bluuebunny/crossref_metadata_2025_split huggingface dataset:
P3-14-11: Comparison of Two Nomograms To Predict Pathologic Complete Response to Neoadjuvant Chemotherapy – Evidence That HER2 Positive Tumors Need Specific Predictors.
Authors: Rouzier R , Frati A , Coutant C , Bezu C , Antoine M , Uzan S , Chopier J , Chereau E , Gligorov J 
Published: June 2012

Abstract
Aim: The purpose of this study was to compare two different nomograms to predict pathologic complete response (pCR) to preoperative chemotherapy in an independent cohort of 200 patients with breast cancer. The first model was the MDACC nomogram published in 2005 and the other one was a nomogram based on the number of preoperative courses, Ki-67 and steroid hormone receptors expression published by Colleoni et al. in 2010
Patients and methods: Data from 200 patients with breast carcinoma treated with preoperative chemotherapy and operated at Tenon Hospital from 2001 to 2009 were collected. We calculated pCR rate predictions with the two nomograms and compare those predictions with outcome. Patients received between 4 and 8 course of anthracycline/taxanes based chemotherapy. More than 90% of patients with HER2 positive tumors received concomitant trastuzumab with taxanes. Model performances were quantified with respect to discrimination (evaluated by the areas under the receiver operating characteristics curves (AUC)) and calibration.
Results: In the entire population, the AUC for the MDACC nomogram and the Colleoni nomogram were respectively 0.74 and 0.75. Both of them underestimated the pCR rate (p=0.02 and 0.0005). When excluding patients treated with trastuzumab, the AUC were 0.78 for both of them with no significant difference between the predicted and the observed pCR (p=0.14 and 0.15). When analyzing the specific population treated with trastuzumab as preoperative treatment, the AUC for the MDACC nomogram and the Colleoni nomogram were respectively 0.52 and 0.53.
Conclusion: The MDACC and the Colleoni nomograms are accurate to predict the probability of pCR after preoperative chemotherapy in HER2 negative population but did not correctly predict pCR in HER2 positive patients who received trastuzumab. This suggests that response to preoperative chemotherapy including trastuzumab is biologically driven and that a specific nomogram or predictor for HER2 positive patients has to be developed.
Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-14-11.
P3-14-24: A Phase II Trial of TS-1 and Docetaxel Followed by 5-FU/Epirubicin/Cyclophosphamide (FEC) as Preoperative Treatment in Women with Stage II/III Breast Cancer.
Authors: Hayashida T , Jinno H , Sakata M , Takahashi M , Kitagawa Y 
Published: June 2012

Abstract
Background: TS-1 is an oral anticancer drug, composed of tegafur, gimestat and otastat potassium in a molar ratio of 1:0.4:1, based on the biochemical modulation of 5-Fuorouracil. In vivo xenograft model showed a synergistic effect of combination chemotherapy with TS-1 and docetaxel through down-regulation of dihydropyrimidine dehydrogenase (DPD). (Suto et al, Oncology Reports 2006) The sequential combination of anthracycline and taxane is a standard of care in the preoperative setting, as well as the adjuvant setting. This study's purpose was to determine the efficacy of TS-1 and docetaxel followed by fluorouracil/epirubicin/cyclophosphamide (FEC) in the preoperative setting.
Patients and Methods: Patients with stage II/III breast cancer received 4 cycles of TS-1 (TS-1 80 mg/m2 on days 1–14 and docetaxel 40 mg/m2 on day 1 every 3 weeks), followed by 4 cycles of FEC (fluorouracil 500 mg/m2, epirubicin 100 mg/m2, cyclophosphamide 500 mg/m2 on day 1 every 3 weeks). The tumor objective response was assessed by both caliper and ultrasound. Primary endpoint was the pathological complete response (pCR) rate. Secondary endpoints were the clinical response rate, adverse drug reactions, and the breast conservation rate.
Results: From November 2009 to December 2010, 39 patients were enrolled and 38 patients were assessable for clinical and pathologic responses. The overall response rate (caliper) was 81.5% and ultrasound response rate was 76.3%. The pCR rate was 13.2% (5/38). The breast-conserving rate in patients whose tumor size was 3 cm or smaller was 92.9%, and 54.5% of patients whose tumor size was larger than 3 cm underwent breast-conserving surgery. The ratio of Ki-67 positive cell and progesterone receptor (PgR) status were significantly correlated with pCR. Grade 4 neutropenia was observed in a patient (2.6%) at the first cycle. The most common grade 3/4 non-hematologic adverse event was hand-foot syndrome, observed in 3 patients (7.9%). One patient (2.6%) showed grade 3 diarrhea.
Conclusions: These results indicate that the sequential combination of TS-1 and docetaxel followed by FEC is a well-tolerated, effective neoadjuvant treatment for stage II/III breast cancer.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-14-24.
P3-18-05: Triple-Negative Breast Cancer: Stem Cells, Cancer and New Treatment Strategy.
Authors: Yin S , Xu L , Reddy KB 
Published: June 2012

Abstract
Background: Women with triple negative breast cancer (TNBC) have a worse prognosis compared with other breast cancer subtypes. Hormonal or Herceptin-based therapies were found to be ineffective because of the loss of target receptors such as ER, PR and HER-2 amplification. TNBC has increased recurrence and poor survival, despite initial response to neoadjuvant chemotherapy. The aim of this study was to determine whether current drug treatment(s) eliminates bulk of tumor cells, but it has a minimal effect on cancer stem cells (CSCs) leading to tumor recurrence.
Materials and Methods: Cancer stem cells were identified by ALDH-positive population by fluorescence-activated cell sorting. CSCs ability to form mammospheres, are generated in nonadherent and nondifferentiated conditions. Apoptosis was assessed using the Cell Death Detection ELISAPLUS kit. Xenograft tumor formation was evaluated using Nod/Scid mice. Cancer stem cell levels in human TNBC and ER+, PR+ and HER2− tumors were obtained by CD44+/ CD24low/− immunohistochemistry.
Results: TNBC tumors express relatively higher levels of cancer stem cells compared to ER-positive, PR-positive and HER2−negative breast tumors. We studied the effects of PARP inhibitors (AZD2281 and BSI-201), paclitaxel, docetaxel, cisplatin and cisplatin plus TRAIL on CSCs derived from CRL-2335 and MDA-MB-468 TNBC cells in vitro. The in vitro data indicates that cisplatin plus TRIAL treatment was most effective in eliminating CSCs compared to PARP inhibitors, cisplatin, paclitaxel and docetaxel. Treatment with cisplatin plus TRAIL also inhibits Wnt-1 signaling and its downstream target, β-catenin, phospho β-catenin, cyclin D1, increased apoptosis, reduced proliferation and mammosphere formation. Inhibition of Wnt-1 by siRNA significantly reduced the ability of CSCs to form mammospheres compared to control. However, maximum effect was seen in cisplatin plus TRAIL treated cells. The combination of cisplatin/TRAIL also inhibited the growth of CRL-2335 Xenografts. Discussion: We demonstrate for the first time that cisplatin plus TRAIL is highly effective in reducing CSCs compared to PARP inhibitors, docetaxel or paclitaxel in TNBC cells in vitro. Inhibition of Wnt-1 signaling pathways by siRNA or cisplatin plus TRAIL reduces cell proliferation and increases apoptosis in CSCs. Taken together the data suggests that cisplatin plus TRAIL treatment has the potential of providing a new strategy for improving the therapeutic outcome in TNBC patients.
Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-18-05.
P4-01-02: Endocrine Resistance: Mechanism, Tumorigenic Capacities, and New Therapeutic Strategies.
Authors: Morrison GD , Fu X , Ithimakin S , Rimawi MF , Wicha MS , Osborne CK , Schiff R 
Published: June 2012

Abstract
Background
Endocrine therapy is the most effective therapies for estrogen receptor positive (ER+) breast cancer, but resistance is still a major clinical problem. Tumor initiating cells (TICs) in drug resistant tumors are believed to be part of the contributing factors for tumor relapse. Here we focused on the TICs and how they are related to HER2 expression and its downstream factors in acquired endocrine resistance models.
Materials and Methods: We have established a panel of endocrine resistant cell models through long-term treatment with tamoxifen (Tam, 10−7M, &gt;6 months). Resistance to tamoxifen treatment in cells (TamR) was confirmed by growth curve assays (Celigo, Cyntellect Inc., San Diego, CA). In vitro mammosphere assays were applied to assess the mammosphere forming ability of parental and resistant cells. Aldefluor assays (StemCell Technology) were performed to measure the aldehyde dehydrogenase (ALDH-1) expressing cell population as a marker of TICs. Flow analysis was performed to evaluate HER2 expression in the ALDH subpopulations of both parental and TamR cells. Immunofluorescence staining was further used to detect the expression of ER, HER2, phosphorylated STAT3, and β-catenin in sorted ALDH+/− subpopulations.
Results: We found that endocrine resistant cells have a higher mammosphere forming ability than their parental counterparts. In addition, we showed that the ALDH+ subpopulation was significantly upregulated in all TamR derivative lines.

Further analysis of the ALDH populations indicated that HER2, as well as downstream survival factors such as pSTAT3s727 and β-Catenin, are upregulated in the ALDH+ populations as compared to the ALDH-population, especially in TamR derivatives.
Discussion: Our data suggest that deregulated HER or other growth factor receptor signaling can potentially lead to the enrichment of TICs as a potential contributor to endocrine resistance. We will further investigate the role and molecular signaling of TICs in endocrine resistance of breast cancer, as well as whether and to what extent more potent anti-HER inhibitors can improve endocrine sensitivity and circumvent resistance.
Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-01-02.
P4-01-04: Effects of CYP2D6 Phenotype and Drug Adherence on Tamoxifen Metabolite Levels.
Authors: Sousa BA , Nasim S , Cole DE , Wong BY , Hill K , Pritchard KI , Trudeau M , Verma S , Dent R , Petrella T , Gandhi S , Ueng S , Vandenberghe U , Romaschin A , Warner E 
Published: June 2012

Abstract
Background
Although the activity of cytochrome P450 2D6 (CYP2D6), the enzyme responsible for conversion of tamoxifen (TAM) to its most important active metabolite endoxifen, varies significantly with genotype. Routine genotype testing in patients on TAM has recently been discouraged. Conflicting results in publications regarding the prognostic utility of this test remain unexplained. Confounding factors could be lack of predicted correlation between CYP2D6 genotype and TAM active metabolites, or variability of patient compliance
Methods: Consecutive breast cancer patients on TAM were asked to enroll in a study to examine the relationship between CYP 2D6 phenotype, patient-reported treatment adherence, and TAM metabolites levels. Patients were genotyped for CYP2D6 polymorphisms using long-range PCR allele-specific amplification and single-nucleotide primer extension assay. From the genotypes, four phenotype groups were defined: Ultra rapid Metabolizer (UM), Extensive Metabolizer (EM), Intermediate Metabolizer (IM) and Poor Metabolizer (PM). Plasma was collected after at least 6 weeks of TAM (20 mg daily). The parent drug TAM, as well as 4-hydroxy N-desmethyl tamoxifen (endoxifen), 4 hydroxy tamoxifen (4OHtam) and N-desmethyl-tamoxifen (NDtam), were determined by Liquid Chromatography tandem mass-spectrometry (LC-MS/MS). Patients also completed a questionnaire about ethinicity, side effects, concurrent medications and tamoxifen adherence. Correlation between metabolite/TAM ratio and phenotype was tested by Spearman correlation test. Relationship between metabolite levels and adherence was tested by Wilcoxon rank sum test. Chi square test was used to compare proportions.
Results: Of the 100 patients enrolled there were 62 Caucasians, 25 Asians, 4 Africans and 6 Unknown. We found a strong correlation between ratio of endoxifen/TAM and phenotype (p &lt;.0001) (Table 1) Over a 2 week period 68 never missed a TAM dose, 25 missed 1–2 times, 2 missed 3–5 times and 2 &gt; 5times (2 missing data). In EM group we found significantly lower levels of TAM (p &lt;.0001), NDtam (p=.008), 4OHtam (p=.003) in less adherent patients. A trend to decreased levels was also shown for endoxifen (p=.081). No associations were found between adherence or phenotype activity and side effects.

Conclusions: Our data suggests the predicted association between endoxifen levels and genotype. However, non-adherence may have a significant confounding effect. Prospective studies to evaluate the prognostic impact of CYP2D6 variants for patients on adjuvant tamoxifen should be done but results could be confounded by variable drug adherence if this is not measured concurrently.
Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-01-04.
P4-01-12: The Unfolded Protein Response (UPR) and Pro-Survival Autophagy Contributes to Antiestrogen Resistance in Breast Cancer.
Authors: Clarke R , Shajahan AN , Cook KL , Hickman FE , Facey CO 
Published: June 2012

Abstract
In 2011, most of the 192,000 newly diagnosed cases of invasive breast cancer will be estrogen receptor alpha positive (ER+). Endocrine therapy, administered as an antiestrogen, e.g., Tamoxifen (TAM) or Faslodex (FAS; Fulvestrant; ICI 182,780) or an aromatase inhibitor (AI), e.g., Letrozole or Exemestane is the least toxic and most effective means to manage the hormone-dependent breast cancer in such patients. However, resistance to endocrine therapy remains a significant clinical problem. Previously, we have shown that antiestrogen resistant breast cancer cells over-express X-Box Binding Protein 1 (XBP1), a transcription factor that belongs to the basic region/leucine zipper (bZIP) family. XBP1(U) and XBP1(S) variants result from an unconventional splicing of the XBP1 mRNA by IRE1alpha. In the unfolded protein response (UPR), endoplasmic reticulum proteins (IRE1alpha, ATF6 and PERK) sense cellular stress to regulate the accumulation of unfolded proteins. Initially a compensatory mechanism allowing cells to recover normal endoplasmic reticulum function, prolonged UPR may induce cell death; this is often dependent upon which arm of the UPR predominates. XBP1 is an obligate component in both the IRE1alpha and ATF6 arms of the UPR. In this study, we show that in MCF7/LCC9 [FAS resistant; TAM cross-resistant] cells, there is an increase in UPR signaling as detected by increased expression of BiP/GRP78. Transient expression of XBP1(S) in MCF7/LCC1 [antiestrogen sensitive] cells show decreased sensitivity to FAS that correlated with increased levels of both basal and FAS-induced autophagy as measured by cleaved LC3BII protein fragment, GFP-LC3 activity, and reduced expression of p62/SQSTM1. Furthermore, we show that MCF7/LCC9 cells survive by activating pro-survival autophagy through UPR that is regulated by the transcription factor c-MYC. An inhibitor of c-MYC, 10058-F4, synergized with FAS to inhibit proliferation in MCF7/LCC9 cells by preventing pro-survival autophagy and increasing apoptosis. Thus, our study shows that antiestrogen resistant breast cancer cells evade cell death by activating XBP1-mediated UPR that results in c-MYC-mediated prosurvival autophagy.
Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-01-12.
P4-01-14: Fulvestrant Regulates Epidermal Growth Factor (EGF) Ligands and Induces EGF Receptor Activation in MCF-7 Breast Cancer Cells.
Authors: Zhang X , Diaz MR , Yee D 
Published: June 2012

Abstract
Selective Estrogen Receptor Modulators (SERMs) 4 Hydroxy Tamoxifen (4-HT) and fulvestrant (fulv) inhibit estrogen receptor a (ER) positive breast cancer growth. We have observed that treatment (24 to 48 hours) of fulv in MCF-7 cells induced a 190kDa tyrosine phosphorylated band, which was blocked by EGFR and HER-2 tyrosine kinase inhibitors erlotinib and lapatinib. Immunoprecipitation showed EGFR, HER-2, and HER-3 were all phosphorylated after fulv treatment. 4-HT and estradiol did not cause this phosphorylation. No changes in receptor level after fulv treatment were noted. Downstream MAPK signaling was also blocked by erlotinib and lapatinib. Fulv induced activation of EGFR was ER dependent, since fulv treatment in C4-12, an ER negative cell line derived from MCF-7 cells, did not induce EGFR activation. Co-treatment with estradiol and fulv prevented EGFR activation. To explore the possibility that fulv enhanced ligand expression, we collected conditioned media (CM) from MCF-7 cells after 48 hours of treatment. pEGFR was lost when CM was removed, but recurred within 30 minutes. Cycloheximide abolished the ability of fulv to activate EGFR suggesting autorcine production of EGFR ligands was induced by fulv. To detect specific EGFR ligands, we used qPCR to measure various EGFR ligand mRNA levels in MCF-7 and C4-12 cell at 1, 4, 24 and 48 hours after fulv treatment. TGF-α and HB-EGF mRNAs were upregulated over 48 hours, which correlated well with pEGFR activation. 4-HT did not affect mRNA levels of these ligands. In contrast, amphiregulin (AREG) mRNA levels were substantially reduced 48 hours after fulv treatment. A similar trend was seen for AREG mRNA levels in 4HT treated cells but to a much lesser extent. There was no change in any EGFR ligand mRNA levels in ER negative C4-12 cells. These qPCR data suggested differential regulation of EGFR ligands by fulv treatment contributed to EGFR family member activation and was dependent on ER expression. Upon fulv treatment, levels of ER were diminished with decreased detection of S167 and S118 ER phosphorylation sites. Monolayer cell growth analysis showed that while fulv treatment in SFM reduced MCF-7 cell numbers compared to control treatment, erlotinib plus fulv significantly reduced cell numbers below the level of cells maintained in fulv alone. In conclusion, we show that fulv, but not E2 or 4-HT, activates EGFR family members accompanied by upregulation of ligands. Since SERM resistance has been associated with EGFR family member activation, differential control of EGFR ligand gene expression by ER may contribute to the development of fulvestrant resistance in breast cancer.
Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-01-14.
P4-05-04: Arachidonic Acid-Induced Elevated Expression of 5-Lipoxygenase Is Linked to Metastatic Migration of Breast Cancer Cells.
Authors: Roy D , De Chatterjee A , Pal R , Roychowdhury S , Das S 
Published: June 2012

Abstract
Background: In invasive breast cancers, cancerous cells spread outside the ducts of the breast and metastasize to lung and other tissues. Although the rapid arachidonic acid (AA) metabolism and concomitant increase of eicosanoid molecules are involved in the proliferation and invasion of breast cancer cells, the exact mechanism by which AA metabolites regulate these phenomena are not well understood. Here, we show that leukotriene B4 (LTB4), one of the metabolites of AA, which is produced by the action of 5-lipoxygenase (5-LOX), causes the proliferation and metastatic migration of breast cancer cells.
Material and Methods: MDA-MB-231 (invasive breast cancer cells) and MCF7 (non-invasive breast cancer cells) were used in this study. Briefly, cells were treated with AA (100 μM) and nordihydroguaiarectin acid (NDGA, 10 μM), and the secreted eicosanoids were characterized by HPLC. Immunofluorescence microscopy was performed to elucidate the expression and intracellular localization of 5-LOX. The metastatic migration was analyzed by wound-healing assays.
Results: Our results suggest that while MDA-MB-231 cells produce high levels of PGE2 and PGD2, MCF7 cells synthesize excess HETE compounds (HETE5 and HETE8). Interestingly, MDA-MB-231 cells, when stimulated with AA, show the increased syntheses of LTB4 (∼3 fold) and decreased PGE2 and PGD2 (∼2 fold). In MCF7, on the contrary, AA treatment reduced the syntheses of all eicosanoids. Furthermore, the expression of 5-LOX in MDA-MB-231 cells was also increased by ∼2 fold. We observed that AA promotes cell migration in MDA-MB-231 cells, which could be blocked by NDGA, a generic inhibitor of LOX enzymes. Because LTB4 production and its binding with BLT receptors are linked to IL-8 secretion, we measured the level of IL-8 synthesized by both MCF7 and MDA-MB-231 cells. The results suggest that MDA-MB-231 cells secrete excess IL-8 (∼2 fold) when stimulated with AA, suggesting that LTB4 and IL-8 interactive pathways are important for cancer metastasis.
Discussion: Our results demonstrate that LTB4 synthesis is linked to the metastatic migration of MDA-MB-231 cells, and that it therefore should be considered as a target for developing new drugs for the treatment of invasive breast cancers.
Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-05-04.
P4-06-04: Detection of HER2 Gene Amplification in Circulating Tumor Cells and Disseminated Tumor Cells by Fluorescence In Situ Hybridization Using OncoCEE™.
Authors: Krishnamurthy S , Bischoff FZ , Mayer JA , Wong K , Pham T , Kuerer HM , Lodhi AK , Bhattacharyya A , Hall CS , Lucci A 
Published: June 2012

Abstract
BACKGROUND: The status of HER2 gene amplification in circulating tumor cells (CTCs) and disseminated tumor cells (DTCs) might provide useful information for monitoring response to trastuzumab therapy, and may provide a basis for consideration of trastuzumab in patients with HER2 negative primary tumors who have HER2 positive CTCs and/or DTCs. The majority of techniques utilized for detection of minimal residual disease are limited in their ability to allow detailed phenotypic and genotypic evaluation of the cells. We report the utility of a microfluidic platform (OncoCEE™,Biocept, San Diego) for detecting HER2 gene amplification in CTCs and DTCs in patients with non-metastatic breast cancer.
METHODS: Peripheral blood (10ml) and bone marrow (BM) (1-2ml) were collected from patients with clinical stage I-III breast cancer in acid citrate dextrose solution (BD, Franklin Lakes, NJ) and anti-clumping reagent (OncoCEE-Sure™). Mononuclear cells were recovered using a Percoll density gradient method, incubated with a mixture of 10 primary capture antibodies (Abs), introduced into CEE™ microchannels, stained with fluorescent anti cytokeratin (CK) and CD45 abs and finally processed for fluorescence in situ hybridization (FISH) using probes specific to centromere 17 (spectrum green) and HER2 (spectrum arrange). The ratio of HER2:CEP17 &gt;2.2 in any CK+/CD45- and CK-/CD45- cell was regarded as positive for HER2 gene amplification.
RESULTS: Peripheral blood and/or BM from 78 patients (65 BM; 70 blood; 57 matched blood and BM) with T1NO (39), T1N1 (8), T2N0 (12), T2N1 (2), T2N2 (1), T2N3 (3), T3N0 (2), T3N1 (2), T3N2 (1), T4N0(5), T4N1 (3), with HER2+ (n=12) and HER2− (n=58) primary invasive breast tumors were studied. The 12 patients with HER2+ primary tumors had HER2+ DTCs in 3/12 (25%) and HER2+ CTCs in 1/9 (11%) cases respectively. HER2+ DTCs and HER2+ CTCs occurred in 12/55 (24%) and in 4/63 (6%) of the patients with HER2− primary breast tumors. HER2+ CTCs and DTCs occurred simultaneously in only 2 patients and in either blood (3) or BM (13) in the remaining patients.
CONCLUSION: 1. The cell enrichment and extraction microfluidic technology (OncoCEE™) provides a sensitive platform for evaluation of HER2 gene amplification of CTCs and DTCs.
2. HER2+ primary tumors were associated with either HER2+ CTCs or DTCs in 25% of the patients.
3. HER2+ CTCs or DTCs occurred in 28% of patients with HER2−primary tumor.
4. Discordant HER2 status was contributed mainly by HER2+ DTCs occurring in HER2 - primary tumors.
5. The clinical significance of evaluating the status of HER2 gene amplification in CTCs and DTCs in the management of patients with breast cancer needs to be evaluated prospectively in larger clinical trials.
Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-06-04.