bluuebunny/crossref_metadata_2025 huggingface dataset:
Hyperbilirubinemia in Lapatinib Treated Patients Is Associated with Gilbert's Syndrome UGT1A1 Polymorphism.
Authors: Spraggs C. , Budde L. , Briley L. , Bing N. , Newstat B. , Rappold E. , Mooser V. , Cardon L. , Stein S.
Published: December 2010

Background: Lapatinib, an oral EGFR/HER2 tyrosine kinase inhibitor, is approved in combination with capecitabine for the treatment of patients with advanced or metastatic HER2-positive breast cancer who have been treated previously with an anthracycline, taxane and trastuzumab. Treatment associated elevations in transaminases and bilirubin have been observed in lapatinib clinical trials. A common TA repeat polymorphism in the promoter region of uridine diphosphoglucuronyl transferase (UGT1A1*28, TA7 allele) reduces UGT1A1 enzyme levels and has been associated with hyperbilirubinemia for a range of drugs. We sought to determine the effect of the UGT1A1*28 variant on bilirubin elevation in lapatinib treated patients.Patients and Methods: Association between the UGT1A1 TA repeat polymorphism and total bilirubin levels (TBL) was examined in lapatinib treated patients using data pooled from 12 trials in advanced and metastatic breast cancer. A total of 899 lapatinib treated (as monotherapy or combined with chemotherapy) patients provided a germline DNA sample and were available for this analysis. UGT1A1 TA repeat genotype was determined by PCR amplification and sequencing. For case-control analysis, patients were defined as having normal TBL at baseline, with on-treatment TBL ≥1.5 and ≤1 times upper limit of normal for cases and controls, respectively, and at least 13 weeks of treatment for controls.Results: The UGT1A1 TA repeat polymorphism strongly correlated with lapatinib associated hyperbilirubinemia. A significant association (P=1.03x10-5, 60 cases and 396 controls) was observed. For hyperbilirubinemia cases, 21/60 (35%) were carriers of two TA7 alleles and 28/60 (47%) were carriers of one TA7 allele. Overall, when compared with other genotypes, the odds ratio (95% confidence interval) of the TA7/TA7 genotype for developing hyperbilirubinemia was 10.7 (5.3-21.6).Conclusion: These data suggest that UGT1A1*28 accounts for the major proportion of total bilirubin elevation observed during lapatinib treatment. This is consistent with an underlying predisposition to Gilbert's syndrome. Given the life-threatening nature of advanced and metastatic breast cancer and the benign clinical course of Gilbert's syndrome, these data support treatment continuation with lapatinib for patients with mild to moderate, isolated, indirect bilirubin elevation, when accompanied by appropriate ongoing safety evaluation (eg, bilirubin fractionation, ALT evaluation) to exclude liver injury risk in these patients.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 1112.
Significance of Viral Reactivation in Hepatitis B Virus Carriers with Early Breast Cancer Receiving Anthracycline- or Taxane-Containing Chemotherapy.
Authors: Kim S. , Ro J. , Park I. , Lee K. , Jeong J.
Published: December 2010

Background :It is known that cancer patients in healthy hepatitis B viral (HBV) carrier status are prone to develop hepatic dysfunction during cytotoxic chemotherapy. The objectives of this study were to determine the frequency of healthy HBV carrier status and subsequent viral reactivation resulting in hepatic dysfunction during cytotoxic chemotherapy, and its significance in early breast cancer patients.Methods :Among 3,433 patients with operable breast cancer diagnosed at the National Cancer Center, Korea between January 2001 and March 2009, 3,337 patients were tested for HBsAg. Retrospectively, medical records were reviewed for 139 (4.2%) patients who were positive for HBs Ag.Results :Of 139 patients, 112 patients received anthracycline or taxane based neoadjuvant (n=25) or adjuvant (n=87) combination chemotherapy. Prophylactic lamivudine therapy was administered in 33 patients (29.5%).Thirty of 112 (26.8%) patients developed reactivation of HBV with deterioration of hepatic function during chemotherapy. The rate of reactivaton was not different with or without further taxane therapy (43% vs. 44%, p=0.408).Although there was no significant difference in the rate of chemotherapy delay between two groups, the incidence of early termination of planned chemotherapy was higher in B viral reactivation group (23.3% vs. 4.9%, p=0.008). Within a median follow-up duration of 39 mo (range 1 mo to 95 mo), the rate of disease recurrence was comparable between two groups (10% vs. 18.3%, p=0.390).Lamivudine prophylaxis decreased the rate of HBV reactivation without statistical significance (15.1% vs. 35.4%, p=0.101). Despite more patients without prophylaxis developed G 3/4 AST/ALT elevation (40% vs. 84%, p=0.068), the majority of patients recovered to G 1/2 AST/ALT level within 12 months with anti-viral or supportive treatment (0% vs. 14.3%, p=1.00). One patient without lamivudine prophylaxis developed fulminant hepatic failure after 3rd cycles of neo-adjuvant chemotherapy and received living donor liver transplantation. After that, she received radical mastectomy and maintained adjuvant anti-hormonal therapy.Conclusions :Although lamivudine prophylaxis in HBV healthy carrier decreased the rate of viral reactivation during neo- or adjuvant chemotherapy, the majority of patients irrespective of prophylaxis recovered hepatic function in a year with conservative treatment. Early detection and intensive supportive care are as important as anti-viral prophylaxis in HBV healthy carrier when treated with cytotoxic chemotherapy.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 1113.
Prospective Study of Ice Gel Pack as Less Expensive Alternative for Prevention of Skin and Nail Toxicity in Patients with Breast Cancer Receiving Docetaxel.
Authors: Matsumoto K. , Hino C. , Fukuda K. , Hamamoto M. , Tanioka M. , Hirokaga K. , Maekawa Y. , Negoro S. , Takao S.
Published: December 2010

Background: Docetaxel (DTX)-induced nail and skin toxicity can be prevented by frozen glove. We investigate whether we can use cooling gel pad (CGP), which is available less than 10 dollars in Japan, as less expensive alternative.Method: Patients (pts) with breast cancer receiving DTX as preoperative or post operative chemotherapy were eligible for this study. Two CGPs, which were refrigerated for eight hours beforehand, were used to cover patient's hand from 15 minutes before to 15minutes after DTX administration. To reduce discomfort from icing, pts were wearing cotton glove during CGP usage. If pts felt CGP as too cold during the cooling process, they were allowed to remove CGP tentatively until they felt comfort. Another hand was not covered as the control. Nail and skin toxicity were assessed at each cycle by CTCAEver.3.0 and documented by photography. Fisher's exact probability test was used. Satisfaction and discomfort of CGP covering was assessed by pts using two item questionnaires. All patients gave written informed consent. The protocol was approved by institutional review board.Result: From October 2006 to September 2008, 50 pts were enrolled to this study. All the patients were female, and received DTX 75mg/m2 in every three weeks schedule for four cycles. Most of patients received DTX after anthracycline-containing regimen. Both nail (10% v 66%, p<0.0001) and skin (0% v 34%, p<0.0001) toxicity was reduced in CGP covered hands. Nail toxicity was grade (G) 0 in 90% v 34%, G1 in 10% v 42%, G2 in 0% v 22%, and G3 in 0% v 2%, for CGP covered and the control hand, respectively. Skin toxicity was G0 in 100% v 66%, G1 in 0% v 4%, G2 in 0% v 30% in CGP covered hand and the control hand, respectively. About the timing of toxicity occurrence, “after the first cycle” was most common, 13 pts (39%) in nail change and 7 pts (41%) in skin toxicity. In the questionnaires, 94% were satisfied with CGP covering.Discussion: The iced CGP cover method reduced the incidence and severity of nail and skin toxicity associated with DTX, even if patients wear cotton glove and remove CGP for a while, in case of excess coldness. This method has no adverse effect, and patients were satisfied with the effect. The iced CGP may be considered as a new, effective and less expensive method for prevention of nail and skin toxicity caused by DTX.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 1114.
Abnormalities of 53BP1 in Basal-Like Breast Cancer.
Authors: Aly A. , Yang Q. , Bilal E. , Yao M. , Bhanot G. , Toppmeyer D. , Haffty B. , Ganesan S.
Published: December 2010

Background: Abnormalities in the DNA repair mechanism have been proposed to play a central role in the development of breast cancers. However the specific mechanism underlying DNA repair defects in sporadic breast cancer are mostly unknown. As 53BP1 plays a critical role in DNA repair response and has been identified as a potential tumor suppressor, its expression in basal-like breast cancer cell lines and clinical samples was investigated.Experimental Design: mRNA expression of 53BP1 was investigated in publicly available gene expression data sets. Protein expression of 53BP1 was evaluated in a large set of clinically annotated breast cancer specimens by immunohistochemistry. Analysis of 53BP1 and related proteins was performed in a set of human and mouse breast cancer cell lines.Results: Gene expression analysis demonstrated that a subset of basal-like breast cancer had decreased expression of 53BP1. Analysis of over 400 breast cancers in a tissue microarray showed that loss of 53BP1 staining was almost exclusively found in tumors that lacked ER, PR and HER2 expression; this "triple-negative" phenotype is characteristic of BLC. Approximately 42% of "triple-negative" breast cancers have abnormally low levels of 53BP1, while <2% of tumor that express either ER or HER2 lack 53BP1 expression. Tumors that have low 53BP1 expression are associated with worse distant metastasis free survival when compared to tumors with normal 53BP1 expression. A set of breast cancer cell lines with abnormal 53BP1 expression was also identified. 53BP1 abnormalities were present in some tumors and cell lines with known BRCA1 mutations.Discussion: These data suggest that a subset of basal-like breast cancers lack normal 53BP1 expression and imply that these tumors may have profound defects in DNA repair/check point function that could be exploited therapeutically. As 53BP1 abnormalities were found in some tumors with known BRCA1 mutations, this suggests that loss of 53BP1 may cooperate with abnormalities in the BRCA1-related DNA repair pathways in the pathogenesis of basal-like breast cancers. Data regarding the molecular mechanisms underlying 53BP1 abnormalities in breast cancer cell lines and samples will be presented.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 1122.
Estrogen and IGF-I Independently Down-Regulate Critical Repressors of Breast Cancer Growth.
Authors: Casa A. , Casa A. , Malik S. , Malik S. , Lazard Z. , Kuiatse I. , Tsimelzon A. , Creighton C. , Hilsenbeck S. , Lee A. , Lee A. , Lee A. , Oesterreich S. , Oesterreich S. , Oesterreich S.
Published: December 2010

Background: Estrogen receptor and insulin-like growth factor (IGF) signaling pathways are important for both normal mammary gland development and breast cancer pathogenesis. Despite this evidence, cross-talk between these two critical mitogenic and survival pathways remains poorly understood, particularly at the level of downstream gene transcriptional networks.Methods: We performed microarray analysis on ER-positive MCF-7 breast cancer cells following stimulation with either estradiol (10nM) or IGF-I (10nM) for 3hr or 24hr. We defined an E2-IGF gene expression signature and examined the effect of the gene signature on time to recurrence in ER+ tumors in the publicly available NKI dataset. Q-RT-PCR was used to validate the effects of IGF-I and E2 on mRNA of repressed genes. To examine gene regulation further, MCF-7 cells were incubated with various signaling inhibitors and gene expression measured.Results: We found 183 probesets to be co-regulated by E2 and IGF-I after 3hr of stimulation and 454 probesets to be co-regulated at the 24hr time point. Patients with tumors that had the same set of genes up- and down-regulated had a poor outcome (p=7.04E-07). E2-IGF co-regulated genes showed a significant enrichment for down-regulated genes (p<0.001). The top 10% of down-regulated genes contained four (BLNK, SOCS2, CCNG2, and ING4) that have previously been shown to have tumor suppressor function and/or show loss in human breast tumors. We confirmed that E2 and IGF-I indeed repressed levels of these candidate genes. Blockade of ER by ICI 182780 (ICI) completely reversed E2-mediated repression but had little or no effect on IGF-mediated repression of these genes. Similarly, blockade of IGF-IR was able to completely reverse IGF-I-mediated repression of all four genes, but had essentially no effect upon E2-mediated repression. Inhibition of the PI3K pathway with LY294002 (20μM) affected short-term (3hr) down-regulation of BLNK and SOCS2 by both E2 and IGF-I, and CCNG2 by IGF-I. By 24 hours, LY completely reversed both E2 and IGF-I mediated repression of CCNG2, and significantly reversed both E2 and IGF-I repression of BLNK and SOCS2. Blockade of MEK1/2 with U0126 (10μM) had no effect on short-term repression by E2 or IGF-I, though after 24hr it affected both E2 and IGF-I repression of BLNK and SOCS2.Conclusions: E2 and IGF-I co-regulate a set of genes that affect breast cancer outcome. E2 and IGF-I down-regulate several critical tumor suppressor genes. While the down-regulation is independent of each other at the level of ER and IGF-IR, for some genes, there is convergence on the PI3K pathway for repression.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 1126.
Let-7 Inverts the Chemoresistance of Breast Tumor-Initiating Cells.
Authors: Yu F. , Wu J. , Gong C. , Su F. , Song E.
Published: December 2010

Background: Cancers may arise from rare self-renewing highly tumorigenic “stem” cells, which are thought not only to be the source of the tumor, but also to be responsible for resistance to cancer therapy and subsequent tumor recurrence. Our preliminary data strongly suggest that let-7 is a master regulator of BT-IC self-renewal and multipotency. It is likely that let-7 regulates many other genes/pathways during BT-IC differentiation. However, we do not know whether let-7 might also regulate other putative aspects of “stemness” such as chemoresistance.Material and Methods: Quantified qRT-PCR was used to test the expression of let-7a in chemoresistant breast tumor tissue from patients, as well as the chemosensitive samples. The location of let-7a in primary breast cancer samples from patients with chemoresistance were compared to the samples without chemoresistance by in situ hybridization analysis.SK-3rd Cells, which is a cell line enriched in breast tumor-initiating cells, were cultured in suspension to form mammosphere which could be induced to differentiation with adhesive culture. Lentivirus-mediated let-7a transduction was used to increase the expression level of let-7a.Let-7 target gene expression (HRAS and HMGA2) was determined by qRT-PCR and western blot. The response of BT-ICs to chemotherapy was determined by MTT, Annexin V and Tunel.Results: The expression level of let-7a microRNA in chemoresistant patients were significantly lower than the patients without chemoresistance (P<0.05). The let-7a mimics can effectively enhance the expression of let-7a in mammospheric SK-3rd cells, and reduced the expression level of H-RAS and HMGA2 proteins, which are the targets of let-7. Mammospheric SK-3rd cells tranfected with let-7a mimics had a lower proliferation rate and a higher apoptosis rate under epirubicin treatment.Conclusion: Lack of expression of let-7 regulates BT-IC resistance to chemotherapy. Therefore, let-7a may be an effective maker to predict the response of tumor to chemotherapy.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 1136.
Increased Nuclear Localization of Transcription Factor Y-Box Binding Protein 1 Accompanied by Up-Regulation of P-Glycoprotein in Breast Cancer Pretreated with Paclitaxel.
Authors: Fujita T. , Nagura N. , Koshikawa K. , Nishimura H. , Ito K. , Izumi H. , Kohno K. , Fujimori M.
Published: December 2010

Purpose: The Y-box binding protein 1 (YB-1) regulates expression of P-glycoprotein (P-gp) encoded by the MDR1 gene. There have been no previous studies regarding the involvement of YB-1 in the development of resistance to paclitaxel. The present study was performed to examine how paclitaxel affects the localization and expression of YB-1 in breast cancer.Experimental Design: We evaluated the expression and localization of YB-1 and P-gp in breast cancer tissues obtained from 27 patients before and after treatment with paclitaxel. The effect of paclitaxel on localization of cellular YB-1 was examined by using GFP-YB-1. Interaction of YB-1 with the Y-box motif of the MDR1 promoters was studied by EMSA. The effects of paclitaxel on MDR1 promoter activity were examined by luciferase assay.Results: Of 27 breast cancer tissues treated with paclitaxel, nine (33%) showed translocation of YB-1 from the cytoplasm to the nucleus together with increased expression of P-gp during the course of treatment. Twelve breast cancer tissues (44%) showed neither translocation of YB-1 nor increased expression of P-gp. Nuclear translocation of YB-1 was correlated significantly with increased expression of P-gp (P=0.0037). Confocal analysis indicated that paclitaxel induced nuclear translocation of green fluorescent fused YB-1 in MCF7 cells. Furthermore, binding of YB-1 to the Y-box of MDR1 promoter was increased in response to treatment with paclitaxel. In addition, MDR1 promoter activity was significantly up-regulated by paclitaxel in MCF7 cells (p< 0.001).Conclusions: The results of the present study suggested that YB-1 may be involved in the development of resistance to paclitaxel in breast cancer.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 1141.
Methylation Status of Promoters 0K, 0N and a Newly Identified Promoter Regulate ERβ1 Expression in Breast Cancer.
Authors: Al-Nakhle H. , Smith L. , Hughes T. , Cummings M. , Hanby A. , Shaaban A. , Burns P. , Speirs V.
Published: December 2010

Gene expression is regulated at multiple levels, including transcriptional silencing by methylation. ERβ1 is downregulated in breast cancer compared to normal breast and mechanisms surrounding this are unclear. Two novel ERβ mRNA isoforms with distinct 5'-UTRs have been identified, ERβ-0K-1 and ERβ-0N-1. The aim of study was to examine whether methylation at these promoter regions plus a novel promoter identified by our group were involved in ERβ1 regulation (figure 1).Bisulfite modification and direct sequencing analysis were performed for promoters 0K, 0N, and a novel mini CpG island upstream of ERβ exon1 in two ERβ1- and one ERβ1+ breast cancer cell lines. Distinct methylation patterns were observed. Promoter 0N was completely methylated in BT20, partially methylated in MDAMB453 and unmethylated in T47D, while the mini CpG island was methylated in all 3 cell lines. In contrast, promoter 0K was unmethylated. Furthermore, a negative correlation between ERβ1 mRNA expression and the methylation status of promoter 0N was observed in breast cells. To further investigate whether methylation of the ERβ1 promoter was responsible for the loss of ERβ1 expression, BT20 and MDAMB453 cells were treated with either 5-aza-dC, TSA or both, concentrations of which had been previously optimised for each cell line. In BT20 cells which had complete methylation of 0N, both agents were required for induction of ERβ1 and ERβ-0N-1 but not ERβ-0K-1 expression, both of which are part of the 5'UTR region. However in MDAMB453 treatment with 5-aza-dC was sufficient to induce ERβ1 and ERβ-0N-1 expression with no additional re-expression seen with TSA and no effects on ERβ-0K-1. This suggests that promoter 0N is responsible for driving the transcription of ERβ1. On going work is confirming these observations in clinical samples. Our results suggest that promoter 0N plays an important role in regulation of ERβ1 mRNA expression in breast cancer. Our results add to growing literatures which demonstrate ERβ1 is regulated at multiple levels in breast cancer and that DNA methylation is an important mechanism for silencing ERβ1 gene expression.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 1148.
Gene Promoter Hypermethylation in Tumors and Matched Affected Lymph Nodes of Breast Cancer Patients.
Authors: Van der Auwera I. , Renard I. , Van den Eynden G. , Limame R. , Trinh X. , van Dam P. , Van Marck E. , Van Laere S. , Vermeulen P. , Dirix L.
Published: December 2010

Gene promoter hypermethylation plays a pivotal role in breast carcinogenesis and breast cancer progression. In this study, we investigated 6 gene promoters (RASSF1A, HIN-1, RARβ2, TWIST, CCND2 and APC) for aberrant DNA methylation by real-time methylation-specific PCR in primary tumors and matched affected lymph nodes from 30 patients with breast cancer. These data were used to explore associations between gene promoter hypermethylation and clinicopathological features, including tumor cell proliferation, angiogenesis and hypoxia marker expression.Overall, gene promoter methylation levels (ratios of DNA copy numbers for methylated and reference genes) were higher in primary tumors than in matched lymph nodes. Notably, we found significant correlations between methylation levels in primary tumors and matched lymph nodes for 5 of 6 genes (P<0.05). An identical gene promoter methylation status (high or low according to the median methylation level) in primary tumors and matched lymph nodes was observed in 67% to 83% of cases. No significant associations between risk factors and gene promoter hypermethylation of the 6 genes was observed, except for the degree of spreading into nearby lymph nodes, which associated with high RASSF1A methylation levels in tumors (P=0.020) and high CCDN2 methylation levels in lymph nodes, and for HER2 expression, which was most frequently observed in tumors with low HIN-1 methylation levels (P=0.039). HIN-1 methylation levels in tumors negatively correlated with the fractions of proliferating tumor and endothelial cells determined by CD34/Ki-67 immunohistochemistry in both primary tumors (r=-0.594, P=0.001 and r=-0.619, P<0.001) and in lymph nodes (r=-0.412, P=0.024 and r=-0.400, P=0.029). Furthermore, high HIN-1 methylation levels in primary tumors tended to associate with breast tumors lacking expression of the hypoxia marker carbonic anhydrase IX in tumor cells (P=0.063) and associated stroma (P=0.086). CCND2 methylation levels in affected lymph nodes positively correlated with the ratios of the fractions of proliferating tumor cells to the fractions of proliferating endothelial cells observed in both primary tumors (r=0.418, P=0.002) and in lymph nodes (r=0.563, P=0.002).In summary, gene promoter hypermethylation in primary breast tumors and matched lymph nodes are highly correlated. Further studies with larger numbers of samples are necessary to investigate the prognostic or predictive potential of these observations. For two genes, HIN-1 and CCDN2, we observed correlations between gene promoter methylation levels and tumor cell proliferation. HIN-1 and CCDN2 are frequently lost in breast cancer and are both involved in cell cycle regulation. The biological significance of these findings needs to be further explored.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 1149.
Gene-Set Enrichment Analysis (GSEA) of Microarray Data in Patients with HER2-Positive Breast Cancer Treated with Preoperative Chemotherapy Including Trastuzumab.
Authors: Shimizu C. , Shimizu C. , Mogushi K. , Tamura K. , Koizumi F. , Ando M. , Kinoshita T. , Tanaka H. , Fujiawara Y.
Published: December 2010

Background: Identification of molecular predictors of response is important in the aspect of personalization of treatment, especially in the era of molecular targeted drugs. Genome-wide analysis such as microarray may be useful in exploring genes related to mechanism of action or resistance to trastuzumab-based treatment.Patients and Methods: 17 HER2-negative and 13 HER2-positve patients with primary breast cancer were enrolled onto this prospective study. HER2 status was determined by HercepTest (3+) or (2+) with gene amplification. The patients were prospectively treated with preoperative chemotherapy (4 cycles of 5FU/epirubicin/cyclophosphamide 500/100/500 mg/m2, q3w, followed by 12 cycles of weekly paclitaxel 80mg/m2. All HER2-positve patients were administered trastuzumab 2mg/kg weekly with paclitaxel. Gene expressions were measured using Affimetrix GeneChip Human Genome U133 Plus 2.0 array in mRNA samples extracted from pretreatment core-needle biopsy specimens. GSEA (www.broad.mit.edu/gsea/msigdb/) were performed to explore gene sets differentially expressed between tumors that achieved pathological complete response (pCR) and those did not (non-pCR). 386 gene sets corresponding to human chromosome and 1454 gene sets named by gene ontology term were examined.Results: The 5 gene sets significantly differentiated HER2-positive and HER2-negative tumors with nominal p-value of <.03 and false-discovery rate (FDR) q-value of <.25 included 3 gene sets located in 17q chromosome including 17q11, indicating the validity of the microarray experiment and GSEA in a relatively small study. Six HER2-positive tumors achieved pCR. We identified 22 and 112 gene sets with nominal p-value of <0.01 and <0.05 that differentiated pCR and non-pCR in HER2-positive tumors, respectively. The top 22 gene sets included gene sets related to regulation of phosphorylation, receptor complex, cell proliferation, cytoplasmic vesicle, regulation of apoptosis and immune response. Other significant gene sets were those related to cell adhesion, signal transduction, cell proliferation and protein homodimerization. We could not identify gene sets which was differentiately expressed in tumors with pCR and non-pCR with FDR q-value of <.25 probablely because of the sample size.Conclusion: Although the results are still exploratory, GSEA seems to be helpful in narrowing down the gene sets from wide range of pathways in molecular network even in a small study if the selected gene sets are biologicaly robust.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 1167.
Outcomes of Women Who Were Premenopausal at Diagnosis of Early Stage Breast Cancer in the NCIC CTG MA17 Trial.
Authors: Goss P. , Ingle J. , Martino S. , Robert N. , Muss H. , Shepherd L. , Pritchard K. , Livingston R. , Davidson N. , Perez E. , Cameron D. , Whelan T. , Palmer M. , Tu D.
Published: December 2010

Background: MA17 showed that adjuvant letrozole after 5 years of tamoxifen markedly reduced the risk of recurrence in women with ER+ early stage breast cancer and improved overall survival in women presenting with node +ve disease. Most trials of early adjuvant aromatase inhibitor therapy required women to be postmenopausal at diagnosis to be eligible. We report here on a subset of women in the MA17 trial who were premenopausal at initial diagnosis and in whom subsequent menopause, prior to randomization, may have influenced their outcome on extended adjuvant letrozole.Methods: Women randomized to MA17 were divided into 2 groups: 1) pre-menopausal (< 50 years of age but underwent bilateral oophorectomy at time when tamoxifen treatment started or < 50 years of age at the start of tamoxifen treatment but became amenorrheic during adjuvant chemotherapy or tamoxifen treatment) and 2) post-menopausal. Disease-free survival (DFS) from time of randomization for women in these two groups were summarized by Kaplan-Meier curves and compared based on log-rank test and Cox model.Results: 889 women were identified as premenopausal and 4,277 as postmenopausal. Treatment, age, race, duration of tamoxifen, prior adjuvant chemotherapy, and mastectomy were independently associated with menopausal status on multivariate analysis. The interaction between treatment and menopausal status was statistically significant for DFS (p=0.02), indicating that women diagnosed with pre-menopausal breast cancer had significantly greater benefit (Hazard ratio=0.25, 95% CI: 0.12-0.51) with letrozole treatment in terms of DFS than those with post-menopausal status (hazard ratio=0.69 with 95% CI from 0.52 to 0.91). In comparison with women treated with placebo, pre-menopausal women on letrozole had a significantly higher incidence of arthralgia (24 vs. 16%; p=0.004) but a lower incidence of vaginal bleeding (10 vs. 16%; p=0.01). Compared with placebo post-menopausal women receiving letrozole had a higher incidence of hot flashes/flashes (55 vs. 50%; p=0.001), arthralgia (25 vs. 21%; p=0.002), myalgia (15 vs. 12%; p=0.007) and alopecia (5 vs. 3%; p=0.003).Conclusions: Letrozole after tamoxifen was more effective in improving DFS in women who were premenopausal at primary diagnosis than those who were postmenopausal. Letrozole was well tolerated in premenopausal women. These data indicate that women who are premenopausal at the time of diagnosis but become postmenopausal anytime before, or during, adjuvant tamoxifen should be considered for extended adjuvant therapy with letrozole.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 13.
Progesterone Receptor (PR) Promoter Methylation – Role as a Predictive and Prognostic Marker in Breast Cancer.
Authors: Pathiraja T. , Jelinek J. , He R. , Shetty P. , Hartmaier R. , Margossian A. , Hilsenbeck S. , Issa J. , Oesterreich S.
Published: December 2010

Estrogen receptor α expression is a well known and clinically utilized predictor of endocrine treatment response. In addition, loss of expression of the progesterone receptor (PR), an estrogen induced gene, has been shown to be associated with resistance to endocrine therapy. PR is known to be epigenetically regulated by DNA methylation, but only a few studies have correlated PR DNA methylation with its expression in breast cancer. Previous studies are contradictory and have been limited by small sample size. There is also evidence that differential expression of the two PR isoforms, PRA and PRB, might contribute to breast tumorigenesis, and altered hormone response. Whether PRA and/or PRB DNA methylation could predict response to endocrine therapy remains an open question. Further, there are no published studies so far testing whether PRA and/or PRB methylation have prognostic significance.Our objective was 1) to determine whether there was an association between PRA and/or PRB methylation levels with total PR expression in breast tumors, and 2) to determine if PRA and/or PRB methylation would have predictive and/or prognostic value. For this study, we utilized tumor DNA from patients with ER-positive breast tumors which were treated with adjuvant tamoxifen after surgery (n=500), and from patients treated with surgery only (n=500). PR expression was determined by ligand binding assay, and PRA and PRB promoter methylation was measured by bisulfite pyrosequencing.Both PRA and PRB methylation were significantly (p<0.0001) associated with PR expression, in an inverse relationship. Low PR expression was significantly (p=0.02) associated with worse overall survival (OS), and there was a trend towards shorter disease free survival (DFS) in the tamoxifen treated group. There was no association between PR expression and survival in the untreated group. PRA methylation was significantly (p<0.0001) associated with PRB methylation. Intriguingly, increased PRA methylation was significantly associated with shorter DFS (p=0.047) and OS (p=0.0067) in tamoxifen treated patients, and with shorter OS (p=0.008) in the untreated patients. In contrast, there was no significant association between PRB methylation and survival in either tamoxifen treated or untreated patients.Our results show that PR expression is significantly associated with PR methylation. The data suggest that PRA methylation is a predictive marker for tamoxifen response and also a prognostic marker for breast cancer progression. This is the largest study so far to elucidate the association of PRA and PRB methylation with PR status, response to tamoxifen, and tumor prognosis, and we believe that it provides useful insights into the role of PR methylation in breast tumorigenesis.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 2002.
Delay in Treatment Does Not Predict Outcome in Inflammatory Breast Cancer.
Authors: Levine P. , Schenfeld J. , Hoffman H. , Young H.
Published: December 2010

Aim: Inflammatory breast cancer (IBC) is an extremely aggressive form of breast cancer that fortunately has proven to be increasingly controlled by modern multimodal chemotherapy. Many doctors consider the immediate use of chemotherapy as critical to the outcome, and delays in treatment due to diagnostic issues have on occasion led to law suits. One such law suit led us to examine the relative importance of delay in diagnosis and other potential prognostic factors in a series of well documented IBC patients.Methods: 134 patients reported to the North American IBC Registry were evaluated for prognostic factors related to disease outcome. Relevant information included the time intervals between onset of symptoms and first medical visit, first medical visit to onset of chemotherapy, and progression free survival (PFS). Other parameters included initial response to chemotherapy, cell surface receptors and extent of disease at diagnosis. Patients were categorized according to the extent of clinical breast involvement and the presence or absence of dermal lymphatic invasion (DLI).Results: Patients with a delay of diagnosis of 60 days or less had a mean PFS of 4.41 years (N=62) while those with a delay >60 days had a mean PFS of 4.58 years (N=58). Initial response to chemotherapy was an important predictor of outcome: 31 patients with a complete clinical response had a median PFS of 5.12 years vs. 4.38 years for 71 patients that had a partial clinical response and 4.01 years for 15 patients with no significant response or progression of disease. The detection of dermal lymphatic invasion (RR: 2.77; 95%CI: 1.4-5.46) and clinical metastases at diagnosis (RR 2.81. 95% CI:1.16-6.80) were also significantly associated with a poorer outcome. The presence or absence of hormone receptors and HER2-neu markers were not predictive of outcome.Conclusion: Delay in initiating chemotherapy was not a significant factor in determining outcome of IBC. Initial extent of disease and response to chemotherapy were the two most imnportant factors in predicitng outcome of treated patients.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 2012.
Steroid Receptor RNA Activator Protein (SRAP): A Potential New Prognostic Marker for Estrogen Receptor-Positive/Node-Negative/Younger Breast Cancer Patients.
Authors: Yan Y. , Yan Y. , Skliris G. , Skliris G. , Penner C. , Chooniedass-Kothari S. , Chooniedass-Kothari S. , Cooper C. , Nugent Z. , Fristenski A. , Hamedani M. , Blanchard A. , Myal Y. , Murphy L. , Murphy L. , Leygue E. , Leygue E.
Published: December 2010

Purpose: The steroid receptor RNA activator (SRA) is a functional RNA suspected to participate in the mechanisms underlying breast tumor progression. This RNA is also able to encode for a protein, SRAP, whose exact function remains to be determined. Our aim was to assess, in a large breast cancer cohort, whether levels of this protein could be associated with outcome or established clinical parameters.Experimental Design: Following antibody validation, we have assessed SRAP expression by tissue-microarray (TMA) analysis of 372 tumors with known steroid receptor and node status. Clinical follow-up was available for all the corresponding patients. Immunohistochemical scores were independently determined by two investigators and averaged. Statistical analyses were performed using standard univariate and multivariate tests.Results: SRAP levels were significantly (Mann-Whitney rank sum test, P<0.05) higher in estrogen receptor-alpha positive (ER+, n = 273), in progesterone receptor positive (PR+, n= 256) and in older patients (age ≥ 65 years, n = 183). When considering ER+ tumors, PR+ tumors, or young patients (< 65 years), patients with high SRAP expression had a significantly (Mantel-Cox test, P < 0.05) worse breast cancer specific survival (BCSS) than patients with low SRAP levels. SRAP also appeared as a very powerful indicator of poor prognostic for BCSS in the subset of ER+, node negative and young breast cancer patients (Cox regression analysis, n = 60, BCSS Hazard Ratio=13.937, P<0.0001).Conclusion: Our data suggest that SRAP might be a new predictor of breast cancer specific survival in younger breast cancer patients with ER+/node negative tumors.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 2017.
The 76-Gene Signature Defines High-Risk Patients That Benefit from Adjuvant Tamoxifen Therapy.
Authors: Zhang Y. , Sieuwerts A. , McGreevy M. , Graham C. , Cufer T. , Paradiso A. , Harbeck N. , Span P. , Hicks D. , Crowe J. , Tubbs R. , Thomas B. , Lyons J. , Sweep F. , Schmitt M. , Schittulli F. , Golouh R. , Talantov D. , Wang Y. , Foekens J.
Published: December 2010

Background: A significant proportion of ER-positive tumors exhibit resistance to endocrine therapy. Clinical studies in LNN/ER+ patients show that the absolute distant disease-free survival benefit of tamoxifen therapy is only about 9%, with a relative benefit of about 30%. These benefits from treatment may, however, differ substantially according to e.g. patient's age and the tumor level of ER and it is therefore of importance to identify biomarkers that allow identification of those breast cancer patients who may actually benefit from adjuvant tamoxifen. This study was to assess the benefit from adjuvant systemic tamoxifen therapy in breast cancer risk groups identified by the previously established prognostic 76-gene signature.Material and Methods: In 300 lymph node-negative (LNN) estrogen receptor-positive (ER+) breast cancer patients (136 treated with adjuvant tamoxifen, 164 having received no systemic adjuvant therapy), distant metastasis-free survival (DMFS) as a function of the 76-gene signature was determined in a multicenter fashion.Results: In 136 tamoxifen-treated patients, the 76-gene signature identified a group of patients with a poor prognosis (hazard ratio (HR), 4.62, P = 0.0248). These patients showed a 12.3% absolute benefit of tamoxifen in 10-year DMFS (HR, 0.52, P = 0.0318) compared with untreated high-risk patients. This represented a 71% increase in relative benefit compared with the 7.2% absolute benefit observed for all 300 patients without using the gene signature. In the low-risk group there was no significant 10-year DMFS benefit of tamoxifen.Discussion: In summary, the 76-gene signature defines high-risk patients who benefit from adjuvant tamoxifen therapy. Although we did not study the value of chemotherapy in this study, low-risk patients identified by the 76-gene signature have a prognosis good enough that chemotherapy would be difficult to justify. The prognosis of these patients is sufficiently good, in fact, that a disease-free benefit for tamoxifen therapy is difficult to prove, though benefits in terms of oco-regional relapse and a reduction in risk for contralateral breast cancer might justify ormonal therapy in these patients.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 2021.
Biopsy Confirmation of Metastatic Disease in Breast Cancer: Results from a Large Prospective Study.
Authors: Amir E. , Freedman O. , Simmons C. , Miller N. , Geddie W. , Dranitsaris G. , Maung H. , Napolskikh J. , Lax M. , Clemons M.
Published: December 2010

Purpose: Decisions about systemic treatment of women with metastatic breast cancer are often based on estrogen receptor (ER), progesterone receptor (PgR), and HER2/neu status of the primary tumour. This study prospectively investigated concordance in receptor status between primary tumour and distant metastases at various stages of progression and assessed the impact of any discordance on patient management.Methods: Biopsies of suspected metastatic lesions were obtained from patients and analyzed for ER/PgR and HER2/neu. Receptor status of metastases was compared to the primary tumour. The treating physician completed questionnaires before and after biopsy to determine whether the biopsy result lead to a change in treatment plan.Results: One hundred and sixteen women were enrolled and 102 underwent biopsy. Of these 52 (51%) were newly diagnosed with metastatic disease, 17 (17%) had received one line of metastatic treatment and 35 (34%) had received 2 or more lines of treatment in the metastatic setting. 86/102 (84%) of samples were sufficient analysis; of these 5/86 biopsies (6%) showed benign disease and one biopsy (1%) confirmed a different malignancy (indolent lymphoma). Changes in hormone receptor status were observed in 33%. Among these, ER discordance was seen in 11%, PgR discordance in 27% and discordance in both ER and PgR in 5%. Loss of PgR was the most common change in hormone receptor status (25%). HER2/neu showed 4% discordance. Three patients (3%) gained and one patient (1%) lost HER2/neu expression. Biopsy results led to a change of management in 12% of patients. Patients with newly diagnosed metastatic disease were more likely to show discordance than those previously treated in the metastatic setting. Among triple negative primary tumours, no changes in receptor expression of metastases were seen.Conclusions: This is the largest prospective biopsy study we are aware of. Results demonstrate the presence of substantial discordance in receptor status between primary tumour and metastases. The number needed to biopsy to alter immediate management was 8.5, although biopsy information could also be useful in planning subsequent treatments. Tissue confirmation should therefore, be considered in all patients with suspicion of metastatic recurrence or progression.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 2023.
Pleomorphic Calcifications, Tumor Markers, and Response to Therapy.
Authors: DeLeonardis C. , Lodhi A. , Le-Petross H. , Singh B. , Krishnamurthy S. , Lucci A.
Published: December 2010

IntroductionMany breast cancers are initially identified by pleomorphic calcifications (PC) on mammography (MG). We hypothesized that presence of PC correlates with primary tumor markers and with response to systemic therapy.MethodsMammograms of women with Stage I-III breast cancer were evaluated for the presence of PC. Only patients with biopsy proven invasive carcinoma in association with PC were included. Primary tumor markers (ER, PgR, and HER2) were determined in the standard manner at the time of core biopsy and correlated with presence or absence of PC on MG. We also evaluated cyclooxygenase-2 (COX2), based on data showing a role in tumor progression and possible interaction with HER2 and/or ER. Primary tumors were immunostained with an anti-COX2 monoclonal antibody, and staining of ≥ 5% or more of tumor cells was considered as positive. For response to therapy analyses, complete pathologic response (pCR) was defined as lack of any residual invasive disease in primary tumor and regional lymph nodes. We used chi-square and Fischer's exact test for the statistical analysis.ResultsWe prospectively enrolled 239 patients. Median age was 51.2 years and the median follow up was 26 months. Pleomorphic calcifications were more often found in HER2 positive patients (22/37) vs. HER2 negative (71/202), P = 0.005, as well as in COX2 positive patients (28/57), P = 0.044. There were no significant associations between PC and ER or PgR. PC were less prevalent in triple receptor negative patients (11/55)[20%] as compared to receptor positive patients (84/188), P = 0.001. We found PC predicted DCIS to be associated with invasive cancers (27/51)[53%] compared to those who showed no DCIS associated with their invasive breast cancer (57/157) [36%] (P=0.035).When evaluating response to therapy, patients with PC on MG were more likely to have a pCR (6/18) [33%] as compared to those who showed absence of pleomorphic calcifications (2/34)[ 6%], following neoadjuvant chemotherapy, P= 0.015.ConclusionsPleomorphic calcifications were significantly associated with HER2 and COX2 positive primary tumors. Patients with PC were also more likely to have a pCR after systemic therapy. Information regarding PC on MG may be helpful in predicting tumor marker status and response to therapy.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 2027.
Effect of 21-Gene Recurrence Score Results on Treatment Recommendations in Patients with Lymph Node-Positive, Estrogen Receptor-Positive Breast Cancer.
Authors: Oratz R. , Chao C. , Skrzypczak S. , Kim B. , Kim B. , Broder M.
Published: December 2010

Background: Results of recent Oncotype DX® studies indicate that the Recurrence Score (RS) can identify node-positive, estrogen receptor-positive (N+/ER+) patients who may not benefit from chemotherapy and have a low risk of distant recurrence with hormonal therapy alone. Based on these results, the Oncotype DX® breast cancer assay has been ordered for N+/ER+ patients. There are no data, however, on the impact of the assay on adjuvant treatment planning in N+ patients. We performed a survey to characterize the effect of the Oncotype DX® breast cancer assay on adjuvant treatment recommendations in N+/ER+ breast cancer.Methods: U.S. medical oncologists (N=1,017) who ordered Oncotype DX® for at least 1 patient with N+/ER+ breast cancer were contacted and asked to complete a web-based survey regarding the single most recent N+/ER+ patient for whom the Oncotype DX® assay was ordered. The survey was developed through cognitive interviews with four medical oncologists, and the protocol was institutional review board approved.Results: We analyzed surveys from 160 individual physicians for 160 N+/ER+ patients. Physicians were in community practices (71%) more often than in academic practice settings, they were equally dispersed geographically, and they had a median of 11 years (range, 1–45) of practice experience. The median patient age was 61 years (range, 34–82), and 79% were postmenopausal. T1, T2, or T3 disease was reported in 62%, 35%, and 3% of patients, respectively (unknown in 1 patient). One, two, three, or 4+ nodes were reported in 69%, 18%, 6%, and 3%, respectively (unknown in 8 patients). Fifty-four percent of N+/ER+ patients had an RS <18; 38% had an RS of 18–30; and 8% had an RS ≥31. Before obtaining the RS, chemotherapy was planned in 89 patients (56%), endocrine therapy was planned in 46 patients (29%), there was no recommendation for 22 patients (14%), and other treatments for 3 patients (1%). After obtaining the RS, recommended treatment changed from hormonal therapy plus chemotherapy to hormonal therapy alone in 48 patients (35%) and from hormonal therapy to hormonal therapy plus chemotherapy in 14 patients (10%).Discussion: These findings indicate that for N+/ER+ patients, the RS score is used more often (but not exclusively) in patients with 1–3 positive nodes and T1 or T2 stage disease. Treatment recommendations for these patients with N+ disease were frequently changed by the RS results, with an overall reduction in the recommendation for chemotherapy treatment.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 2031.