bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P2-09-05: Risk Group Selection in Primary Breast Cancer According to ASCO Recommended Biomarkers Onkotype DX and uPA/PAI-1: First Experience from Prospective Multicenter WSG Plan B Trial
Authors: Degenhardt T , Gluz O , Thomssen C , Vetter M , Kates R , Warm M , Shak S , Augustin D , Kusche M , Untch M , Paepke S , Uleer C , Dünnebacke J , Nitz U , Harbeck N
Published: April 2011

Background: Both the Oncotype DX multi-gene assay and invasion factors uPA/PAI-1 are guideline-recommended (ASCO, AGO) biomarkers for decision support regarding adjuvant chemotherapy in primary breast cancer (BC). Material and Methods: The West German Study Group (WSG) Plan B trial (planned n=2,448) is evaluating anthracyline-free adjuvant chemotherapy (6x TC) vs. standard 4xEC-4xDOC in HER2- negative BC with 0-3 positive lymph nodes, Oncotype DX is used as selection criterion where pts with RS> 11 are randomized to one of the two chemotherapy arms and pts with RS ≥11 treated with hormonal therapy alone. uPA/PAI-1 (both low vs. either/both high), measured by ELISA, is obtained as an optional risk factor. Results: By June 2010, 1064 patients had been randomized in Plan B (96 recruiting centers). In 153 patients (27 centers), uPA and PAI-1 had been measured; for 131 (84 N0, 47 N+) of these, Oncotype DX Recurrence Score® (RS) results were also available. When considered as continuous variables, RS was weakly positively correlated (Spearman's coefficient) with uPA (rs=0.18, p=0.04) and with PAI-1 (rs=0.23, p=0.01). When considered as risk categories/(Table 1), there was a weak concordance between RS and uPA/PAI-1, using either the standard RS cutoff points (18 and 30) or the TAILORx trial cutoff points (11 and 25).

Table T: Association between RS and uPA/PAI-1 categories

Assignment of high risk was most strongly concordant between uPA/PAI-1 and RS (RS > 25: 22/25 patients had high uPA/PAI-1; RS >30: 14/15 patients had high uPA/PAI-1). This high-risk concordance extends to N0 patients (RS>25: 17/19 N0 pts had high uPA/PAI-1; RS>30: 11/12 pts had high uPA/PAI-1). Discussion: For the first time, risk groups in primary breast cancer according to both Oncotype DX and uPA/PAI-1 have been compared. These preliminary data show that the high RS group seems highly concordant with the prospectively assessed invasion markers uPA/PAI-1. However, within low and intermediate RS, uPA/PAI-1 could still identify a substantial collective at risk; low uPA/PAI-1 could define a clinically relevant low-risk collective within risk groups that would be classified as “intermediate” according to the multigene assay Oncotype DX. Additional recruitment and outcome assessment of the ongoing multicenter WSG Plan B trial will address the clinical significance of these findings.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-09-05.
Abstract P2-09-07: Elevated Pretreatment Serum CA9 (CAIX) (Carbonic Anhydrase 9) Predicts Reduced Progression-Free and Overall Survival in Trastuzumab-Treated Metastatic Breast Cancer
Authors: Anyanwu UI , Hou HY , Shrinvastava V , Evans M , Leitzel K , Ali SM , Koestler W , Fuchs E-M , Brown-Shimer S , Ramirez J , Hamer P , Carney W , Kalu R , Repich K , Rizvi S , Lipton A.
Published: April 2011

Background: Approximately one-half of HER2-positive breast cancer patients will respond to first-line trastuzumab-containing therapy. However, in those patients with an initial trastuzumab response, most will progress within a year with acquired resistance. Since trastuzumab treatment is also now used in the HER2-positive adjuvant breast cancer setting, trastuzumab resistance will continue to be a vexing clinical problem, and better predictive and prognostic biomarkers are urgently needed. Methods: Serum HER2, tissue inhibitor of metalloproteinase-1 (TIMP-1), urokinase-type plasminogen activator (uPA), CA9, Insulin-like growth factor-1 receptor (IGF-1R), VEGF-165, and endoglin were measured using ELISA assays in 60 metastatic breast cancer patients before starting first-line trastuzumab-containing therapy. The HER2, TIMP-1, uPA, CA9, IGF-IR, and VEGF-165 ELISAs were from Oncogene Science/Siemens Healthcare Diagnostics, Cambridge, MA; and the endoglin ELISA was from R&D Systems, Minneapolis, MN. Progression-free survival (PFS) and overall survival (OS) were analyzed using the Kaplan-Meier method and Cox modeling with continuous pretreatment serum biomarker variables.

Results: Pretreatment serum HER2 (p= 0.008), TIMP-1 (p< 0.0001), uPA (p= 0.006), endoglin (p= 0.010), and CA9 (p <0.0001) were all significant as univariate continuous biomarkers for predicting PFS to first-line trastuzumab-containing therapy, but VEGF (p=0.94) and IGF-IR (p=0.11) were not. In multivariate analysis for PFS with all 7 biomarkers, only serum CA9 (p= 0.002) was a significant independent covariate. For OS, pretreatment serum HER2 (p= 0.018), TIMP-1 (P<0.0001), uPA (P<0.0001), endoglin (0.006), CA9 (P<0.0001) were all significant as univariate continuous biomarkers for prognosis, but VEGF (p=0.69) and IGF-IR (p=0.098) were not. In multivariate analysis for OS with all 7 biomarkers, only serum CA9 was a significant independent prognostic covariate (p=0.001), and TIMP-1 trended significant (p=0.069). Conclusions: Elevated pretreatment serum CA9 (a marker of hypoxia) predicts reduced progression-free survival and overall survival in metastatic breast cancer patients treated with first-line trastuzumab-containing therapy. These serum biomarkers deserve further study in larger trials of HER2-targeted breast cancer treatment. Supported by a grant from Komen for the Cure.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-09-07.
Abstract P2-09-09: Loss of BTG2 Expression in Breast Cancer Predicts Tamoxifen Resistance
Authors: Takahashi M , Hayashida T , Sakata M , Jinno H , Kitagawa Y.
Published: April 2011

Background: The B-cell translocation gene-2 (BTG2) belongs to a class of proteins known as the Tob and BTG anti-proliferative protein family. It was shown that estrogen and progesterone suppress BTG2 mRNA for the development of mammary gland. (Kawakubo et al., Oncogene 2004) Expression analysis of BTG2 protein in estrogen receptor (ER) positive human breast cancer was revealed the loss of nuclear expression in 46% of tumors and showed a significant inverse correlation with cyclinD1 expression. (Kawakubo, Hayashida et al., Cancer Res 2006) According to the Oncomine database online, loss of BTG2 expression significantly correlated with increased recurrence rate after tamoxifen monotherapy. Based on these results, we hypothesized that BTG2 expression might be a novel biomarker for predicting the clinical outcome in tamoxifen treatment and analyzed if BTG2 expression affects the sensitivity against tamoxifen both in vitro and in vivo.

Method and Results: The cohort of 60 patients treated with adjuvant tamoxifen monotherapy was available online. (Ma et al., Cancer Cell 2004) Comparison of disease-free survival (DFS) of patients whose laser-captured tumors showed positive BTG2 expression versus patients with loss of BTG2 expression revealed an univariate association in the dataset. (log-rank probability, p=0.029) Multivariate analysis indicated that BTG2 resulted independent prognostic factor for DFS (HR, 0.69; 95% CI, 0.495 to 0.963; p=0.029).

In vitro cellular proliferation assay to evaluate the cytotoxicity induced by tamoxifen against ER positive cell lines revealed that endogenous BTG2 positive cell line, MCF7 and T47D, showed drug sensitivity, whereas MDA-MB468 expressing loss of BTG2 showed tamoxifen resistance. (MCF7; IC50=7.35uM, T47D; =3.79uM, MDA-MB468; >25.0uM) Tetracycline-inducible BTG2 expression model in MCF7 was developed to assess the cytotoxicity by tamoxifen treatment under the condition of BTG2 overexpression in vitro and in vivo. The cellular proliferation was strongly inhibited by the concomitant administration of tetracyclin and tamoxifen in vitro. The cell line was injected into the mammary fad pad of immunodeficient mice and these were administered water including tetracyclin and/or tamoxifen pellet at the time when the xenografted tumor grew to be over 125mm3 for 3 weeks. The transgene expression was confirmed by both RT-PCR and immunohistochemistry. Tumor growth ratio was significantly suppressed in the concomitant administration of tetracyclin and tamoxifen in comparison with single treatment of tamoxifen or of tetracyclin. Consistently, tumor weight and Ki67 expression were also significantly suppressed in the mice administered both tamoxifen and tetracyclin.

Conclusion: Despite further validation studies need to be conducted, BTG2 expression may be useful biomarker to identify patients appropriate for tamoxifen treatment.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-09-09.
Abstract P2-09-11: Response of Invasive Lobular Cancers to Neoadjuvant Therapy
Authors: Mukhtar RA , Hwang ES , Livasy C , Esserman LJ.
Published: April 2011

Background

Invasive lobular carcinoma (ILC) makes up 10-15% of all breast cancer. While neoadjuvant therapy (NAC) is increasingly used in locally advanced invasive cancers, some evidence suggests that ILC does not respond to NAC. The I-SPY trial has shown an association between expression of high risk gene signatures and increased pathologic complete response (pCR) to NAC. We hypothesized that compared to invasive ductal carcinoma (IDC), ILC would have decreased expression of high risk gene signatures, and decreased rates of pCR. Methods

We analyzed data from 221 patients in the I-SPY trial, a prospective, multicenter neoadjuvant breast cancer trial with serial pathology, imaging, and expression arrays. A dedicated breast pathologist at each site reviewed histology, and e-cadherin staining was performed centrally on cases identified as ILC. We compared rates of wound healing signature activation, NKI (Netherlands Cancer Institute) profile status, and proportion of pCR in ILC compared to IDC. Other comparisons were the presence of p53 mutations, molecular subtype, and MRI phenotype. We used the chi-squared test, Fisher's exact test, and Student's t-test for comparisons. Results

Of 221 patients, histology on core biopsy was ductal in 195 (88%), lobular in 18 (8%), and mixed ducto-lobular in 8 (3.6%); the last cases were included in the IDC group. ILC and IDC cases were similar by tumor size, age at diagnosis, and race. Expression array data were available on 12 of 18 (67%) ILC and 149 of 203 (73%) IDC. The proportion of p53 mutations, basal subtype and NKI high risk did not differ significantly. However, ILCs were significantly less likely to express the activated wound healing signature compared to IDCs (50% vs 80%, p = 0.03) and had different imaging phenotypes, with most ILCs appearing multinodular or diffuse on MRI (p = 0.028). Three of 18 (17%) ILCs had a pCR, compared to 55/197 (28%) IDCs. Of the 18 ILC, 11 of 15 tested were e-cadherin negative (73%). Of 18 ILCs, 16 (89%) were hormone receptor (HR) positive, and comparison to HR+ IDC showed similar rates of pCR in the two groups (19% pCR in HR+ ILC vs 14% pCR in HR+ IDC). There was no significant difference in expression of the activated wound healing signature when comparing ILCs to HR+ IDC. Conclusions

There is significant heterogeneity within ILC, such that no one molecular subtype or genetic expression signature identifies all ILCs. The rates of pCR among ILC are similar to those seen in HR+ cancers. Although larger studies are needed to confirm these findings, we saw no evidence that ILC differs from IDC in response to NAC.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-09-11.
Abstract P2-09-16: Effect of Continuous Statistically Standardized qPCR and IHC ER and PR on Disease-Free Survival in NCIC CTG MA.12
Authors: Chapman J-AW , Nielsen T , Ellis MJ , Bernard P , Chia S , Gelmon K , Pritchard KI , Le Maitre A , Shepherd L , Bramwell V.
Published: April 2011

Background: Inter-laboratory comparability of ER and PR assay results has been problematic for decades. Statistical standardization has been proposed as an adjunct following good laboratory quality control. We demonstrate this process for ER and PR centrally assessed by qPCR and IHC. Methods: NCIC CTG MA.12 is a placebo-controlled trial of tamoxifen therapy following adjuvant chemotherapy in premenopausal women with early breast cancer. 672 women were accrued, 505 of whom had locally determined ER and/or PR positive tumours. ER was centrally assessed by qPCR and IHC for respectively 385 and 392 patients, and by PR, for 389 and 376. The effects on DFS of continuous 1) log2 (qPCR) and 2) ln (IHC % positive stain) in statistically standardized units were investigated for all women with all lab results, and in the subgroup with IHC ER+ and/or PR+ tumours.

DFS is defined as the time from randomization to recurrence or death. Multivariate examinations were with adjusted Cox regression, with adjustment by stratification factors nodal status and chemotherapy type; ER and PR had forced inclusion in models, while trial therapy and baseline patient characteristics were added in step-wise mode (p<=0.05). Unvariate tests were with adjusted log-rank statistic and Kaplan-Meier plots for ER and PR (ER and PR 0; standardized cut-points for ER and PR with positive stain of <= 1.0; > 1.0, <= 0.0; > 0.0, <= 1.0; >1.0). Results: Median follow-up for the MA.12 trial was 9.7 years. Univariately, qPCR ER was not associated with DFS (p=0.19), while both qPCR PR (p=0.09) and IHC ER (p=0.08) had weak evidence of association, and IHC PR had p=0.04. Multivariately, patients randomized to tamoxifen had significantly better DFS (p=0.002-0.005) in all patients assessed and in those with ER and/or PR > 0; ER and PR by qPCR and IHC were not associated multivariately with DFS (P>0.05). ER and PR were not associated with DFS (P>0.05) for those who received tamoxifen, while qPCR ER and/or PR, and IHC ER were inconsistently associated with DFS for those on the placebo arm.

Conclusions: In this subgroup of centrally assessed MA.12 patients, trial therapy was significantly associated with DFS. Similar non-significant multivariate results were obtained multivariately with statistically standardized, continuous qPCR and IHC ER and PR. We plan to further investigate adjunctive statistical standardization in larger NCIC CTG trials.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-09-16.
Abstract P2-09-18: High VEGFR1 and VEGFR3 Protein Expression Is Associated with Improved Response to the Combination of Paclitaxel (P) and Bevacizumab (Bev) Therapy in Patients with HER2-Negative Metastatic Breast Cancer (MBC)
Authors: Papadimitriou C , Kourea H , Papazisis K , Christodoulou C , Res E , Papakostas P , Petraki K , Bafaloukos D , Razis E , Fountzilas G.
Published: April 2011

Background: Treatment with weekly P and 2-weekly Bev has been established as an active 1st line treatment in patients with MBC. However, robust predictive or prognostic biomarkers for this treatment have not been identified as yet.

Patients and Methods: In this retrospective analysis, we evaluated the activity of this combination in patients with HER2-negative MBC. Further, we explored the role of a panel of biomarkers on patients’ outcome. VEGF-A, VEGF-C, VEGFR1, VEGFR2, and VEGFR3 were centrally assessed by immunohistochemistry (IHC) in 70 tissue blocks. P was administered either weekly 90 mg/m2 x12 with 2-weekly Bev 10 µg/kg (86 patients) or 3-weekly 175 mg/m2 x6 with 3-weekly Bev 15µg/kg (38 patients). Bev was administered until progression in the majority of the patients.

Results: The ORR did not differ significantly between the weekly or 3-weekly P schedule (55.8% vs 55.3%). In contrast to the published literature with P monotherapy, median PFS was significantly longer in the 3-weekly compared to the weekly P schedule (20.4 months vs 10.2 months, p=0.029). Median survival has not been reached yet. 2-year survival was 76.1% and 67.3%, respectively (p=0.085). High VEGFR1 and VEGFR3 protein expression, assessed by IHC, was associated with higher response rates (p=0.010 and p=0.039, respectively).

Conclusions: This retrospective analysis confirmed the activity of P and Bev in patients with HER2-negative MBC. Protein expression of VEGFR1 and VEGFR3 was found to be of predictive value. These findings need to be validated in a larger cohort of patients treated with this combination. VEGF polymorphisms (VEGF-2578 and VEGF-1154) are currently assessed in peripheral blood DNA from our patients for possible predictive value.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-09-18.
Abstract P2-09-27: Predictive Factors of Complete Axillary Downstaging after Neoadjuvant Chemotherapy in Breast Cancer Patients
Authors: Vidal M , Perez-Garcia JM , Muñoz E , Saura C , de Mattos L , Gomez P , Cortes J , Baselga J , Bellet M.
Published: April 2011

Background: The axillary status after neoadjuvant chemotherapy (NACT) is an important prognostic factor. Complete axillary downstaging (CAD) occurs in up to 20-30% of the patients, however, sentinel node biopsy after NACT is not recommended in patients with initial axillary nodal involvement and complete axillary response after NACT. Our aim was to investigate predictive factors of CAD after NACT to identify patients who could avoid an unnecessary axillary dissection. Patients and methods: From January 2004 to December 2008, 256 consecutive patients were diagnosed with invasive breast cancer and received NACT and breast surgery at our institution. Patients were eligible for this analysis if they met the following inclusion criteria: clinically and/or cytologically proven metastatic axillary nodes; complete axillary response after NACT; and complete axillary dissection after systemic treatment. CAD was defined as pN0 (isolated tumor cells and micrometastasis were excluded). Univariate and multivariate analyses were performed to assess the association between CAD and either tumor characteristics or intrinsic cancer subtypes.

Results: 84 patients were included. Median age was 51.64 years. 75 (89,3%) received anthracyclines-and taxanes-based CT and 11 (13%). 38 (45,2%) had cytologically proven metastatic axillary nodes at diagnosis. According to intrinsic cancer subtypes, 22 (26%) were classified as HER2 positive, 17 (20,2%) as triple negative (ER, PR, and HER2 -), 6 (7,1%) as luminal A [estrogen receptor positive and/or progesterone receptor positive and (Ki67 < 20% and histological grade 1 or 2)], and 23 (27,4%) as luminal B [estrogen receptor positive and/or progesterone receptor positive and (Ki67 ≥20% or histological grade 3)]. In the univariate analysis, HER2 positivity (p=0,002) and neoadjuvant treatment with trastuzumab (p=0,019) were significantly associated with CAD. In the multivariate analysis only HER2 status maintained statistical significance (p=0,003). Among luminal A, luminal B, and triple negative tumors, complete axillary response after NACT did not significantly predict CAD.

Conclusions: In our cohort of breast cancer patients, HER2 status is an independent predictive factor of CAD after NACT. Clinical trials, probably according to specific breast cancer subtypes, are required to identify patients with initial axillary metastatic involvement who could avoid an axillary dissection after systemic therapy.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-09-27.
Abstract P2-09-32: Estrogen Receptor-Related Genes Are Important Predictors of Response to Neo-Adjuvant Chemotherapy in Breast Cancer
Authors: Chen Y , Chen C , Xu Q , Ye X , Meng X , Morgin B , Wu J.
Published: April 2011

Background: Clinical data suggest a contribution of estrogen receptor (ER) to chemotherapy response. However, ER status alone is not perfectly predictive, and there is an urgent need for more reliable markers of chemotherapy responsiveness. In this study, we set out to find novel ER-related genes in predicting chemotherapy response by microarrays, hoping to provide more robust multi-variable prediction to chemotherapy. Material and Methods: One hundred and ten patients with stage II-III breast cancer were included, receiving preoperative four cycles of weekly PCb (paclitaxel plus carboplatin) regimen. Pretreatment gene expression profiling was performed with gene microarrays on core biopsy specimens. 55 training cases were used for marker discovery and to identify any ER-related genes that might be associated with response (“training cases”), and the other 55 patients were available as an independent validation set (“validation cases”) to test whether the ER-related genes do predict pCR in protein levels by immunohistochemistry (IHC). Results: In the training cases, the yield of total RNA was insufficient to assay in 16 patients and 8 chips failed in the quality check (QC) process. Therefore, gene expression profile from 31 samples were available for this study. The overall pCR rate in the 86 patients was 19.7% (n=17, 6 cases of pCR in training set and 11 pCR in validation set). In the training set, by Significance Analysis of Microarrays (SAM), 300 probes (231 genes) were identified as differentially expressed between pCR versus non-pCR when fold change > 2. There were 20 differentially expressed genes, setting fold change > 3. Among these 20 genes, TFF1, ESR1, GATA3, TFF3 were found as ER-related. In 55 independent validation cases, univariate analysis indicated that clinical variables and ER-related genes, including ER, PR, GATA3, TFF1 and TFF3 were all significantly associated with pCR (Pearson x2, P<0.001, P=0.013, P<0.001, P<0.001, P<0.001). Multivariate logistic regression model showed that ER (P=0.016) remained significantly associated with complete remission. In spearman analysis, these 5 ER-related genes were significantly correlated with each other (P<0.05). It was found that pCR rate was as high as 80% (4/5) when these 5 factors were all negative. In contrast, these 5 factors were all positive in 7 of 9 PD/SD patients.

Discussion: ER combined with ER-related genes expression status may provide important predictive value of response to neoadjuvant chemotherapy and allows to identify a subgroup of patients who might highly benefit from chemotherapy and who might be resistant to it.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-09-32.
Abstract P2-09-33: Validity of Bone Marker Measurements for Monitoring Response to Bisphosphonate (BP) Therapy with Zoledronic Acid (ZOL) in Metastatic Breast Cancer (mBC): Useful or Not?
Authors: Aktas B , Kasimir-Bauer S , Lehmann N , Kimmig R , Kuemmel S.
Published: April 2011

BACKGROUND: Bone is the most common site of metastasis in breast cancer. The detection relies on imaging technology which is costly and can only be performed to a certain degree. Bone markers are noninvasive, inexpensive and potentially might serve as predictive and prognostive surrogate endpoints in detecting bone metastases and response to BPs. This study analyzes the value of the serum markers of type I collagen synthesis (PINP) and degradation (ICTP) as markers of bone metastases in mBC patients (pts) receiving ZOL. The results will be correlated with the levels of CEA and Ca 15-3 as well as the clinical response. MATERIALS AND METHODS: 38 mBC pts (mean age was 60 years: range, 35-86), with confirmed bone metastases who were received chemotherapy and/or hormonal therapy and ZOL iv q 4 wks were participating in our study. 5 ml blood were collected at the time of study start and q 3 months for one year for the analysis of PINP, ICTP, CEA and Ca 15-3 using radioimmunoassays and ECLIA System, respectively. Imaging of bone metastases was performed at the same time points. In this first analysis, data were correlated with the number of bone metastases at study start.

RESULTS: At study baseline, PINP, ICTP, CEA and Ca 15-3 were analyzed in all pts. In 28/38 pts (73%) more than 3 bone metastases were confirmed by imaging and 10/38 pts (27%) presented with 3 or less bone metastases. The median value for ICTP was 6 µg/l (interquartile range 3.9-12.7) and for PINP 58.7 µg/l (43.5-164). Serum PINP and ICTP concentrations were significantly different when pts were stratified into > 3 bone metastases (median value ICTP 6.3 µg/l and PINP 75.3 µg/l) and 3 or less bone metastases (median value ICTP 4.1 µg/l and PINP 47 µg/l), respectively (P<0.05, see Figure 1 for ICTP and 2 for PINP for logarithmized values). No significant differences were seen when tumor marker levels were correlated with the number of bone metastases (P>0.2). Statistical analysis for follow-up measurements will be available after having completed sampling for all 38 pts in 5 months. Up to now, 135 samples of all patients have been analyzed and there seems to be an association between the number of bone metastases, serum levels for PINP/ICTP and response to therapy.

CONCLUSION: In contrast to serum tumor markers, the identification of PINP and ICTP permit to estimate the number of bone metastases and seems to be a useful tool for monitoring mBC pts undergoing BP therapy with ZOL for treatment of bone metastases. These results have to be confirmed in a larger prospective trial.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-09-33.
Abstract P2-09-38: Role of Oncotype 21 Gene Assay and Adjuvant! Online in Breast Cancer Adjuvant Treatment Decisions
Authors: Khalil DN , Schneider JG
Published: April 2011

Background: Based upon its validation as both a prognostic and predictive tool, the Oncotype 21 gene RS has been broadly adopted into clinical practice, but little has been reported on how RS actually influences clinical decision making in practice.

Methods: A retrospective chart review was performed on all patients for whom a RS was requested within a single faculty practice since the RS became commercially available in 2005. Data were collected to identify: (1) reasons for requesting RS; (2) reasons for RS cancellations; (3) traditional recurrence risk factors enabling alternative prognostic calculations including the Nottingham Prognostic Index and Adjuvant! Online (AOL) web based instrument; and (4) actual adjuvant treatment decisions. Clinician interviews were performed to confirm RS influence on clinician practice and to compare hypothetical and actual treatment decisions.

Results: 109 RS were requested by five different clinicians. 87 RS were obtained on ER+, node-negative patients. The 10-year relapse risk predicted by RS versus that predicted by AOL is plotted in Figure 1 where the actual adjuvant treatment decision is coded and lines are introduced to separate low, intermediate and high risk groups. In patients with a low RS for whom a treatment decision was made, only 1 of 17 (6%) with an intermediate AOL relapse rate, and 4 of 21 (19%) with a high AOL relapse rate, were treated with chemotherapy. These data suggest that a favorable RS recurrence rate influenced the decision not to initiate chemotherapy when the AOL-predicted relapse rate was less favorable. Two distinct areas of “clinical equipoise” were identified where the combination of AOL and RS risks left clinicians ambivalent about the use of adjuvant chemotherapy (AOL 23-26 with RS 11-15, and AOL 29-43 with RS 5-11). Apparent inconsistencies between RS and adjuvant treatment decisions as well as alternative reasons for obtaining RS were identified and reviewed with the treating clinicians. Conclusions: This single institution experience indicates appropriate use of RS in adjuvant ER+ node-negative breast cancer. The predominant effect in this group was to reduce the utilization of adjuvant chemotherapy in patients with intermediate and high relapse risk predicted by conventional prognostic factors. Figure available in online version.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-09-38.
Abstract P2-14-11: Vitamin D Deficiency in Post-Menopausal Breast Cancer Survivors
Authors: Friedman CF , DeMichele A , Su HI , Feng R , Kapoor S , Desai K , Mao JJ.
Published: April 2011

Background: Breast cancer patients on aromatase inhibitors (AIs) are at an increased risk for decreased bone density and fractures. Given the important role vitamin D plays in bone metabolism, vitamin D deficiency may place these patients at an additional risk. Our objective was to examine the prevalence of and risk factors for vitamin D deficiency in post-menopausal breast cancer survivors (BCS) with current or past AI use. Materials and Methods: We conducted a cross-sectional study. Vitamin D levels were obtained utilizing the Diasorin radioimmunoassay using sera. Multiple logistic regression analyses were performed to identify independent risk factors associated with vitamin D deficiency. Results: In our cohort of 391 post-menopausal women with stage I to III breast cancer on AI therapy, the mean vitamin D level was 35 ng/mL, S.D. 12.49. Vitamin D levels were normally distributed in the cohort. Thirty five percent of women were vitamin D deficient, defined as a level less than 30 ng/mL. In our cohort, 73.4% of the participants had vitamin D supplementation documented in their chart; 26.8% of these women were still vitamin D deficient despite supplementation. Adjusting for demographic and clinical variables, non-white patients were more likely to be vitamin D deficient, adjusted odds ratio (AOR) of 2.00, 95% confidence interval (CI) 1.14-3.51, p=.004. Overweight (AOR 3.07, 95% CI 1.77-5.33, p<.001) and obese (AOR 2.88, 95% CI 1.64-5.06, p<.001) women were also at higher risk. In a second model, adjusting for the same covariates and chart-documented vitamin D supplementation, vitamin D supplementation was associated with lower risk for vitamin D deficiency (AOR .29, 95% CI .17-.48, p<.001). The risk for non-white women (AOR 2.19, 95% CI 1.22-3.89, p=.009) and obese women (AOR 3.21, 95% CI 1.79-5.78, p<.001) were increased after adjusting for vitamin D supplementation. Discussion: Hypovitaminosis D is common in BCS, and those who are non-white or have a BMI greater than 25 are at a higher risk of deficiency. When adjusting for vitamin D supplementation, the discrepancy between normal weight patients and obese patients became even more pronounced, suggesting that standard vitamin D supplementation may not be sufficient for these patients. In light of data linking hypovitaminosis D and bone health, oncologists should consider routine testing with particular attention to non-white or overweight patients, and repleting those patients who are deficient.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-14-11.
Abstract P2-16-01: Overall Survival Data from SOLTI-0701: A Multinational, Double-Blind, Placebo-Controlled, Randomized Phase 2b Study Evaluating the Oral Combination of Sorafenib and Capecitabine in Patients with Locally Advanced or Metastatic HER2-Negative Breast Cancer
Authors: Gomez P , Roché H , Costa F , Segalla JGM , Pinczowski H , Ciruelos EM , Cabral Filho S , Van Eyll B , Baselga J.
Published: April 2011

Background: Sorafenib (SOR) is an oral multikinase inhibitor with antiangiogenic and antiproliferative activity. We previously reported data from the primary analysis of SOLTI-0701 demonstrating a progressionfree survival (PFS) benefit with the oral regimen of SOR + capecitabine (CAP) vs placebo (PL)+CAP in patients with advanced breast cancer (BC). Here we describe overall survival (OS), a secondary endpoint of SOLTI-0701.

Methods: SOLTI-0701 is a multinational (Brazil, France, Spain) phase 2b study in HER2-negative patients with locally advanced or metastatic BC and ≥1 prior chemo regimen for advanced BC. Patients were randomized (1:1) to receive CAP (1000 mg/m2 po, twice daily [BID], 14 of every 21 days) with PL or SOR (400 mg po, BID). The primary endpoint was PFS. The sample size was estimated at 220 patients based on patient accrual projections and the estimated rate of PFS events. OS analysis was planned after 120 events.

Results: A total of 229 patients were enrolled (115 SOR+CAP and 114 PL+CAP) from August 2007 to December 2008. Treatment arms were balanced for age (mean 55.2 y), ECOG status 0 (68%) and 1 (30%), stage IV disease (91%), and visceral metastases (75%). In the primary analysis, median PFS was 6.4 mo for the SOR arm vs 4.1 mo for the PL arm (hazard ratio 0.58; 95% CI, 0.41-0.81; 1-sided P=0.0006); the most common Grade 3/4 toxicity was hand-foot skin reaction/syndrome (45% vs 13%, respectively); discontinuation of treatment due to adverse events was infrequent (15% vs 7%, respectively). OS data are expected by the 3rd quarter of this year and will be presented, as well as updated safety data. Conclusions: The oral combination of SOR+CAP significantly improved PFS in patients with advanced BC. The regimen was tolerable with a manageable toxicity profile. An ongoing phase 3 registration trial is evaluating the combination of SOR+CAP for advanced BC. The secondary endpoint of OS will better characterize the potential role of SOR in advanced BC as randomized controlled trials thus far have yet to demonstrate any survival benefit with antiangiogenic agents.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-16-01.
Abstract P2-16-06: Final Overall Survival Results, Including Analysis of Patients with Triple-Negative Disease and Aged ≥70 Years, from the ATHENA Study Evaluating First-Line Bevacizumab-Containing Therapy for Locally Recurrent/Metastatic Breast Cancer
Authors: Pritchard KI , Thomssen C , Pierga J-Y , Cortes-Funes H , Biganzoli L , Guan Z , Manikhas G , Jendiroba D , Uziely B , Boyle F , Jagiello-Gruszfeld A , Van Arkel C , Tung Y , Dunne A-A , Smith IE.
Published: April 2011

Background: First-line bevacizumab combined with chemotherapy significantly improves efficacy versus chemotherapy alone in HER2-negative locally recurrent/metastatic breast cancer (LR/mBC). ATHENA, a multinational, open-label, single-arm study, further evaluated first-line bevacizumab in combination with standard chemotherapy in patients treated in routine oncology practice.

Patients and methods: Eligible patients had HER2-negative LR/mBC, ECOG performance status 0-2, and had received no prior chemotherapy for LR/mBC. They received bevacizumab 10 mg/kg q2w or 15 mg/kg q3w in combination with taxane-based chemotherapy (or other non-anthracycline chemotherapy) until disease progression, unacceptable toxicity, or patient withdrawal. The primary endpoint was safety; secondary endpoints included time to progression (TTP) and overall survival (OS). Results: Between September 2006 and March 2009, 2264 patients began therapy with bevacizumab, combined with single-agent paclitaxel (34%), single-agent docetaxel (33%), taxane-based combination therapy (11%), non-taxane-containing therapy (10%), or sequential chemotherapy (switching chemotherapy regimen before disease progression while continuing on bevacizumab; 12%). After final data cut-off in July 2010, the median duration of follow-up was 20.1 months (range 0.1-43.6 months). Bevacizumab therapy was continued for ≥1 year in 473 patients (21%). The safety profile and median TTP were very similar to those previously reported after median follow-up of 12.7 months and are consistent with findings from randomized phase III trials. Final OS results in the overall population and in clinically important subgroups are reported below for the first time.

Conclusion: Median OS in the overall population is consistent with OS results reported from three randomized phase III trials in the first-line setting (25.2-30.2 months). OS results in subsets of patients with TN disease or aged ≥70 years suggest that bevacizumab-containing therapy is an effective option for these patients, who typically have fewer treatment options available.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-16-06.
Abstract P2-16-08: Administration of the Novel Antiangiogenic PTC299 in Combination with Aromatase Inhibitors Is Feasible and Shows Antitumor Activity in Metastatic Breast Cancer
Authors: Tiersten A , Schneider BP , Dickler M , Volm M , Speyer J , Novik Y , Lehman R , Callahan LA , Darby CH , Miller LL , Miao H.
Published: April 2011

Background: PTC299 is an investigational new drug that suppresses tumor growth by selective post-transcriptional inhibition of tumor VEGF expression. PTC299 demonstrates antitumor activity in both hormone-receptor-positive (HR+) and-negative (HR-) breast cancer xenograft models and eliminates established tumors in aromatase-overexpressing human breast cancer xenografts when given alone or in combination with letrozole.

Methods: This study is assessing the feasibility of PTC299 administration in combination with aromatase inhibitors (AIs) in women with HR+ metastatic breast cancer and natural or induced post-menopausal levels of ovarian function. Patients already receiving or newly initiating a standard dose of an AI are coadministered 100 mg BID of oral PTC299 continuously until disease progression.

Results: To date, the study has enrolled 24 women with a median [range] age of 57 [26-82] years, ECOG PS of 0 (n=12) or 1 (n=12), to receive PTC299 together with letrozole (n=11), anastrozole (n=6), or exemestane (n=7) for a median [range] duration of ∼16 [3-36] weeks. Combination therapy has been generally well tolerated; AEs have been predominantly Grade 1-2, have usually not been PTC299-related. The most frequent AEs have been headache (25%) and hot flashes (25%). No PTC299 dose reductions or discontinuations due to AEs have occurred. Measures of PTC299 plasma exposure have shown ∼2-fold accumulation, with values reaching steady state by -21 days of treatment; trough values have exceeded those maximally active in xenograft models. Relative to baseline, therapy has been associated with on-study reductions in circulating VEGF concentrations and decreases in tumor perfusion as assessed by DCE-MRI. Antitumor activity includes 3 CRs by PET/CT, 1 PR by PET/CT, and 3 PRs by PET scan. To date, 7/24 (29%) patients have received ≥24 weeks of treatment and 10/24 (42%) patients continue on treatment.

Conclusions: Combining PTC299 100 mg BID with standard AI treatment appears feasible. Safety, exposure, pharmacodynamic, and antitumor activity observations have been encouraging. Supported by a grant award from the Department of Defense

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-16-08.
Abstract P2-17-01: Phase II Study of Bevacizumab in Combination with Docetaxel and Capecitabine for the First-Line Treatment of Patients with Locally Recurrent or Metastatic Breast Cancer
Authors: Musolino A , Bisagni G , De Matteis A , Nuzzo F , Ardizzoni A , Crinò L , Gamucci T , Passalacqua R , Gnoni R , Boni C.
Published: April 2011

Background: Docetaxel (T; Taxotere) with capecitabine (X) is active against metastatic breast cancer (MBC); bevacizumab (B) has demonstrated efficacy with taxanes in the first-line setting. This study was conducted to assess the efficacy and safety of TX-B in patients (pts) with MBC. Patients and methods: In this single-arm, multicenter phase II study, pts received first-line bevacizumab 15 mg/kg and docetaxel 60 mg/m2 on day 1, plus capecitabine 900 mg/m2 twice per day on days 1-14 every 21 days. The treatment was administrated for 3 cycles and in case of objective response or stable disease at that time, pts were treated with 3 additional cycles. More courses of chemotherapy were administered at Investigator's discretion. Bevacizumab was continued until progressive disease, patient refusal, or unacceptable toxicity. Primary end point was progression-free survival (PFS) and secondary end points were tumor response rate (RR), overall survival (OS), and toxicity.

Results: We report data from the first 30 pts enrolled. Median age was 54 (37-72). Eight (27%) pts had triple-negative disease, while 18 (60%) were hormone-receptor positive. HER2 status was recognized as negative in 25 (84%) pts. TX-B was administered for a median of seven cycles. Five complete and 13 partial responses were observed (overall RR 60%). Median response duration was 12 months. Median OS and PFS were 26 and 11 months, respectively. Grade 3/4 adverse events included tromboembolism (10%), neutropenia (23%), hand-foot syndrome (13%), stomatitis (10%). The median TX doses administered per cycle were 60 mg/m2 and 660 mg/m2, respectively. Ten (33%) pts required dose reductions of docetaxel, while capecitabine dose was reduced in 15 (50%) pts. Results of all the 80 assessable pts entered in the study will be presented at the meeting.

Conclusion: TX-B demonstrated significant activity with an acceptable toxicity profile. Maintenance therapy with B is possible for a long period of stable tumor disease.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-17-01.
Abstract P2-19-01: Modulating Response to Phosphoinositol-3-Kinase/mTOR Inhibition in ER Positive Breast Cancer Cell Lines after Acquired Resistance to Estrogen Deprivation
Authors: Sanchez CG , Crowder RJ , Phommaly C , Ellis MJ.
Published: April 2011

Background: We have recently shown that acquired resistance to aromatase inhibitors (AI) can, in some instances, be reversed low-dose estradiol treatment until progression followed by repeat treatment with an aromatase inhibitor (Ellis, M. JAMA 302:774, 2009). We have also demonstrated that simultaneous inhibition of phosphoinositol-3-kinase (PI3K) and ER (though estrogen deprivation) activates cell death through apoptosis, which is reversed by estradiol, revealing a link between ER activation and the efficacy of PI3 kinase inhibition (Crowder, R. Cancer Res 69:3955, 2009). Consistent with this concept we have also shown that long term estrogen deprived (LTED) cells are co-resistant to both classes of agent (SABCS 2009 Abs#3131). We therefore studied ways to overcome resistance to PI3 kinase inhibitors in LTED cell lines using second line endocrine therapy strategies that could be offered to patients with AI resistant advanced disease.

Methods: ER+ MCF7 and T47D (PIK3CA mutant) cells were cultured in phenol-red free media containing charcoal-stripped serum for 12 months to create long term estrogen deprivation (LTED) variants that mimicked clinical endocrine therapy resistance. A variant of the MCF7 LTED line that regained estrogen-dependent proliferation was created by re-exposure to estradiol for 4 months (MCF7 LTED-R). The effects of the PI3K isoform inhibitor BKM120, the mTOR inhibitor RAD001 and the dual PI3K/mTOR inhibitor BGT226 in MCF7 LTED and T47D LTED were assessed in combination with estrogen deprivation and fulvestrant. MCF7 LTED-R cells were treated with PI3 kinase pathway inhibitors after acute estradiol withdrawal. Apoptosis was measured using TUNEL flow cytometry. Results: MCF7 LTED were resistant to all three PI3 kinase inhibitors, but combination treatment with fulvestrant restored apoptosis to levels seen with parental MCF7 cells after acute estrogen-deprivation. Acute estrogen deprivation in MCF7 LTED-R cells was also effective in restoring an apoptotic response to BGT226. T47D LTED cells, which, unlike MCF7 cells, have markedly down-regulated ER expression, become more sensitive to PI3 kinase monotherapy than the parental T47D cell line, particularly in response to BKM120, and fulvestrant did not increase the apoptotic response.

Conclusions: The MCF7 cell results suggest that PI3 kinase inhibitors will be effective in endocrine therapy resistant advanced breast cancer if these agents are administered with appropriate second line endocrine strategies. Potentially efficacious approaches include fulvestrant combinations and second-line aromatase inhibition, but only after restoration of AI sensitivity with estradiol. The T47D experiments suggest that in the setting of ER+ tumors that lose ER upon disease progression PI3 kinase inhibitors may be very effective.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-19-01.
Abstract P2-19-04: Anti-Jak2 and Breast Cancer Activity of NIM811, a Non-Immunosuppressive Cyclosporine A Analog
Authors: Clevenger CV , Zheng J , Harrington K , Zeng X.
Published: April 2011

Cyclophilin A (CypA) is a member of the immunophilin family of peptidyl prolyl isomerases (PPI) that catalyze the cis-trans interconversion of proline imide bonds of peptides. In this role, cyclophilins have been found to function as signaling switches, regulating the activity of receptors, kinases and transcription factors. The PPI activity of CypA is inhibited by the immunosuppressive drug cyclosporine A (CsA), and in turn the CypA-CsA complex inhibits calcineurin-mediated NFAT activation. Our laboratory has recently demonstrated that CypA is necessary for the prolactin (PRL)-induced activation of Jak2/Stat5 signaling and gene expression (Cancer Res 68:7769, 2008). In addition, we have also shown that CsA inhibits the in vitro and in vivo growth and progression of both ER+ and ER-breast cancer cell lines. Here, we show that the non-immunosuppressive CsA analog NIM811 blocked PRL-stimulated activation of the Jak2/Stat5 pathway, and the kinases AKT and MAPK in T47D cells. NIM811 also inhibited ER+, ER-, and Her2+ breast cancer cell proliferation, survival, motility and anchorage independent growth in a dose-dependent manner. NIM811 inhibited PRL induced gene expression, such as CISH and Cyclin D1. NIM811 also blocked the PRL-induced association between Stat5 and cMyb, and at higher doses, triggered PARP cleavage. In summary, these results indicate that non-immunosuppressive cyclophilin inhibitor NIM811 has significant potential as a novel for breast cancer therapeutic.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-19-04.
Abstract P2-19-05: Inhibitor Screening Utilizing Human Kinase Multiplex Arrays
Authors: Sahli N , Finkel D , Lindberg M , Buehl C , Stoesz S , Campos-Gonzalez R , Brumbaugh K.
Published: April 2011

Protein kinase pathways show dysregulation in cancer, usually displaying increased activation. Kinases are typically influenced by extracellular signals, and many kinases are now considered to be therapeutic targets. Upon phosphorylation, kinases regulate signaling pathways that effect cellular processes. Individual cell lines, including cancer cells, express different kinase activation patterns. Previously, we have shown the phospho-tyrosine kinase profile of MDA-MB-453 cells, most notably the Erb family of Receptor Tyrosine Kinases, to be hyperactivated in this breast cancer cell line. Using a new kinase multiplex plate-based array, we have seen similar activation of multiple intracellular kinases, such as Akt and Erk. As a result, the MDA-MB-453 cells were selected for treatment with a kinase inhibitor library (Enzo, Catalog # BML-2832-0100). While several of the inhibitors block kinase activation, other inhibitors block proteins that are upstream from their kinase target. After incubation with the inhibitor library, a multiplex kinase array was utilized to screen the effects of the inhibitors on the MDA-MB-453 cells. In contrast with MCF-7 cells, MDA-MB-453 breast cancer cells showed a basal hyperactivation of the Akt and GSK-3β kinases. The activation of Akt could be selectively inhibited by SB-203580, SB-202190, ML-7, and BML-265 inhibitors. The results obtained in the multiplex assay were verified by singleplex ELISA for Akt. Hence, the data collected with the multiplex assay provide a rapid analysis of the inhibitors’ effects on defined signaling pathways. This allows for a faster identification of kinase inhibitors that may affect cancer cells.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-19-05.