade 3 / 4 toxicity. However, considering the use of primary G-CSF prophylaxis in the TAC arm, and the fact that the incidence of febrile neutropenia was higher during taxane containing chemotherapy in the AC-T arm, the difference might (partly) disappear if primary G-CSF prophylaxis would be used in the sequential arm. Primary G-CSF prophylaxis may be considered during docetaxel if used sequentially after anthracycline-containing chemotherapy.

Support: Unrestricted grants from sanofi-aventis NL BV and Amgen BV.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-14-02.
P3-14-09: A Phase II Preoperative Study of Dasatinib, a Multi-Targeted Tyrosine Kinase Inhibitor, in Locally Advanced “Triple-Negative” Breast Cancer Patients.
Authors: Rimawi MF , Rodriguez AA , Yang WT , Gonzalez-Angulo AM , Nangia JR , Wang T , Speers C , Mills G , Hilsenbeck SG , Brown PH , Chang JC
Published: June 2012

Background: We previously reported that kinases (Src, Yes-1, cKIT, Abl, and EPH4) were druggable in triple negative breast cancer (TNBC). In this clinical trial, we sought to translate these findings by treating TNBC patients with dasatinib, a multi-targeted kinase inhibitor against these targets.

Methods: Women with stage II-III TNBC were eligible. Patients received dasatinib at 100 mg daily for 3 to 4 weeks before standard-of-care definitive surgery and chemotherapy. Biopsies were performed at baseline, week 1, and at the time of surgery. A cohort of patients had positron emission mammography (PEM; baseline and at 2–3 weeks of dasatinib therapy). This study was designed to detect an increase in clinical response rate from 10% to 25%, using a Simon optimal two stage design, with one-sided alpha=5% and power=80%. At least 3 responses out of 22 patients were needed to proceed to the second stage.

Results: 22 patients were enrolled (Table 1). Median tumor size was 7.0 cm (range 2.4-25 cm). Adverse events were modest, mainly grade 1–2 (headache: 45%, abnormal LFTs: 55%, GI: 23%, fatigue: 18%). One patient had a myocardial infarction 24 hours after starting dasatinib. Out of 22 patients, 2 (9%) had a clinical partial response after 3–4 weeks of therapy, 15 had stable disease (68%), while 5 had progressive disease (23%). Of the 8 patients who received paired PEM imaging, metabolic responses were observed in 2 patients (25%). Conclusion: A short course of dasatinib led to clinical responses in 2 out of 22 patients with TNBC, and the study did not proceed to second stage. Since TNBC is a heterogeneous disease, biomarker studies including sequencing of candidate genes like B-RAF for inactivating mutations might enable selection of those TNBC patients who could benefit from dasatinib.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-14-09.
P3-14-12: Local Control of Primary Breast Cancer Treated with Radical Radiotherapy Alone after Neoadjuvant Chemotherapy.
Authors: Makris A , Li SP , Ravichandran D , Ostler PJ , Pittam M
Published: June 2012

Introduction: The aim of this retrospective study was to evaluate the local recurrence rates in a cohort of patients who achieved a complete clinical response (cCR) to neoadjuvant chemotherapy and did not have surgery.

Materials and Methods: 148 women who achieved a cCR to neoadjuvant chemotherapy were identified from a prospectively maintained database (1995-2011) of 667 patients. 122 patients went on to have surgery (either wide local excision or mastectomy) followed by radiotherapy. In 26 patients (median age 49, range 35–72 years; T2-T4, N0-N3, M0), no surgery was performed but all received radical radiotherapy. Surgery was avoided due to either physician or patient choice. Recurrence was defined as first relapse of disease, either local (ipsilateral breast and/or axilla) or distant.

Results: All 26 patients who avoided surgery had neoadjuvant chemotherapy with 20 patients (77%) receiving anthracycline-based (FEC, FAC, ECF), 5 (19%) MMM and 1 (4%) CMF chemotherapy. The median number of cycles was 6 (range 4–8). Chemotherapy was followed by radical external beam radiotherapy to the breast +/− supraclavicular fossa and axilla (median dose delivered, 60Gy in 2Gy fractions). All were identified as operable at diagnosis including 3 patients who had supraclavicular lymphadenopathy. All 26 patients achieved a final cCR in the breast to chemotherapy. 21 patients had imaging with mammography and/or ultrasonography to assess radiological response at the end of neoadjuvant chemotherapy, of which 19 had a complete response and 2, a partial response. After a median follow-up of 144 months, 10/26 (38%) patients experienced local disease recurrence (2 also had distant recurrence) and 4/26 (15%) patients with distant metastases only. Patients with local recurrence only, went on to have a mastectomy whilst those with distant disease received systemic therapy. There were 10 deaths, 9 of which were breast cancer related (33%).

Conclusions: Local recurrence rates were high in patients achieving a cCR following neoadjuvant chemotherapy and who avoided surgery. Our practice has subsequently changed to include clip insertion and surgical excision on completion of chemotherapy. With increasing pathologic complete response rates to more active chemotherapy schedules (including taxanes +/− herceptin), it is being proposed that surgery could be avoided in selected patients. Our study shows that caution should be exercised.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-14-12.
P3-14-17: Paclitaxel, Carboplatin, and Trastuzumab in a Neoadjuvant Regimen for HER2−Positive Breast Cancer: The TRAIN Study.
Authors: Sonke GS , Mandjes IA , Holtkamp M , Schot M , Oosterkamp HM , Wesseling J , Vrancken Peeters M-JT , Rodenhuis S , Linn SC
Published: June 2012

Background: Treatment with trastuzumab is highly active in HER2 positive breast cancer, although cardiotoxicity is a well known side effect. The cardiotoxicity of trastuzumab may be aggravated by combined treatment with anthracyclines. Consequently, treatment with trastuzumab is often delayed pending the administration of anthracyclines. Both in vitro and in vivo data suggest that trastuzumab synergizes with a range of chemotherapeutic drugs, including taxoid drugs and carboplatin. In addition, prolonged pre-operative treatment leads to higher pathologic complete remission (pCR) rates. We report the results of a fase 2 trial integrating trastuzumab at the start of a non-anthracycline containing weekly paclitaxel-carboplatin based neo-adjuvant chemotherapy regimen in HER2−positive breast cancer. Patients and methods: One-hundred patients with stage II or III breast cancer, including inflammatory disease, with HER2 overexpression (immunohistochemistry and/or in situ hybridization) were treated with 24 weekly administrations of paclitaxel 70 mg/m2, carboplatin AUC=3 mg.ml-1.min, and trastuzumab 2 mg/kg (loading dose 4 mg/kg). In cycles 7, 8, 15, 16, 23, and 24 only trastuzumab was given. The primary end point was pathologic complete response (pCR) in both breast and axilla. The trial was preceded by an initial pilot cohort of 55 patients treated with the same regimen.

Results: Final efficacy and safety results of all patients included in the phase 2 trial will be reported at the meeting. In the pilot cohort of 55 similarly treated patients, 33% had stage II disease, 69% was clinically node positive, and 49% was ER and PgR negative. Twenty-four patients (41%) had a pCR in breast and lymph nodes. pCR in ER negative patients was 67%, pCR in ER positive patients was 21%. The most commonly reported grade 3/4 toxicities were neutropenia (30%) and thrombocytopenia (27%). Dose reduction was required in 24% of the patients. Grade 3/4 left ventricular systolic dysfunction was not observed.

Conclusion: A weekly carboplatin-paclitaxel-trastuzumab neo-adjuvant regimen is highly active in HER2 positive breast cancer with a good safety profile. A subsequent multicenter phase 3 trial will compare this regimen to a similar 16 week regimen preceded by 4 cycles of anthracyclines plus cyclophosphamide.

The study protocol was developed at the joint ECCO-AACR-EORTC-ESMO Workshop on Methods in Clinical Cancer Research in Flims, Switserland.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-14-17.
P4-01-16: The Influence of CYP2D6 Genetic Polymorphisms on Variability of Tamoxifen Metabolism in the Lebanese Breast Cancer Population.
Authors: Tfayli A , Zgheib N , Holail J , Habbal M-Z , Halawi R , El-Saghir N , Salem Z
Published: June 2012

Introduction The selective estrogen receptor modulator tamoxifen is commonly used for the treatment and prevention of breast cancer. Tamoxifen needs to be converted by the highly polymorphic enzyme, CYP2D6, to its active metabolites (4-hydroxytamoxifen and endoxifen) to exert its anti-estrogenic effect. Several clinical trials have shown that CYP2D6 poor metabolizers have decreased benefit from tamoxifen compared to extensive metabolizers. Large ethnic variations have been noticed in the frequency and effect of CYP2D6 polymorphisms.

Objectives To assess for the frequency of CYP2D6*3, *4, *5 (nonfunctional alleles with *5 being a gene deletion), *41 (reduced functional allele) as well as CYP2D6*x N (allele copy number) in a sample of Lebanese participants receiving maintenance doses of tamoxifen for early stage breast cancer, and to assess the effect of these polymorphisms on tamoxifen metabolism in the same subjects.

Methods 111 (median age= 48.7± 8.6years) Lebanese breast cancer women were recruited. Blood for isolation of DNA and measurement of tamoxifen metabolites was collected. CYP2D6 genotyping was performed using real-time PCR. Plasma levels of tamoxifen were being measured using HPLC.

Results The allele frequencies of CYP2D6*3, *4, and *41 were 0.9, 15.9 and 12.1 respectively. 18.9% of patients had more than 2 CYP2D6 copies and 9% had only 1 copy. Tamoxifen plasma concentration didn't significantly vary among the 4 phenotypic groups, although a trend was observed whereby PMs had the highest tamoxifen plasma concentrations and UMs had the lowest tamoxifen plasma concentrations (Tamoxifen median plasma concentrations in μg/L: PM=290, IM=210, EM=190 and UM=170). Furthermore, there was a statistically significant variation of Z-tamoxifen plasma concentrations among the phenotypic groups when PMs and IMs were grouped together: Z-tamoxifen median plasma concentrations in μg/L: PM+IM=130, EM=90 and UM=62 (P=0.03).

Conclusion This is the first study in the Middle East that evaluates the frequencies of CYP2D6 polymorhisms in a Lebanese breast cancer population and the effect of these polymorphisms on tamoxifen metabolism. We have shown that CYP2D6 phenotype is significantly correlated with Z-tamoxifen levels. PMs and IM group had the highest Z-tamoxifen plasma concentrations; in contrast UMs had the lowest Z-tamoxifen plasma concentrations. Further recruitment and follow up are being performed to assess for the role of these polymorphisms in disease outcome.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-01-16.
P4-01-20: Specific Anti-Proliferative Profile of Endoxifen for MCF-7 Cell Compared to 4-OH Tamoxifen.
Authors: Saji S , Hirose M , Hayashi S-I , Kuroi K
Published: June 2012

Background: Endoxifen and 4-hydroxy tamoxifen are both active metabolites of tamoxifen and exert anti-tumor activity for ER+ breast cancer. However, only small numbers of paper reported biological profile of endoxifen.

Methods: Cells were incubated with various concentration of endoxifen ( (Z)-4-Hydroxy-N-desmethyl Tamoxifen (<10% with (E)-form)) for 6 days and viability (anti-proliferative index) was assessed by cell counting kit-8. For seeking the influence of ERβ, two independent ERβ expressing clones of MCF-7 cells were established by stable transfection of ERβ vesctor to MCF-7. Western blotting was done with ERα, ERβ, PgR and HER2 antibodies.

Results: IC50s of endoxifen for MCF-7 were 100 nM in estradiol (E2) deprivation condition and 500 nM in the presence of 1nM E2. These are significantly higher than those of 4-OH tamoxifen, which are 10 nM without E2 and 50 nM with E2, respectively. From the previous reports, plasma endoxifen concentrations in breast cancer patients were ranged from 20 to 200 nM according to CYP2D6 phenotype. Anti-proliferative index of 20 nM endoxifen for MCF-7 was 0.7 (indicating 30% reduction in proliferative activity), and 200 nM endoxifen was 0.5 in the absence of E2. These were 0.95 (20 nM) and 0.75 (200nM), respectively, when incubated with 1 nM E2. 10 nM 4-OH tamoxifen, which is mean concentration in patient plasma, showed about 0.6 anti-proliferative index with no regards to E2 condition. Without E2, endoxifen reduced ERα expression as previously reported, however, endoxifen increased ERα amount in the presence of E2 accompanied with decrease of HER-2 expression. This alternation of ERα and HER2 expression were identical to those observed using 4-OH tamoxifen. There was a weak tendency that ERβ expressing MCF-7 clones showed favorable response to endoxifen compared to parent MCF-7, but this was not significant.

Conclusion: Endoxifen is active agent for MCF-7 cell, however, it requires 10-fold higher concentration compared to 4-OH tamoxifen to achieve IC 50. Difference of anti-proliferative effect between 20 nM and 200 nM of endoxifen are relatively small, and it might result in minor difference for clinical response, as expected from recent clinical studies of CYP2D6. This is the first time to show that endoxifen has opposite effect to ERα protein level depending on the presence or absence of E2. ERβ expression did not enhance activity of endoxifen in contrast to previous report.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-01-20.
P4-01-22: Cyclin D1 and Its Prognostic Value in Planning of Endocrine Therapy for Women of Postmenopausal Age with Breast Cancer.
Authors: Skvortsov V , Manikhas A , Manikhas G , Oganesyan A , Raskin G
Published: June 2012

Background: Nowadays tamoxifen still remains the primary drug in breast cancer endocrine therapy. However, its application is limited due to the resistance of tumor cells. The search of adequate biomarkers is one of the most actual problems in prognosis of effectiveness of tamoxifen adjuvant therapy. The most perspective biomarker is cell cycle regulator cyclin D. Besides its central role in cell cycle regulation cyclin D1 modulates the estrogen receptors activity and can influence on the effectiveness of treatment with antiestrogenes and aromatase inhibitors.

The objective of our work was to evaluate the effectiveness of tamoxifen in adjuvant therapy of hormone-receptor-positive breast cancer in women of postmenopausal age with cyclin D1 expression.

Material and Methods: To evaluate the effectiveness of tamoxifen adjuvant application in women with cyclin D1-positive T1-4N0-3M0 breast cancer we have researched 2 retrospective groups of 70 patients that have been on regular medical check-up for a period of 5 years or that had previously undergone treatment. On the basis of the archive histological material we have revealed cyclin D1 in tumor cells.

Results: The expression of cyclin was classified in 4 levels: negative, low, moderate and high. Patients with lack of cyclin D1 expression or with low quantitative value (according to our data less than 30%) have no neoplastic process progression throughout the 5 years of tamoxifen adjuvant therapy. These women were both with axillary lymph nodes affection and without it (+N; -N). On the contrary, women with moderate and high cyclin D1 expression (more than 30%) had a relapse of tumor and distant metastasis is prognosed in 5 years of observation (p=0,005). Moderate level of cyclin D1 expression was observed in 45 (64%) patients and 28 (62%) of them had progression with metastasis in bones, 9 (20%) metastasis in soft tissues, 2 (4%) metastasis in lungs and 6 (14%) relapse in postoperative scar. High expression was revealed in 25 (35%) women and 17 (68 %) of them had bone affection, 7 (28%) soft tissue metastasis and 2 (4%) tumor relapse in postoperative scar. Lung metastasis were not observed. The average period of tumor relapse and progression of neoplastic process in patients with cyclin D1-positive breast cancer is 20 months. Discussion: The results of our work suggest that women with hormone-receptor-positive cyclin D1-negative breast cancer on early stages have more prolonged non-relapse period during the tamoxifen adjuvant therapy. Patients with cyclin D1-positive breast cancer are less sensitive to tamoxifen treatment and in adjuvant regime should receive therapy with other effective equivalent drugs (aromatase inhibitors).

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-01-22.
P4-02-09: Estrogen-Related Receptor a Mediates Insulin-Like Growth Factor-I Dependent Migration in Breast Cancer Cells.
Authors: Fettig-Anderson LM , Oh AS , Yee D
Published: June 2012

The orphan nuclear estrogen-related receptors (ERR) have homology to steroid hormone receptors. Unlike classical steroid hormone receptors, ERRs do not have known ligands but are transcriptionally activated upon binding to nuclear receptor coregulators such as PGC1α, PGC1β, and PPRC1. ERRs regulate energy metabolism and are accordingly expressed in tissues that require high energy production. ERRs recognize their own ERRE (estrogen-related receptor response element) gene promoters, but can also activate transcription via ERE (estrogen receptor response element) promoters, representing the ability of ERRs to cross participate in ERα regulatory pathways in the absence of estrogen. Of the three family members (α,β,γ), ERRα is the most highly expressed in breast tissue. ERRα expression was found to be increased in ERα negative breast cancers and associated with poor prognoses. ERRα and its coactivators can increase the expression of a critical mediator of breast tumor angiogenesis and metastasis, VEGFA. We have previously shown that VEGFA is regulated by insulin and IGF receptor activation and hypothesized that ERRα may be a mediator of this finding. We therefore explored whether ERRα and its coactivators are necessary for IGF-I/insulin dependent regulation of VEGFA and cell migration. ERRα was expressed in breast cancer cell lines, but no correlation was observed with breast cancer subtypes. Reducing endogenous ERRα levels by shRNA reduced IGF-I dependent migration in MDA-MB-231 cells. These results were confirmed using an ERRα specific inverse agonist (XCT-790), which also inhibited MDA-MB-231 cell migration. Modulating ERRα mRNA levels via shRNA resulted in decreased VEGFA mRNA levels in MCF7L cells and increased VEGFA mRNA levels in MDA-MB-231 cells, while the inverse agonist XCT-790 caused an increase in VEGFA mRNA levels in both cell lines. Reducing ERRa levels did not affect the expression of the IGF1R, InsR, IRS-1, IRS-2, PI-3K-Akt-mTOR dependent signaling pathway. Neither ERRα protein nor mRNA levels were modulated by IGF-I or insulin. We conclude that ERRα is important for mediating IGF-I dependent migration, but have shown that ERRα is not directly regulated by the IGF signaling. We will next explore the role of ERRα's coregulators (PGC1α, PGC1β, and PPRC1) in the regulation of VEGFA by the IGF/insulin pathway given their central role in regulating ERRα's function.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-02-09.
P4-02-10: The Relationship between Estrogen Receptor Gene Polymorphism and Mammographic Density in Postmenopausal Women.
Authors: Baldisserotto FDG , Elias S , Silva IDCG , Nazario ACP
Published: June 2012

Background Assess the relation between the presence of PVUII and XBAI polymorphisms in the estrogen receptor alpha gene and mammographic density in postmenopausal women.

Methods For the present analysis, 189 postmenopausal women who had never used hormonal therapy and who did not have clinical or mammographic features were selected. Based on the ACR-BIRADSâ 2003 classification, the mammographic density was determined by three independent readers (two subjective ratings and one computerized - Adobe Photoshop â 7.0 software). Blood samples were available to extract DNA according to KIT GFX â protocol. PCR-RFLP (Polymerase Chain Reaction - Restriction Fragment Length Polymorphism) was then used to identify the polymorphisms.

Results There was a high degree of agreement among the three readers to determine the mammographic density (Kappa>0.75). Sixty women (32%) had dense breasts and 129 (68%) had non-dense breasts. The PVUII polymorphism was found in 132 (69.8%) of 189 women, while the XBAI was found in 135 (71.4%) of women. Parity (p=0.02) and body mass index (p<0.0001) were associated with mammographic density. It was observed that for the XBAI polymorphism, women with two mutated alleles were approximately 2.5 times more likely to be classified in dense breasts group (p=0.003) and the presence of both wild alleles was associated with fibroglandular tissue replacement by fat (p=0.02).

Conclusions There was no significant association of the PVUII polymorphism in the estrogen receptor alpha gene with mammographic density (p=0.34). However, the XBAI polymorphism was observed at a higher mutated homozygous frequency in women with dense breasts and there was an increased frequency of wild-type homozygous and heterozygous women with fat-replaced breasts (p=0.01).

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-02-10.
P4-07-07: Circulating Tumor Cells Predict Survival in Non-Metastatic Breast Cancer.
Authors: Lucci A , Krishnamurthy S , Bhattacharyya A , Lodhi A , Hall C , Singh B , Anderson A , Bedrosian I , Kuerer H
Published: June 2012

Background: Circulating tumor cells (CTCs) predict outcome in metastatic breast cancer (BC), but their prognostic significance in non-metastatic BC is unclear. This study determined whether CTCs could be identified in non-metastatic BC patients and 2) if CTCs predict relapse-free and overall survival.

Methods: Clinical stage I-III BC patients seen at a single tertiary cancer center provided informed consent to participate in an IRB-approved study involving collection of blood (7.5 ml x 2 tubes) before systemic therapy and at the time of surgery for their primary BC. CTCs were detected using the Cell SearchTM system. A positive result was defined as the presence of two or more cells per 7.5 ml blood since a threshold for positivity has not been established in primary BC. Statistical analyses were done using STATA-IC 11 software.

Results: We prospectively evaluated 291 patients. Mean age was 54 years, and median follow-up was 30 months. 54% (157) had T1 tumors, 36%(104) T2, 6% (18) T3, and 4%(12) had T4 disease; 37% (107/289) were clinically node-positive (LNs) by axillary ultrasound and FNA. Two or more CTCs were identified in 10% (29) patients. Seventy-seven percent (225) patients were hormone receptor positive, 11% (32) were HER2 positive while 16% (48) expressed no receptors. Sixteen percent (48) were tumor grade 1, 50% (143) were grade 2 and 34% (98) were grade 3. Systemic chemotherapy and/or endocrine therapy were administered in 80% (233/289) of patients. Sixty-eight percent (167/244) of patients were post-menopausal. Additionally, there was no significant correlation between CTCs with primary tumor size, lymph node status, estrogen or progesterone receptor status, HER2 amplification or tumor grade. Thirty-eight percent (6/16) of all relapses occurred in patients with 2 or more CTCs (HR: 4.48 (95% C.I. 1.61−12.45), Logrank P 0.002) while 40% (4/10) of all deaths occurred in patients with 2 or more CTCs (HR: 4.54 (95% C.I. 1.27−16.25), Logrank P 0.011).

Conclusions: CTCs were a significant predictor of disease-free and overall survival in non-metastatic breast cancer.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-07-07.
P4-07-18: Prospective Assessment of Circulating Tumor Cells and Serum Markers for PFS Prediction in Metastatic Breast Cancer.
Authors: Bidard F-C , Hajage D , Bachelot T , Delaloge S , Brain E , Campone M , Cottu P , Beuzeboc P , Mathiot C , Pierga J-Y
Published: June 2012

Purpose: Circulating tumors cells (CTC) have been recently proposed as a new dynamic blood marker whose positivity at baseline is a prognostic factor and whose changes under treatment are correlated with progression-free survival (PFS) in metastatic breast cancer patients. However, serum markers levels are also used for the same purpose, and no clear comparison as been reported to date.

Patients and methods: The IC 2006–04 enrolled prospectively 267 metastatic breast cancer patients treated by first line chemotherapy and confirmed that CTC levels are an independent prognostic factor for PFS and Overall survival (OS). A pre-planned endpoint was to compare prospectively the positivity rates and the value of CTC (CellSearch®), of serum tumor markers (CEA, CA 15–3, CYFRA 21.1), and of serum non-tumor markers (LDH, ALP) at baseline and under treatment for PFS prediction, independently from the other known prognostic factors, using univariate analyses and concordance indexes.

Results:

Table 1 shows the incidence of each of the 6 blood markers. Assessing all the 6 markers retrieved 90% of patients with at least one elevated marker at baseline. Interestingly, a combination of two markers (CA 15–3 and CYFRA 21.1, often used in lung cancer) retrieved 86% of patients with at least one marker elevated at baseline.

All 6 markers were correlated with poor performance status, high number of metastatic sites and with each other. Each marker was associated, when elevated at baseline, with a significantly shorter PFS in univariate anlaysis.

Serum marker changes during treatment, assessed either between baseline and week 3 or between baseline and week 6–9, were significantly associated with PFS, as reported for CTC. Concordance indexes comparison showed no clear superiority of any of the serum marker or CTC for PFS prediction.

Conclusion: In the largest prospective CTC study in metastatic breast breast cancer, we previously reported that CTC count, but not serum markers, is an independent prognostic factor for PFS and overall survival. However, for the purpose of PFS prediction by measuring blood marker changes during treatment, currently available blood-derived markers (CTC and serum markers) had globally similar performances. Besides CEA and CA 15–3, CYFRA 21.1 is commonly elevated in metastatic breast cancer and has a strong prognostic value.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-07-18.
P4-08-05: Diagnostic Significance of Exosomal miRNAs in the Plasma of Breast Cancer Patients.
Authors: Kwong A , Ng EKO , Leung CPH , Chan V , Li R , Wong CLP , Ma ESK
Published: June 2012

Background and Aims: Emerging evidence that microRNAs (miRNAs) play an important role in cancer development has opened up new opportunities for cancer diagnosis. Recent studies demonstrated that released exosomes which contain a subset of both cellular mRNA and miRNA could be a useful source of biomarkers for cancer detection. Here, we aim to develop a novel biomarker for breast cancer diagnosis using exosomal miRNAs in plasma. Methods: We have developed a rapid and novel isolation protocol to enrich tumor-associated exosomes from plasma samples by capturing tumor specific surface markers containing exosomes. After enrichment, we performed miRNA profiling on four sample sets; (1) Ep-CAM marker enriched plasma exosomes of breast cancer patients; (2) breast tumors of the same patients; (3) adjacent non-cancerous tissues of the same patients; (4) Ep-CAM marker enriched plasma exosomes of normal control subjects. Profiling is performed using PCR-based array with human microRNA panels that contain more than 700 miRNAs.

Results: Our profiling data showed that 15 miRNAs are concordantly up-regulated and 13 miRNAs are concordantly down-regulated in both plasma exosomes and corresponding tumors. These account for ∼25% (up-regulation) and ∼15% (down-regulation) of all miRNAs detectable in plasma exosomes. Our findings demonstrate that miRNA profile in EpCAM-enriched plasma exosomes from breast cancer patients exhibit certain similar pattern to that in the corresponding tumors. Based on our profiling results, plasma signatures that differentiated breast cancer from control are generated and some of the well-known breast cancer related miRNAs such as miR-10b, miR-21, miR-155 and miR-145 are included in our panel list. The putative miRNA biomarkers are validated on plasma samples from an independent cohort from more than 100 cancer patients. Further validation of the selected markers is likely to offer an accurate, noninvasive and specific diagnostic assay for breast cancer.

Conclusions: These results suggest that exosomal miRNAs in plasma may be a novel biomarker for breast cancer diagnosis.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-08-05.
P4-09-06: The Prognostic Value of Tumour-Stroma Ratio in Triple Negative Breast Cancer.
Authors: Moorman AM , Vink R , Heijmans HJ , van der Palen J , Kouwenhoven EA
Published: June 2012

Background: Triple-negative cancer constitutes one of the most challenging groups of breast cancer given its aggressive clinical behaviour, poor outcome and lack of targeted therapy. Until now, profiling techniques have not been able to distinguish between patients with good and poor outcome. Recent studies suggested an important role for stroma in tumour growth and progression. In colorectal-, oesophageal- and breast cancer, the tumour-stroma ratio was found to be of prognostic value.

Objective: to evaluate the prognostic value of the tumour-stroma ratio in triple-negative breast cancer.

Methods: During the period January 2004–2008, 124 consecutive triple negative breast cancer patients treated in our hospital were retrospective evaluated. Routine Haematoxylin-Eosin (H&E) stained histological sections were evaluated by two investigators (kappa 0.735) for stroma percentage, growth pattern (pushing margin), necrosis and amount of lymphocytic infiltrate. Patients with less than 50% stroma were classified as stroma-low and patients with ≥ 50% stroma were classified as stroma-high.

Results: of 124 triple-negative breast cancer patients, 50 (40%) had a stroma-high and 74 (60%) had a stroma-low tumour. Survival analysis revealed a 5 years relapse free period (RFP) of 85% in the stroma-low and 45% in the stroma-high group. Overall survival (OS) was 89% for stroma-low and 65% in the patients with a stroma-high tumour. Both RFP and OS were significantly worse in patients with stroma-high tumours compared to stroma-low. In a multivariate cox-regression analysis, tumour stroma remained an independent prognostic variable for RFP (HR 2.39; 95% CI 1.07−5.29; p=0.033) and OS (HR 3.00; 95% CI 1.08−8.32; 0.034) when corrected for other clinical-pathological variables. In addition, tumour-stroma proved to be a strong prognostic variable when compared to nodal status, tumour grade and tumour size, with respectively a HR of 2.39 (95% CI 1.37−6.26) versus HR 2.18 (95% CI 1.06−4.48), HR 2.10 (95% CI 0.89−4.91) and HR 0.53 (95% CI 0.70−3.91) for RFP with comparable numbers for OS. Conclusion: Tumour-stroma ratio is a strong independent prognostic variable in triple-negative breast cancer. It is easy to determine, reproducible (kappa 0.735) and can be easily incorporated into routine histological examination. This parameter optimizes risk stratification and could be target for future therapies.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-09-06.
P4-09-26: Prognostic Significance of Tissue Inhibitor of Metalloproteinase-1 (TIMP-1) in Breast Cancer.
Authors: Dechaphunkul A , Phukaoloun M , Kanjanapradit K , Graham K , Ghosh S , Santos C , Mackey JR
Published: June 2012

Background: Despite advances of systemic treatment in breast cancer guided by hormonal status and HER2 amplification, new prognostic and predictive factors are still warranted to optimize treatments among these patients. Tissue Inhibitor of Metalloproteinase-1 (TIMP-1), a physiologic inhibitor of Matrix Metalloproteinases (MMPs), can act in both pro- and anti-tumoral effects. The prognostic significance of TIMP-1 in breast cancer is still controversial. This study aims to determine the prognostic significance of TIMP-1 in breast cancer. Material and Methods: One-hundred and seventy-six primary breast cancers from women with early stage disease treated with standard adjuvant therapy were analyzed by gene expression microarrays and immunohistochemistry for TIMP-1. Immunohistochemical analysis was independently reviewed by two pathologists.

Results: At the optimized cut-off point, patients with high TIMP-1 RNA levels had a significantly shorter time to relapse, with a hazard ratio (HR) of 1.6 (p = 0.039), but without significant differences in overall survival (HR 1.29, p = 0.37). Although cytoplasmic overexpression of TIMP-1 protein was not correlated with early relapse (HR 1.2, p = 0.35), high expression was associated with shorter overall survival (HR 1.73, p = 0.027). In multivariate analysis, when considering stage, histologic grade, hormonal and HER2 status, TIMP-1 RNA levels remained independently prognostic for early relapse (HR 1.68, p = 0.04).

Discussion: Elevated TIMP-1 RNA levels are independently prognostic for early recurrence, whereas protein overexpression of TIMP-1 is correlated with short overall survival in primary breast cancer.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-09-26.
P4-10-02: A Meta-Analysis of the Association of Blood Levels of Vitamin-D and the Risk of Breast Cancer.
Authors: Amir E , Carlsson L , Seruga B , Ocana A , Goodwin PJ
Published: June 2012

Background: A considerable body of literature has examined the association of vitamin-D with breast cancer risk and the potential role in its prevention. Geographic studies show higher incidence of breast cancer in patients residing at high latitudes. Other data linking vitamin-D deficiency to breast cancer risk are inconsistent.

Materials and Methods: A literature based meta-analysis was conducted. Odds ratios (OR) for breast cancer based on blood levels of 25-hydroxy or 1,25-hydroxy vitamin-D were computed and pooled. Analysis was conducted separately for studies where blood levels were taken before (group A) or after (group B) breast cancer diagnosis.

Results: Thirteen studies were identified. Nine studies were included in group A and 4 studies included in group B. For group A, there was no significant association between lower vitamin-D levels and breast cancer risk (pooled OR = 1.09, 95% confidence intervals 0.99−1.20, p=0.08). For group B, there was a highly significant association between lower vitamin-D levels and breast cancer (pooled OR = 2.81, 95% confidence intervals 1.70−4.65, p<0.001). The test for interaction between groups was highly significant (p<0.001). When all studies were pooled, the OR was 1.38 (95% confidence intervals 1.13−1.70, p=0.002).

Conclusion: When measured before breast cancer diagnosis, blood levels of vitamin-D are not associated with breast cancer risk. Breast tumors have been shown to differentially express vitamin-D hydroxylase. Therefore, any association of vitamin-D and breast cancer in studies measuring blood levels after breast cancer diagnosis may be confounded by reverse causation bias.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-10-02.
P4-10-08: Risk Factor Profiles of Women with DCIS and Invasive Ductal Breast Cancers.
Authors: Schnabel F , Chun Kim J , Billig J , Cimeno A , Mehta P , Guth A , Hochman T , Shapiro R , Hiotis K , Siegel B , Axelrod D
Published: June 2012

Background There are well established risk factors for breast cancer, including elements of personal and family history. Ductal carcinoma in situ (DCIS) and invasive ductal carcinoma (IDC) have long been viewed as representing two points on the spectrum of one disease. There is limited and conflicting information in the literature regarding risk factor profiles for DCIS as compared with IDC. The purpose of this study was to investigate the relationship of established risk factors in a population of women newly diagnosed with pure DCIS and IDC. Methods

The Breast Cancer Database at NYU Langone Medical Center was queried for women who were diagnosed with pure DCIS and IDC from 1/2010-3/2011. Variables of interest included: age, family history of breast cancer (FHBC), BRCA1/2 status, age at menarche and menopause, parity, age at first birth, breast feeding, body mass index (BMI), history of atypical hyperplasia or lobular carcinoma in situ (ADH, ALH, LCIS), stage, ER/PR status, and method of presentation. Wilcoxon non-parametric tests, Chi-Square tests, and Fisher's Exact Tests were used to evaluate differences among DCIS and IDC patients.

Results Of the 593 women identified in this study, 140 (24%) had pure DCIS and 453 (76%) had IDC. The median age at diagnosis was 59 years. There were 9 (1.5%) BRCA1/2 mutation carriers. Of these, 7 had IDC and 2 had DCIS. The majority of patients with IDC were stage I (67%) and ER/PR+ (73%). The majority of patients with DCIS were also ER/PR+ (71%). In our cohort, the majority of DCIS (83%) was mammographically detected versus 47% of IDC cases. There were no statistically significant differences in breast cancer risk factors between the two groups.

Conclusions In contrast to some previously published work, the risk factor profile in our cohort was similar for patients with IDC and DCIS. In this population, the majority of cases of DCIS were detected mammographically, underscoring the importance of mammography in early detection of breast cancer. There is increasing interest in identifying a population of women with DCIS who might never progress to invasive disease. Our data suggests that this population is unlikely to be distinguished by its risk factor profile. Genetic and molecular characteristics are more likely to provide the criteria by which this subgroup of patients is identified.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-10-08.