bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P1-04-06: Insertional mutagenesis identifies HACE1 as a HER2/Neu Cooperating Breast Cancer Tumor Suppressor Gene
Authors: Goka E , Miller P , Baker K , Stark G , Lippman ME
Published: August 2014

The development of human breast cancer is a multi-step process leading from normal epithelium to fully transformed breast cancer. Early genetic changes result in hyperplasias which evolves into ductal carcinoma in situ (DCIS) and culminates as invasive ductal carcinoma. The transition from DCIS to invasive disease has been implicated as the key transition in breast cancer progression. Specific genomic, transcriptomic, and proteomic alterations responsible for this transition process have yet to been elucidated.

HER2/Neu, a member of the EGFR family of receptor tyrosine kinases, is associated with poor clinical outcome. HER2/Neu overexpression/amplification is commonly seen (50–60%) in non-invasive lesions but is significantly less common (20–30%) in invasive and metastatic breast carcinoma. This indicates that HER2/Neu gives normal epithelium a proliferative advantage but additional genetic alterations are required for full malignant transformation. This idea is supported by the fact that transgenic mice that overexpress HER2/Neu develop spontaneous mammary tumors but only after a prolonged latency period.

Recent forward genetic approaches use insertional mutagenesis to randomly integrate a strong promoter into the genome, activating downstream gene transcription that can lead to either gain or loss-of-function mutations based on the integration site. We used this insertional mutagenesis approach to induce anchorage-independent growth of isolated murine HER2/Neu over-expressing mammary epithelial cells. We identified HECT domain and ankyrin repeat containing E3 ubiquitin protein ligase 1 (HACE1) as a putative breast tumor suppressor protein whose loss leads to the formation of anchorage-independent colonies.

Loss of HACE1 expression is commonly seen in breast cancer patients [as well as other tumor types] supporting the role of HACE1 as a breast tumor suppressor gene. Knockdown of HACE1 in human mammary epithelial cells (HMECs) results in the acquisition of anchorage-independent growth. Moreover, knockdown of HACE1 in established breast cancer cell lines that express moderate levels of HACE1 results in enhanced cloniginicity.

Therefore, we propose that the loss of HACE1 can potentially be used as a predictive marker for breast cancer progression. Additional studies investigating the mechanism of HACE1 action may also identify therapeutic targets that counteract HACE1 loss.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P1-04-06.
Abstract P1-04-07: The mRNA Expression of DAP1 in Human Breast Cancer: Correlation with Clinicopathological Parameters
Authors: Wazir U , Jiang WG , Sharma AK , Mokbel K
Published: August 2014

BACKGROUND: This pilot study is the first to focus on the potential role for death-associated protein 1 (DAP1) in human breast cancer.

MATERIAL AND METHODS: mRNA expression of DAP1 in breast cancer tissues (n = 127) and normal background tissues (n = 33) were determined using quantitative polymerase chain reaction (qPCR) and correlated with clinicopathological data accumulated over a 10-year follow-up period. Furthermore the effects of DAP1 knockout in breast cancer cell lines were investigated.

RESULTS: The expression of DAP1 mRNA was demonstrated to decrease with increasing Nottingham Prognostic Index (NPI2 vs. NPI3, p = 0.0026), and TNM stage (TNM1 vs. 4, p = 0.0039). Lower DAP1 expression levels were significantly associated with local recurrence (p = 0.02) and distant metastasis (p = 0.001).

The knockout of the DAP1 gene in MCF-7 cell lines resulted in a significant decrease in the mRNA expression of P-21 and DELE compared with controls, p = 0.04). However, the knockout of DAP1 had no effect on the expression of caspase 8 and 9.

CONCLUSIONS: This study demonstrates an inverse association between DAP1 mRNA levels and tumour stage and clinical outcome in breast cancer. This may be suggestive of a relationship between oncogenesis and the autophagy pathway meriting further investigation. These results also indicate that the pro-apoptotic function seems to be estrogen-dependent but independent of caspase 8 and 9. The DAP1 pro-apoptotic pathway may involve P-21 and DELE proteins.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P1-04-07.
Abstract P1-04-09: mTORC1 and Rictor expression in human breast cancer: correlations with clinicopathological parameters and disease outcome
Authors: Wazir U , Kasem A , Sharma AK , Jiang WG , Mokbel K
Published: August 2014

BACKGROUND: mTOR (the mammalian target of rapamycin) plays a key role in regulation of cellular metabolism, growth, and proliferation. It forms two multi-protein complexes known as complex 1 (mTORC1) and 2 (mTORC2). Raptor and Rictor are the core proteins for mTORC1 and mTORC2 respectively. There is a growing body of evidence that mTORC1 is up-regulated in many cancers and plays a role in carcinogenesis. The aim of the study was to investigate the mRNA expression of mTOR and Rictor in human breast cancer and examine the relationship between their expression and clinicopathological parameters.

METHODS: Specimens of breast cancer (BC) tissues (N = 150) and normal tissues (N = 31) underwent RNA extraction and reverse transcription. mTOR and Rictor transcript levels were determined using real-time quantitative PCR. Expression levels were analyzed against tumor size, grade, nodal involvement, TNM stage, and clinical outcome over a 10 year follow-up period.

RESULTS: Significantly higher mRNA transcript levels of mTOR were found in the breast cancer specimens compared to normal glandular tissue (p = 0.0018). The expression of mTOR mRNA was demonstrated to increase with increasing NPI (53 for NPI1 to 219 for NPI3) and tumor grade (37 for grade 2 vs. 159 for grade 3, p = 0.047). mTOR expression was found to be higher in ductal tumors compared with non-ductal tumors (p = 0.0014). The patients who developed recurrent disease or died from breast cancer had higher expression levels than those who had been disease-free after a median follow-up period of 10 years (p = 0.17). Higher expression levels were significantly associated with worse overall survival (p = 0.01).

In contrast, higher levels of Rictor mRNA expression were found in normal breast tissue, lower NPI stage (NPI1 vs. 2: p = 0.03) and lower tumor grade (grade 1 vs. grade 3: p = 0.01). Patients with higher Rictor expression had a significantly better overall (p = 0.037) and disease-free (p = 0.048) survival.

CONCLUSIONS: mTOR expression was found to be significantly higher in BC specimens compared to normal breast tissue. Higher transcript levels were significantly associated with unfavorable pathological parameters and adverse clinical outcomes. The core protein of the less predominant mTORC2 was associated with favorable pathological parameters and clinical outcome. Taken together these observations are consistent with mTORC1 role in the anti-apoptosis pathway and suggest that selective inhibitors of mTORC1 are likely to be a more effective therapeutic strategy than dual inhibitors in human breast cancer.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P1-04-09.
Abstract P1-07-04: Comparison of HER2 expression by immunohistochemistry (IHC) using automated imaging system and fluorescence in situ hybridization (FISH). A retrospective analysis of 2853 cases
Authors: Collins R , Xiang D , Christie A , Leitch M , Euhus D , Rao R , Haley B , Sarode V
Published: August 2014

Background: Accurate assessment of HER2 status is critical for selecting patients who will benefit from trastuzumab therapy. There is still no consensus regarding the optimal method to assess HER2 status. Computerized image analysis has been shown to provide a more accurate and objective way for quantification of HER2 expression by IHC than manual evaluation. It has been suggested that the use of image analysis may help to resolve some of the discrepancies between IHC and FISH assay.

We compared the results of HER2 expression by IHC using automated image analysis with fluorescent in situ hybridization (FISH) assay.

Design: Testing for HER2 expression by IHC and FISH was performed on 2853 specimens at UT Southwestern Medical Center between the years 2002 to 2011. Quantification of IHC HER2 expression was done by image analysis and scored as positive (>2.0), borderline (1.5 to 2.0) and negative. (<1.5). The PathVysion kit was used for FISH assay to evaluate HER2 amplification. Ratios >2.0, 1.8 to 2.0 and <1.8 were defined positive, borderline and negative amplification respectively.

Results: IHC compared to FISH

Conclusion: Despite improvements in IHC testing, the FISH assay may be a better method for determining HER2 status. Factors such as tissue fixation, scoring methods and choice of antibodies may contribute to the lower specificity of IHC.

In the amplified group, the gene amplification ratio correlated with protein expression, being highest in the IHC positive cases and lowest in those that were negative.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P1-07-04.
Abstract P1-07-14: Enabling biomarker validation in breast cancer molecular subtypes: sensitivity and specificity of array-based subtype classification in 983 patients
Authors: Györffy B , Lanczky A
Published: August 2014

Background: The diverse breast cancer molecular subtypes have different clinicopathological characteristics and outcomes. Here, we assessed the correlation between microarray-defined and author-reported molecular subtypes using publicly available breast cancer datasets.

Methods: GEO was searched for datasets where the authors determined molecular subtypes. The molecular subtype was re-calculated for each patient using the StGallen guidelines by assessing the microarray-based expression of ER, HER2 and MKI67 (Basal: ER negative + HER2 negative, Luminal A: ER positive + HER2 negative + low MKI67, HER2 enriched: HER2 positive + ER negative, Luminal B: ER positive + HER2 positive and ER positive + HER2 negative + high MKI67). The thresholds used were 500 for ER (probe set 205225_at), 4800 for HER2 (216836_s_at) and 470 for MKI67 (212021_s_at). Sensitivity and specificity were calculated for each subtype separately.

Results: Molecular subtype was published in all together six datasets (GSE1456, GSE21653, GSE25066, GSE20711, GSE31519 and GSE17907) for 983 patients. In these, 380, 306, 169 and 128 were Basal, Luminal A, Luminal B and HER2 enriched, respectively. The microarray-based molecular subtype determination resulted in a sensitivity of 0.70, 0.55, 0.63 and 0.38 and a specificity of 0.96, 0.87, 0.67 and 0.99 for Basal, Luminal A, Luminal B and HER2 enriched subtypes, respectively. Finally, the option to filter for molecular subtypes was implemented into our online biomarker validation platform at www.kmplot.com.

Discussion. Microarray data provided highest sensitivity and specificity to independent subtype classification for Basal tumors, while the luminal B subtype displayed the highest discordance. Our registration-free online service enables the validation of gene expression based biomarkers in each subtype separately.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P1-07-14.
Abstract P1-10-01: Curcumin suppresses MMP-9 expression via inhibition of PKCα/MAPKs and NF-κB/AP-1 activation in MCF-7 cells
Authors: Kim SK , Kim YW , Youn HJ , Jung SH
Published: August 2014

Background: Curcumin is a polyphenol derived from the plant turmeric (Curcuma longa), which is commonly used as a spice. It was recently reported for its anticancer effect on several types of cancer cells in vitro, however the molecular mechanisms of this anticancer effect are not fully understood. In the present study, we investigated the effect of curcumin on 12-O-tetradecanoylphorbol-13-acetate(TPA)-induced matrix metalloproteinase-9(MMP-9) expression and cell invasion in MCF-7 breast cancer cells.

Materials and Methods: The effect of curcumin on MCF-7 cell viability was determined using MTT assay. The cells invasion was demonstrated by the Matrigel-coated transwell assay. Western blot analysis was performed to detect the effect of curcumin on the expression of MMP-9. MMP-9 mRNA levels were analyzed by real-time PCR. NF-κB and AP-1 DNA binding was analyzed by EMSA.

Results: Our results showed that curcumin inhibits TPA-induced MMP-9 expression and cell invasion through suppressing NF-κB and AP-1 activation. Curcumin strongly repressed the TPA-induced phosphorylation of p38 and JNK and also inhibited TPA-induced translocation of PKCa from the cytosol to the membrane, but did not affect the translocation of PKCδ.

Conclusion: It is concluded that curcumin inhibits the TPA-induced MMP-9 expression and cell invasion through the suppression of the PKCα/MAPK/NF-kB/AP-1 pathway in MCF-7 breast cancer cells. Accordingly, curcumin may have the therapeutic potential in restricting breast cancer metastasis.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P1-10-01.
Abstract P1-13-06: Breast cancer survival advantage with oral bisphosphonates use
Authors: Rennert Gad , Rennert Hedy S , Pinchev Mila , Landsman Keren , Lejbkowicz Flavio
Published: August 2014

Withdrawn by Author

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P1-13-06.
Abstract P1-14-09: Immunohistochemical classification of intrinsic subtypes as a predictive biomarker of pathological compelete response in breast cancer patients treated with preoperative chemotherapy.
Authors: Nagayama A , Jinno H , Takahashi M , Hayashida T , Hirose S , Kitagawa Y
Published: August 2014

Background: Analysis of gene expression arrays has resulted in the recognition of several fundamentally different subtypes of breast cancer. These subtypes have different natural histories and different responses to systemic and local therapies. However, widespread use of gene expression profiling is limited because of its expense and of technical difficulty. Alternatively, this classification has also been reproduced in immunohistochemical studies. The objective of this study was to evaluate the clinical utility of immunohistochemical classification of breast cancer intrinsic subtypes in the prediction of pathological compelete response (pCR) in a cohort of breast cancer patients receiving the sequential combination of taxane followed by anthracycline as preoperative chemotherapy.

Materials and methods: A prospective database of 150 women with stage II/ III breast cancer, who received neoadjuvant chemotherapy, was analyzed. The regimens were either four cycles of docetaxel (40 mg/m2 on day 1 every 3 weeks) with S-1 (80 mg/m2 on days 1–14 every 3 weeks) followed by four cycles of 5-fluorouracil (500 mg/m2), epirubicin (100 mg/m2) and cyclophosphamide (500 mg/m2) (FEC) or four cycles of docetaxel (60 mg/m2 on day 8 every 3 weeks) with capecitabine (1650 mg/m2 on days 1–14 every 3 weeks) followed by four cycles of FEC. Subtypes were classified into luminal A (lumA) (ER+ and/or PgR+, HER2−, Ki67<14%), luminal B (lumB) (ER+ and/or PgR+, HER2−, Ki67>14%), triple-positive (TP) (ER+ and/or PgR+, HER2+), HER2 (ER−, PgR−, HER2+), and triple-negative (TN) (ER−, PgR−, HER2−) by using immunohistochemically stained specimen obtained by core needle biopsy. Absence of invasive tumor cells in the breast at the time of surgery was defined as pCR.

Results: We found 34 women in lumA, 54 in lumB, 15 in TP, 8 in HER2, 33 in TN. The median age in each group was 53, 51, 50, 57 and 53 respectively (p = .397). The mean tumor size was 3.2cm, 3.5cm, 4.4cm, 3.9cm and 3.6cm respectively (p = .158). The clinical response rate measured by physical examination (calliper) was 80.6%, 93.9%, 86.7%, 100% and 82.4% (p = .378). pCR rate was 5.9%, 14.8%. 13.3%, 50%, 21.2% respectively (p = .031). The multivariate analysis showed ER negative, PgR negative and intrinsic subtype (TN) were the significant predictive factors of pCR (p = .027, p = .003, p = .004).

Conclusion: This study indicated that immunohistochemical classification of intrinsic subtype might be a useful predictive biomarker of pCR in breast cancer patients treated with preoperative chemotherapy.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P1-14-09.
Abstract P1-15-10: Chemotherapy-Induced Neutropenia in Breast Cancer Patients Receiving Sequential Anthracycline and Taxane Chemotherapy: An institutional experience
Authors: Staudigl C , Seifert M , Tea M-K , Pfeiler G , Fink-Retter A , Fritzer N , Singer CF
Published: August 2014

Background: Sequential regimen of anthracyclines and taxanes are commonly used in patients with early stage breast cancer (BC) and curative treatment intent. Due to the associated myelotoxicity, internationally accepted guidelines recommend granulocyte-colony stimulating factors (G-CSF) as primary or secondary prophylaxis, dependent on chemotherapy (CT) and patient-associated risk of febrile neutropenia. As part of a quality control initiative at our center, we have analyzed rates of grade 1 or 2 (<2.0 to >1.0 × 109/L) and 3 or 4 (<1.0 to <0.5 × 109/L) neutropenia, febrile neutropenia (<0.5 × 109/L and single oral temperature ≥38.3°C/100.4°F), CT dose reductions, anti-infective use, and hospitalizations in cycles with pegfilgrastim prophylaxis.

Methods: Patients (pts) with BC stages I to III received four three-weekly (q21) cycles of adjuvant or neoadjuvant EC (Epirubicin 90 mg/m2 + Cyclophosphamide 600 mg/m2 q21) followed by four cycles of D (Docetaxel 100 mg/m2 q21) plus primary or secondary pegfilgrastim prophylaxis. Chart records were collected and analyzed retrospectively for patients treated between 2009 and 2012 at our institution for incidence of neutropenia and for the presence of neutropenia-associated risk factors. Only cycles with pegfilgrastim prophylaxis were considered for this analysis.

Results: 100 consecutive patients were identified who had overall received 200 EC cycles followed by 251 D cycles under G-CSF prophylaxis with pegfilgrastim. Pts age mean: 51,57 (SD:11,23), BMI mean: 25,8 (SD:5,03) were well controlled in both sequences. Overall, neutropenia was significantly higher for EC (p = 0.001), however higher grade neutropenia (Grade3 and 4) were comparable (p = 0.200). No statistically significant correlations were found between neutropenia and age or body mass index.

Conclusions: Sequential use of EC followed by D in combination with prophylactic pegfilgrastim results in lower den expected FN rate [1]. In contrast to most published trials EC leads to significantly more neutropenias than D. However, the use of pegfilgrastim reduces both EC and D associated neutropenias grade 3 and 4 to a similar extent.

References:

1. Aapro, M.S., et al., 2010 update of EORTC guidelines for the use of granulocyte-colony stimulating factor to reduce the incidence of chemotherapy-induced febrile neutropenia in adult patients with lymphoproliferative disorders and solid tumours. Eur J Cancer. 47(1): p. 8–32.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P1-15-10.
Abstract P2-04-02: Identification of genes associated with breast cancer micrometastatic disease in bone marrow using a human-in-mouse xenograft system
Authors: Aft R , Li S , Mudalagiriyappa C , Dasgupta N , Watson M , Fleming T , Ellis M , Pillai S
Published: August 2014

Disseminated tumor cells (DTCs) found in the bone marrow (BM) of breast cancer patients portend a poor prognosis and are thought to be the intermediaries in the metastatic process. Study of these cells has been limited due to their scarcity. To develop a clinically relevant model to characterize hese cells, we have employed a human in mouse (HIM) xenograft model for propagating, isolating, and molecularly characterizing DTCs. Human breast adenocarcinomas were prospectively collected from 5 patients and implanted into humanized NOD/SCID mouse mammary fat pads. BM was collected from the long bones at varying passages of the tumors and analyzed for human-specific gene expression by qRT-PCR and gene expression microarray. Human-specific gene expression of SNAI1, GSC, FOXC2, KRT19, and STAM2, presumably originating from disseminated tumor cells, was detectable in the BM of all mice that had developed metastatic disease to other solid organs, but was not detectable in xenotransplanted mice that did not develop metastatic disease. Comparative gene expression microarray analysis of the HIM primary tumor, the corresponding BM from mice with metastatic disease, and BM from control mice identified additional patterns of gene expression enriched in BM-associated DTCs which included several genes associated with epithelial-mesenchymal transition, aggressive clinical phenotype, and metastatic disease development in primary human tumors. We have found that BM DTCs can be detected using the HIM xenograft model and have identified unique patterns of gene expression associated with BM DTCs, which may provide further insight into the biology and therapeutic vulnerability of metastatic tumor cell populations in breast cancer patients.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P2-04-02.
Abstract P2-05-05: Circulating HER2 extracellular domain (ECD) predicts a poor prognosis for metastatic breast cancer patients
Authors: Wang X-j , Shao X-Y , Xu X-H , Chen Z-H , Wang S , Cai J-F , Huang J , Huang P , Lou C-J , Ling Z-Q , Han MX
Published: August 2014

Background: Amplification or overexpression of HER2 has been shown to play an important role in approximately 30% of breast cancers and is strongly associated with increased recurrence and a worse prognosis. The HER2 extracellular domain (ECD) may be cleaved and shed from the surface of breast cancer cells and serum HER2 ECD levels can be detected by enzyme-linked immunosorbent assay (ELISA). In this study, we explored the relationship between circulating HER2 ECD and tissue HER2 status, then we also examined its predictive value in a cohort of metastatic breast cancer patients.

Methods: Two hundred and seven metastatic breast cancer patients from March 2009 to July 2011 were involved in this prospective study at Zhejiang Cancer Hospital. The patients were identified histopathologically as having breast cancer by pathologists and staged mainly based on the pathology, the clinical manifestation, and the imaging findings of CT and MRI according to the classification system of the International Union Against Cancer. Serum HER2 ECD levels were measured by ELISA. Tissue HER2 was determined by IHC and FISH in tumor samples, respectively.

Results: The level of serum HER2 ECD was at least more than 15 ng/ml in 31.4% (65/207) metastatic breast cancer patients, 39.1%(43/110) in HER2-positive patients and 23.4%(22/94) in HER2-negative cases, respectively, P = 0.017. We also found that high serum HER2 ECD levels (≥15 ng/ml) were significantly associated with elevated serum CEA (52.1% v 21.5%, OR 3.978, 95%CI 2.138–7.401, P = 0.000), CA125 (48.5% v 23.5%, OR 3.064, 95%CI 1.650–5.691, P = 0.000), CA153 (53.2% v 17.1%, OR 5.48, 95%CI 2.897–10.366, P = 0.000), LDH (53.3% v 23.1%, OR 3.798, 95%CI 2.011–7.173, P = 0.000) and AKP (51.2% v 26.5%, OR 2.911, 95%CI 1.442–5.879, P = 0.002), respectively. For the effect of HER2 ECD on the site of relapse, still there were statistically significant correlation between increased serum HER2 ECD levels and liver involvement (42.7% v 24.0%, OR 2.358, 95%CI 1.294–4.297, P = 0.005), brain metastasis (50.0% v 26.7%, OR 2.744, 95%CI 1.397–5.391, P = 0.003) and visceral involved (37.9% v 14.8%, OR 3.511, 95%CI 1.548–7.961, P = 0.002), respectively. While serum HER2 ECD levels were not related to age, BMI, tumor size, lymph node involvement and hormone receptors status, respectively (P>0.05).

Conclusions: Monitoring the circulating levels of the HER2 ECD in patients with metastatic breast cancer provides a real-time assessment of tumor burden and indicates poor prognosis, and may provide important information for reassessment of HER2 in HER2-negative metastatic breast cancer.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P2-05-05.
Abstract P2-05-10: 64Cu-DOTA-trastuzumab positron emission tomography imaging of HER2 in women with advanced breast cancer
Authors: Mortimer JE , Conti P , Shan T , Carroll M , Kofi P , Colcher D , Raubitschek AA , Bading JR , Miles J
Published: August 2014

Background: We propose to utilize64Cu-DOTA-trastuzumab with PET imaging to assess the in vivo expression of HER2 in women with advanced breast cancer. We hypothesized that the uptake of HER2 in normal tissues compromises the quality of radiographic images and that the pre-administration of trastuzumab prior to injection of64Cu-DOTA-trastuzumab would improve image quality. Two different doses of cold trastuzumab (5 mg and 50 mg) were tested. Once the dose of cold trastuzumab was established in women with documented HER2 positive metastatic disease, we initiated a second study of64Cu-DOTA-trastuzumab PET imaging in women whose cancers were HER2 1+ and 2+ by immunohistochemical staining.

Methods: Patients with biopsy confirmation of recurrent disease located outside the breast and axilla considered for study. Complete staging workup included CT of the chest, abdomen, and pelvis, bone scintigraphy and18F FDG PET. At lease 1 non-hepatic site of metastasis that was > 2 cm separate from the biopsy site was also required. HER2 status was assessed by both IHC and FISH. In the first study all patients were required to have HER2 positive disease and could not have received anti-HER therapy for > 2 months. Immediately prior to injection of64Cu-DOTA-trastuzumab, a cold dose of either 5 mg or 50 mg of trastuzumab was administered.64Cu-DOTA-trastuzumab PET imaging was performed at 24 and 48 hours.

Results: Eight women with HER2 positive metastatic breast cancer have undergone64Cu-DOTA-trastuzumab PET imaging: 2 received a pre-64Cu-DOTA-trastuzumab PET dose of 5 mg trastuzumab and 6 received 50 mg.64Cu-DOTA-trastuzumab PET effectively visualized and provided uptake measurements for presumably HER2+ metastatic lesions in bone, liver, lung and, to a lesser degree, lymph nodes. The 50 mg dose resulted in approximately 75% decrease in liver uptake of64Cu and improved the visualization of hepatic metastasis. We have initiated the second phase of the study utilizing the pre-administration of 50 mg trastuzumab in women with HER2 1+ and 2 + disease. To date we have imaged one patient.

Conclusion:64Cu-DOTA-trastuzumab PET imaging is feasible and image quality is improved with the pre-administration of 50 mg trastuzumab. Enrollment of women with IHC 1+ and 2+ disease continues. This work was supported by the Department of Defense grant # 1024511.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P2-05-10.
Abstract P2-06-04: Impact of genomic testing on chemotherapy utilization
Authors: Riley L , Nakijima H , Balinger K , Anderson D
Published: August 2014

Genomic testing for breast cancer has become a common practice. The two main tests at our institution are Oncotype DX® and MammaPrint®. Each test has unique features and criteria. We sought to compare the practice pattern between these tests and compare them to patients who did not undergo any genomic testing.

Methods: This is a retrospective review of all breast cancer patients between May 2008 and August 2011. Data was primarily thorough our tumor registry database. In some cases individual charts were also reviewed. Results: We analyzed 1093 breast cancers in 1064 patients. The average age was 62. The stage at diagnosis was 19% DCIS, 41% stage I, 24% stage II, 11% stage III and 5% stage IV. Of the 1093 cancers, 70 (6%) had an Oncotype test performed, 180 (16%) had a MammaPrint test done, 4 (0.4%) had both test, and 839 (77%) had no genomic test performed. Patients with an Oncotype test were more likely to be stage I than those with a MammaPrint (66% vs 48%). Additionally, patients who had a MammaPrint test more often received chemotherapy when compared to either the Oncotype test or no genomic testing.

With a mean follow-up of 1.5 years, there have been 28 recurrences (0 in the Oncotype group, 5 in the MammaPrint group, and 23 in the no genomic testing group). All five of the MammaPrint recurrences were high risk and received adjuvant chemotherapy. Of the 23 recurrences in the no genomic testing group, eight patients were node negative and could have been considered appropriate for genomic testing, 6 of these eight did not have adjuvant chemotherapy.

Conclusions: There are distinct practice pattern differences between Oncotype DX® and MammaPrint® utilization. MammaPrint testing was ordered more often in higher staged patients and was associated with an increased use of chemotherapy. This may relate to MammaPrint's previous requirement for fresh tissue that typically occurs before the final nodal status is known. This difference should diminish with the recent availability of MammaPrint on FFPE tissue. All patients who underwent genomic testing and developed a recurrence had been treated with adjuvant chemotherapy. In contrast, patients who did not have a genomic test and experienced a recurrence may have benefited from genomic testing.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P2-06-04.
Abstract P2-10-23: Lymphovascular Invasion (LVI) and Overall Survival in Node-negative and Node-positive Breast Cancer Patients: A Meta-analysis.
Authors: Sahebjam S , Diaz-Padilla I , Ocana A , Seruga B , Amir E
Published: August 2014

Introduction: LVI is an important prognostic factor in patients with lymph node-negative invasive breast cancer. The prognostic value of LVI in lymph node-positive disease or in other subgroups remains unclear. Here we present a meta-analysis of studies assessing the impact of LVI on overall survival (OS) in different subgroups of breast cancer patients.

Methods: A published data meta-analysis was conducted. Studies reporting hazard ratios (HR) for the association of LVI with OS in a multivariable model were included. Those not reporting HR or utilizing only univariable analyses were excluded. HR and 95% confidence intervals (CI) were extracted or computed and pooled using the DerSimonian and Laird random effects model. Subgroup analyses were conducted for the differential effect of lymph node involvement, estrogen receptor (ER) status, extent of LVI and decade in which the study was conducted (pre 1990, 1990–1999 and 2000 onwards).

Results: Twenty studies comprising 40,417 patients were included in the analysis. There was marked heterogeneity in the effect size between studies (Cochran Q p < 0.001, I2=62%). When all studies were pooled, LVI was significantly associated with worse OS (HR:1.70 95% CI 1.49–1.95, p < 0.001). Among studies selecting patients by lymph node status, the association between LVI and OS was similar for lymph node positive and negative patients (HR:2.03 and 2.41 respectively, subgroup difference p = 0.56). There were no significant differences based on ER-status (subgroup difference p = 0.75) or decade of study (subgroup difference p = 0.21). Pooled assessment of the effect of extent of LVI was not possible as this parameter was not consistently reported.

Conclusion: LVI is associated with similarly detrimental OS regardless of lymph-node or ER status. Improvements in adjuvant therapies over the past 20 years do not appear to have diluted the impact of LVI on OS. The marked heterogeneity in the effect size may result from differences in the extent of LVI.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P2-10-23.
Abstract P2-11-12: Proactive approach to risk reduction and early detection of breast cancer related lymphedema
Authors: Fu MR , Guth A , Kleinman R , Cartwright F , Haber J , Axelrod D
Published: August 2014

Advances in surgery and post-operative treatment continue to reduce the incidence of late effects of cancer treatment such as lymphedema. Yet, many breast cancer survivors still face considerable post-operative challenges as a result of developing lymphedema. The purpose of this study was to preliminarily evaluate a nursing educational and behavioral program “The Optimal YOU” to enhance lymphedema risk reduction, focusing on preventing inflammation-infection, promoting lymph drainage, and maintaining optimal Body Mass Index (BMI) as well as preoperative and subsequent follow-up assessment of limb volume changes and symptoms.

A quasi-experimental design with repeated-measures was used. The Study outcomes included lymph volume changes by infra-red perometer, lymphedema-related symptoms, and BMI by Bioimpedance at pre-operative baseline, post-operative, 6-month, and 12-month follow-up. A total of 147 patients were recruited and received the program, 141 patients completed the study (4% attrition rate).

Majority (over 90%) of patients maintained preoperative limb volume and improved their symptom experience at 12-month follow-up. During the study period, only 7 patients (5%) had measurable lymphedema. At 12-month follow-up, among the 7 patients who developed measurable lymphedema, 2 patients' limb volume returned to pre-operative level without compression therapy only by maintaining exercises of The Optimal YOU to promote daily lymph flow. About 80% of patients maintained or improved their preoperative BMI.

Implementation of this nursing educational and behavioral intervention is effective to enhance lymphedema risk reduction and early detection. Findings of the study provided evidence to support the emerging policy change in lymphedema care, shifting treatment-focus care to proactive risk reduction.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P2-11-12.
Abstract P2-13-08: Comparison of Compliance to Anti Estrogen Therapy in Patients with Early Breast Cancer followed at Tertiary Centers versus Through Family Physicians and Primary Surgeons: A Practice Review
Authors: Alkhayyat SS , Younus J , Mirza FN , Stitt L
Published: August 2014

Background: Poor adherence or non-compliance to pharmacologic therapies in chronic diseases is a major clinical problem. Adherence to adjuvant anti-estrogen therapies among patients with early breast cancer (EBC) is variable as reported in different clinical trials. Cancer centres, at present, frequently refer patients with EBC on adjuvant anti-estrogen therapy back to their surgeons and family doctors for follow up. In order to find any difference in compliance for such patients, we reviewed patients followed by their primary surgeons and family doctors (peripheral cohort) for adherence and compared them to patients regularly followed in the cancer center (central cohort).

Patients and Methods: Women with EBC receiving anti-estrogen therapy were identified from breast cancer database at London Regional Cancer Program LRCP). A standardized telephone interview was conducted with patients. We assumed that adherence in the central cohort will be 20% higher than the peripheral cohort. Patient are considered adherent if they took more than 80%, Non-adherent if they took less than 50% and semi-adherent if they took 50–80% of the prescribed medication.

Results: We recruited a total of 160 patients (80 patients in each cohort). Seventy seven (96.3%) patients in central cohort were compliant and 76 (95%) patients were compliant in the peripheral cohort. The HR was 0.7 (p >.999)

Conclusion: From our retrospective review, we did not observe any significant difference in the adherence to adjuvant anti-estrogen therapy in patients with EBC. The present process of discharging such patients to surgeons and family physicians for follow up does not seem to affect their compliance.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P2-13-08.
Abstract P2-14-02: Preclinical Evaluation of Enzalutamide in Breast Cancer Models
Authors: Cochrane DR , Bernales S , Jacobsen BM , D'Amato NC , Guerrero J , Gómez F , Protter AA , Elias AD , Richer JK
Published: August 2014

Background: The vast majority of breast cancers express the androgen receptor (AR) with ∼80% of ER+ and 30% to50% of ER− tumors being positive for AR by immunohistochemistry (IHC). Approximately 30% of breast cancer patients with estrogen receptor positive (ER+) tumors do not respond to tamoxifen or aromatase inhibitors and AR signaling has been implicated as a possible mechanism of this insensitivity. AR overexpression has also been associated with resistance to either tamoxifen or aromatase inhibitors in cell line models.

Hypothesis: ER+ breast cancers can switch from estrogen to androgen-dependent growth as they become resistant to traditional endocrine therapies and AR can serve as a therapeutic target in AR+ breast cancers, irrespective of ER status.

Methods: The proliferative effect of dihydrotestosterone (DHT) and estradiol (E2) in the presence enzalutamide (formerly MDV3100) was examined in vitro and in xenograft studies using models of both ER+ and ER− breast cancer that express AR. Furthermore, AR and ER were examined by IHC in clinical specimens from breast cancer patients treated with neoadjuvant exemestane and adjuvant tamoxifen. Tumor tissue was compared to adjacent normal breast epithelium.

Results: In ER+ models, bicalutamide and enzalutamide both inhibited DHT-mediated proliferation, while enzalutamide uniquely inhibited E2-mediated proliferation. In vivo, enzalutamide significantly reduced estrogen- and androgen-mediated growth of MCF7 (ER+/AR+) xenograft tumors. Remarkably, enzalutamide demonstrated an anti-proliferative effect comparable to tamoxifen. in vitro, enzalutamide inhibits E2-stimulated tumor cell proliferation and E2 bound ER activation of genes such as PR and SDF-1, despite no observed binding of enzalutamide to ER alpha or beta. In ER−/AR+ MDA-MB-453 xenografts, enzalutamide treatment resulted in decreased nuclear AR localization, increased apoptosis and tumor growth inhibition.

Conclusions: Enzalutamide demonstrated significant anti-tumor activity in preclinical models of breast cancers that express AR, regardless of ER status. When enzalutamide opposes DHT, it functions by excluding AR from the nucleus, thereby reversing the anti-apoptotic effect of AR. In contrast, when opposing E2, it causes a decrease in tumor cell proliferation. Profiling of ER+ xenografts is underway to further elucidate the mechanism by which this occurs.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P2-14-02.
Abstract P2-14-06: A phase II trial of low dose estradiol in postmenopausal women with advanced breast cancer and acquired resistance to an aromatase inhibitor.
Authors: Howell SJ , Seif MW , Armstrong AC , Cope J , Wilson G , Welch RS , Misra V , Ryder D , Blowers E , Palmieri C , Wardley AM
Published: August 2014

Background High dose estrogen (HDE) is an effective but toxic treatment for postmenopausal women with advanced breast cancer (ABC). In vitro, prolonged periods of estrogen deprivation (ED) sensitises cancers to the inhibitory effects of estrogen. We hypothesised that the profound ED seen with third generation aromatase inhibitor (AI) would sensitise cancers to LDE which would be better tolerated.

Methods Single arm phase II study in postmenopausal women with measurable ER+ ABC and demonstrated clinical benefit (CB) with a third generation AI. Treatment: estradiol valerate 2mg daily. Primary endpoint: CB rate (CBR) was correlated with baseline and dynamic LH/FSH levels and on-treatment estradiol levels. Secondary endpoints included TTF, PFS, toxicity and QoL. If LDE was effective, retreatment with the AI on which the cancer had progressed prior to study entry was offered on progression. If LDE was ineffective HDE was offered.

Results 21/50 patients were recruited before early closure due to slow accrual. 19 were assessable for efficacy and toxicity (1 ineligible; 1 no LDE). CBR was 5/19 (26%; 95%CI 9.1%,51.2%). Median TTF 2.8months (range 0.5–29.8). CBR durations were 11.1, 16.8, 17.3*, 19.8, 29.8 months (*censored). 4 stopped treatment early due to toxicity (G4 hypercalcaemia/ G2 vaginal bleeding (VB) plus G4 hyponatraemia/ G2 mucositis/ G2 headaches) 1 with SD at 3 months and 3 before response assessment. Other common toxicities were all G1/2 and mainly limitted to nausea (8/19), breast pain (7/19) and tumour flare (7/19). VB occurred in 8/11 without prior hysterectomy. Baseline LH correlated with CBR by logistic regression (p = 0.01). Of 3 retreated with the same AI post LDE; 1 had PR, 1 SD ≥6months and 1 PD. One woman received HDE after failure of LDE and achieved a PR.

Conclusion LDE is an effective and largely well tolerated treatment in women with acquired resistance to AI. VB is common without prior hysterectomy. Rechallenge with the same AI post LDE seems effective and may offer an extra line of endocrine therapy. This should be tested in future trials.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P2-14-06.
Abstract P3-02-08: Is repetition of the contralateral mammogram of patients referred from breast cancer screening for unilateral findings necessary?
Authors: Castro C , Schipper R-J , van Roozendaal L , van Goethem M , Lobbes M , Smidt M
Published: August 2014

Introduction - The Netherlands started a nationwide breast cancer screening program in 1989, including women from the age of 50 until 75. Screening mammograms are performed two yearly in a mobile unit and consist of a bilateral two-view mammogram of the breast (mediolateral oblique and craniocaudal views).

If indicated, patients are referred to a breast clinic for diagnostic analysis. This work-up consist of among others repeating the bilateral two-view mammogram. Since the screening is digitalized, repeating of at least the mammogram of the non suspicious side might be unnecessary.

The aim of this study is to determine the additional value of repeating the contralateral mammogram in patients referred for a suspicious unilateral lesion.

Material and methods – 395 patients were referred from breast screening program to our institution for unilateral findings between October 2009 and August 2011. In all patients a bilateral mammogram was repeated and analyzed by an experienced breast radiologist. In the case of breast cancer a breast magnetic resonance imaging (MRI) was performed for preoperative staging. Anonymised data concerning the date of registration of the screening mammogram, the referred side (left/right or bilateral), age, screening's BI-RADS classification, breast density, biopsy results and breast MRI results were collected.

Results - Of the 395 patients referred for a suspicious unilateral finding, a malignancy on the referred side was observed in 144 patients (36.5%). In five patients bilateral breast cancer was detected. In one patient no malignancy was detected on the referred side, though on the contralateral side. Three of these six contralateral malignancies were directly mammographically detected. All six malignancies were detected with preoperative breast MRI.

Conclusion - Repetition of the two-view mammogram of the contralateral side in patients referred with a unilateral suspicious finding seems unnecessary, since all contralateral malignancies were depicted on the preoperative staging breast MRI recommended according to the EUSOBI-guidelines. Omission of the contralateral mammogram could lead to reduction of associated health care costs, radiation exposure, and patient discomfort caused by the exam.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P3-02-08.
Abstract P3-02-13: ANALYSIS OF INFRASTRUCTURE FOR MAMMOGRAPHY SCREENING IN THE STATE OF GOIAS, BRAZIL, 2010
Authors: Freitas-Junior R , Correa RS , Peixoto J-E , Rodrigues DCN , Rahal RMS
Published: August 2014

Objective: Assess the infrastructure for mammography screening in the state of Goiás, Brazil, taking into consideration the quality of the image generated by the equipment and the dose received by the patients, according to the type of technology (conventional and digital mammography).

Methods: Cross-sectional study in which the observational units were the diagnostic imaging services that were in operation in 2010. In an in loco visit, we collected information on the equipment and performed tests to assess quality of the image, accuracy values for tube tension, quality of X-ray beam, and dose at the simulator entrance. The percentage of mammography equipments in conformity with the parameters evaluated was calculated, and the mean glandular tissue dose (Dg) was estimated based on the dose at the simulator entrance. To assess quality of the image and the radiation dose, we used a standard breast simulator (5.3 cm thickness and 50% glandular tissue).

Results: In 2010, 102 services were in operation, using 106 equipments. Among these, 100 (94.3%) – 64 conventional and 36 digital mammography equipments – had their performance and dose assessed. The visualization of fibers (p = 0.044) and the dose at the simulator entrance (p = 0.042) were the parameters that presented differences between the types of technology.

The mean dose at the simulator entrance was 9.75 mGy (± 2.46) in conventional and 14.93 mGy (± 8.39) in digital equipments (p < 0.001); 12% of the conventional and 50% of the digital equipments presented Dg above the limit established by the European guidelines.

Conclusion: The infrastructure for mammography screening in the state of Goias, Brazil, in 2010 presented acceptable quality standard according to international guidelines. However, the radiation doses were heterogeneous between the two types of technology, with a predominance of higher doses in digital equipments.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P3-02-13.
Abstract P3-05-09: The role of TGF-beta receptor type 3 in breast cancer progression
Authors: Jovanovic B , Ashby WJ , Zijlstra A , Pietenpol JA , Moses HL
Published: August 2014

Breast cancer is a diverse group of diseases. Gene expression profiling has identified luminal A, luminal B, Her2-like, Normal breast-like and Basal-like as major classifications of breast cancer diseases. These correlate to the overall patient survival with the basal-like subtype, clinically termed as triple-negative breast cancers (TNBC), demonstrating the poorest overall outcome. The poor patient outcome is primarily due to absence of hormonal receptor targets ER, PR and HER2 which are the major drug-targets for currently available therapies. Considering that TNBC lacks well-defined molecular targets, our group pursued extensive genomic molecular and biological analysis of over 500 tumor tissues from TNBC patients. Ultimately, this led to the discovery of six, TNBC subtypes; among which are Mesenchymal-Stem Like (MSL) and Mesechymal (M). These two subtypes share a unique biological driver, the TGF-beta pathway.

To gain insight into the role of the TGF-β pathway signaling in TNBC we have performed gene expression analysis on all six TNBC subtypes. A major finding was that among all TGF-β pathway-associated genes, TGF-beta receptor type 3 (TBR3) is most differentially expressed in MSL and M tumor subtypes of TNBC. Based on these findings, we are hypothesizing that TBR3 is required for maintenance of tumorigenicity in MSL and M subtypes of the triple-negative breast cancer.

In order to test our hypothesis we have screened 15 TNBC cell lines for both RNA and protein levels of TBR3 and validated that TBR3 is differentially expressed in M and MSL subtypes. Furthermore, we have manipulated TBR3 expression in M and MSL TNBC cell lines and tested their ‘biologies’ using both an in vitro and in vivo model systems. Preliminary results indicate that upon silencing TBR3 expression, cell motility as well as ability to cluster together in 3D culture system is significantly changed. In addition, our xenograft mouse model also demonstrated a significant change in tumor onset and growth.

We are interested in validating the prognostic and functional contribution of TBR3 to the breast cancer progression. The results of this study will provide insight into the potential use of the TGF-β signaling axis for prognostic or therapeutic utility in breast cancer patients.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P3-05-09.