bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P2-06-12: HER2 mRNA Expression Is Influenced by Estrogen Receptor Status and PI3K Mutational Status
Authors: Bloom KJ , Choppa P , Kyshtoobayeva A , Barry T , Goldstein N.
Published: April 2011

Molecular profiling has suggested that ER+/HER2+ breast cancer is a distinct entity from ER-/HER2+ breast cancer and confers a worse prognosis in systemically untreated patients. It has recently been reported that genes associated with PI3K/AKT pathway activation predict pathologic complete response in patients receiving trastuzumab-chemotherapy in ER+/HER+ but not in ER-/HER2+ tumors. Past studies have indicated that 30-40% of breast cancers harbor a PI3K mutation, predominantly in exons 9 and 20. We analyzed 100 HER2+ breast cancers stratified by ER status to confirm the differing level of HER2 mRNA expression relative to ER status and to determine the association, in any, to the presence of a PI3K mutation.

Methods: Fifty ER+ and 50 ER-breast cancers previously assessed as HER2+ (HER2:CEP-17 ratio ranging from 2.40-16.20, PathVysion Kit, Abbott) were pulled from the archives. ER was assessed by IHC, (6F11, NovaCastra) using a clinically validated method in accordance with the new CAP/ASCO guidelines. Molecular analysis was performed on paraffin embedded tissues. Tumor areas of interest were identified and selectively removed from the section. RNA and DNA were isolated from the collected tissue. Real-time quantitative RT-PCR was used to evaluate the expression of the HER2 gene from the RNA fraction. The expression values were calculated as a ratio of HER2 to the reference gene. The DNA was added to an allele specific PCR to assess 7 of the most common mutations in exons 9 and 20 of the PI3K gene. All data was generated on an AB 7900 HT real-time platform.

Results:

Quantitative HER2 mRNA Results

Discusssion: The level of HER2 mRNA expression differs significantly based on the expression of ER. This is further accentuated by the presence of a PI3K mutation, increasing the level of HER2 mRNA expression is ER-patients and decreasing the level of HER2 mRNA expression in ER+ patients. This observation needs to be further explored and may have profound implications, if as current data suggests, the subset of patients most responsive to trastuzumab therapy are the ER+, PI3K activated tumors.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-06-12.
Abstract P2-06-15: The Use of ADH-5 Stain as a Surrogate Marker for Molecular Classification of Basal and Luminal Breast Cancers
Authors: Turner BM , Yeh I-T
Published: April 2011

BACKGROUND: Immunohistochemical (IHC) analyses of protein expression have recently been used as surrogate assays for the molecular classification of breast cancers. Specifically, ER+/PR+/HER2- tumors are typically considered luminal-type breast cancers, ER-/PR-/HER2+ tumors equate to HER2 over-expressing tumors, whereas ER-/PR-/HER2- (triple negative, TN) tumors are basal-like breast cancers. A newly available cocktail of antibodies called ADH-5 allows for the simultaneous evaluation of both luminal and basal keratins. In this study, we examine the expression of luminal and basal keratins in breast cancers stained with ADH-5, particularly in a subgroup of basal keratin positive tumors. METHODS: All invasive breast cancers from 12/2009-6/2010 in a single pathology laboratory were studied by ADH-5 antibody cocktail consisting of two luminal keratin (LK) markers (CK7 and CK18), and two basal keratin (BK) markers (CK5 and CK14), as well as a myoepithelial marker, P63 (Biocare, Concord, CA). ER, PR, HER2 and Ki-67 were also evaluated by IHC and scored in all tumors. The group included 81 breast cancers and 3 metastatic tumors from the breast. ADH-5 staining was evaluated in areas of invasive carcinoma, and intensity and proportion of staining by BK and LK were scored. Mean statistics and ratios were evaluated and compared for each of the scored stains using the student t-test (stt) and chi squared analysis (cs), respectively.

RESULTS: Of the 84 cancers, 20 (24%) showed some staining for BK, and 63 (75%) showed only LK staining. One case (1%) showed no staining for either BK or LK. Of the BK+ cases, 19/20 (95%) showed some degree of LK staining as well. Only one BK+ tumor lacked luminal keratin positivity (5%). Co-expression both BK and LK was seen in the same cells, though many cells were more strongly positive for one keratin type. The BK+ tumors were significantly more likely to be TN (p = 0.5, cs), and all TN tumors showed some BK staining. Four ER+/PR+/HER2- tumors expressed low levels of BK; two additional ER+/PR+/HER2+ tumors also expressed BK. ER-, PR-and TN tumors tended to show a significantly greater proportion of BK staining and decreased amount of luminal staining than ER+, PR+ and non-TN tumors.

DISCUSSION: ADH-5 stain demonstrates co-expression of luminal keratins in basal keratin positive breast cancers, including all triple negative breast cancers in our study. In addition, some ER+/PR+/HER2- tumors and ER+/PR+/HER2+ (considered luminal A and B, respectively, by current usage of ER/PR/HER2 as surrogate markers) actually express basal keratins, bringing up the issue of whether these tumors should be considered luminal or basal-like. The ADH-5 stain may be helpful as a surrogate molecular marker for the classification of breast cancers.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-06-15.
Abstract P2-07-07: Ehm2 Influences the Aggressiveness of Breast Cancer Cells through Regulation of MMP-9 Expression, and Aberrant Expression of Ehm2 Correlates with Disease Progression in Breast Cancer
Authors: Yu H , Ye L , Zhang Y , Mansel RE , Jiang WG.
Published: April 2011

Background. Ehm2 is a member of NF2/ERM/4.1 superfamily and has been indicated in cell membrane/cytoskeletal interaction and cellular motility. In the present study, we investigated the expression pattern of Ehm2 in breast cancer and the impact of Ehm2 on the biological behavior of breast cancer cells.

Methods. A cohort of breast cancer specimens (normal, n=33; cancer, n=127) was tested for Ehm2 expression, using both quantitative real time PCR and immunohistochemical staining. A hammerhead ribozyme transgene was employed to knockdown Ehm2 expression in breast cancer cells. The effect on growth, cell matrix adhesion, motility and invasion by Ehm2 knockdown was then examined using in vitro assays. Results. Immunohistochemical staining and quantitative real time PCR revealed an increased expression of Ehm2 in breast cancer. High levels of Ehm2 transcripts correlated with disease progression, metastasis and poor prognosis. Disease free survival of the patients with lower level of Ehm2 was 135.8 (125.1-146.5, 95%CI) months, significantly longer compared with 102.5 (78.7-126.4, 95%CI) months of patients with higher level of Ehm2 expression, P=0.039. Knockdown of Ehm2 had inhibitory effects on both in vitro growth and invasion of breast cancer cells. Flow cytometric analysis showed that knockdown of Ehm2 resulted in an increase (>10%) of apoptotic cells overthe control cells. Furthermore, it was shown that knockdown of Ehm2 significantly decreased the protein and mRNA expression of matrix metalloprotease MMP9. Zymograhpy revealed a reduced activity of MMP9 in breast cancer cells after knockdown of Ehm2.

In conclusion, Ehm2 is able to promote invasion of cancer cells via regulating MMP9, and can also protect them from apoptosis. This may have important implication in the correlation between increased Ehm2 expression and poor prognosis and metastasis in breast cancer.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-07-07.
Abstract P2-08-01: XPD Single Nucleotide Polymorphisms and Copy Number Variation in Puerto Rican Women and Breast Cancer Risk
Authors: Pacheco-Torres AL , Colon C , Matta J , Dutil J.
Published: April 2011

Background: The nucleotide excision repair (NER) pathway is the primary mechanism for the repair of bulky and helical distorting adducts. Previous studies suggest that polymorphisms in the XPD gene may contribute to the susceptibility of an individual to develop cancer. However, this data have not been validated in the population of Puerto Rico and most studies were limited to assessing the role of single nucleotide polymorphisms (SNPs). Copy number variations (CNVs) are segments of DNA that have a variable copy number compared to a reference genome. Our objective was to characterize the role of the SNPs and CNVs in the XPD gene and the susceptibility for breast cancer in Puerto Rican women. Material and Methods: Extracted DNA from healthy controls (n=252) and breast cancer patients (n=152) was used to perform TaqMan allelic discrimination for Single Nucleotide Polymorphisms (SNPs) rs13181, rs1799793 and rs238407 of the XPD gene. The XPD genomic region was screened for the presence of copy number variations using a Taqman CNV genotyping assay in exon 10 of the gene.

Results: Interestingly, the control population was in Hardy-Weinberg disequilibrium for the SNPs rs1799793 and rs238407, suggesting the presence of copy number variation (CNV) in the XPD genomic region. In our study population, we found that the copy number around XPD ranges from one copy to more of 10 copies. The distribution of the CNV genotypes between controls and cases was statistically significant (p=0.021). Individuals carrying zero copy of the gene were more likely to develop breast cancer than individuals with more than two copies (Odds ratio: 2.6, 95% confidence interval 5.134 — 1.336, P<0.005). Significance: This study is expected to contribute to the identification of individuals that are at higher risk of breast cancer, prior to the development of the disease. This will aid in prevention and treatment decisions in an individual's breast cancer management strategies.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-08-01.
Abstract P2-09-01: Serial [18F] FDG-PET after the 2nd Cycle of Preoperative Chemotherapy Is Predictive for Pathological Complete Response in Stage II/III Breast Cancer
Authors: Jung S-Y , Kim S-K , Kwon Y , Kim E-A , Ko KL , Park IH , Lee KS , Kang KW , Noh D-Y , Shin SH , Jeong JS , Lee S , Kim SW , Kang H-S , Ro J.
Published: April 2011

Purpose: One of substudies of the prospective trials aimed to evaluate the usefulness of serial [18F] 2-fluoro-2-deoxy-D-glucose-positron emission tomography ([18F] FDG-PET) for predicting pathological complete response (pCR) in stage II/III breast cancer with preoperative chemotherapy (PST).

Methods: Serial PET was undertaken in 57 breast cancer patients enrolled in three different neoadjuvant trials: 35 patients from a phase II study with paclitaxel/gemcitabine/trastuzumab with ClinicalTrial.gov NCT 00532857, 9 patients from a phase Ib study with paclitaxel/gemcitabine/lapatinib with ClinicalTrial.gov NCT 01133912, and 13 patients from a phase Ib with paclitaxel/gemcitabine/sunitinib with ClinicalTrial.gov NCT0 1070706. All patients received 6 cycles of PST followed by surgery and radiotherapy. We assessed the peak standardized uptake value (SUVp) in the primary tumor at the baseline and after the 2nd cycle (37 patients) or after completion (20 patients) of 6 cycles of PST, and calculated the reduction rate (RR) of the SUVp. Pathological response was classified into pCR and non-pCR. To compare the mean of SUVp and RR of SUVp between different response groups, two-way tables and chi-square tests were used

Results: Fifteen (40.6%) of 37 patients who took repeat PET after the 2nd PST and 15 (75%) of 20 patients after completion of PST achieved a pCR with overall pCR rate of 52.6% in the primary tumor. In patients with repeat PET after the 2nd PST, post-treatment SUVp and RR of the SUVp in primary tumors were significantly different by the pathological response (post-treatment SUVp, 1.54 ± 0.63 in pCR vs 2.54 ± 1.06 in non-pCR, P=0.002; RR of the SUVp, 79.2% ± 11.9% in pCR vs 68.9% ± 15.4% in non-pCR, P=0.03). However, in patients with repeat PET after completion of PST, there were no statistical differences of these values (post-treatment SUVp, 1.09 ± 0.63 in pCR vs 1.29 ± 0.36 in non-pCR, P=0.42; RR of the SUVp, 83.7% ± 14.0% in pCR vs 67.5% ± 21.1% in non-pCR, P=0.17)

Conclusions: This study demonstrated that repeat PET after the 2nd cycle of PST, not after completion of PST could predict pCR in stage II/III breast cancer with preoperative chemotherapy. Acknowledgement NCC Grant #0910320.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-09-01.
Abstract P2-09-02: Validation of a Novel Staging System for Disease-Specific Survival in Breast Cancer Patients Treated with Neoadjuvant Chemotherapy
Authors: Mittendorf EA , Jeruss JS , Tucker SL , Kolli A , Newman LA , Gonzalez-Angulo AM , Buchholz TA , Sahin AA , Cormier JN , Buzdar A , Hortobagyi GN , Hunt KK.
Published: April 2011

Purpose We previously described a novel breast cancer staging system for assessing prognosis after neoadjuvant chemotherapy based on pretreatment clinical stage (CS), estrogen receptor status (E), grade (G) and post-treatment pathologic stage (PS). This CPS+EG staging system assigned and summed points for each factor, allowing for better determination of breast cancer-specific survival than CS or PS alone. The current study was undertaken to validate this staging system using internal and external cohorts.

Methods We identified 804 patients treated with neoadjuvant chemotherapy at our institution from 2003-2005 who were not part of the original determination of the CPS+EG staging system and an external cohort of 165 patients treated at another institution. Clinicopathologic characteristics, treatment regimens, and patient outcomes were assessed. Outcomes were stratified by CPS+EG score in order to test whether the 5-year disease-specific survival (DSS) in each CPS+EG group was numerically as predicted.

Results The 5-year DSS for the internal cohort was 77% (95% CI: 72-82) at a median follow-up of 3.4 years (range, 03-5.9). The 5-year DSS for the external cohort was 86% (95% CI: 7 -91) at a median follow-up of 4.7 years (range, 0.5-10.5). The ability of the CPS+EG score to distinguish outcomes based on good or poor prognosis was confirmed in both the internal and external cohorts. In both cohorts, application of the CPS+EG staging system facilitated more accurate categorization of patients into prognostic subgroups than CS or final PS as defined by the American Joint Committee on Cancer (AJCC) staging system.

Disease-Specific Survival Outcomes Based on the CPS+EG Staging System

Application of the CPS+EG to all three cohorts combined, demonstrated 5-year DSS ranging from 23% to 99% versus 5-year DSS ranging from 61% to 92% based on presenting CS or 58% to 95% based on post-treatment PS.

Conclusion The current study validates the CPS+EG staging system in independent patient cohorts. We recommend that biologic markers and response to treatment be incorporated into revised versions of the AJCC staging system for patients receiving neoadjuvant chemotherapy.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-09-02.
Abstract P2-09-06: Meta-Analysis of the Decision Impact of the 21-Gene Breast Cancer Recurrence Score in Clinical Practice
Authors: Hornberger J , Chien R
Published: April 2011

Background: The 21-gene Recurrence Score® (RS) is a clinically validated assay that predicts the likelihood of chemotherapy benefit and the risk of distant recurrence for individual patients with estrogen receptor positive (ER+), early stage breast cancer (ESBC). The 21-gene RS is included in the published guidelines of the National Comprehensive Cancer Network and American Society of Clinical Oncology as an adjuvant-treatment decision tool. Recently, university-led studies have reported approximately 17-27% decision impact of 21-gene RS in actual practice. We reviewed the published literature and conducted a meta-analysis of the impact of the 21-gene breast cancer RS on treatment recommendations and decisions in clinical practice.

Method: Published cohort studies that reported the change in the recommendation or actual use of adjuvant chemotherapy (CTX) before and after the RS were included. Patients must be identified as ER+, LN-ESBC. Outcomes evaluated were: treatment decision change from 1) CTX plus hormone therapy to hormone therapy only or 2) hormone therapy-only to CTX plus hormone therapy. Actual treatment change was used when available. Reductions in the relative and absolute CTX use associated with the 21-gene breast cancer RS were computed with Review Manager 5

Version 5.0.24. (Copenhagen: The Nordic Cochrane Centre, the Cochrane Collaboration, 2008).

Results: 7 published studies, with a total of 913 patients were included in the meta-analysis. 62% (566/913) of patients were recommended or received CTX prior to testing with the 21-gene RS. After RS testing, 29% percent (268/913) of patients were recommended or received CTX. The absolute reduction of CTX use associated with RS testing was 30% (95% CI [-40%, -21%]) The relative risk ratio is computed as a ratio, weighted by the sample size of each study, of (1) proportion of patients treated with CTX plus hormone therapy after 21-gene RS and (2) proportion of patients treated with CTX plus hormone therapy before 21-gene RS. The estimated relative ratio in CTX use before and after 21-gene assay is 49% (95% CI [41%, 58%]). One limitation of this analysis is the heterogeneity in the study designs of the published studies. Of the 7 available studies, 1 was a prospective physician survey; 6 were retrospective chart reviews. A second limitation of the study is in the consistency of reported outcomes. 4 studies reported the use of CTX after 21-gene breast cancer RS, whereas the remaining 3 reported the physicians’ treatment recommendations. Lastly, the meta-analysis does not capture the important benefit of the RS for patients who switch from no CTX to CTX when a high RS is obtained. Conclusion: The meta-analysis shows consistent overall reduction in AC with the adoption of the 21-gene breast cancer RS.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-09-06.
Abstract P2-09-10: Feasibility Assessment of Pharmacogenomic Predictors Developed from Breast Cancer Cell Lines To Predict Breast Cancer Patient Pathological Response in Neoadjuvant Chemotherapy
Authors: Shen K , Rice S , Gingrich D , Wang D , Tian C , Ding Z , Brower S , Ervin P , Huang S , Gabrin M , Song N.
Published: April 2011

Background: Studies of developing pharmacogenomic predictors from cancer cell lines to predict cancer patient clinical response and outcome to chemotherapy have yielded both positive and negative results. The variability in these results may arise from the noise inherent of microarray technology as well as the small sample size of in cell line studies. Therefore, it is greatly needed to evaluate the feasibility of using cell lines to develop pharmacogenomic predictors of patient pathological response. Material and Methods: Thirty breast cancer cell lines were exposed to 10 concentrations of paclitaxel-5-fluorouracil-doxorubicin-cyclophosphamide (TFAC) to measure in vitro chemosensitivity. Two independent and publicly available microarray datasets (Hoeflich and Neve) on these breast cancer cell lines together with the chemosensitivity results were used to identify pharmacogenomic predictors from each dataset. Two independent clinical trials (Hess and Popovici) with publically available data, having 130 and 100 patients respectively, were used as test sets to validate the accuracy of the pharmacogenomic predictors. All patients received TFAC as neoadjuvant therapy and the gene expression profiles of patients were assessed before receiving chemotherapy treatment. The patient's pathological complete response (pCR) was determined after treatment to evaluate the chemotherapy efficacy.

The pharmacogenomic predictors developed from each of the cell line studies were evaluated for their ability to predict patient pCR in each of the clinical trials using the supervised principle component regression method.

Results: The pharmacogenomic predictors identified from the Hoeflich and Neve cell line data (training sets) predicted pCR of the patients in the two clinical trials (test sets) with 64%-68% accuracy, 70%-87% sensitivity, and 60%-67% specificity when 100 genes were selected as pharmacogenomic predictors (Table 1).

Conclusions: The four independent prediction results generated in this study demonstrate the feasibility of using breast cancer cell lines to identify pharmacogenomic predictors of response to chemotherapy treatment for breast cancer patients. Future studies will examine the use of drug responses from primary cultures of patient tumors to develop pharmacogenomic predictors of breast cancer patient responses to chemotherapy treatment.

Table 1. Prediction of breast cancer patients’ pCR by pharmacogenomic predictors derived from breast cancer cell lines.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-09-10.
Abstract P2-09-12: Phosphoproteomic Pathway Profiling of Breast Cancers for Biomarker and Target Discovery for Personalized Therapy
Authors: Kay C , Mullen P , Langdon SP , Harrison DJ , Faratian D.
Published: April 2011

Introduction: The control and activation of signaling pathways has been shown to influence response to targeted therapy (PTEN and PI3K for trastuzumab, KRAS and MAPK for cetuximab). Single tissue biomarkers (eg HER2-receptor status) are often poor surrogates for the activation state of downstream pathways such as MAPK and PI3K. We have used phosphoproteomic profiling in order to reclassify breast tumors for therapeutic response prediction.

Methods: Eleven cell lines were profiled using phosphoproteome arrays (250 targets) and reclassified using unsupervised clustering. Sensitivity to therapy predicted by pathway activation status was correlated with sensitivity to therapies in vitro measured by the AlamarBlue cell viability assay. Target specificity was assayed using in-cell western blotting. Phosphoprotein clusters were validated in 107 breast tumors by reverse phase protein arrays (RPPA), and the expression of targets validated in independent cohorts of cancers by quantitative immunofluorescence (AQUA) on tissue microarrays.

Results: Phosphoproteomic profiling revealed 3 main clusters of breast tumors with overlap with existing molecular phenotypes (luminal, basal, and mixed) and different profiles of pathway activation (Cluster A: anti-apoptotic, Cluster B: growth factor-dependent, Cluster C: cell cycle-dependent). Good to excellent correlations between pathway activation and in vitro sensitivity to targeted therapies were shown for several pathways and individual molecules within pathways, including components of NFkB, STAT, and cell cycle. These biomarkers were shown to be present in clinical cohorts by RPPA and AQUA, and related to molecular phenotype. Conclusions: Phosphoproteomic profiling can be used to reclassify tumors in order to make a priori predictions about therapeutic targeting. These clusters exist in real clinical specimens and are therefore also promising biomarkers for targeted therapy.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-09-12.
Abstract P2-09-22: Predictive Significance of the Proportion of ER-or PgR-Positive Tumor Cells in Neoadjuvant Chemotherapy for Operable HER2-Negative Breast Cancer
Authors: Osako T , Nishimura R , Okumura Y , Hayashi M , Arima N.
Published: April 2011

Background

Estrogen receptor (ER) and progesterone receptor (PgR) status are predictive for clinical and pathological response in neoadjuvant chemotherapy for operable breast cancer. However, it remains unclear how the proportion of ER-or PgR-positive staining tumor cells affect responses to neoadjuvant chemotherapy. The purpose of this study was to examine the correlation of the proportion of ER-or PgR-positive staining tumor cells with the clinical and pathological responses to neoadjuvant chemotherapy for operable HER2-negative breast cancer. Patients and Methods

From April 2002 to May 2010, 101 patients received neoadjuvant chemotherapy containing epirubicin and taxane in our clinic. We assessed menopausal status, tumor size, lymph node status, histological grade, ER, PgR, p53 and Ki-67. In this study, we defined the positivity of ER and PgR as more than or equal to 1% of the positive staining tumor cells according to ASCO/CAP announcement in April 2010. Furthermore, patients with ER-positive tumor were divided into two groups at 30% of ER positive staining tumor cells. Results

A clinical response was seen in 82% of all patients and a pathological complete response (pCR) was seen in 17%. Of the 101 patients, 60 (59%) had ER-positive tumors and 38 (63%) of the patients had tumors with more than 90% of ER-positive staining cells. Also, 52 (51%) had PgR-positive tumors. Fourteen (34%) of the 41 patients with ER-negative tumors achieved a pCR and 15 (31%) of the 49 patients with PgR-negative tumors achieved a pCR. Patients with more than 30% of ER-or more than 1% of PgR-positive staining tumor cells did not achieve a pCR. There was no significant correlation of pCR with menopausal status, tumor size, grade and Ki-67. In univariate analysis, a pCR was associated with ER status (p=0.0001), PgR status (p=0.0001), and chemotherapy regimens (p=0.014). Multivariate analysis revealed that ER status was a significant factor for pCR, and patients with ER-negative tumors were 20.1 times more likely to achieve a pCR than those with more than or equal to 30% tumors staining tumor cells (p=0.005; 95% confidential interval, 2.5-16.5).

Conclusion

We demonstrated a predictive significance of the proportion of ER-or PgR-positive tumor cells in neoadjuvant chemotherapy for operable HER2- negative breast cancer. ER-negativity (<1%) is significantly predictive to achieve a pCR in multivariate analysis. Conversely, it seems very possible that patients with more than 30% ER-or more than 1% PgR-positive staining tumor cells do not achieve a pCR. It is important to take the proportion of ER-and PgR-positive staining tumor cells into consideration in the treatment of breast cancer.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-09-22.
Abstract P2-09-34: ERK Phosphorylation in Fibroblasts within Breast Cancer Stroma Is Associated with an Improved Tamoxifen Response
Authors: Busch S , Ryden L , Stål O , Jirströ K , Landberg G.
Published: April 2011

Background. Extra-cellular signal-regulated kinase (ERK) 1/2 is an integral part of the mitogen-activated protein kinase (MAPK) pathway, transmitting signals involved in cell proliferation and cell death upon binding of extracellular growth factors to their receptors. Whereas its role in tumour cells has been linked to key events in cell transformation, its role in tumour-associated fibroblasts, a component of the tumour stroma, is largely unknown.

Material and methods. Tissue microarrays (TMAs) of two cohorts including invasive breast cancer patients were stained for ERK phosphorylation level (pERK) and SMA-alpha expression (SMAa) which were scored and analysed in regard to clinico-pathological, prognostic and molecular markers and recurrence-free survival.

Results. In both clinical materials we show a significant correlation between stromal ERK phosphorylation and the marker for activated fibroblasts SMAa. SMAa expression was not linked to treatment-predictive information but instead had prognostic qualities (P=0.034, multivariate Cox regression). By analysing the unique randomised tamoxifen trial including patients receiving no adjuvant treatment we show for the first time that ERK phosphorylation in cancer-associated fibroblasts (CAF) was associated with an improved outcome upon tamoxifen treatment. In contrast, patients having ERα-positive tumours but lacking ERK phosphorylation in the stroma did not respond to tamoxifen treatment (P=0.015, multivariate interaction analyses).

Discussion. This suggests that only a subpopulation of activated fibroblasts, defined by phosphorylation of ERK, is linked to a response to tamoxifen treatment illustrating the importance of the stroma for monitoring and potentially mediating treatment effects in premenopausal breast cancer.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-09-34.
Abstract P2-09-36: Role of ABCB1 Polymorphisms as Predictive Markers in Patients with HER-2 FISH Positive Metastatic Breast Cancer Who Were Treated with Taxane Plus Trastuzumab First Line Chemotherapy
Authors: Kim J-W , Im S-A , Kim JH , Ham HS , Han H-S , Kim J-S , Kim T-M , Han S-W , Oh D-Y , Lee S-H , Kim D-W , Cho N , Moon WK , Kim T-Y , Park IA , Heo DS , Bang Y-J.
Published: April 2011

Background: ABCB1 polymorphisms could predict treatment results of taxane therapy in several malignancies. FCGR2A and FCGR3A polymorphisms were associated with clinical outcomes in several diseases after treatment with monoclonal antibody drugs which had antibody-dependent cell-mediated cytotoxicity activity. These polymorphisms could be possible predictive markers after taxane plus trastuzumab (TH) chemotherapy in patients with HER-2-positive metastatic breast cancer (MBC).

Methods: Fifty-seven patients with HER-2 FISH positive MBC who received TH chemotherapy as the 1st-line treatment were enrolled. We analyzed 5 polymorphisms using DNA from peripheral blood mononuclear cells: ABCB1 1236C>T (rs1128503), ABCB1 2677G>T/A (rs2032582), ABCB1 3435C>T (rs1045642), FCGR2A 131H/R (rs1801274), and FCGR3A 158V/F (rs396991), then correlated them to treatment results of patients.

Results: Among 57 patients, 22 patients (38.6%) received weekly paclitaxel plus trastuzumab, 26 patients (45.6%) tri-weekly paclitaxel plus trastuzumab, and 9 patients (15.8%) tri-weekly docetaxel plus trastuzumab. After a median follow-up of 30.6 (range, 0.6-75.9) months, median progression-free survival (PFS) was 15.1 (95% confidence interval (CI), 10.3-19.8) months. ABCB1 2677T allele carriers had longer PFS than the others (42.1 (95% CI, 12.7-71.4) months vs. 13.0 (95% CI, 10.6-15.4) months; p=0.037) along with a tendency toward higher response rate (RR) (86.4% vs. 76.0%; p=0.470) and longer overall survival (OS) (54.7 (95% CI, 43.0-66.4) months vs. 38.9 (95% CI, 18.1-59.7) months; p=0.057). In addition, ABCB1 3435CC genotype carriers had shorter PFS than the others (13.0 (95% CI, 10.8-15.2) months vs. 19.1 (95% CI, 0.0-38.5) months; p=0.039) along with a tendency toward lower RR (78.6% vs. 100%; p=0.567) and shorter OS (38.9 (95% CI, 19.7-58.1) months vs. 54.7 (95% CI, 43.0-66.4) months; p=0.093). ABCB1 1236C>T, FCGR2A 131H/R, and FCGR3A 158V/F were not significantly associated with RR, PFS, and OS. None of these polymorphisms were associated with any grades of hematologic or cardiac toxicities.

Conclusions: Our results support that ABCB1 2677G>T/A and 3435C>T may have predictive roles after the 1st-line TH chemotherapy in patients with HER-2-positive MBC. In contrast, ABCB1 1236C>T, FCGR2A 131H/R, and FCGR3A 158V/F could not predict response after TH treatment.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-09-36.
Abstract P2-09-39: Multi-Gene Predictors Developed from Breast Cancer Cell Lines To Predict Response to Chemotherapy: A Validation Study on US Oncology Study 02-103
Authors: Shen K , Pusztai L , Qi Y , Symmans WF , Song N , Rice SD , Gabrin MJ , O'Shaughnessy JA , Holmes FA.
Published: April 2011

PURPOSE: Multi-gene predictors (MGPs) of response to multidrug chemotherapy regimens were developed using an in vitro chemoresponse assay in which cell lines were exposed to chemotherapy. The goal of this study was to assess the predictive value of these MGPs using clinical breast cancer patient gene expression data from a clinical trial. METHOD: US Oncology 02-103 was a phase II trial in which women with stage II/III breast cancer were treated with neoadjuvant chemotherapy consisting of four cycles of fluorouracil/epirubicin/cyclophosphamide (FEC) followed by four cycles of docetaxel/capecitabine (TX). Most HER-2 positive patients also received trastuzumab. MGPs of FEC, TX and TFEC (docetaxel/fluorouracil/epirubicin/cyclophosphamide) sensitivity were developed using in vitro assay results from breast cancer cell lines exposed to these drug combinations and publicly-available gene expression data for the same cell lines. MGPs were not developed for trastuzumab treatment. Area under the receiver-operator curve (AUC) was used to evaluate the performance of the three MGPs’ to predict patient pathologic complete response (pCR). Patients who did or did not receive trastuzumab were evaluated separately. Validation was performed blindly and the predictors were applied without knowledge of patient clinical outcome. RESULTS: Eighty-six patients had genomic data available and were included in this analysis. The predictive performance of the FEC, TX and TFEC MGPs were AUCs of 0.72, 0.69, and 0.73, respectively, in the patients who received FEC-TX chemotherapy without trastuzumab (n=61). Within this group, higher AUCs were observed in ER-negative patients compared to ER-positive patients (0.69, 0.72, 0.74 vs. 0.64, 0.54, 0.62, respectively). The prediction accuracies were low (AUCs = 0.43, 0.56 and 0.43) for patients who received trastuzumab together with chemotherapy (n=25) as expected, indicating that the MGPs may have the potential to be regimen-specific.

CONCLUSION: Cell line-derived MGPs of multidrug chemotherapy regimens showed promising performance in this blinded validation study, particularly among patients with ER-negative breast cancers. Further clinical data are needed to confirm this finding.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-09-39.
Abstract P2-10-01: The Accuracy of BRCA1/2 Mutation Prediction Models in Different Ethnicity and Gender: Experience in a Chinese Cohort
Authors: Kwong A , Wong CHN , Suen DTK , Choi CLY , Wong CLP , Law FFB , Kurian A , Ma ESK , West DW , Ford JM.
Published: April 2011

Background: Risk models (BRCAPRO, Myriad, Couch and Shattuck-Eidens, BOADICEA) are well established in Caucasian and African American cohorts to estimate the probability of BRCA1/2 mutation. Few studies have suggested its performance limitation in Asian cohorts. Most studies did not account for gender specific prediction. The aim of the study is to evaluate the performance of these models in a Chinese cohort who have breast/ovarian cancer at a pre-genetic test setting. Methods: Four risk assessment models, Boadicea, BRCAPRO, Myriad, Couch and Shattuck-Eidens, were used to perform risk calculations to 217 non-BRCA carriers (198 females and 19 males) and 32 BRCA carriers (28 females and 4 males. Sensitivity, specificity and area under the receiver operator characteristic (ROC) curve were calculated for each model to evaluate for calibration, discrimination and accuracy in BRCA mutation prediction stratified by gender.

Results: The mean in prediction score in all models were statistically significantly higher in female BRCA mutation carriers. However, there were no statistically difference in mean prediction score between BRCA carriers and non-carriers in all models for male patients. BRCAPRO slightly over-estimated the total numbers of BRCA1 and BRCA2 female carriers (13 vs. 11 and 20 vs.17), but underestimated the number of BRCA2 male carriers (2.8 vs. 4). While Myriad underestimated both the total numbers of BRCA1/2 male (3.1 vs. 4) and female (25.6 vs. 28) carriers. Boadicea did the closest estimation for both male (3.2 vs. 4) and female (1.5 vs. 11 for BRCA1 and 16.3 vs. 17 in BRCA2). BRCAPRO showed the greatest ROC area for BRCA1 (93%), BRCA2 (73%) and BRCA1/2 (79%) combination mutation prediction and highest sensitivity at conventional thresholds of 10% and 20% in female patients (71.4% vs.60.7%). Boadicea had the greatest ROC area for BRCA2 and BRCA1/2 combination mutation prediction and the same sensitivity at conventional thresholds of 10% and 20% in male patients (92% vs. 93%).

Conclusion: All 4 models could perform reasonably well in female patients, but not in male patients. Boadicea has the best performance in male and female Chinese cohort overall. Whereas when comparing females alone, BRCAPRO is most accurate.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-10-01.
Abstract P2-10-04: Characterization of Mutations and Sequence Variations in Breast Cancer Susceptibility Gene 2 (BRCA2) in a Group of Breast Cancer Patients in Sri Lanka
Authors: De Silva S , Tennekoon KH , Karunanayake EH , Amerasinghe I , Angunawala P , De Silva W.
Published: April 2011

Background: Germline mutations in tumour suppressor genes BRCA1 and BRCA2 confer susceptibility to breast and ovarian cancers. Breast cancer is a leading cause of death in women aged 40-60 years in Sri Lanka. Incidence though low in Sri Lanka, compared to the developed world has almost doubled during the last 15 years. BRCA1 mutations and sequence variants in a group of Sri Lankan breast cancer patients have been characterized recently. Here we report mutations and sequence variants of BRCA2 using the same cohort.

Methods: A total of 129 patients comprising of 75 familial breast cancer patients (mean age at diagnosis is 44.64 ± 12.7 years) and 54 sporadic breast cancer patients (mean age at diagnosis 49.13 ± 11 years) were analyzed for BRCA2 sequence variants and mutations. 20 healthy controls were used to compare the results of the patients. Genomic DNA from blood samples was isolated and all exons except exon 11 were screened using Single Strand Conformation Polymorphism (SSCP). Samples in which abnormal bands were detected were confirmed by direct sequencing using MegaBACE 1000 automated DNA Sequencer. Exon 11 will be subjected to direct sequencing without SSCP Technique. Representative samples from exons which did not show abnormal bands were also subjected to direct sequencing.

Results: To date twelve sequence variants have been found in this study. One reported sequence variant: c.203G>A/exon2 (44 familial and 39 sporadic cases), one novel silent mutation: c.969C>T/exon9, one novel missense mutation: c.971C>G/exon9, and one reported intronic variant: IVS8-1G>C/exon9 (in one familial case each) were detected. In exon 10, three reported missense mutations: c.1093A>C (10 familial cases), c.1342A>T/C (58 familial cases) and c.1352C>T (4 familial cases), two novel silent mutations: c.1191A>C (5 familial cases) and c.1353C>T (1 familial case) and one reported silent mutation: c.1593A>G (5 familial cases) were characterized. In exon 14, one novel silent mutation: c.7452A>G (20 familial cases and 28 sporadic cases) and one reported silent mutation: c.7470A>G (one familial case) were identified. Discussion: Among the BRCA2 exons screened to date, seven six reported sequence variants and five novel mutations have been detected. Out of them one pathogenic mutation (IVS8 1G>C/exon9) found in one familial case and in the same patient two other novel missense and novel silent mutations were detected (c.971C>G/exon9 and c.969C>T/exon9). Studies are in progress to screen the remaining exons of BRCA2 gene in this cohort of breast cancer patients. Data obtained will help in developing an economical screening test for BRCA2 mutations in Sri Lanka. Supported by Sida/Secretariat for Research Cooperation Grant for Molecular Biology and Biotechnology

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-10-04.
Abstract P2-11-02: Mapping Mutations: Importance of Family Geographic Origin in Portuguese High-Risk Breast/Ovarian Cancer Families for BRCA Mutation Screening
Authors: Fragoso S , Machado P , Santos S , Rodrigues P , Parreira J , Bento S , Luís A , Vaz F.
Published: April 2011

Background: Hereditary breast/ovarian cancer is mainly related with BRCA1/2 mutations. The complete screening of both genes is long and expensive due to their large number of exons and the inexistence of hotspots. Besides our most frequent BRCA mutation, c.156_157insAlu in BRCA2, the Portuguese population has many other mutations in these genes, described over the years. Aim: to evaluate the importance of screening specific mutations, found in BRCA positive families, in other probands with the same geographic origin. Patients and methods: 407 high risk breast/ovarian cancer patients were screened for c.156_157insAlu in BRCA2 (the Portuguese founder mutation), BRCA1/2 point mutations and large rearrangements. All probands were questioned about their family origin. Mutations detected in families were studied in new probands with the same geographic origin. Results: we were able to draw a map with the geographic origin of our 87 BRCA positive families. Most of our BRCA families have origin in the centre of Portugal. While searching probands with the same geographic origin as others previously detected, we found three additional mutations without screening the whole genes: 1 family from Madeira island; 1 family from the south (Algarve) and 1 non-affected individual, without previous family testing, that was positive for c.156_157insAlu in BRCA2 (her family had origin in an area of high prevalence of this mutation). Our maps also allowed the linking of several families with the same mutation and family origin, but apparently unrelated. Conclusion: Mapping geographic origin of BRCA1/2 mutations may help in the quick identification of new BRCA1/2 carriers and, in some cases, reduce the screening time. Most importantly it permits the linking of apparently unrelated families. This link permits a more accurate estimation of BRCA1/2 positive families and a better understanding of all at-risk family members.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-11-02.
Abstract P2-12-02: How Common Is Sexual Dysfunction among Women with Early Stage Breast Cancer?
Authors: Dent SF , Frechette DAM , Verma S , Clemons MJ , Wheatley-Price PF , Gertler SZ , Song X , Asmis TR , Graham NA , Paquet L.
Published: April 2011

Background: Despite the high prevalence of sexual problems (50—90%) among women with early stage breast cancer (EBC), relatively little is known about the prevalence of clinically significant post-systemic treatment sexual dysfunction (SD) and its impact on quality of life (QOL). According to the American Psychiatric Association (APA), SD consists of sexual problems causing marked personal distress. While 40% of healthy US women report sexual problems, only 12% experience SD. No study has applied the APA criterion to document the prevalence of SD in women with EBC who have completed systemic therapy. The goal of this study is to address this gap and to examine some correlates (anxious predispositions and menopausal symptoms) of SD and the impact of SD on QOL. Methods: Post-menopausal women receiving cancer therapy for EBC or early colon cancer were approached for this study. Sexual problems were evaluated with the Female Sexual Function Index while sexual distress was assessed with the Female Sexual Distress Scale. QOL was measured with FACT-B, using its endocrine symptoms subscale (ES) to assess menopausal symptoms. Spielberger State-Trait Anxiety Inventory measured anxious predispositions. Participants completed questionnaires upon completion of adjuvant chemotherapy but prior to initiation of hormonal therapy (estrogen sensitive EBC). SD was assessed using the APA classification. Results: Between January 2009 and May 1 2010, 70 EBC patients entered this study. The proportion of women (mean age 61) reporting 1 or more sexual problems was high (93%) and included problems with 1) frequency (66%) and level (65%) of sexual desire, 2) frequency (66%), level (52%) and satisfaction (43%) of arousal, 3) frequency (21%) of vaginal dryness and difficulty becoming lubricated (47%) during sexual activity, 4) satisfaction with their sexual life (30%) and 5) pain during intercourse (18%). However, the APA criterion identified only 30% of patients as having SD. Multiple logistic regression showed that anxiety predispositions and menopausal symptoms predicted SD (p< .01). Women classified as having SD had higher anxiety scores (No SD=34 SD=41, p<.05) as well as higher levels of menopausal symptoms (i.e, lower ES scores; No SD= 62 SD=54, p<.01). QOL was negatively impacted by SD (r=-0.42, p<.01). Discussion: Despite the fact that an overwhelming majority of patients had sexual difficulties, the APA distress criterion identified far fewer patients (30 %) with SD. Nevertheless, we found a high prevalence rate of SD, which was 2.5 times higher (%2 < .01) than that reported in women without a cancer diagnosis. Women with anxious predispositions and high menopausal symptoms may be at risk for SD and may benefit from early interventions to prevent SD.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-12-02.
Abstract P2-13-01: Effects of 12-Week Resistance Training on Strength, Fatigue Syndrome and Quality of Life with Breast Cancer Patients during Chemotherapy
Authors: Baumann FT , Leskaroski A , Knicker A , Krakowski-Roosen H , Bloch W , Harbeck N.
Published: April 2011

Introduction: Up to 100% of all breast cancer patients are suffering from fatigue during chemotherapy [Mock 2001, Jacobsson 1998, Irvine 1994]. Causality as well as the treatment of this non-uniform symptom complex is yet unclear. Exercising studies show a positive effect of endurance training on the development of fatigue [Dimeo 2003]. The study situation regarding feasibility and efficiency of pure resistance training with breast cancer patients with fatigue is still unsatisfactory [De Backer 2009, Courneya 2007].

Methods: In this prospective, controlled, randomized study 32 female patients were recruited 6 weeks after operation. 20 probands (IG) accomplished a 12-week resistance training (2 x per week, 60 min each with 75% of the maximum voluntary capacity) during adjuvant chemotherapy. The control group (CG, n=12) did not train. The measurements took place before and after the intervention and maximally isometric and isocinetic strength was tested with the Isomed 2000. Quality of life was evaluated with the EORTC QLQ-C30 inclusive module BR23 and fatigue symptoms by the MFI questionnaires.

Results: No exacerbations could be observed during the study. Strength (p=0.029), fatigue (p=0.048) and different ranges of the quality of life significantly improved in IG, while in CG fatigue and quality of life worsened. 4 out of 5 ranges of the fatigue diagnostic (f.e. General Fatigue, p=0.01) as well as nearly all ranges of the quality of life evaluation were significantly different between IG and KG. In addition, muscle strength (of the arm concerned) showed significant differences between the groups (p=0.032).

Discussion: It could be shown for the first time, that maximum strength testing as well as resistance training with breast cancer patients were feasible already 6 weeks post-OP. Positively, the increase in muscle power, the reduction of fatigue symptoms and the improvement of the quality of life occurred despite concurrent cytotoxic treatment. We rate the high acceptance of the training program as a success. Based on the positive results and the past studies [De Backer 2009, Courneya 2007] we recommend the integration of strength training into the early rehabilitation of breast cancer patients. We note that this study will be completed by December 2010 yet further studies have to be conducted, in order to better understand the effects of different training intensities. Literature:

Courneya et al.: JCO 2007; 25: 4396-4404

Dimeo et al.: Sup Care in Canc 2003; 11:S623-628

Irvine et al. Cancer Nurs 1994; 17: 367-378

Jacobsson et al. J Pain Symptom Manage 1999;18 (4): 233-242

Mock V. Cancer Suppl 2001; 92 (6): 1699-1707

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P2-13-01.