bluuebunny/crossref_metadata_2025 huggingface dataset:
P4-07-08: Subsets and Molecular Signatures of Circulating Tumor Cells in Breast Cancer Brain Metastasis.
Authors: Marchetti D , Zhang L , Wetzel M , Zaidi T , Ridgway L , Schoeber W , He W , Groves MD , Katz RL
Published: June 2012

Background: Circulating tumor cells (CTCs) represent the “seeds” of intractable brain metastatic breast cancer (BMBC); however, properties of CTCs targeting the brain remain elusive. For example, the FDA-approved CTC platform (CellSearch™, Veridex, LLC) detects only CTCs positive for epithelial cell adhesion molecule (EpCAM) and cytokeratins (CKs), but is unable to capture any other CTC subtypes or analyze biomarkers of brain-homing CTCs. We hypothesized that profiling CTCs from BMBC patients might result in the identification of brain-colonizing CTC signatures. Materials and Methods: We employed CellSearch™ and a novel technology that uses analysis of specific antigenic markers by immunofluorescence, coupled with detecting gene amplification by fluorescence in situ hybridization on the same cells; and quantification of the signal via automated scanning (FICTION; BioView Duet-3™ system).

Results: We established that our approach was feasible by performing CTC analyses on peripheral blood mononuclear cells isolated from BMBC patients or patients not possessing overt metastatic disease. We detected a differential gene amplification for human epidermal growth factor receptor1 and 2 (EGFR and HER2, respectively). Second, the number of EpCAM-positive CTCs visualized by the BioView™ platform was at least three orders of magnitude higher than one obtained from CellSearch™ CTC analyses using the same specimen. Third, we identified the presence of CTCs positive for CKs but negative for EpCAM. Conversely, high levels of prometastatic heparanase, in conjunction with the expression of aldehyde dehydrogenase-1 (ALDH1), a known cancer stem-cell marker, were detected in CTCs from BMBC patients; with a correlation between heparanase, ALDH1, and high EGFR amplification. Finally, extensive flow cytometric/FACS analyses validated the presence of CTC subsets negative for EpCAM and CD45, a hematolymphoid marker, however enriched for heparanase/ALDH1 expression.

Discussion: These findings indicate that the BioView™ platform not only captures more EpCAM-positive CTCs than CellSearch™ but also allows the detection of novel CTC subtypes possessing varying EpCAM levels. Importantly, they suggest that profiling CTC subtypes in patients with BMBC can be relevant towards the discovery of BMBC founder CTCs. Work is ongoing to further characterize these CTC subtypes, and to assess their abilities to metastasize to brain in xenotransplantation studies using immunodeficient mice.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-07-08.
P4-07-10: Circulating Tumor Cells in Newly Diagnosed Inflammatory Breast Cancer.
Authors: Mego M , Giordano A , De Giorgi U , Hsu L , Lucci A , Dawood S , Woodward WA , Ueno NT , Valero V , Andreopoulou E , Hortobagyi GN , Reuben JM , Cristofanilli M
Published: June 2012

Background: Inflammatory breast cancer (IBC) is one of the most aggressive forms of breast cancer and is associated with special clinical and biological features. Circulating tumor cells (CTC) are an independent prognostic factor in metastatic breast cancer for progression-free and overall survival. Previously we showed that metastatic IBC patients treated with first and subsequent lines of chemotherapy had a lower number of CTC when compared with CTC levels of non-IBC patients; hence the prognostic value of CTC in mIBC patients is limited. In the present study, we investigated the prognostic value of baseline CTC in newly diagnosed IBC patients. Patients and methods: This retrospective study included 84 newly diagnosed IBC (37 stage III B or C and 47 stage IV) patients treated with neoadjuvant therapy or first line chemotherapy between January 2004 and July 2009 at the MD Anderson Cancer Center. The median age was 55 years (range, 23–78). Twenty-four (28.8%) patients were HER2 positive, 36 (42.9%) hormone receptor positive and 33 (39.3%) were triple receptor negative. CTC were detected and enumerated using the CellSearch® system before patients started chemotherapy. Progression free survival (PFS) and overall survival (OS) were calculated from the date of CTC measurement and estimated by the Kaplan-Meier product limit method.

Results: At baseline, 64 (76.2%) patients had at least ≥1 CTC and 29 (34.5%) ≥5 CTC. Proportions of patients with stage III IBC with ≥1 CTC and ≥5 CTC were lower than those of newly diagnosed stage IV IBC patients (67.6% vs. 86%; p = 0.12 and 24.3% vs. 42.6%; p = 0.11, respectively). At a median follow up of 22.4 months (range: 4.8 - 58.3 months), 48 (57.1%) patients experienced disease progression and 26 (31.0%) had died. Patients with < 5 CTC had similar PFS (p = 0.36) and OS (p = 0.80) to those of patients with ≥5 CTC, respectively. We investigated the prognostic value of CTC ≥ 1 in stage III IBC patients, as well. There was no difference in PFS (p = 0.71) and OS (p = 0.15) in patients with CTC = 0 vs. those of patients with CTC ≥ 1, respectively. Moreover, we correlated CTC levels with prognosis, using different CTC threshold, but we observed no correlation between baseline CTC and patients’ outcome (Table 1). Conclusions: CTC can be detected in a large proportion of patients with newly diagnosed IBC; however, baseline CTC was not prognostic for PFS and OS. Collective dissemination of cancer cells via lymphovascular tumor emboli and/or epithelial-mesenchymal transition accompanied by inadequate detection of CTC due to lack of sensitivity of current assays may account for limited prognostic value of CTC in newly diagnosed IBC. Nevertheless, sample size and study power could influence study results as well.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-07-10.
P4-07-11: Circulating Tumor Cells after Neoadjuvant Therapy Predict Outcome in Stage I to III Breast Cancer.
Authors: Gainer S , Krishnamurthy S , Bhattacharyya A , Lodhi A , Hall C , Kuerer H , Bedrosian I , Anderson A , Singh B , Lucci A
Published: June 2012

Introduction: Circulating tumor cells (CTCs) predict outcome in metastatic breast cancer, but their significance is unclear in non-metastatic patients. Furthermore, it is unclear if the presence of CTCs after completion of neoadjuvant chemotherapy (NACT) predicts worse outcome. The purpose of this study was to determine if CTCs after NACT predicts worse outcome.

Methods: Clinical stage I-III breast cancer patients seen at a single tertiary cancer center provided informed consent to participate in an IRB-approved study involving collection of blood (7.5 ml x 2 tubes) at the time of surgery for their primary breast cancer. CTCs were detected using the Cell SearchTM system. A positive result was defined as the presence of one or more cells per 7.5 ml blood since the threshold for positivity has not been established in non-metastatic breast cancer. Statistical analyses used chi-square and Fischer's exact test.

Results: One hundred and twenty patients were prospectively enrolled. Median age was 50 years and median follow-up was 33 months. Eight percent of patients had T1 disease, 35% T2, 18% T3, and 39% T4. Fifty-three percent of patients (63/120) had hormone receptor positive disease. Thirty-two percent of patients (38/120) were HER-2 positive. Thirty percent (36/120) were triple negative. Seventy-eight percent (91/120) had lymph node positive disease. Two or more CTCs were present in 9% of patients (11/120). Of the 11 patients who died, 3 had 2 or more CTCs (P=0.08). Of the 20 who relapsed, 6 had 2 or more CTCs (P=0.0019).

Conclusions: Presence of two or more CTCs after NACT predicted worse relapse free survival in patients with stage I-III breast cancer.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-07-11.
P4-07-15: Insulin-Like Growth Factor Receptor I (IGFIR) Expression in Circulating Tumor Cells (CTCs) of Patients with Early and Metastatic Breast Cancer (BC).
Authors: Spiliotaki M , Markomanolaki H , Kokotsaki M , Kallergi G , Mavroudis D , Georgoulias V , Agelaki S
Published: June 2012

Background: Components of the insulin-like growth factor system are deregulated and IGFIR overexpression is commonly observed in BC, however the expression of the receptor in CTCs of patients with BC has not been studied.

Methods: IGFIR expression was assessed in CTCs from 65 patients with early and 101 with metastatic breast cancer before the start of adjuvant and first-line chemotherapy, respectively. Peripheral blood mononuclear cells’ (PBMCs) cytospins were stained with a monoclonal A45-B/B3 anti-cytokeratin (CK) antibody and IGFIR β-subunit anti-rabbit antibody. IGFIR(+)CK(+) CTCs were detected using immunofluorescent microscopy (Ariol system). Phosphorylated IGFIR was also evaluated after staining with phospho-IGFIR-β (Tyr1131)/Insulin Receptor-β (Tyr1146) antibody.

Results: CK(+) CTCs were detected in 23 (35.3%) patients with early and 48.5% with metastatic BC. IGFIR(+) CTCs were identified in all (100%) patients with early and in 38 (78%) out of 49 patients with metastatic BC (p=0.014). Eighteen (78%) out of 23 patients with early disease had exclusively IGFIR(+) CTCs and 5(22%) had both IGFIR(+) and IGFIR(−) CTCs. There were no patients in whom exclusively IGFIR(−) CTCs were identified. The respective values in patients with metastatic disease were 19 (39%), 19 (39%) and 11 (22%) (p=0.004, compared to early disease). The median percentage of the expression of IGFIR(+) and IGFIR(−) CTCs per patient was 100% (range 25–100) and 0% (range 0–75), in early and 68% (range, 0–100) and 32% (range 0–100), in metastatic disease, respectively (p=0.003). A total of 222 and 386 CTCs were detected in patients with early and metastatic disease, respectively. IGFIR expression was observed in 84% and 64% of all detected CTCs in adjuvant and metastatic patients, respectively (p<0.001). Phosphorylated IGFIR has been evaluated in PBMCs’ cytospins from 6 patients with adjuvant and 6 with metastatic disease. Interestingly, among 18 CTCs detected in early BC, all expressed phosphorylated IGFIR, whereas, among 34 CTCs identified in metastatic patients, none expressed the phosphorylated form of the receptor. The evaluation of phosphorylated IGFIR is ongoing and updated results will be presented.

Conclusions: IGFIR expression is commonly observed in CTCs of patients with BC. IGFIR expression is detected in a higher proportion of patients and in a higher proportion of the total number of CTCs identified in early compared to metastatic disease. The above observations suggest that metastatic progression is associated with loss or downregulation of IGFIR expression in breast cancer.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-07-15.
P4-07-21: A Comparison of Two Methods for the Detection of Circulating Tumor Cells (CTCs) in Patients (pts) with Early and Metastatic Breast Cancer (BC): RT-PCR for Cytokeratin (CK) −19 mRNA Versus the CellSearch System.
Authors: Politaki E , Agelaki S , Apostolaki S , Perraki M , Hatzidaki D , Lianidou ES , Mavroudis D , Georgoulias V
Published: June 2012

Background: Different methods are available for the detection of CTCs in pts with BC, however, the variable performance of these assays, the heterogeneity of CTCs, and the possible treatment-induced alteration of the markers evaluated, imply that no ideal method currently exists. We compared the efficiency of two methods to detect CTCs in pts with BC.

Patients and Methods: Blood was obtained from 200 pts with early and 164 with metastatic BC before the start of adjuvant or first-line chemotherapy, respectively. Different aliquots of the same sample were evaluated by RT-PCR for CK-19 mRNA and by the CellSearch System. Blood samples were available after the end of adjuvant or first-line therapy in 99 and 93 pts, respectively. CTCs in 23 and 7.5 ml of blood were enumerated by CellSearch, in adjuvant and metastatic pts, respectively. Cut-off values of ≥1 and ≥2 CTCs/23 ml and ≥2 and ≥5 CTCs/7.5 ml were used. Twenty ml of blood were obtained for mRNA extraction in all RT-PCR experiments.

Results: In early BC, 18.0% of samples were positive prior to therapy by RT-PCR and 37.0% (CTC≥1) and 16.5% (CTC≥2) by CellSearch. No significant correlation was shown for the detection of CTCs between methods. Overall (positive and negative) concordance was 62% for CTC≥1 and 73.5% for CTC≥2 (Chi-Square, p=0.161 and p=0.307). Post-chemotherapy, the positivity rate was 33.6% (CTC≥1) and 18.8% (CTC≥2) by CellSearch and 11.6% by RT-PCR (Spearman, R=-0.031, p=0.761). Overall agreement was 60.6% (CTC≥1) and 71.4% (CTC≥2) (Chi-Square, p=0.771 and p=0.708). In the metastatic setting, 37.8% of the samples were positive by RT-PCR, and 50% (CTC≥2) or 32.3% (CTC≥5), by CellSearch (Spearman, R=0.373, p=0.0001). Overall agreement was 70.0% for CTC≥5 (Chi-Square, p=0.0001). Post-chemotherapy, 21% were positive by RT-PCR and 13.7% and 8.4% by CellSearch for CTC≥2 and ≥5, respectively (Spearman, R=0.194, p=0.063). Overall concordance for CTC≥5 was 79.6% (Chi-Square, p=0.017). Agreement was also observed for CTC≥2, pre-and post-chemotherapy (Chi-Square, p=0.0001 and p=0.010). The concordance rates in the adjuvant and metastatic settings are shown in Table 1.

Conclusions: A high concordance between the two methods was detected in metastatic but not in early disease. Patient follow-up will determine the clinical relevance of each individual assay or their combination in the assessment of patient prognosis.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-07-21.
P4-09-14: Differential Expression of the Akt1 Isoform in Mammary Ductal Carcinoma.
Authors: Sanders AJ , Mansel RE , Jiang WG
Published: June 2012

Introduction:

AKT1 is an oncogenic protein and controls numerous cellular processes in both normal and cancerous cells. There are three isoforms, Akt1 (PKBα), Akt2 (PKBβ) and Akt3 (PKBγ). Frequently, regulation of the Akt pathway is disrupted in cancerous cells and interest within the scientific community has focused on the use of this pathway in treating cancer. However, recent studies have also implicated that Akt1 may act to limit invasive migration of breast cancer cells suggesting the potential of this isoform to have both pro- and anti- tumorigenic roles. In the current study we examined the expression profile of Akt1 in a cohort of ducal carcinoma of the breast. Methods:

Quantitative transcript analysis was used to assess the expression levels of Akt1 in a range of ductal carcinoma of the breast (n=85) and normal samples. Expression levels were used in conjunction with clinical and pathological data, as well as the clinical outcomes of the patients (median follow-up 120 months).

Results:

Expression of Akt1 levels in the mammary ductal carcinoma cohort demonstrated a lower level of transcript expression in the lower grade and stage cancers. Levels of Akt1 were found to be significantly lower in grade 1 ductal breast carcinomas compared to the combined higher grade 1 and 2 carcinomas (ductal carcinoma grade 1 median = 3.7 vs ductal carcinoma grade 2,3 median = 33.6, p = 0.01). In addition to this, median transcript levels of Akt1 were found to increase with increasing TNM stage of the ductal carcinoma (Median values of Akt1 in ductal carcinoma TNM 1 = 5.9, TNM 2 = 63.6, TNM 3 = 533 and TNM 4 = 13680). This enhanced expression of Akt1 in higher TNM stages, compared to TNM stage 1, was found to be significant in most cases (TNM1 vs TNM2, p = 0.03; TNM1 vs TNM3, p = 0.15; TNM1 vs TNM4, p = 0.03).

Conclusion:

Variable expression of Akt1 was seen throughout the mammary ductal carcinoma cohort and correlated with a number of prognostic factors. Akt1 transcript levels were found to be significantly reduced in the low grade 1 and TNM1 stages when compared to the higher grades and TNM stages. Our current data suggests that low levels of Akt1 may be associated with better clinical patient prognosis in mammary ductal carcinoma.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-09-14.
P4-09-18: Australian Decision Impact Study: The Impact of Oncotype DX Recurrence Score (RS) on Adjuvant Treatment Decisions in Hormone Receptor Positive (HR+), Node Negative (N0) and Node Positive (N+) Early Stage Breast Cancer (ESBC) in the Multidisciplinary Clinic (MDC).
Authors: de Boer RH , Baker C , Speakman D , Mann B
Published: June 2012

Introduction: There has been increasing interest in more accurately defining the need for chemotherapy in patients with HR+ ESBC. Oncotype DX® has been shown to provide prognostic and predictive information beyond traditional histopathological factors. Prior clinical impact studies have focused on decisions made by individual practitioners. In Australia, the MDC has become the standard forum for determining treatment decisions. In this study, we have assessed how the RS influences adjuvant systemic treatment decisions in the MDC at 4 separate hospitals. Methods: Patients with unifocal HR+, HER2−ve ESBC, N0 and N+ (1-3 positive nodes, including micrometastases and isolated tumour cells) and no contraindication to adjuvant chemotherapy (CTx) or endocrine therapy (H) were eligible. A total of 150 patients will be entered. Systemic adjuvant treatment recommendations were made and recorded in the MDC, both before and after testing with Oncotype DX. The primary endpoint was the overall change in the treatment recommendation. Other endpoints included confidence in recommendations, assessment of impact of the RS, and treatment the patient actually received. Results: Currently, RS results on 84 patients are available. Patient age range is 33–82 years. There are 56 (67%) N0 pts and 28 (33%) N+. Overall, 37 (44%) women had low, 36 (43%) intermediate and 11 (13%) high RS. The RS changed the initial treatment recommendations in 20 (24%) patients. Of the 34 patients initially recommended CTx+H, there was a shift to H alone for 12 (35%), whilst of the 50 patients initially recommended H alone, 8 (16%) shifted to CTx+H. A shift in treatment recommendations was reported for 8/37 (22%) pts with low, 8/36 (22%) with intermediate and 4/11 (36%) with high RS. 13/56 (23%) of N0 pts had a change in recommendation, whilst 7/28 (25%) N+ pts changed (table 1). 5 patients have decided against the final MDC recommendations: 3 chose CTx+H after recommendation of H (RS of 13,19 and 21 respectively), and 2 declined a CTx+H recommendation (RS of 19 and 25). Conclusion: Early results of our Australian study suggest an impact of Oncotype DX RS on adjuvant treatment decision making in the MDC setting. Updated results on the full 150 patients will be presented at the meeting.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-09-18.
P4-09-19: PCNA+ Tumor Associated Macrophages Are Associated with M1 and M2 Gene Expression, and Confer Poor Prognosis in the Absence of Anti-Tumor Immune Environment.
Authors: Mukhtar R , Wolf D , Tandon V , Zhu J , Lenburg M , van't Veer L , Campbell MJ
Published: June 2012

Background

Tumor associated macrophages (TAMs) promote breast tumor progression through the production of angiogenic factors, stromal breakdown factors, and the suppression of adaptive immunity. TAMs are recruited from the circulation to the tumor site, and can undergo a spectrum of phenotypic changes, with two contrasting activation states described in the literature: the M1 anti-tumoral and M2 pro-tumoral phenotypes. We previously identified a population of PCNA+ TAMs associated with high grade, hormone receptor (HR) negative tumors with poor outcomes. We hypothesized that high PCNA+ TAMs would be associated with expression of M2 related genes.

Methods

We used immunohistochemistry to measure PCNA+ TAM levels (double positive for PCNA and CD68) in 135 invasive breast cancer cases from the I-SPY 1 Trial, a prospective neoadjuvant trial with serial core biopsies and gene array data. We developed gene-sets representing M1 related, M2 related, and anti-tumoral immune response (represented by cytotoxic T cells and MHC Class II) genes based on literature review. We compared PCNA+ TAM levels, expression of these gene-sets, and outcomes.

Results

Higher than mean PCNA+ TAM counts were associated with increasing grade (p < 0.001), HR negativity (p < 0.001), and decreased recurrence free survival (RFS, p = 0.05). Among subjects who had a pathologic complete response (pCR), there was no difference in RFS between those with high versus low PCNA+ TAMs. Among subjects without pCR, those with high PCNA+ TAMs had significantly worse RFS than those with low PCNA+ TAMs (p = 0.0028). In the 95 subjects with both PCNA+ TAM results and gene expression arrays available, high PCNA+ TAM levels were associated with more M1 than M2 related genes. The gene-set representing anti-tumoral immune environment was not by itself associated with RFS. However, those subjects with both high PCNA+ TAMs and the absence of anti-tumoral immune response gene expression had significantly worse RFS than those with high PCNA+ TAMs but the presence of anti-tumoral immune related genes (p = 0.01).

Conclusions

High PCNA+ TAMs had different effects on outcomes depending on tumoral immune environment. Instead of being purely M2 macrophages, PCNA+ TAMs likely represent a heterogeneous mixture of TAMs with different polarization states. Additional markers are needed to distinguish anti-tumoral from pro-tumoral PCNA+ TAMs.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-09-19.
P4-09-24: Correlation of Aurora Family Member Expression with Clinical Breast Cancer Prognosis.
Authors: Annakesavan A , Sanders AJ , Harding KG , Jiang WG
Published: June 2012

Introduction:

Aurora-A, Aurora-B and Aurora-C are a family of protein kinases which have been identified as key regulators of the mitotic cell division process. Members of this group have been linked with pro-tumorigenic effects and have been reported as being up-regulated in certain cancers. Our current study examines the expression profile of the Aurora-A, Aurora-B and Aurora-C members in a breast cancer cohort in relation to a number of predictive factors.

Materials and Methods:

The expression of Aurora-A, Aurora-B and Aurora-C was examined in a breast cancer cohort using quantitative real time PCR (normal samples n = 34, tumour samples n = 109) and immunhistochemistry (IHC). Expression of the aurora members was related to clinical parameters such as staging and Nottingham prognostic index (NPI).

Results:

Examination of Aurora-A levels in the breast cancer cohort revealed an association of Aurora-A expression with NPI grouping. High levels of Aurora-A levels were seen in NPI-1 and NPI-2 stages (NPI-1 median = 66.0, NPI-2 median = 64.3), with expression reducing significantly at the higher NPI-3 group (NPI-3 median = 20.3; p = 0.029 vs NPI-2 and p = 0.009 vs NPI-1 respectively). Additionally, Aurora-A levels were found to be significantly lower in grade 3 cancers compared to grade 1 (median grade 1 = 86.5 vs median grade 3 levels = 52.6, p = 0.04). In contrast to this, examination of Aurora-C levels in the cohort identified a relatively low level of Aurora-C in NPI-1 and NPI-2 group (median values NPI-1 = 88, NPI-2 = 34.4) compared to a substantial increase in Aurora-C expression in NPI-3 (median value = 1602, p = 0.05 vs NPI-1 and p = 0.06 vs NPI-2). Aurora-B expression levels did not exhibit any significant differences within the breast cancer cohort.

Conclusions:

Differential expression of Aurora-A, Aurora-B and Aurora-C were observed within the breast cancer cohort. Higher Aurora-A expression was associated with low NPI groups and grades and potentially with better patient prognosis whereas lower levels were seen in the poorer prognostic NPI-3 group and higher grades. An opposite trend was seen in Aurora-C where lower levels were associated with the NPI-1 and NPI-2 groups and significantly higher levels were seen in the NPI-3 group. Aurora-B levels appeared to have no predictive function. This data suggests that together Aurora-A and Aurora-C levels, within a patient may be useful for predicting patient outcome.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-09-24.
P4-09-29: A Nomogram To Predict Prognosis in Node-Negative Breast Carcinoma.
Authors: Mazouni C , Spyratos F , Romain S , Fina F , Bonnier P , Ouafik L , Martin P-M
Published: June 2012

Background: Currently, the usefulness of conventional markers for breast carcinoma is limited to assess individual outcome. A statistical model was developed to improve the prognostic accuracy using multiple conventional and emerging prognostic biomarkers.

Methods: A total of 305 node-negative breast carcinomas who underwent surgery (+/− radiotherapy) but no adjuvant treatment was selected. Putative prognosis factors including age, tumor size, ER, PR, SBR grading, urokinase plasminogen activator (uPA), plasminogen activator inhibitor type 1 (PAI-1) and thymidine kinase (TK) were evaluated. The developed model was internally validated using Harrell's concordance index and calibrated. An external validation of the new model is warranted.

Results: Age (p<0.001), PR (p= 0.02), and PAI-1(p=<0.001) were included in the Cox regression model predicting overall survival at 5-years. Internal validation revealed a concordance index of 0.711 to 0.694 before and after calibration.

Conclusion: A nomogram can be used to predict probability survival curves for individual breast cancer patients and the effect of treatment options can be evaluated using these models.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-09-29.
P4-09-33: The Prognostic Significance of Metaplastic Carcinoma of the Breast – A Comprehensive Comparison Study with Common Breast Cancers.
Authors: Lai H-W , Kuo S-J , Chen D-R , Chen S-T
Published: June 2012

Purpose : Metaplastic carcinoma of the breast (MCB) is a rare histological subtype of breast cancer with incidence less than 1%. Due to its rarity, the clinical characteristics and prognostic significance of MCB compared with other common breast cancer, such as infiltrating ductal carcinoma (IDC) and infiltrating lobular carcinoma (ILC) are not clear and controversial between different reports. We performed a multi-institutional cases collective comparison study to evaluate the clinical characteristics and prognostic status with IDC and ILC.

Materials and methods : Cases of MCB were enrolled from 4 medical centers in Taiwan. Forty-five MCB patients combined with 1777 IDC and 53 ILC patients from cancer registry database of Changhua Christian Hospital (CCH) comprised the current study. The patients’ demographic data, tumor characteristics and prognosis of MCB were analyzed and compared with IDC, and ILC. To further clarify the prognostic difference between MCB and IDC, a case control analysis was performed to minimize the effect of tumor size, lymph node status and other clinicopathologic factors. The disease free survival (DFS) and overall survival (OS) between groups were compared.

Results : Compared with IDC, MCB was associated with older age, larger tumor size, lesser lymph node positive rate, higher distant metastasis, higher tumor grade, lower ER positive tumor, and higher triple negative breast cancer subtype (TNBC). Compared with IDC, MCB was associated with worse OS (p=0.031), but no difference in DFS (p=0.071). While MCB was not statistically different from ILC in both DFS and OS (p=0.132 and 0.289, respectively). To eliminate the influence of other prognostic factors, 135 cases controlled IDC with comparable clinicopathologic parameters were enrolled to compare with those 45 MCB patients.

Compared with cases controlled IDC group, the MCB patients remained showed poorer OS (p=0.040), but not different in DFS (p=0.439).

Conclusion : MCB was associated with poorer OS compared with IDC, and this poorer prognosis was related to tumor behavior rather than clinicopathologic factors.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-09-33.
P4-09-34: Luminal Infiltrating Lobular Carcinoma of the Breast: Clinical and Prognostic Features.
Authors: Valdiviezo NI , Vega E , Castañeda C , Flores C , Mendiola CD , Manso LM , Dorta M , De Velasco GA , Cortés-Funés H , Ghanem I , Ciruelos EM
Published: June 2012

Background

Infiltrating lobular carcinoma (ILC) and infiltrating ductal carcinoma (IDC) are the two most common histologic types of invasive breast cancer. Previous studies have demonstrated distinctive clinical and biologic characteristics, however some features of ILC are yet uncertain like the relationship with the particular subtypes according to the gene expression profile.

The purpose of our study is to analyze how the patients with ILC differ from IDC regarding of percentage of luminal subtype, the tumor factors, treatment and patterns of recurrence.

Patients and Methods

We assessed 1190 breast cancer patients treated at the Hospital Universitario 12 de Octubre between 1995 and 2006 categorized as having ILC or IDC pure types.

Results: One thousand one hundred ninety tumors were included, being ILC 223 patients (18, 7%), IDC 967 patients (81, 3%) and 200 patients (10%) were luminal HER2+. The current analysis is limited to the 990 patients classified as luminal HER2−.

Median age was 59, 2 years for ILC and 58, 8 for IDC, with no significant differences among the groups. Compared to IDC, ILC showed a larger tumor size (≥T3: 15, 2% vs. 4, 8%, ***p<0.01), lesser nodal involvement (35% vs 42, 7% p 0.048) and lower Ki67 expression (62, 6% vs. 52, 2% p=0.01). The proportion of luminal A subtype was greater in ILC (56, 3% vs. 46, 7% p=0.01). Mastectomy was more frequently performed for ILC (61, 2 % vs. 47, 7%, p=0.01). Regarding systemic therapy, 127(62,4%) ILC patients and 450 (57, 2%) IDC patients received adjuvant chemotherapy (QT). That was anthracycline-based in most cases (47, 2% for ILC and 46,4% for IDC). Adjuvant hormonal treatment was prescribed equally among both groups (196, 96% ILC and 739, 94% IDC), being the most commonly used drug ***tamoxifen.

With a median follow-up of 94 months, 45 ILC patients (22%) and 159(20, 2%) IDC relapsed. No differences were observed in DFS (p=0, 81) or OS (p=0, 47).

The most frequent metastatic site was bone (33% for ILC and 40, 8% for IDC), followed by local relapses (17, 7% for ILC vs. 17% for IDC) and liver (15, 5% for ILC vs. 15, 7% for IDC). ILC was associated with an increased incidence of peritoneal/mesenteric metastases (8 pts 17, 7% vs. 1pts 0,6% ***p<0.01) and a lower incidence of lung relapses (8, 8% vs. 19, 4 % pts p<0.05).

Conclusion: Luminal HER2− Infiltrating lobular carcinoma carries distinct clinicopathological features. In our study, ILC had similar prognosis as IDC.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-09-34.
P4-10-06: Evaluation of BRCAPro Risk Assessment Model in Patients with Ductal Carcinoma In Situ.
Authors: Muse KI , Elsayegh N , Gutierrez-Barrera AM , Kuerer H , Valero V , Litton JK , Hortobagyi GN , Arun BK
Published: June 2012

Background: The BRCAPRo model is used to predict a patient's likelihood to possess a BRCA1 or BRCA2 gene mutation based upon personal and family history. Ductal carcinoma in situ (DCIS) is considered a non-invasive condition which can progress to an invasive breast cancer if left untreated. Currently, DCIS is not specifically accounted for in the BRCAPro model, thereby causing DCIS to be weighted in the same manner as an invasive breast cancer diagnosis. Historically, a diagnosis of DCIS has been entered as having developed into an invasive breast cancer ten years later. However, there are no standard guidelines of how DCIS should be entered. We sought to determine if there were any differences in how DCIS was treated in the BRCAPro model to predict the more effective method in calculating the BRCAPro. Methods: Women with pure DCIS, who were referred for genetic counseling and underwent genetic testing, were included in the study. The likelihood of carrying a BRCA mutation was calculated using the BRCAPRO model (Version 5.1). Patient characteristics which were entered into the BRCAPro model include: tumor markers (estrogen receptor-ER and progesterone receptor-PR), history of oophorectomy prior to diagnosis, family history of 1st and 2nd degree relatives with breast and ovarian cancer, race and Ashkenazi Jewish ancestry. Each patient's BRCAPro risk estimate was calculated and compared by entering DCIS at the presenting age of diagnosis and by adding 10 years to the age of diagnosis. Descriptive statistics and a student's t-test were used to compare BRCAPro estimates between the two groups. Results: Ninety-five patients with pure DCIS underwent genetic counseling and testing. The average age of DCIS diagnosis was 45 years (range 26–65). Of the 95 DCIS patients included in the study 21% (n=20) tested positive for a BRCA gene mutation (8 BRCA1 and 12 BRCA2), 77% (n=74) test negative and 0.01% (n=1) had a variant of uncertain significance. Overall, DCIS patients who tested positive for a BRCA mutation had a higher BRCAPro (40%) than patients who tested negative (12%) when presenting age of diagnosis was assessed. When 10 years was added, the BRCAPro estimate was still higher amongst BRCA positive patients (28%) than BRCA negative patients (8%). The mean BRCAPro probability when DCIS was entered at presenting age of diagnosis was 18% (0.1−95.4) versus 12% (0.1−89.9) when calculated 10 years later. Conclusion: In our cohort there was no significant difference in BRCAPro probability whether DCIS was entered at the presenting age of diagnosis or 10 years later (p=0.1). However, future studies are needed to determine the most effective method to incorporate DCIS into the BRCAPro model in order to determine those individuals who may or may not be at increase risk to possess a BRCA gene mutation.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-10-06.
P4-10-10: Breast Cancer Risk Factors among Asian Versus Caucasian Women with BRCA1/2 Mutations.
Authors: de Bruin MA , Kwong A , Goldstein BA , Lipson JA , Ikeda DM , McPherson LA , Sharma B , Kardashian A , Schackmann EA , Kingham KE , Mills MA , West DW , Ford JM , Kurian AW
Published: June 2012

Background

Prior research suggests that the prevalence and penetrance of BRCA1/2 mutations may be different in Asians compared with Caucasians. Such differences in penetrance could be related to genetic, as well as hormonal, reproductive and lifestyle factors.

Methods: Chinese and Caucasian BRCA1/2 mutation carriers were recruited from genetics clinics in Hong Kong and California. We compared personal and family history of cancer, as well as hormonal, reproductive and lifestyle exposures between racial groups. Breast density was estimated using BIRADS breast composition categories on mammogram. We analyzed DNA samples for single-nucleotide polymorphisms (SNPs) that may modify BRCA-associated cancer risk, and compared the prevalence between groups. A multivariable analysis of potential modifiers of cancer risk is underway.

Results: Forty-two Chinese women from Hong Kong and forty-nine Caucasian women from California were enrolled. BRCA1 mutations were more common in Caucasian women (63% vs. 43%, P=0.05), while BRCA2 mutations were more common among Chinese women (57% vs. 33%, P=0.02). More Chinese women had a personal history of breast cancer (91% vs. 53%) and ovarian cancer (12% vs. 4%). In contrast, significantly more Caucasian women had a family history of breast and ovarian cancer. On average, Caucasian women had higher parity, breastfed more, had less dense breasts on mammogram, and more often underwent prophylactic oophorectomy, all of which are protective against breast cancer in the general population. However, Caucasian women also consumed more alcohol and had higher average BMI. Risk associated alleles in RAD51, MAP3K1 and TOX3/TNRC9 were more common in Chinese women; risk associated alleles in FGFR2 and RASSF1 were similar in frequency between groups. Results of the multivariable analysis are pending and will be presented.

Discussion: We found notable differences between Chinese and Caucasian women in the proportion of BRCA1 versus BRCA2 mutations, and in the distribution of hormonal, reproductive and lifestyle factors, and SNPs associated with breast cancer risk. Such variations in risk-modifying factors may contribute to differences in BRCA mutation penetrance. Enhanced understanding of racial differences in BRCA1/2 mutation epidemiology may inform targeted cancer screening and prevention strategies.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-10-10.
P4-11-02: Insulin-Like Growth Factor-1 (IGF-1), Insulin-Like Growth Factor Binding Protein-3 (IGFBP-3) and Lobule Type among Women in the Nurses' Health Study II (NHS II).
Authors: Collins LC , Rice MS , Shen D , Connolly JL , Schnitt SJ , Tamimi RM
Published: June 2012

In a previous analysis of women enrolled in NHSII, we found that among women diagnosed with benign breast disease (BBD), those with predominant type 1/no type 3 lobules were at lower risk of subsequent breast cancer compared to women with other lobule types. Additionally, studies in animal models suggest that higher levels of IGF-1, a polypeptide hormone involved in the proliferation/differentiation of normal mammary epithelium, may inhibit involution of breast lobules. However, the interaction between IGF-1 levels and lobule types in determining breast cancer risk has not been previously evaluated. Therefore, we examined the association between IGF-1 levels and lobule type among women with BBD.

Methods: We conducted a cross-sectional study among 484 women in NHSII with biopsy-confirmed BBD between 1993–2001 who had blood samples available for determining levels of IGF-1 and IGFBP-3. A pathologist, blinded to exposure status, classified lobule type on biopsy slides according to the number of acini per lobule (type 1 < 12; type 2∼50; type 3∼80 acini). Lobule type was classified into (1) predominant type 1/no type 3 lobules or (2) other lobule types. Multivariate logistic models were used to assess the associations between plasma IGF-1, IGFBP-3, and IGF-1/IGFBP-3 levels with lobule type. Models were adjusted for age, IGF-1 batch and additional potential confounders in secondary analyses.

Results: In univariate analyses, older age at biopsy, higher body mass index, postmenopausal status, nulliparity, and lower IGF-1 levels were associated with predominant type1/no type 3 lobules (p<0.05). In multivariate logistic models adjusting for age, higher IGF-1 levels were associated with a decreased risk of predominant type 1/no type 3 lobules (OR quartile 4 vs. quartile 1 = 0.35, 95%CI: 0.15−0.81). Greater IGF-1/IGFBP-3 ratio was also associated with a decreased risk of predominant type1/no type 3 lobules (OR quartile 4 vs. quartile 1 = 0.24, 95%CI: 0.10−0.57).

These associations persisted, though were slightly attenuated, in models adjusting for additional potential confounders.

Conclusion: Higher IGF-1 levels and greater IGF-1/IGFBP-3 ratios are associated with a decreased risk of predominant type 1 lobules/no type 3 lobules among women with BBD in the NHSII. Whether this association contributes to the mechanism by which IGF-1 confers an elevated breast cancer risk requires further investigation.

Acknowledgements: This work was supported by T32 CA09001-35 CA124865, R01 CA050385, and the Breast Cancer Research Foundation

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-11-02.
P4-14-02: Risk Factors for Early Lymphoedema Do Not Predict Late Development of Lymphoedema.
Authors: Kilbreath SL , Beith J , Refshauge KM , Lee M-J , Ward LC
Published: June 2012

Background: Previously, we investigated whether an upper limb 8 week exercise program commencing 1 month after surgery was associated with higher incidence of LE. As part of that study, we investigated the risk factors for development of LE at 12 month follow-up, which was equivalent to 15 m post surgery. As LE fluctuates during the first year, our secondary aim was to identify risk factors for LE at the conclusion of the intervention and at 6 m follow-up.

Methods: Women (n=160) were recruited who had undergone breast and axillary surgery within the previous month for unilateral early stage breast cancer. Potential predictors were measured at approximately one month postoperatively after which women were randomised to the exercise or control group. Possible predictors of LE included age, BMI, side of surgery, treatments for breast cancer, physical measures of shoulder range of motion and strength, inter-limb bioimpedance ratio at each previous measurement occasion and the treatment group to which women were randomised. The outcome measure for LE was based on the inter-limb impedance ratio. Presence of LE was determined using established cut-offs for LE. The relationship of each variable to the outcome measures (i.e. presence of LE) was assessed using t-tests and correlations. Significant variables (P < 0.25) were further examined with a multivariate logistic regression model.

Results: At least 10% of women developed LE: following intervention (N=15), and at 6m (N=15) and 12 m (N=20) follow-up. Risk factors for LE varied across the 3 time points (Table 1). At 12 m follow-up, the only significant risk factor was surgery on the non-dominant side. In contrast, the risk factors for LE following the intervention were allocation to the control group and commencement of taxane-based chemotherapy. At 6 m follow-up, risk factors for LE were having LE following the intervention and receiving taxane-based chemotherapy.

Conclusion: Fluctuation of LE over the first year is associated with different risk factors. For example, fluid accumulation is a common side effect of taxanes and it was therefore not surprising that there was a transient increase in LE. However, overall, we speculate that the higher risk for LE after 1 year is because of lack of use of the non-dominant affected arm. It is use of the upper limb which is likely the critical issue in this first year. Exercise was preventative while it was being undertaken; we question whether women maintained the level of exercise once the intervention concluded.

Acknowledgement: Cancer Council NSW; NBCF Australia

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-14-02.
P4-15-02: Clinical and Epidemiological Correlates of Elevated Distress Thermometer Scores in Breast Cancer Patients.
Authors: Powers K , Pappas L , Buchmann L , Anderson L , Gauchay L , Rich A , Agarwal J
Published: June 2012

Objectives: Distress is prevalent in breast cancer patients and can be detrimental to quality of life, performance status, treatment adherence, and satisfaction with medical care. The National Comprehensive Cancer Network (NCCN) developed the Distress Thermometer (DT) as a quick and efficient self-assessment tool for screening distress in cancer patients. While surveys estimate that between 20–40% of patients with cancer have significant levels of distress, fewer than 10% are identified and treated. Given time and monetary constraints, it is important to refine screening criteria to identify patients with elevated risk for distress. In this study, we identify clinical and epidemiological factors that are associated with an increased likelihood of elevated DT scores (≥4 and ≥7) in breast cancer patients.

Methods: We assessed 229 consecutive female patients with the DT at their initial consultation for breast cancer at the Huntsman Cancer Hospital between September 2007 and December 2008. The DT screening tool measures a global level of distress using a visual analogue scale from 0–10 in the shape of a thermometer, with zero identified as “No Distress” and ten labeled as “Extreme Distress.” The DT screening tool also includes a checklist of common emotional, family, physical, practical, and spiritual concerns with instructions for the paitent to indicate which of those concerns contributed to the distress they experienced within the past week. We chose a score ≥4 as our cutoff for a positive screen for “distress” and a score ≥7 as our cutoff for a positive screen for “extreme distress/depression” based on previous studies. Variables included in the analyses were: age, employment status, race/ethnicity, personal history of depression, family history of breast cancer, marital status, estrogen and progesterone receptor status, stage of cancer, time since diagnosis, and recurrence. Descriptive statistics and logistic regression models were used to determine associations between DT and patient data. Results: Emotional and physical concerns were associated with scores ≥4 and scores ≥7. Spiritual concerns were significantly associated with patients reporting scores ≥7. Patients who were non-Caucasian, unemployed, had a prior history of depression, who presented for recurrent disease, or who had been recently diagnosed had a higher likelihood of scores ≥4 and scores ≥7.

Conclusions: The likelihood of scoring ≥4 and ≥7 on the DT screening tool is highest during the first 30 days after receiving a breast cancer diagnosis. Four groups of patients should be targeted for aggressive screening: patients with a prior diagnosis of depression, patients presenting with recurrent disease, unemployed patients, and non-Caucasian patients. Interventions should address physical, emotional and spiritual concerns.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-15-02.
P4-16-01: Proteomic Analysis of Patient Plasma Identifies Parathyroid Hormone Related Protein (PTHrP12-48) as a Potential Biomarker of Breast Cancer Bone Metastasis.
Authors: Washam CL , Byrum SD , Leitzel K , Suhail AM , Lipton A , Suva LJ
Published: June 2012

Background: Bone metastasis is a devastating and often incurable phase of breast cancer progression that significantly compromises patient morbidity and mortality. Despite the well-characterized issues associated with tumor progression, there is currently no reliable method to detect or predict which patients have or are at increased risk for developing bone metastasis. In this study a validated plasma-based proteomic profile for the diagnosis of breast cancer bone metastasis was developed and a highly discriminatory protein component of the profile identified as a novel fragment of parathyroid hormone related protein (PTHrP).

Materials and Methods: Plasma samples were collected from a total of 110 breast cancer patients. The training set consisted of 38 breast cancer patients with clinical evidence of bone metastasis and 38 with no evidence of a bone metastasis from time of diagnosis to clinical outcome. The independent validation cohort consisted of 34 breast cancer patients with an unknown bone metastasis classification. The training set was analyzed using surface enhanced laser desorption time-of-flight mass spectrometry (SELDI-TOF MS) and 13 discriminatory protein peaks that contributed to a homogeneous partitioning of the training set (n = 76) in multiple statistical, bioinformatics, and machine learning modeling scenarios were identified and used to construct a RandomForest classifier for the detection of breast cancer bone metastasis (sensitivity: 100%, specificity: 100%).

Results: The diagnostic profile identified was then independently confirmed in a blinded fashion using the validation cohort (sensitivity: 97%; specificity: 82%). Importantly, the most discriminatory protein peak (m/z 4260.92 Da) in the plasma of bone metastasis patients was subsequently identified using specific immunodepletion, tryptic peptide mapping and peptide sequencing as a unique proteolytic PTHrP fragment, PTHrP(12-48). Ongoing studies are determining the biogenesis and biological activity of this unique PTHrP peptide. Discussion: Our discovery of PTHrP(12-48) as a novel plasma biomarker of breast cancer bone metastasis validates our plasma proteomic approach and provides the first direct evidence for the existence of a circulating PTHrP biomarker in breast cancer patient plasma that is associated with bone metastasis.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P4-16-01.