bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P5-01-04: Preoperative PET-CT Staging of Axillary Node Positive Breast Cancer
Authors: Ni Mhuircheartaigh NM , Kell M , Flanagan FL , Hargaden GC , Fenlon HM , Gorey TF , Stokes M , Salman RR , Smith CS.
Published: April 2011

Purpose: 18F-FDG PET CT is a powerful diagnostic tool, which has enhanced systemic staging in oncology. Our aim was to evaluate the utility of performing staging PET-CT on patients with biopsy proven metastatic axillary lymph nodes that had clinically occult distant metastatic disease and to assess any change in management arising from it. Materials & Methods: All patients with newly diagnosed breast cancer undergo axillary ultrasound and cytology of suspicious lymph nodes, and those with positive cytology undergo pre operative PET-CT. Nodes were classified on ultrasound according to assessment categories R1-R5. PET-CT images were acquired with a Siemens Biograph High Resolution 16 slice PET/CT scanner using full dose CT and intravenous contrast. Results: Eighty two patients were identified with positive axillary cytology following ultrasound biopsy and underwent PET-CT; 28/82 patients (34%) had extra axillary disease. In total 30 patients (37%) had a change in management due to the PET-CT findings. 2 patients had more extensive surgery than previously planned, 7 patients (8%) had an extended radiation field and 21 patients (26%) were treated for distant metastases. Discussion: The detection of metastatic disease beyond the axilla has important implications for treatment options and survival. We detected clinically overt metastatic disease beyond the axilla and altered management in 37% patients due to the PET/CT findings. On this basis we recommend PET-CT for this preoperative cohort of patients with node positive breast cancer.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P5-01-04.
Abstract P5-01-13: The Clinical Role and Cost Effectiveness of Breast-Specific Gamma Imaging (BSGI) in the Management of Patients with Negative or Indeterminate Mammographic and Sonographic Findings and an Unresolved Diagnostic Concern
Authors: Ward BH , Bohm-Velez M , Kieper D , Chang TS , Straka MR.
Published: April 2011

Background: BSGI is a molecular imaging technique increasingly used to aid in the management of patients with negative or indeterminate mammograms and an unresolved clinical concern such as indeterminate densities, new vague calcifications, bloody nipple discharge, focal breast pain or palpable masses; especially in those of elevated risk and patients with dense breast tissue.

Purpose: This study examines the role and cost effectiveness of BSGI in providing additional information when conventional imaging is indeterminate and greater confidence is required to improve diagnostic accuracy.

Methods: This is a retrospective analysis of patients in our practice for whom BSGI was recommended due to an unresolved clinical concern following negative (BI-RADS 1 or2), indeterminate (BI-RADS 0 or 3), or discordant findings on mammography (MMG) or ultrasound (US). For all modalities, the radiologist had the patient's history and imaging studies available to them at the time of interpretation. BSGI studies were classified as negative (BI-RADS 0-3) or positive (BI-RADS 4-5). Biopsy or follow up imaging was performed as recommended by the radiologist. Pathology or 6 month follow up imaging was used as the gold standard and were classified as negative (benign pathology or no evidence of malignancy on follow up imaging) or positive (biopsy proven cancer).

Results: There were 95 patients with 102 regions of concern, yielding 15 biopsy proven malignant lesions and 87 benign findings on pathology or follow up imaging. BSGI was positive in 12 malignant and 33 benign lesions and negative in 54 benign lesions, yielding a sensitivity of 80% and a specificity of 62%. The negative predictive value of BSGI in this population was 95%.

Conclusion: BSGI contributed significantly to patient management by detecting cancer in 12 of the 95 patients (12.6%). The Medicare reimbursement rate for BSGI in our area is $203.26. Therefore the cost per cancer diagnosis in this population was approximately $1,608.35. If these patients had been sent for breast MRI, $1,026.38 Medicare locally, the cost per diagnosis would rise to $8,129.07. BSGI is a useful, cost effective, complimentary tool in the diagnostic work up of patients when a diagnostic concern remains after negative or indeterminate conventional imaging.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P5-01-13.
Abstract P5-03-03: DNA Methylation of the Estrogen Receptor (alpha) Promoter Is Higher in Triple Negative Breast Cancers Compared to the ER Negative Sub-Type
Authors: Prabhu JS , Wahi K , Correa M , Manjunath S , Rao RN , Shastry SB , Tirumalai SS.
Published: April 2011

Background: The incidence of Estrogen Receptor alpha (ER) negative breast cancer and triple negative cases in India is twice that of Western Caucasians. The underlying causes for this two fold difference are unclear. DNA methylation of the ER promoter is one of the mechanisms of gene silencing. Recent data from the West indicate that the ER promoter is weakly methylated in a majority of patients independent of ER status. Data from an Indian cohort found 66% of ER negative breast tumors to be methylated at the ER gene. However, the technique used, Methylation specific PCR (MSP) suffers from a number of disadvantages such as false priming and low specificity. We have checked the methylation status of ER promoter A by a quantitative method and correlated it with clinicopathological parameters such as ER, progesterone receptor (PR) and Her2 protein status.

Materials and Methods: Tumor sections from 51 cases of primary invasive carcinomas were used for DNA extraction and bisulphite conversion. We adopted a quantitative method, Methylight assay for estimating the methylation status of the ER gene. Percentage Methylation Ratio (PMR) was computed and graded according to previously published standards. Methylation specific PCR was done for all these samples for comparison. Grade and Lymph node (LN) status were assessed. Immunohistochemistry was done to examine the status of ER, PR and Her2 protein.

Results: 16.6% of the ER negative tumors were moderately methylated (PMR score 5-50) at the ER promoter while none of the ER positive tumors were methylated to the same extent. Among the Triple Negative (TN) tumors in our cohort, 25% were moderately methylated. MSP tends to overestimate the methylated tumors and indicated that 43.5% of the ER negative tumors and19.2% of ER positive tumors were methylated. Discussion: A subgroup of ER negative and triple negative Indian patients have enhanced methylation of the ER gene. However this does not account for the doubling of ER negative incidence. Our demographics of a large pre-menopausal population might explain some part of this increase. Methylation of ER promoter was highest in TN tumors. This finding hints towards epigenetic dysregulation being a trait of TN tumors. The possibility of reversing the ER status from negative to positive in the clinic has raised an interest in the mechanism by which ER is lost.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P5-03-03.
Abstract P5-03-07: Adiponectin Deficiency Accelerated Breast Cancer Development and Promoted Cholesterol Accumulation in Mammary Tumors of MMTV-Polyomavirus Middle T Antigen Mice
Authors: Liu J , Xu A , Wang Y.
Published: April 2011

Breast cancer is the most common cancer in women. Visceral obesity and body weight gain are associated with the increased risk of postmenopausal breast cancer. Adiponectin is an adipokine possessing anti-tumor activities against various types of obesity-related cancers. The present study has established a MMTV-polyomavirus middle T antigen (PyVT) transgenic mouse deficient with adiponectin (ADN) expression. Compared to the control mice [PyVT(+/−)ADN(+/+)], the development of mammary tumors in mice lacking this adipokine [PyVT(+/−)ADN(-/-)] was significantly accelerated. While the serum total cholesterol was progressively decreased from week 11 onwards, the cholesterol content in tumor tissues was elevated in both types of mice. The ratio between serum LDL-cholesterol and HDL-cholesterol was significantly higher in PyVT(+/−)ADN(-/-) mice than that in PyVT(+/−)ADN(+/+) mice. Adiponectin was able to inhibit cholesterol uptake in both MDA-MB-231 human breast cancer cells and the primary tumor cells isolated from PyVT transgenic mice. These data demonstrated that the anti-breast cancer activity of adiponectin was at least in part attributed to its suppressive effects on cholesterol accumulation in mammary tumors.

This work is supported by Research Grants Council of Hong Kong (Project no.777908M). Adiponectin knockout mice were kindly provided by Dr. Lawrence Chan at Baylor College of Medicine, who generated these mice with the support of the US National Institutes of Health grant HL-51586.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P5-03-07.
Abstract P5-03-08: Overexpression of AIB1 mRNA in Breast Cancer Inversely Correlates with Estrogen Receptor Positivity
Authors: Lee A , Lee K , Chae BJ , Song BJ , Jung SS , Kang CS.
Published: April 2011

Background: The AIB1 is a steroid receptor coactivator that promotes the transcriptional activity of nuclear receptors such as estrogen receptor. It is overexpressed at the mRNA level in up to 60% of primary breast carcinomas.

Methods: In situ hybridization to examine AIB1 mRNA expression was performed in 113 breast carcinomas in the Catholic university of Korea, from Jan. 2004 to Dec. 2008. We investigated the prognostic significance of AIB1 and the correlation of immunohistochemical staining results of estrogen receptor(ER), progesterone receptor (PR), and Her2/neu. Results: AIB1 mRNA overexpression showed in 55 of 113 (48.7%) breast carcinomas. AIB1 mRNA expression was significantly associated with T stage (P=0.002), but not associated with age, lymph node metastasis, histologic type and histologic grade. High expression of AIB1 mRNA was inversely correlated with expression of ER (P=0.011, R=-0.241) and not significantly associated, but also adversely related, with expression of PR (P=0.101, R=-0.157). AIB1 mRNA overexpression was associated with positivity of Her2/neu (P=0.001, R= 0.311). Survival and recurrent free analysis did not reveal significant association with AIB1 expression. Conclusion: AIB1 mRNA overexpression could be involved in breast cancer by mechanism not totally dependent on steroid receptor cofactor, and may involved other estrogen independent growth pathway such as Her2/neu expression.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P5-03-08.
Abstract P5-04-02: Tumor Associated High Endothelial Venules (HEV) Predict Lymphocyte Infiltration and Breast Cancer Patients Clinical Outcome
Authors: Garrido I , Martinet L , Filleron T , Leguellec S , Rochaix P , Girard JP.
Published: April 2011

Even if lymphocyte presence within tumor tissues and its impact on cancer patient prognosis have been well documented past few years, the type of tumor vessels governing infiltration of effector cells into tumors remain to be determined. High endothelial venules (HEV) are specialized vessels for lymphocyte recruitment physiologically present in secondary lymphoidorgans also found in chronic inflammation that may impact lymphocyte recruitment and cancer patient's clinical outcome. We recently identified HEV like vessels in the peritumoral stroma of human solid tumors including melanomas, breast, colon, ovarian, and lung carcinomas. In order to better analyse the functional consequence of HEV presence within human solid tumors, we quantified HEV and tumor infiltrating lymphocytes by immunohistochemistry on serial tumor sections of 146 breast cancer patients. We demonstrate that HEV like vessels density within tumor stroma is an important predictor of CD3+, CD8+ T and B lymphocyte tumor infiltration suggesting an active role played by these vessels in peripheral blood lymphocyte migration into tumors. The breast tumor infiltrating lymphocyte phenotype associated with HEV presence was further characterized by large scale flow cytometry analysis in 30 freshly operated breast tumors. Breast tumors with a high density of HEV (HEVhi) have increased numbers of effector memory T cells (CD45RA-CD62L-) that display an activated mature phenotype (CD69+, Granzyme A+, Granzyme B+, perforin+). T cells with a naïve (CD45RA+CD62L+) and a central memory (CD45RA-CD62L+) phenotype are also specifically increased within HEVhi tumors as compared to HEVlo tumors. Finally, we analyzed the prognostic value of HEV in a retrospective cohort of 146 invasive breast cancer patients and demonstrate that HEV density within tumor stroma is an independent predictor of good clinical outcome. Indeed, HEVhi patients have a significantly longer metastasis free survival (MFS) (p< 0,004), disease free survival (DFS) (P<0,012) and overall survival (OS) (P<0,023) than HEVlo patients. HEV like vessels, through the recruitment of high number of T lymphocytes and B lymphocytes from the periphery could limit the dissemination of primary breast tumors and prevent metastatic recurrence. Therefore, signals and cells that stimulate HEV differentiation within tumor stroma represent important solid tumors therapeutic target to modulate immune infiltration and patient's clinical outcome.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P5-04-02.
Abstract P5-05-03: FOXP3 Regulates Metastatic Spread of Breast Cancer Via Control of Expression of CXCR4 Chemokine Receptor
Authors: Overbeck-Zubrzycka D , Ali S , Kirby JA , Lennard TW.
Published: April 2011

Background: The transcription factor FOXP3 can regulate T cell migration by inhibiting expression of CXCR4, the receptor for the chemokine CXCL12. Increased expression of CXCR4 by breast cancer cells can drive metastatic migration towards sites which express CXCL12. Intracellular trafficking of FOXP3 to the nucleus is required in order for this protein to function. We hypothesised that FOXP3 gene is inactivated in breast cancer causing failure of nuclear localisation with subsequent increase in CXCR4 and potential for metastasis.

Methods: The expression patterns of FOXP3 and CXCR4 were measured at mRNA (PCR, real-time PCR) and protein (immunohistochemistry, immunofluorescence, flow cytometry) levels. A hundred sections of benign breast tissue, cancers, lymph node metastases and seven breast cell lines were examined with traditional and digital microscopy techniques. The results were correlated with pathological and clinical prognostic indicators of breast cancer.

Results: “Normal” breast epithelial cells (both patient-derived tissues and laboratory cultured cell lines) expressed FOXP3 in their nuclei and at the same time fail to express CXCR4. Breast cancer cells significantly overexpressed CXCR4 (P<0.05), whereas FOXP3 expression was significantly decreased (P<0.05) and confined to the cytoplasm with negligible nuclear expression [Figure 1]. Metastases expressed significantly less FOXP3 and more CXCR4 than primaries (P<0.05). Transfection of cancer cells with wild-type FOXP3 cDNA expressing plasmid restored physiological nuclear FOXP3 expression. Discussion: We have found failure of FOXP3 nuclear localisation in breast cancer cells and an inverse correlation between this failure and CXCR4 expression [Figure 2]. Increased CXCR4 expression may adversely affect signalling pathways in favour of metastasis. Expression of wild type FOXP3 reverses this suggesting that breast cancer cells have mutations in the FOXP3 gene resulting in cytoplasmic localisation of the protein. Our findings suggest that disruption of FOXP3 nuclear localisation due to dysfunctional FOXP3 transcription factor may be a novel explanation for breast cancer invasion and metastases.

Figures available in online version.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P5-05-03.
Abstract P5-05-12: Heparan Sulfatase 2 Regulates Breast Cancer Tumorigenesis and Promotes Invasive Ductal Lesions
Authors: Khurana A , McKean H , Mcguire J , Roberts L , Goetz MP , Shridhar V.
Published: April 2011

Ductal carcinoma in situ (DCIS) of the breast is a heterogenous group of proliferative cellular lesions that have the potential to become invasive. Very little is known about the molecular alterations involved in the progression from DCIS to invasive ductal carcinoma (IDC). Heparan endosulfatases HSulf-1 and -2 catalyze or edit sulfate moieties on heparan sulfate proteoglycans and have been implicated in modulating heparin binding growth factor signaling. Several studies have suggested that while expression of HSulf-1 is down-regulated and is considered a tumor suppressor gene, expression of HSulf-2 is over-expressed in primary breast tumors and cell lines. However, the role of HSulf-2 in breast tumorigenesis is poorly understood. Here we have evaluated the effect of HSulf-2 in an in vivo model of breast cancer progression (MCF10DCIS.com). These cells form comedo type DCIS and progress to IDC when transplanted in immune-deficient mice and therefore are an ideal model to study breast cancer progression.

Methods: Stable clones depleted of HSulf-2 in MCF10DCIS.com cells were generated and injected into mouse mammary fat pads with non-targeted control shRNA clones (NTC) as controls. HSulf-1, HSulf-2, smooth muscle actin immunostaining and H&E staining was perfromed at weeks 3, 5 and 7. Matrixmetallo Protease (MMP) expression by real time PCR was evaluated at Weeks 3 and 5 in both groups.

Results: A significant reduction in tumor growth was observed in HSulf-2 depleted clones compared to NTC clones (p=<0.01). HSulf-2 immunohistochemical staining showed distinct pattern of staining localized to ductal lesions as well as stromal (myoepithelial and myofibroblasts cells). In contrast, the closely related family member, HSulf-1 expression was restricted largely to stromal and myoepithelial layer in ductal lesions. While HSulf-2 staining remained intense from weeks 3 to 7, HSulf-1 expression was diminished during the same time period. In NTC clones, ductal like lesions were observed by week 3 with disruption of the basement membrane and appearance of invasive phenotype by weeks 5 and 7. In contrast, HSulf2 knockdown xenografts showed retention of ductal like structures even at week 5 and retention of basement membrane (p=<0.05). Additionally marked necrotic areas were also observed in HSulf-2 silenced clones. Previous studies have shown that the malignant epithelium induces the development of the stroma necessary for the development of invasive phenotype. Consistent with this observation, we demonstrated decreased MMP expression in xenografts derived from HSulf-2 depleted clones compared with NTC xenografts (p=<0.05). Conclusions: Our data suggest that HSulf-2 may play an important role in the transition from DCIS to an invasive phenotype (IDC) potentially by up regulating MMP expression and activity.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P5-05-12.
Abstract P5-05-13: HER2-Induced MMP-1 Expression Is Mediated by HER1 Though AR, EGF, and TGF Independent of HER3 through HRG in Breast Cancer Cells
Authors: Jung HH , Park YH , Ahn JS , Im Y-H.
Published: April 2011

introduction. However, one of plausible explanation for resistance to trastuzumab is associated with HER dimerization. Furthermore, HER3 seems to be the preferred dimerization partner when signaling occurs through the PI3K pathway and, as such, is emerging as a key target for inhibition of HER signaling. Recently, we reported that HER2 could induce enhanced MMP-1 expression. In addition, this up-regulation of MMP1 expression was shown to be mediated MAPK pathway synergized with ER. Heregulin (HRG) acts as a preferential ligand for HER3 and/or HER4 receptors. Upon binding, HRG activates the EGFR/HER2 receptors via dimerization with HER3 and/or HER4. In the other words, HRG-β was reported to promote MMP-7 expression via HER2-mediated AP-1 activation in MCF-7 cells. In addition, MMP-9 is associated with HRG-≥1/HER2 aggressive phenotype of breast cancer.

Based on these backgrounds, we conducted this study to test the role of HER3 in breast cancer cells in relation to the expression of HER2 and MMP-1.

Our result showed that MMP-1 and MMP-13 expressions were up-regulated via EGF and amphiregulin (AR) independent of HRG in HER2 transfected MCF-7 and SK-BR-3 breast cancer cells through Western immunoblotting and quantitative RT-PCR. Enhanced MMP-1 and MMP-13 expressions were markedly decreased by treating HER2 targeting agents in AR, EGF, and TGF-β stimulating cell lines, while these findings were not observed in HRG1-β and HRG1-β stimulating cell lines in quantitative RT-PCR. In addition, these findings were reversed by treating siEGFR and siHER2, not by siHER3. Consequently, we could confirm the downstream pathways were mediated by MMP-1 AP-1 not by HRG1-β, nor HRG1-β with EMSA.

This study showed that HER2-induced MMP-1 expression was mediated by HER1 though AR, EGF, and TGF independent of HER3 through HRG in breast cancer cells. This result implies that the dimerization responsible for MMP-1 up-regulation by HER2 is HER1/HER2, not HER2/HER3. Further implications of MMP-1 in HER2 overexpressing breast cancer through HER1 are going to following.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P5-05-13.
Abstract P5-06-13: Reduced NIMA Related Kinase 2 (Nek2) Expression Level Enhances Anti-Cancer Drug Sensitivity in the Breast Cancer Cells
Authors: Lee J , Lee K , Gollahon L.
Published: April 2011

Although the anticancer drugs paclitaxel and doxorubicin are commonly used to treat many types of solid tumors, their effectiveness is highly variable due to tumor cell resistance. Therefore, it is important to find mechanisms that can be targeted to increase the sensitivity of cancer cells to antitumor agents such as paclitaxel and doxorubicin. NIMA related kinase 2 (Nek2), a serine/threonine kinase is emerging as an important oncogene because of its regulatory role in mammalian cell mitosis. Thus, regulation of the Nek2 expression levels may prove important as a target for cancer treatment. In this study, we investigated whether drug sensitivity was increased in the triple negative breast cancer cell line MDA-MB-468 by using antisense oligonucleotides (ASOs) and siRNA against Nek2. MDA-MB-468 breast cancer cells transfected with Nek2 siRNA or ASO were exposed to various concentrations of paclitaxel and doxorubicin. Cell viability, cell cycle distribution, and apoptosis were evaluated. We observed that drug susceptibility in these transfected cells was dramatically increased compared with either agent alone. Interestingly, the cell proliferation of the Nek2 ASO transfected breast cancer cells was much lower than the Nek2 siRNA transfected breast cancer cells. Our FACS data showed paclitaxel and doxorubicin treated cells transfected with ASOs arrested in G2/M phase as expected due to Nek2 regulatory function in centrosome duplication. We observed that ASOs and siRNA against Nek2 work with paclitaxel and doxorubicin by arresting cells at G2/M and progressing the cells to apoptosis. Our results suggest that these drugs in combination with Nek2 siRNA or Nek2 ASO treatment greatly improve the sensitivity of cancer cells. Thus, these results provide a possible new strategy for the treatment of breast cancer.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P5-06-13.
Abstract P5-06-14: Ezrin Binding Protein Ebp50 Negatively Regulates HER2 and Alters Tamoxifen Sensitivity
Authors: Gu G , Covington KR , Fernandez NM , Ando S , Fuqua SAW.
Published: April 2011

Background: Targeted therapy with antiestrogens and aromatase inhibitors are the mainstay of treatment for estrogen receptor (ER) a-positive breast cancer; however, resistance to these therapies ultimately occurs and patients recur with resistant disease.

Materials and Methods: Stable overexpression of a shRNA to Ebp50 or full-length Ebp50 was performed in MCF-7 breast cancer cells. Cell responsiveness to tamoxifen (Tam) was evaluated using soft agar and MTT growth assays. The molecular signaling pathways affected by Ebp50 knockdown were explored using immunoblot analysis, and immunoprecipitation assays. Cellular localization was determined using confocal microscopy.

Results: Modulation of Ebp50 levels altered cellular response to Tam treatment. Overexpression enhanced Tam sensitivity, whereas knockdown decreased Tam sensitivity. Thus reduction in Ebp50 levels analygous to that discovered in resistant patients, conferred resistance to the growth inhibitory effects of Tam. Since it is known that Ebp50 can bind to a number of cell surface receptors, we performed binding assays, and found that Ebp50 was able to bind to HER-2 in MCF-7 cells. In addition, this interaction was enhanced with Tam treatment. Enhanced activation of phospho-HER2 and total-HER2 levels, as well as pAKT were seen in the Ebp50 knockdown cells. We expressed deletion mutants of Ebp50 we saw increased levels of cytoplasmic ERa when the ERM domain was deleted. The ERM domain, which is know to be important for the binding cytoplasmic proteins, may thus be a determinant of ERa localization. Ebp50 was also shown to affect ERa transcriptional activity. Discussion: Our data suggest that Ebp50 may be a negative regulator of HER2 signaling, and its loss confered homrone resistance. Ebp50 loss might function to stabilize HER2 levels.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P5-06-14.
Abstract P5-06-15: JNK2 Regulates Mammary Epithelial Cell Differentiation Through Inhibition of p53 and Notch-1 Expression
Authors: Cantrell M , Ebelt ND , Van Den Berg CL
Published: April 2011

The classification of patient tumors by clinical subtype has gained great interest due to the implications for prognosis and treatment. For example, it is known that the five-year survival rate of patients diagnosed with the basal subtype of breast cancer is considerably lower than those with luminal subtypes because they lack expression of common drug targets. For this reason, it is important to elucidate the molecular mechanisms responsible for differentiation of mammary stem cell populations into basal and luminal cell lineages. Data presented in this poster outline a novel pathway whereby a ubiquitously expressed protein kinase, JNK2, influences the differentiation of basal, myoepithelial cells (MEps) while suppressing luminal cell (LEp) populations. Gross anatomical observation of whole-mounted mammary glands from wildtype (jnk2wt) and jnk2-/- (jnk2ko) mice revealed that JNK2 increases the rate of pubertal development. Loss of jnk2 increased the rate of ductal invasion, branching, and terminal end bud (TEB) count. Although a slight increase in proliferation was seen in jnk2ko glands, data suggested that it was not due to growth factor signaling. Instead, it was noticed that jnk2ko glands possess 35% fewer MEps (p=0.0078) with a corresponding increase in LEp populations (p=0.100). LEps are known to proliferate at a higher rate than MEps. The differentiation phenotype was corroborated through 3D culture of primarymammary epithelial cells (MECs). The rate of growth of 3D MEC acini was 31.5% higher in jnk2ko cultures than in jnk2wt (P<0.0001). Notch-1, a well-known regulator of MEC difFerentiation, is present in its active form in a greater proportion of cells within jnk2ko TEBs than jnk2wt (p=0.0118, 0.0209). Further, jnk2ko glands were found to possess greater amounts of Notch-1 protein as well as 10.5 times more expression of the Notch target gene, Hes-1 (p=0.005). These results were corroborated with 3D culture of primary MECs as well as in mammary tumor cells. Inhibition of Notch signaling with GSI, in 3D cultures of primary MECs, normalized growth (p=0.8992) and differentiation to jnk2wt control levels. QPCR results showed that jnk2ko glands possess 6.4 times more notch-1 transcript in vivo (P<0.0001). Luciferase assays comparing activity of the notch-1 wildtype promoter to a notch-1 promoter with mutated p53 response elements revealed a dependence of increased notch-1 promoter activity in jnk2ko cells on the p53 response element. QPCR showed 2.78 times more p53 transcript in jnk2ko mammary glands as compared to jnk2wt. These data suggest that JNK2 regulates MEC differentiation through p53 and its target, Notch-1. These findings are significant because they reveal JNK2 as a possible drug target for novel differentiation therapy of basal type breast tumors.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P5-06-15.
Abstract P5-10-06: TACT2 Randomised Adjuvant Trial in Early Breast Cancer (EBC): Tolerability and Toxicity of Standard 3 Weekly Epirubicin (E) Versus Accelerated Epirubicin (aE) in 129 UK Hospitals (4391 Patients) (CRUK/05/019)
Authors: Cameron D , Barrett-Lee P , Velikova G , Canney P , Moyses H , McDermaid M , Banerji J , Gaunt C , Reynolds C , Wardley A , Bowman A , Bertelli G , Murray N , Bliss JM
Published: April 2011

Introduction: TACT2, a multicentre phase III trial with E-CMF as control (NEAT Poole NEJM 2006), tests 2 hypotheses in a 2x2 factorial design: A) accelerating chemotherapy (CT) offers superior benefits (CALGB9741 Citron JCO 2003); B) the oral 5FU prodrug capecitabine (X) gives similar efficacy but preferential side-effect profile to CMF. Here, focus is on hypothesis A with results for compliance, QL and acute toxicities (physician & patient (pt) reported) during E vs aE.

Materials &

Methods: Dec 2005-08 4391 pts (4371 women, 20 men) with node +ve or high risk node -ve invasive EBC were recruited. Pts were randomised to receive either E (100mg/m2 x 4) q3wk or q2wk aE (100mg/m2 x 4 plus pegfilgrastim, 6mg d2) followed by classical CMF q4wk x 4 or X (2500mg/m2/day x 14) q3wk x 4. Detailed CTCAE toxicity was to be assessed in subset (38 centres, 2137 pts). Of these, 2127 pts received ≥1 cycle CT. 1279 of these pts completed pt-reported outcomes (EORTC QLQ-C30 and pre-defined pt-reported TACT2 toxicities) with 910 completing baseline on time and having end cycle 4 data. P<0.01 classed as significant. Toxicities/which differ between groups are shown where p-values are trend tests across all grades & % grade 3+ are reported. QL assessed via linear regression models.

Results: 33/4391 pts did not start CT. 4259 (97%) completed the initial 4 cycles (E 2143 (96%), aE 2116 (98%) with RDI >85% for E 2058 (93%) and aE 1983 (91%). During cycles 1-4 higher rates of hand-foot syndrome & anaemia were seen with aE vs E (P<0.001). Few were grade 3+ (E 0%, aE 0.9% & E 0%, aE 0.1%) respectively. Lower grade 3+ rates (all P<0.001) were seen with aE vs E for: ANY toxicity (E 27%, aE 18%), leucopenia (E 4%, aE 1%), neutropenia (E 17%, aE 2%) & febrile neutropenia (E 4%, aE 2%). Change from baseline in global QL, role functioning & fatigue was worse with aE vs E (P<0.001); no differences were reported for aE vs E for impact of individual pre-defined TACT2 toxicities on daily activities (Nausea, vomiting, tingling hands and/or feet, diarrhoea, constipation, sore mouth, mouth ulcers).

Conclusion: In TACT2, the largest adjuvant trial reporting acute toxicities for standard vs accelerated E in EBC, serious CTCAE toxicity was less common with aE compared with E, however pts report poorer global QL, fatigue & role function which does not appear to be explained by impact on daily activities due to individual side-effects.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P5-10-06.
Abstract P5-10-07: TACT2 Randomised Adjuvant Trial in Early Breast Cancer (EBC): Tolerability and Toxicity of Standard 3 Weekly Epirubicin (E) Versus Accelerated Epirubicin (aE) Followed by Capecitabine (X) or CMF in 129 UK Hospitals (CRUK/05/019)
Authors: Bliss JM , Canney P , Velikova G , Barrett-Lee P , Moyses H , McDermaid M , Banerji J , Gaunt C , Reynolds C , Agrawal R , Murray P , Clark P , Goodman A , Cameron D
Published: April 2011

Introduction: TACT2, a multicentre phase III trial with E-CMF as control (NEAT Poole NEJM 2006), tests 2 hypotheses in a 2x2 factorial design: A) accelerating chemotherapy (CT) offers superior benefits (CALGB9741 Citron JCO 2003); B) the oral 5FU prodrug X gives similar efficacy but preferential side-effect profile to CMF. Here, focus is on hypothesis B with results for compliance, QL & acute toxicities (physician & patient (pt) reported) during CMF vs. X

Materials &

Methods: 4391 pts (4371 women, 20 men) with node+ve/high risk node-ve invasive EBC were recruited between Dec 2005-08. Treatment was E(100mg/m2 x 4) q3wk vs q2wk aE(100mg/m2 x 4 + pegfilgrastim 6mg d2) followed by classical CMF q4wk x 4 vs X(2500mg/m2/day x 14) q3wk x 4. Detailed CTCAE toxicity was assessed in a subset (38 centres, 2086 pts receiving ≥1 cycle CMF or X). 1279/2086 also participated in substudy of pt-reported outcomes (EORTC QLQ-C30 and TACT2-specific toxicities). 826 had complete dataset (baseline, cycles 4&8). P-values are trend tests across all grades & %grade 3+ are reported. QL assessed via linear regression models adjusted for baseline, end cycle 4 score, & aE vs E. P<0.01 classed as significant

QL in cycles 5-8 was better with X than CMF for global QL& fatigue (P<0.001). Pts reporting clinically meaningful deterioration (>10 points): global QL CMF 106/398 (27%), X 79/428 (18%); fatigue CMF 164/398 (41%), X 122/428 (29%)).

Conclusion: TACT2, the largest adjuvant EBC trial with X, confirms that X has preferential side-effect profile and global QL compared to CMF, with no evidence that prior aE compromised treatment delivery. Dose delivery data are consistent with advanced disease observations that for some pts, 2000mg/m2/day may be correct dose.

Results: 4264 (97%) pts continued on CT beyond cycle 4. 3726 completed all 8 cycles (E-CMF 951 (85%), aE-CMF 938 (86%), E-X 932 (84%), aE-X 905 (83%). For cycles 5-8, %RDI > 85% was 69% after E and 68% after aE. Cycles delivered on time CMF 59%, X 63%; cycles without dose reduction CMF 75%, X 62%. 15 deaths in total within 30 days of CT: 9 on CMF, 6 on X. Worst grade toxicities which differed between CMF & X during cycles 5-8:

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P5-10-07.
Abstract P5-10-09: Comparison of the (Non-)Hematological Toxicity and Compliance of Different Anthracycline Containing Regimen with the Standard AC/EC in Elderly Patients with Primary Breast Cancer
Authors: Linder M , von Minckwitz G , Harbeck N , Janni W , Kümmel S , Kaufmann M , Elling D , Nekljudova V , Loibl S.
Published: April 2011

Background:

There is a paucity on toxicity (tox) data in elderly patients (pts) above 65 years (yrs) treated with anthracycline containing standard chemotherapy (ct) for primary breast cancer. We investigated the tox of standard anthracycline based regimen (AC/EC) and compared those with alternatives like dose-dense monotherapies and combinations (Edd; ADdd), intensified combinations (Canadian FEC) and taxane/anthracycline combinations (TAC) in various age groups. Patients and methods:

In this pooled analysis, individual pts data from 4 German randomized trials (n= 4775) were evaluated regarding tox and compliance. Docetaxel/doxorubicin/cyclophosphamide (75/500/500mg/m2) 3qw (A=TAC), 5-fluorouracil/epirubicin (500/60mg/m2) iv d1+8 followed by cyclophosphamide (75mg/m2) po d1-14 q4w (B=cFEC), doxorubicin (epirubicin)/cyclophosphamide (60(90)/600mg/m2) q3w followed by docetaxel (100mg/m2) or paclitaxel (175mg/m2) q3w (C= AC/EC-D/p), doxorubicin/docetaxel (50/75mg/m2) q2w (D= AD dose dense(dd)) and epirubicin-paclitaxel q2w (120/175 mg/m2) (E= sequence dd) were given. Only the anthracycline containing cycles were investigated. A descriptive analysis was performed on compliance and (non-) hematological tox. Pts were grouped according to their age: < 60yrs [1], 60-64 yrs [2] and >64yrs [3]. Primary G-CSF support was given in A, D, E.

Results:

73.6% were <60 yrs, 15.8% were 60-64 and 10.6% were > 64. A total of 22,306 anthracycline containing cycles were administered, of those 74.6% were given to [1], 15.2% to [2] and 10.0 % to [3]. 41.6% of the pts received TAC; 11.5% cFEC; 29.2% AC/EC-D/P; 8.2% ADoc dd and 9.5% Edd-Pdd.

The incidence of febrile neutropenia (FN) did not significantly differ between the 3 age groups (overall A:10.0%; B:2.9%; C:1.0%; D:4.7%) apart from E with 0.9% ([1]0%; [2]6.4%; [3]0% P<0.001). FN in pts >64 was lower in B (2.4%) than in A (14.4%) and D (5.1%). Diarrhea grade 3+4 increased with age to 8.2% for [3] under TAC (p= 0.01) and mucositis grade 3+4 under E to 12% for >64 (P<0.001). Conclusion:

In total, range and intensity of tox increased with age. Neutropenia did significantly increase with age in standard regimen but not in the dd groups

Rate of dose reduction increased with age in A ([1] 6.1%; [2]6.5%; [3] 11.5; p=0.02) and B ([1]15.7%; [2]26.9%; [3]39.3%; p< 0.001). Similar significant results were observed for discontinuations and hospitalizations in A, B, D. Neutropenia grade 3+4 in [3] was 77% with cFEC, vs 55%with TAC (with primary G-CSF).

Leucopenia Grade 3+4 in %

(D, E). The severity of the non-hematotox differed between age groups and regimen. Monotherapies and sequential regimen seem more favorable in elderly pts. cFEC without G-CSF is not an option in pts >64.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P5-10-09.
Abstract P5-10-15: First Safety Data from a Randomised Phase III (CIBOMA 2004- 01/GEICAM 2003-11) Trial Assessing Adjuvant Capecitabine Maintenance Therapy after Standard Chemotherapy for Triple-Negative Early Breast Cancer
Authors: Lluch A , Gomez H , Ruiz-Borrego M , Bines J , Llombart A , Ramos M , Torres R , Martins-Segalla JG , Torrecillas L , Barrios C.
Published: April 2011

Background: A large adjuvant trial programme is exploring the role of capecitabine (C) in early breast cancer (EBC). CIBOMA 2004-01/GEICAM 2003-11 is a multicenter, international randomised phase III trial that focuses on adjuvant C maintenance therapy after conventional induction chemotherapy in triple-negative EBC. Here we report interim safety data from the trial.

Materials and methods: Patients (pts) with operable, node-positive (or node-negative with tumour diameter ≥1 cm), hormone receptor-negative, HER2-negative EBC (central laboratory confirmation) who have received standard anthracycline-and/or taxane-containing chemotherapy in the (neo)adjuvant setting are eligible. Pts are randomised to receive 8 cycles of C (1000 mg/m2 bid, d1-14 q21d) (Arm A) or observation (Arm B). The primary endpoint is disease-free survival (DFS). Assuming 30% risk reduction in DFS rate at 5 years (from 64.7% to 73.7%, HR of 0.701) with a power of 80% and a two-tailed log-rank test at 0.05, 834 evaluable patients will be needed. With an expected drop-out rate of 5%, 876 pts will be included. We anticipate completing recruitment by the end of 2010. Efficacy analysis will be triggered after 255 events.

Results: To date, 724 pts have been recruited; here we report safety data from the first 278 pts (136 in Arm A and 142 in Arm B). Baseline characteristics are well balanced between the treatment arms.

Baseline Characteristics

In total, 920 cycles of C were administered (median 8.0, range 0-8). All 8 cycles were completed by 74.3% of pts. Median relative dose-intensity of C was 91.1%. The most common grade 3/4 clinical AEs related to C were: hand-foot syndrome (Grade 3 19.1%), diarrhoea (3.0%), vomiting (1.5%), fatigue (1.5%), nail changes (1.5%) and bilirubin elevation (1.5%).

Conclusions: The safety profile of adjuvant C as maintenance therapy is consistent with its known toxicity profile. Safety data from the first 400 randomised pts will be reviewed by an Independent Data Monitoring Committee and used to update this analysis at the meeting.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P5-10-15.
Abstract P5-10-18: Echocardiography on Neonates Exposed to Anthracycline Therapy In Utero for Maternal Breast Cancer
Authors: Cardonick EH , Usmani A , Ghaffar S , Wood D , Levine M.
Published: April 2011

Background: Anthracycline therapy carries a risk for cardiac toxicity in pediatric and adult populations. It is unknown if this risk also applies to fetuses exposed in utero. The objective of this study is to evaluate the cardiac function of neonates exposed to anthracyclines in utero. Materials and Methods: This was an observational study. Pregnant women diagnosed with breast cancer who underwent anthracycline based chemotherapy and delivered a liveborn infant after 24 weeks, were offered enrollment. Patients provided written consent to obtain doses and dates of chemotherapy treatment. Echocardiograms were offered to all newborns exposed as fetuses to anthracyclines. Women chose to perform this echocardiogram locally or at the institution of the primary investigator. Baseline fetal cardiac evaluation was reportedly normal on ultrasound prior to starting chemotherapy.

Chemotherapy Regimens:

Adriamycin/Cytoxan (n=20).

5-Fluorouracil/Adriamycin/Cytoxan (n=3)

There were no cases of exposure to epirubicin or idarubicin.

Mean dosage of Adriamycin was 57/m2 +/−8.0.

Results: Twenty three neonates participated.

Neonatal echocardiograms done at a mean age of 15 +/−26 months, (range 3 days of life to 9.75 years). RV wall, IVS and LV posterior wall diastolic and systolic thickness measurements were within normal limits for age and BSA. Mean shortening fraction was 41+/−10%. (normal 33 +/− 3%). In all 23, AV and semilunar valves were normal. Great arteries were normal in 22/23, with one case of a small pulmonary artery fistula. At 2 years of age, this was not hemodynamically significant. Patent foramen ovale was noted in 4 cases, all in children less than 2.0 months of age. Two children ages 3 days to 1.4 months had a PDA. One child at 1 year of age had an ASD. No structural or functional heart defects were diagnosed in any of these children.

Discussion:

Cardiac function appears to be preserved in neonates exposed to adriamycin chemotherapy in utero.

Any toxic effects on fetal myocytes was not evident in postnatal echocardiograms.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P5-10-18.
Abstract P5-10-20: Breast Cancer Multi Disciplinary Team (MDT)Adjuvant Chemotherapy Decision: Impact of Adjuvant! Online Estimates of 10-Year Risk of Death and Chemotherapy Benefit on the Decision Making Process
Authors: Balakrishnan A , Haysom S , Rivett L , Austin A , Ravichandran D.
Published: April 2011

Introduction: Breast cancer patients in the UK National Health Service (NHS) are cared for by breast cancer multidisciplinary teams (MDT). The wide range of knowledge and experience available in MDT produce a collective expert opinion on the management strategy for individual patients. Adjuvant! Online is an online computer programme that makes quantitative estimates of the effect of adjuvant chemotherapy (CT) on the 10-year risk of death and relapse. We studied whether the Adjuvant! Online produced numerical estimates would influence the adjuvant CT decision making of a breast cancer MDT.

Methods and Materials: We prospectively monitored a breast cancer MDT for a period of 12 months. Once the MDT had finished discussing a patient and adjuvant therapy decisions were made, Adjuvant! Online produced 10-year risk of breast cancer related death and the reduction of this risk that may be achieved by adjuvant CT were presented to the MDT. Any change to the original MDT recommendations as a result of this information was recorded.

Results: 109 post-operative early breast cancer patients that were suitable for input into Adjuvant! Online discussed in 41 MDT's were included in the study. Adjuvant therapy decisions included chemotherapy (n=56), radiotherapy (n=96) and endocrine therapy (n=89). The numerical estimates from Adjuvant! Online only changed the management of one patient, by showing a marginal (3.8%) survival benefit with CT that made the MDT offer chemotherapy to this patient where the original decision was not to.

Conclusions: Most adjuvant therapy decisions are easy to make without the need for numerical estimates of benefit when the MDT's are staffed by experienced breast cancer clinicians. Regular use of Adjuvant! Online or similar aids may not be necessary during the MDT discussion. These could be used on a selective basis where there is difficulty reaching a consensus.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P5-10-20.
Abstract P5-10-21: Maternal and Neonatal Outcomes of Dose Dense Chemotherapy for Breast Cancer in Pregnancy
Authors: Cardonick EH , Gilmandyr D
Published: April 2011

Background: Fear of neutropenia requiring the use of pegfilgrastim has prevented the use of dose dense chemotherapy (dd) during pregnancy. Some physicians/patients may be concerned as well about the ability of the pregnant patient and her neonate to tolerate dose dense chemotherapy during pregnancy. The objective of this study is to compare the outcome of women and their neonates when dose dense chemotherapy is used in pregnancy compared to treatment at least 3 weeks apart. Materials and Methods: A retrospective cohort study of pregnant women receiving chemotherapy for breast cancer were identified from the Cancer and Pregnancy Registry. The timing of chemotherapy in pregnancy was decided by the patient's oncologist in consultation with their patient. Fisher's Exact test comparing patient and neonatal outcome was performed.

Results: 109 pregnant women diagnosed with breast cancer participated. Ten women received dose dense (dd) and 99 conventional chemotherapy (cc). The mean gestational age at delivery for dd group was 35.8 (SD 3.1) weeks and 36.3 (SD 2.7) weeks in the cc group, p=.60. Mean birth weight was 2560 (SD 579)grams in the dd group, 2634 (SD 599) grams in the cc group, p=.642]There were no fetuses with growth restriction in the dd group, compared to 7.0% in the cc group, p=1.0. Of the 10 neonates in the dd group, 10% (1/10) experienced transient neutropenia, none did so in the cc group, p=0.09.

Neonatal Outcomes

GA=gestational age at delivery;IUGR=fetal growth restriction. No significant differences in GA at delivery, Birthweight or neutropenia was found.

There was an 10% (1/10) incidence of maternal neutropenia in the dd group, 5.1% in the cc group, p=.45. Maternal mean follow up was 3.5 (2.6) years. The mean time to recurrence was 17.6 months for dd versus 19.7 months in the conventional group, p=0.84. Overall survival in the dd group was 90%, in the cc group, 86%, p=1.0.

Maternal Outcomes

No significant difference in neutropenia, time to recurrence or survival was found.

Discussion: The small number of women receiving dose dense treatment is a limitation of this study. We found that maternal neutropenia, time to recurrence as well as overall survival was comparable between groups. Neonatal birth weight, gestational age at delivery and incidence of neonatal neutropenia was comparable between groups. Dose dense chemotherapy should be considered in pregnant women if the benefits are demonstrated in the nonpregnant population.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P5-10-21.