bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P6-01-05: Long-Term Diagnostic Accuracy of Fine-Needle Aspiration Cytology Performed in a One Stop Clinic for the Diagnosis of Suspect Nodular Breast Lesions
Authors: Borget I , Vielh P , Canale S , Balleyguier C , Azoulay S , Garbay J-R , Bourgier C , Mathieu M-C , Delaloge S.
Published: April 2011

Background: Fine-needle aspiration cytology (FNAC) has been used extensively in the diagnosis of breast lesions, but false-negative rates are a matter of concern. Immediate onsite evaluation of breast lesions, combining FNAC results with clinical and radiological data allows improving its diagnostic accuracy. The objective of this study was to evaluate the diagnostic accuracy of a large series of FNAC of breast lesions during the first 3-year period of a dedicated one-stop clinic, by comparing the FNAC results with the corresponding definitive histological examination outcome or the results of the radiological follow-up at 18 months.

Method: Data of all consecutive patients (pts) whose lesions were prospectively characterized by ultrasonography (US) and diagnosed by FNAC in the one stop clinic of the Institute Gustave Roussy between May 2004 and March 2007. Histological verification by core-needle biopsy or surgery was systematically performed for lesions classified malignant or suspicious by cytopathology, and for benign lesions, when no perfect concordance between clinico-radiologic features and FNAC results (such as benign FNAC but BI-RAD 5) was found. For non-operated patients, follow-up consisted on biannual US and/or mammography; the results at 18 months were considered. Pts characteristics, radiological findings, cytopathological and histological results were extracted from the hospital computerized prospectively registered medical records. Complete sensitivity (number of carcinomas that were malignant or suspicious by cytopathology) and full specificity (benign lesions at cytology of the total number of benign lesions) were calculated by comparing the cytopathological results to the histological or the long-term follow-up status.

Results: A total of 1822 nodular breast lesions (mean size: 20 mm, BI-RAD ACR 1/2/3/4/5/unknown: 10/96/471/459/777/9) in 1739 pts (mean age: 56 years) were studied. FNAC was US-guided in 1115 lesions (61 %). Lesions were classified by FNAC as malignant in 842 (46%), benign in 771 (42%), suspicious in 154 (9%), and unsatisfactory in 55 (3%) cases. Complete sensitivity (number of carcinomas that were malignant or suspicious by cytopathology) was estimated at 98.5%, whereas full specificity (benign lesions at cytology of the total number of benign lesions) was equal to 94.4%. There were 4 false-positive lesions and 32 false-negative lesions, but all these lesions except one were operated as radiological results were suspicious of malignity.

Conclusions: Breast FNAC performed in a dedicated one-stop clinic with immediate on-site diagnosis presented a low rate of unsatisfactory specimens and a very efficient tool for triaging patients candidates for histological evaluation conversely for follow-up by imaging.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P6-01-05.
Abstract P6-01-09: HER4 Is Downregulated in Lymphnode Metastases Compared to the Paired Primary Breast Carcinoma
Authors: Brügmann A , Jensen V , Garne JP , Nexo E , Sorensen B.
Published: April 2011

Introduction: The Human Epidermal Growth Factor Receptor 4 (HER4) of the EGF receptor family has been characterized in both normal and malignant human breast tissue and HER4 overexpression has been shown to predict prolonged survival compared to HER4 receptor negative disease.

In our study we investigated the HER4 expression in normal breast tissue, primary breast carcinoma and in ipsilateral metastatic axillary lymphnodes at the time of primary breast cancer surgery.

Material and methods: Paired tissue samples from normal breast tissue and primary breast carcinomas were obtained from 169 patients. Out of these a third sample was obtained from 66 patients with metastatic lymphnodes. The primary tumour specimens were sampled uniformly at random. The mRNA expression of HER4 was quantified with real time RT-PCR and expressed relative to the householdgene (HMBS) in arbitrary units (arb.u.).

Results: The mRNA expression of HER4 was significantly higher in breast carcinoma with a mean of 2.26 arb.u. [95% c.i.:1.87 to 2.65 arb.u.] than in the paired sample of normal breast tissue with a mean of 0.82 arb.u. [95% c.i.: 0.47 to 1.16 arb.u.] (p=0.0001). The mRNA expression of HER4 was also significantly higher in breast carcinoma than in the corresponding lymphnode mean 0.75 arb. u. [95% c.i.: 0.43 to 1.07 arb.u.] (p=0.015). There was no significant difference observed in mRNA expression of HER4 between the metastatic lymphnode and the corresponding normal breast tissue (P>0.05). When HER4 expression was compared between the primary tumours that had metastasised and those that had not, there was no significant difference (P>0.05). Conclusion:

The HER4 expression was high in the primary tumour as compared to normal breast tissue and the corresponding lymphnode. In view of previously published relations between a low expression of HER4 and a poor prognosis our results warrant further studies in order to evaluate whether suppression of HER4 in tumour cells could be involved in lymphogenic metastatic spread.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P6-01-09.
Abstract P6-02-01: Meta-Analysis of Ductal Carcinoma In Situ (DCIS) on Core Needle Biopsy — Underestimation and Predictors of Invasion
Authors: Brennan ME , Turner RM , Ciatto S , Marinovich ML , French JR , Macaskill P , Nehmat H.
Published: April 2011

Background: A diagnosis of ductal carcinoma in situ (DCIS) on core needle biopsy (CNB) may represent an underestimate (or understaging) of invasive breast cancer (IBC). This is identified on excision histology in 4%-35% of CNB diagnoses of DCIS. It is of clinical significance as it may prevent informed discussion about management options for IBC and may mean that more than one surgical procedure is required. We systematically review the literature on CNB to report pooled estimates for underestimation of IBC following CNB diagnosis of DCIS and identify preoperative predictors of invasion.

Methods: Studies were identified by searching MEDLINE and were evaluated against predetermined inclusion criteria: data were extracted independently by 2 authors. We calculated study-specific proportions for DCIS underestimates. Using meta-regression (random effects logistic models) we investigated the association between study-level preoperative variables and understaged IBC.

Results: 47 studies met eligibility criteria reporting 6,213 cases of DCIS on CNB with excision histology as the reference standard.

Table 1: Pooled underestimation rates for

There were 1,482 underestimates (DCIS on CNB and IBC on excision); pooled estimate 25.9%(95% CI 22.2, 29.9). Preoperative variables associated with underestimation included ultrasound-guidance (vs stereotaxis), 14Gautomated (vs 11GVAB), high grade CNB lesion (vs nonhigh grade), size >20mm, BIRADS 4/5 (vs BIRADS3), mammographic mass (vs calc only) and palpability.

Conclusion: Using available evidence we estimate that around 26% of CNB DCIS diagnoses represent understaged IBC. We identified preoperative variables associated with higher risk of understaging. This information can be used for preoperative discussion of management options, including possible sentinel node biopsy at the time of excision.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P6-02-01.
Abstract P6-04-03: Unique Recurrent Signature and Signaling Pathway of Asian Triple Negative Breast Cancer
Authors: Kuo WH , Chen HL , Hsiao CK , Chuang EY , Chang KJ.
Published: April 2011

Background: Triple-negative (TN) breast cancer, a lack of understanding of the driving oncogenic mechanisms and thus therapeutic targets has left this an understudied ‘Cinderella’ in breast cancer phase III trials. No publication has been addressed on recurrent signature and pathway signaling of Asia TN breast cancer. The gene expression difference between Eastern and Western breast cancer is also unknown.

Material and Methods: 185 represent breast cancer cases including 43 Luminal A, 31 Luminal B, 35 Her-2 type, 51 TN and remaining 25 unclassified were selected from our tissue bank. Using Agilent microarray chip, we explored gene expression profile of these patients. For direct western and eastern comparison, 195 Caucasian breast cancer gene expression profile using the same Agilent platform were downloaded wedsite. In TN breast cancers, excluding 3 cases with stage IV disease at diagnosis, 11 patients developed distant recurrence during follow up. For these 48 patients, recurrent model was established by two step Cox proportional hazard regression: in first step, 35 candidate recurrent genes were selected from 20173 genes with significant level less than 0.0005. Then, we randomly choose 4 genes from the 35 candidate genes to construct recurrent model to avoid overfitting. TN related signaling pathways were discovered using Ingenuity Pathway Analysis(IPA) and GSEA(Gene Set Enrichment Analysis).

Results: Using non-supervised analysis, we found the eastern and western breast cancers have their own TN cancer clustering phenomenon, obviously in a different cluster.

This represented the eastern and western breast cancer have fundamental differences. The model including PKRAG3, MORC2, GRB7, FAM43A showed the best recurrence prediction power in 52360 combination models(R Square 0.71). This model was cross-validated by Vector machine(SVM) with leave one out setting. The overall accuracy was 87.5%. The sensitivity and specificity were 72.7% and 91.9%. This model was failed to predict recurrence in Luminal or Her-2 Molecular type. With IPA, we found 9 TN related pathway. The top two were cAMP-mediated and Ephrin Receptor signaling. 10 pathways were found from GSEA. Nicotinate, Olfactory transduction and Axon guidance were on the top of the list. Further functional study is on going.

Discussion: This is the first study to compare the gene expression between eastern and western breast cancer on the same gene expression platform. The gene expression profile is fundamentally different between eastern and western TN breast cancers. This phenomenon infers different biological behavior and drug sensitivity may exist among different ethnic groups. We established recurrent prediction model unique for Taiwanese TN breast cancer by four genes with high accuracy. We can develop commercial recurrent kit after further clinical validation.

Figure available in online version.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P6-04-03.
Abstract P6-04-07: Pathway Activation Probability Score Analysis Reveals Patterns of Pathway Deregulation in Inflammatory Breast Cancer
Authors: Van Laere S , Van der Auwera I , van Dam P , Vermeulen P , Dirix L.
Published: April 2011

Background: Inflammatory breast cancer (IBC) is the most aggressive manifestation of primary epithelial breast cancer and differs from non-IBC both from a clinical and a biological perspective. The high metastatic potential of IBC is highlighted by the low 5-year survival rate of 50%. In the past, we demonstrated that IBC is characterized by a specific molecular profile, partly mediated by increased NFkB-activation. In the current study we aim to identify additional molecular alterations responsible for the altered gene expression profile.

Methods: We profiled 46 IBC samples and 56 non-stage matched nIBC samples using the Affymetrix HGU133plus2 platform. Using a set of recently published gene signatures (Gatza et al, PNAS, 2010) we calculated the activation probability scores for 18 distinct pathways (ER, PR, HER2, P53, P63, EGFR, RAS, SRC, TNFa, INFa, INFg, TGFb, STAT3, E2F1, PI3K, AKT, CTNNB and MYC). Unsupervised cluster analysis was performed on the IBC samples only to identify IBC-specific transcriptional heterogeneity. The silhouette-algorithm was used to test the robustness of clustering. Supervised analysis was performed to compare IBC to nIBC. All P-values were FDR-corrected.

Results: Using only IBC samples, unsupervised hierarchical cluster analysis identified four robust sample clusters (Average silhouette width = 0.345; P<0.0001). Interestingly, the primary bifurcation of the resulting dendrogram is determined by INFa-, INFg-and TNFa-signalling. In addition to ER-, PR-, HER2- and P53-signalling, the transcriptional heterogeneity in IBC is also determined by AKT-, RAS-, STAT3- and TGFb-signalling. When comparing IBC to nIBC we identified 12 pathways that are differentially regulated (P<0.05). Out of these 12 pathways, 4 (CTNNB, PI3K, E2F1 and P63) did not reveal any IBC-specific heterogeneity and all of these were hyperactivated in IBC. Interestingly, the PI3K-, E2F1- and P63-pathways form a tightly correlated signaling nodule (Spearman correlation coefficients > 0.65, P<0.001). Conclusions: In this study we demonstrated that IBC is transcriptionally heterogeneous and this heterogeneity is mainly determined INFa-, INFg-and TNFa-signalling. Also, we identified four pathways (CTNNB, PI3K, E2F1 and P63) which are differentially regulated in IBC as compared to nIBC but which do not contribute to the establishment of transcriptional heterogeneity within IBC. Therefore, we reason that these pathways can be added to the molecular repertoire of IBC.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P6-04-07.
Abstract P6-05-01: Triple Negative Breast Cancer in Korea — Distinct Biology with Different Impact of Prognostic Factors on Survival
Authors: Lee JA , Bae JW , Jung Y-H , An H , Lee ES.
Published: April 2011

Background: Patients with triple negative breast cancer(TNBC) are known to have poor prognosis and derive no benefit from endocrine therapy or targeted treatments. Using a database from a multicenter registry in Korea, we present the clinical features and prognosis for TNBC with other subtypes of breast cancer and clinicopathologic variables that influence the 3-year survival of the TNBC patients.

Materials and Methods: From 1993 to 2008, patients diagnosed with breast cancer who were registered to the Korean Breast Cancer Society Registry were analyzed retrospectively. A cohort of 26,767 patients were divided in four groups: luminal A (ER+ and/or PR+, HER2-), luminal B (ER+ and/or PR+, HER2+), HER2 overexpression (ER-, PR-, HER2+), and triple negative (ER-, PR-, HER2-). Clinicopathologic features such as age, tumor size, nodal status, p53, ki-67 expression and survival were evaluated. Results: The luminal A (14437 patients, 53.9%) subtype was the largest in our study sample, as compared with luminal B (3517 patients, 13.1%), ER-/HER2+ (3227 patients, 12%), and TNBC (5586 patients, 21%) subtypes. Compared with luminal A subtype, TNBC correlated with younger age and more aggressive characteristics, such as larger size, more lymph node metastasis, and higher proliferation rate. Moreover, TNBC correlated with poor overall survival and breast cancer-specific survival. The hazard rate showed a peak at 24 months for the TNBC subtype, but after 60 months, the risk was similar to that of the luminal A subtype. Higher T, N stage and histologic grade, and lymphatic and vascular invasion showed poor prognosis in TNBC patients, but on multivariate analysis only histologic grade and ki-67 status were related to poor prognosis. Young age was related to poor prognosis in the luminal A subtype, however, age was not related to prognosis in the TNBC subtype. Of the 5586 TNBC patients, 282 patients (7.11%) expired within 3 years of diagnosis. T and N stage, and grade was significantly associated with prognosis on multivariate analysis.

Discussion: TNBC subtype is characterized by a younger age with poorer outcome. However, younger age is not related to prognosis, and mortality risk decreases to that of the luminal A subtype, which is known to have the best prognosis after a few years. The underlying biology of the TNBC subtype is important, and further studies to discover novel biomarkers to predict prognosis and target treatments for the TNBC subtype are necessary.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P6-05-01.
Abstract P6-05-07: Molecular Classification of Breast Cancer: Clinical Correlation and Outcome in a Venezuelan Population
Authors: Mackarem G , Duran B , Vilchez L , Sierra M , Bracho E , Colina J.
Published: April 2011

Background: Gene expression analysis has defined several subtypes of breast cancer that differ in prognosis and response to therapy. Distribution of clinical, pathological features and outcome was performed among Venezuelan women diagnosed with breast cancer. The objective of this study was to evaluate clinical features and outcome. Material and Methods Of 1004 women treated at the BCTC between January 1, 2000 and April 30, 2010, 624 patients had assessable data for estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER 2/neu). Molecular subtypes were defined as Luminal A (ER+, PR+, HER 2-), Luminal B (ER+, PR+, HER 2+), Basal Like (ER-, PR-, HER 2-) and HER 2/neu (ER+, PR+, HER 2+). Clinical and Pathologic features were available for all 624 patients. Follow-up (FU) for distant recurrence (D.R.) and deaths were available for 514 patients (survival and distant disease free survival). Data was compared for women younger than 50 years with those 50 and older

Results Median FU was 33 months. 268 patients (43%) were 49 years or younger and 356 were 50 years (57%) or older.

Discussion: Distribution of molecular subtypes in Venezuelan women is different than other ethnicities. This study also revealed that both age subgroups in the HER2/neu subtype had the worst outcome followed by patients under 50 in the Basal subtype.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P6-05-07.
Abstract P6-06-02: UGT1A1*28 and Breast Cancer: Increased Risk for Gilbert's Syndrome Patients
Authors: Francisco ARF , Gomes DBD , Souza MMT , Martini RR , Maciel GAR , Ono-Nita SK , Góes JCS , Baracat EC.
Published: April 2011

Introduction: Estrogens are essential for the initiation and development of breast cancer (BC). Among the enzymes involved in conjugation and inactivation of estrogens and its metabolites, there is the UDP-glucuronosyltransferase 1A1 (UGT1A1). Polymorphisms in the promoter region of the UGT1A1 gene are common in patients with Gilbert's syndrome (GS), reducing the enzyme activity by about 80%. Although considered benign, patients with Gilbert's Syndrome (GS) have lower activity of UGT1A1, which may lead to a higher risk for estrogen-dependent cancers, such as breast cancer (BC).

Methods: Case control-study and meta-analysis. Setting: academic tertiary hospital in Brazil. Cases: Women with recently operated BC, no previous neoplasm or immunossuppression. Controls: Healthy women, older than 40 years old, mammography BIRADS 1 or 2 in the last 12 months, no previous neoplasm or immunossupression. Interventions: data collection; sequencing of the UGT1A1 gene. Literature search: PubMed, EMBASE, SciElo, hand search. Languages: English, German, Spanish, Portuguese. Studies: case-control studies, BC patients and healthy subjects, genetic diagnosis of GS.

Results: From August 2008 to November 2009, 89 cases and 56 controls were enrolled. Patients were significantly older than controls (median 52 (46-64) vs 47 (43-50) y/o p=0.013), with comparable body mass index (median 27.55 vs 25.95 p=0.46), age at menarche (median 13 vs 13 p=0.68), age at menopause (median 48 vs 45, p=0.24), age at first full-term pregnancy (median 22 vs 23 p=o.46) and proportion of individuals with first degree-relatives with BC (22% vs 19%, p=0.68). The prevalence of GS was 19.1% in cases and 12.5% in controls (adjusted OR 1.65 CI95% 0.63-4.28 p=0.29), but the study was underpowered to detect differences smaller than 21%. The meta-analysis included 10 studies with 5815 BC patients and 8255 controls. There was a weak association between GS and BC (OR 1.13 CI95% 1.01-1.26 I2=2%), which was stronger for Caucasians (OR 1.14 CI95% 1.01-1,29 I2 = 20%), estrogen-receptor positive BC (OR 1.18 CI 95% 1.02-1.35 I2=0%) and individuals with a family history of BC (OR 1.4 CI95% 1.01-1.95 I2=0%).

Conclusions: In our study, the polymorphism UGT1A1 *28/*28, present in most individuals with Gilbert's syndrome, was an important risk factor for developing breast cancer. Gilbert's syndrome, usually considered benign, may be a risk factor for breast cancer. There have been no studies evaluating the incidence of BC in patients with GS. Prospective studies with patients diagnosed with SG are needed to confirm this hypothesis and assess other patients subgroups.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P6-06-02.
Abstract P6-08-06: C-Reactive Protein Expression and Its Association with Disease Specific Survival and Disease Recurrence in ER Negative Breast Cancer Patients
Authors: Elsberger B , Tannahill C , Orange C , Mallon EA , Wilson C , Doughty JC , Edwards J.
Published: April 2011

Background: The systemic inflammatory response, as evidenced by elevated serum C-reactive protein (CRP) has been reported to be predictive of poor survival, independent of tumour stage, in a variety of solid tumours (1). Previous studies suggest that systemic elevated serum CRP did not predict relapse-free, cancer-specific (DSS) nor overall survival (2). Aim of this study was to examine the relationship between expression of CRP within ER negative breast cancers and patients’ clinical outcome. Methods: Tissue microarray technology was employed to analyse tissue taken from 171 ER negative breast cancer patients at time of surgery. Immunohistochemistry was performed using an antibody to CRP. Expression was assessed using the weighted histoscore method by two independent scorers.

Results: Cohort's median age was 56 years and tumour size was 20mm, 22% were pathologically graded G2 and 74% G3 and 46% were lymph node positive. 60% underwent a mastectomy, 56% received radiotherapy and 67% chemotherapy. 33% had recurrence after a median time of 5.6 years. 47/171 patient were deceased at time of analysis. There was a definitive trend noticed between high nuclear and cytoplasmic CRP expression and DSS (p=0.078 and p=0.173, figure 1a and 1b) and recurrence (p=0.133, p=0.253, figure 2a and 2b). There was a significant association seen with high CRP expression in the surrounding tumoural stroma and DSS (p=0.007) as well as recurrence (p=0.016). Conclusion: Due to small patient numbers only trends indicating that high CRP expression within and surrounding the tumour play an important role in ER negative patients regards poorer survival and earlier recurrence. We are currently expanding our cohort further. References:

(1) Roxburgh CSD (2009). Future Oncol. 2010 Jan;6(1):149-63

(2) Al Murri AM (2007) British Journal of Cancer, 96, 891-895 Figure available in online version.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P6-08-06.
Abstract P6-08-08: Nucleolin as an Intracellular Transporter of Hsp72
Authors: Asea A , Pradeep A , Kaur P , Bempong P , Lillard S.
Published: April 2011

Background: Our previous studies demonstrated that thermal stress induces the release of Hsp72 from cells by a mechanism independent of the classical protein transport pathway. However, the exact mechanism by which Hsp72, a leaderless protein, gains access to the extracellular milieu remains unknown.

Materials and Methods: In this study we use flow cytometry to measure the expression of surface bound Hsp72 in THP1 human monocytic cells. Sucrose gradient ultracentrifugation was also used to isolate and recover intracellular fractions. Western blot analysis was used to probe for intracellular fractions expressing Hsp72, Hsp90, Grp96, CAP2, TLR2, 4, or 7. Baculovirus expression vector system (BEVS) was used to generate highly purified endotoxin free Hsp72. Plasma membrane cross-linking assay was used to identify the interacting proteins of Hsp72 on cell plasma membrane. Laser scanning confocal microscopy was used to demonstrate the plasma membrane localization and internalization of Hsp72 with other interacting proteins. In-gel digestion and LC-mass spectrometry was used to identify unique proteins colocalizing with Hsp72. Results: The data presented in this study suggest that Hsp72 trafficking within the cell and its release is assisted in part by, nucleolin. We further demonstrate that within 60 minutes after first exposure of cells to heat shock treatment, plasma membrane bound Hsp72 is internalized and redistributed into cytosolic compartments. Inhibition of active cell transport by pre-treatment of cells with Cytochalasin B completely abrogated Hsp72 redistribution from the plasma membrane into the cytosol. Cross-linking of plasma membrane bound proteins with Hsp72 followed by Western blot analysis, in-gel digestion and LC-MS/MS analysis revealed seven interacting partners with Hsp72, including Hsp90, nucleolin, gp96, CAP2, TLR2, 4 and 7. Transfection of cells with nucleolin-siRNA completely inhibited baseline and heat shock-induced Hsp72 release. Discussion: Our study for the first time demonstrates that the plasma membrane acts as a reservoir for Hsp72 and confirms that nucleolin plays an important role in Hsp72 trafficking and release.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P6-08-08.
Abstract P6-08-09: Leptin and Estrogen Receptor Expression in Patients Suffered with Breast Cancer and Fibroadenoma
Authors: Peredereeva EV , Lushnikova AA , Lyubchenko LN , Parokonnaya AA , Polevaya EB.
Published: April 2011

Introduction: Leptin is a multifunctional hormone produced by adipocytes. It plays important role in angiogenesis. Induction of cell proliferation, survival and anchorage-independent growth. These leptin activities are mediated through leptin receptor (ObR) that binds leptin molecule and stimulates Jak/STAT 3, ERK ½, cyclin D1 expression and other signal pathways. A recent data show that targeting leptin signaling may reduce mammary carcinogenesis and breast cancer (BC) progression. However, the link between obesity and leptin expression in serum/breast tumor as well as its role in modulation of estrogen receptors (EsR) and HER2/neu expression is not clear. Material and methods: We studied leptin, ObR, EsR, HER2/neu expression in patients with sporadic, familial and pregnancy-associated BC by RT PCR using BC fresh tissue and primers for genes encoding leptin, ObR, EsR-α, β. Leptin level in the patient sera was estimated also by ELISA (Leptin Sandwich DRG, DRG Diagnostics, Germany) followed by comassie staining. The data on immunohistochemical staining of HER2/neu, EsR and PrR were also obtained. In control group were patients with benign fibroadenoma (BFa) and healthy women of comparable age. Results: RT PCR results and immunohistochemistry method are mainly concordant : only 5% (5/29) of data were different. In triplonegative tumors (n=40) leptin overexpression was significantly higher than in other tumor types. Blood sera leptin level was correlates positively with ObR expression. Leptin expression in tumor tissue also correlates with HER2/neu over expression in all BC groups except triplonegative ones: Esr (-), PrR(-), HER2/neu(-). Serum leptin levels in BFa patients was higher (100-300 ng/ml) than in healthy women and the mean leptin level in BC patient of comparable age and body weight. Moreover, leptin serum level was positively correlates with ObR expression in tumors on I-III BC stages and with high body weight index (obesity). Conclusions: The data obtained indicate that leptin/ObR maight involved in BC progression. It indicates that ObR suppression isthe possible way for target BC therapy, especially of triplonegative tumors which do not express HER2/neu, so hormone therapy is not effective for these neoplasia.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P6-08-09.
Abstract P6-08-10: Disease Specific Survival of Breast Cancer Patients Is Not Associated with Expression of Inactive or Even Partially Activated Src Kinase
Authors: Elsberger B , Tan BA , Brown SF , Mallon EA , Edwards J.
Published: April 2011

Background: c-Src is implicated as a regulator of cell proliferation and survival (1). Recent work has demonstrated that c-Src and fully activated Y419Src expression in breast cancer specimens was associated with poor clinical outcome of breast cancer patients (2). c-Src is activated by a number of pathways. Known as the classical activation pathway, dephosphorylation of Y530 is needed to initiate a configuration change of the protein allowing full activation by autophosphorylation of tyrosine site 419. Phosphorylation of the tyrosine residual 530 on the c-terminal tail by Csk tyrosine kinase acts as a negative regulatory protein binding site, keeping Src kinase in a closed confirmation (3).

The aim of this translational study was to assess associations between inactivate and partially activated Src expression to clinic-pathological features of the cohort and if different stages of c-Src activation equally influences breast cancer patients’ disease specific survival. Methods: Tissue microarray technology was used to analyse tissue taken from 165 breast cancer patients taken at time of surgical resection. Immunohistochemistry was performed using anti-Clone 28 and Clone 28 antibody. To investigate those different stages of Src kinase activation, Clone 28 antibody was employed to recognise the semi-active form of Src. To evaluate protein expression of inactive Src kinase, anti-Src family negative regulatory [pY] site (anti-Clone 28= AC28) antibody was utilised for detecting phosphorylated tyrosine site 530. Expression was assessed using the weighted histoscore method by two independent scorers. Results: Using those antibodies, 50% of AC28 and Clone28 expression was observed in the nucleus and 44% AC28 and 46% Clone28 expression in the cytoplasm. Only 3.6% of AC28 was detected in the membrane, compared to Clone28 with 53% in the membrane and 55 % perinuclear. Nuclear expression of AC28 correlated negatively with ER status (chi square P<0.001), whereas cytoplasmic AC28 and Clone28 correlated negatively with membrane c-Src expression (chi square p=0.008, chi square P<0.001). On univariate and multivariate analysis there was no significant association noticed between AC28 and Clone28 expression and disease specific survival at any cellular location.

Conclusions: ER negative breast cancers patients were more likely to express inactive Src in the nucleus. Breast cancer patients with higher cytoplasmic expression of inactive or partially activated Src were more likely not to express c-Src at the membrane (site of activation). Inactive and partially activated Src are not associated with patients’ survival. References:

1) Frame MC. Biochim Biophys Acta 2002; 1602: 114-130.

2) Elsberger B, et. al: Am J Path (2009);175(4):1389-97.

3) Roskoski R Jr. Biochem Biophys Res Commun. 2005 May 27;331(1):1- 14.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P6-08-10.
Abstract P6-09-01: Breast Cancer Incidence and Hormonal Infertility Treatments: A Systematic Review of Population Studies
Authors: Gennari A , Costa M , Puntoni M , De Censi A , D'Amico M , Bruzzi P.
Published: April 2011

Background: With the increasing practice of hormonal infertility treatments in infertile women, both for ovulation induction in anovulatory women and for ovarian hyperstimulation in assisted reproductive technologies, concerns have been raised about the long-term effects of such practice on the subsequent risk of breast cancer. Sterility itself is an independent risk factor, though its effect is not yet clear. In the past years, a number of population-based studies has investigated the possible association between breast cancer incidence and infertility treatments, with inconsistent results. With these premises, we performed a pooled analysis of the association between infertility treatments by any kind of hormonal manipulation and breast cancer incidence, based on published data. Methods: Population cohort studies, evaluating the association between BC incidence and hormonal infertility treatments were identified by MEDLINE search. Standardised Incidence Ratios (SIRs) were pooled across the studies by inverse variance weighting. Subgroup analyses were performed according to the following covariates: lenght of follow up (< 10 yrs vs ≥10 yrs), type of hormonal therapy (clomiphene vs gonadotropins) and type of control group (population based on internal control, ie infertile women). All statistical tests were two-sided. Results: Thirteen eligible cohort studies, including 364.342 women were identified and retrieved. Data on breast cancer incidence and type of fertility treatments were available for all studies. Overall hormonal infertility treatments were associated with a 15% increase in the incidence of breast cancer as compared with untreated women (SIR 1.15, CI 1.09-1.21; p < 0.0001). Breast cancer risk was significantly higher in clomiphene treated women (SIR 1.21; 95%CI 1.10-1.32) than in gonadotropins users (SIR 1.04; 95%CI 0.91-1.20, p for interaction < 0.05). The increase in breast cancer incidence seems to be dependent on the length of follow up (SIR 1.05; 95%CI 0.96-1.14 for <10 yrs versus 1.23; 95%CI 1.16-1.31 for ≥10yrs, p for interaction = 0.03). In the subset of cohort studies using population based estimates as controls, hormonal infertility treatment was associated with an increase in breast cancer risk (SIR 1.22; 95% CI 1.15-1.30); in studies with internal controls, i.e. cohorts of untreated infertile women, a similar breast cancer risk was observed (SIR 1.03; 95%CI 0.93-1.13; p for interaction = 0.09). Conclusions: Overall hormonal infertility treatments are associated with a significant increase in breast cancer incidence. This increase is mainly observed with clomiphene administration, whereas gonadotropins seem not to affect breast cancer risk. Finally, subset analyses according to type of controls suggest that the observed increased risk may be due to the infertility condition itself.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P6-09-01.
Abstract P6-09-08: Patient Preference for Intra-Operative (TARGIT) or External Beam Radiotherapy Following Breast Conservation
Authors: Alvarado M , Connolly J , Oboite M , Moore D , Park C , Esserman L.
Published: April 2011

Background: Recently published data for the TARGIT Trial (a randomized trial comparing Intraoperative Radiotherapy [IORT] for equivalence to External Beam Radiotherapy [EBRT] following lumpectomy for invasive breast cancer for selected low risk patients) shows equivalence for the IORT and EBRT arms at 4 years follow-up[1]. The TARGIT steering committee originally used a 2.5% non-inferiority margin for statistical considerations at trial initiation. We hypothesized that patients would indeed choose IORT even with the possible 2.5% inferior local recurrence rate and can now compare recently published data to patient preference. Methods: We used a validated tradeoff technique to quantify any additional risk patients would accept to receive either treatment. Subjects who were candidates for breast conserving radiation were presented with a slideshow comparing EBRT with IORT, and then asked their preference given hypothetical recurrence risks. Two initial Tradeoff Slides were used to assess whether the initial risk comparison had an effect on patients’ switch points. The first stated both forms of radiotherapy lead to a 10% 10 year risk of local recurrence, while the second stated EBRT leads to a 10% risk versus an IORT risk of 20%. Subsequent tradeoff slides incrementally increased or decreased the rate of recurrence associated with IORT until the subject preference changed.

Results: Data from 81 patients were analyzed. The median additional accepted risk to have IORT was 2% (-9% to 39%). There were two outliers who accepted 34% and 39% additional risk with IORT. Only 7 patients chose to accept additional risk for EBRT; 22 accepted IORT at no additional risk, and the remaining 52 chose IORT with some additional risk. Patients who were grade 2 or 3 (versus 1) were more likely to accept a higher additional risk for IORT. No other factors studied correlated with a higher additional tolerated risk for IORT.

Conclusions: Some patients chose to consider IORT even if experimental and with a higher risk for 10 year local recurrence compared to EBRT. The recetly published results from the TARGIT Trial at 4 years reported a 1.2% (95%CI 0.53-2.71) local recurrence rate for TARGIT and a 0.95% (95%CI 0.39-2.3) local recurrence for EBRT, clearly within the projected margin of 2.5% inferiority. Thus, it may be appropriate to consider offering TARGIT as an informed option on registry trial to selected low-risk patients.

1. Vaidya, J et al. Targeted intraoperative radiotherapy versus whole breast radiotherapy for breast cancer (TARGIT-A trial), Lancet available online June 4, 2010

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P6-09-08.
Abstract P6-10-05: Bilateral Breast Cancer: A Comparative Study with Unilateral Breast Cancer
Authors: Lee Y-m , Ahn S-H
Published: April 2011

Background

The clinical significance of bilateral breast cancer is unclear and its influence on prognosis is controversial, whether contralateral breast cancer occurs synchronous or metachronous. Our purpose is to assess the impact of bilateral breast cancer on prognosis and compared it to unilateral breast cancer. Methods

We included 9,327 patients who diagnosed breast cancer and treated at Asan Medical Center from 1990 to 2007. Bilateral breast cancer was classified as synchronous in patients with a second breast cancer diagnosed within 6 months after the first breast cancer and as metachronous if the second breast cancer occurred more than 6 months after the first. To compare clinical features, we used more advanced-stage tumor in synchronous cases and the first tumor in metachronous cases. The follow-up cut-off date of this study was December 2009 and median value of follow-up was 59 months. This study was performed by medical records review retrospectively. Results

117 (1.3%) had synchronous breast cancer and 114 (1.2%) had metachrnous breast cancer. Except menopausal status, there were no significant differences for clinical features between the patients with unilateral breast cancer and synchronous bilateral breast cancer. Patients with bilateral metachronous breast cancer were significantly younger at the time of first diagnosis and had more often a highly increased familial risk. And women who developed metachronous cancer underwent more often mastectomy for their first tumor.

Patients with synchronous (74.2%) or metachronous bilateral breast cancer (77.7%) had the same risk of death from breast cancer as patients with unilateral breast cancer. (82.7%) (10 years overall survival, p=0.703) Also, disease-free survival was no difference between these three groups. But when we analyzed disease-free survival according to recurrence pattern, patients with metachronous breast cancer had a higher risk of loco-regional recurrence, significantly, compared with other groups. Conclusion

Patients with metachronous breast cancer developed more loco-regional recurrence than patients with unilateral disease or synchronous disease, but this recurrence didn't affect overall survival. Bilaterality of breast cancer is not associated with impaired overall survival.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P6-10-05.
Abstract P6-10-07: Risk-Benefit Framework To Evaluate Gene-Expression Profiling in Early Stage Breast Cancer: A Decision Model Developed in Collaboration with Stakeholders
Authors: Roth JA , Veenstra DL
Published: April 2011

Objectives

Recent comparative effectiveness research recommendations have emphasized that stakeholder involvement in research prioritization and design is critical for ultimate acceptance of results. With this recommendation in mind, an interactive risk-benefit model was developed to evaluate gene-expression profiling (GEP) to guide use of adjuvant chemotherapy in early stage breast cancer compared to clinical guidelines. Methods

A stakeholder collaborative feedback process was utilized to develop a decision model to estimate the comparative benefits and harms of GEP relative to NCCN guidelines, including disease recurrences, adverse events, and quality-adjusted life years (QALYs). The model allows users to specify the GEP, node status, patient cohort age, adjuvant chemotherapy relative risk reduction, and patient chemotherapy preferences in each risk stratum. A Markov process was utilized to estimate long-term clinical outcomes. Based on stakeholder concerns about QALY-based outcomes, alternative outcomes such as disease recurrences and adjuvant chemotherapy serious adverse events were presented. The prognostic and predictive properties of the GEPs were derived from published retrospective analyses of RCTs. Adjuvant chemotherapy utility decrements were derived from trial-based adverse event rates. The base case patient was a pre-menopausal woman of age 44. Results

Preliminary analyses indicate that a 2-risk strata GEP, 3-risk strata GEP, and clinical guidelines would identify 50%, 46% and 92% of women to receive adjuvant chemotherapy, respectively. Based on this level of uptake, relative to a scenario where no patients receive adjuvant chemotherapy, these prognostic approaches would prevent 29%, 36% and 34% of distant recurrences, respectively. This scenario results in 17.4 QALYs, 18.1 QALYs, and 17.7 QALYs, respectively. These findings indicate that depending on the specific GEP examined, testing may lead to a net harm or net benefit relative to clinical guidelines, due to variation in risk of disease recurrence, despite avoidance of chemotherapy and side effects in 42% and 46% of women, respectively. Conclusions

GEP to inform adjuvant chemotherapy treatment decisions in early stage breast cancer could lead to an increase or decrease in QALYs relative to the use of clinical guidelines, depending on the specific GEP examined. Stakeholder feedback has indicated that acceptance of model results will likely depend on its degree of transparency, ease of use and interpretation, and ability to present relevant outcome measures. Ongoing efforts are focused on additional collaboration with stakeholders to align model structure and output with current needs.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P6-10-07.
Abstract P6-11-03: Beyond the Second Line Chemotherapy in Metastatic Breast Cancer: When Stop the Treatment between Science and Conscience
Authors: Bernardo G , Palumbo R , Poggi G , Bernardo A , Teragni C , Frascaroli M , Amatu A , Montagna B , Tagliaferri B , Sottotetti F , Albanese D , Strada MR.
Published: April 2011

Background: Women with metastatic breast cancer (MBC) are increasingly offered third line chemotherapy, but the usefulness of treatment beyond the second line remains a therapeutic challenge. The aim of this study was to determine which benefit could be brought by successive lines of chemotherapy (CT) in patients with MBC and to identify factors affecting outcome and survival.

Patients and methods: This retrospective analysis included 980 women treated with CT for MBC at our Institution over a seven year period (September 1998-September 2006). With overall survival (OS) data updated at December 1, 2008, the median follow-up was 125 months (range 48-192), OS and time to treatment failure (TTF) were calculated according to the Kaplan-Meyer method for each CT line. Cox proportional hazards model was used to identify factors that could influence TTF and OS.

Results: Median OS evaluated from day 1 of each CT line decreased with the line number from 34.8 months (980 patients, first line, range 4-208) to 22.6 months (838 patients, second line), 14.6 months (684 patients, third line), 12.4 months (302 patients, fourth line), 9.4 months (88 patients, fifth line), 8.2 months (45 patients, seven or more lines). Median TTF ranged from 9.2 months to 7.8 and 6.4 months for the first, second and third line, respectively, with no significant decrease observed beyond the third line (median 5.2 months, range 4.8-6.2). In univariate analysis factors positively linked to a longer duration of TTF for each CT line were positive hormonal receptor status, absence of liver metastasis, adjuvant CT exposure, response to CT for the metastatic disease; in the multivariate analysis the duration of TTF for each CT line was the only one factor with significant impact on survival benefit for subsequent treatments (P<0.001).

Conclusions: Our results stress that CT beyond the first line may be beneficial in a significant subset of women treated for MBC. For patients responding to a given CT regimen and asking for further treatment after disease relapse, it appears useful to deliver subsequent active lines to get prolonged TTF and OS benefit. Patients who do not respond to the first two lines of CT should be considered for clinical trials or supportive care.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P6-11-03.
Abstract P6-11-09: Pilot Study of Irinotecan and S-1 for Advanced and Metastatic Breast Cancer
Authors: Tanaka M , Furusawa H , Kamada Y , Sagara Y , Anan K , Miyara K , Kai Y , Wakamatsu S , Uga T , Tamura K , Mitsuyama S
Published: April 2011

Patients with metastatic breast cancer (MBC) are often treated with multiple regimens, and viable options may soon be exhausted. Irinotecan (CPT-11), a topoisomerase 1 inhibitor, is rarely used in this setting, however. Approved for advanced breast cancer in Japan, S-1 is a pyrimidine derivative comprising the combination of tegafur, gimeracil and oteracil. Irinotecan is combined with S-1 (IRIS) to obtain an additive effect, even in the presence of high thymidylate synthase in breast cancer cells. This combination has also been successfully used for gastro-intestinal malignancies in Japan. Method: This study was divided into two steps: step 1, in which the safety profile of the regimen was determined; and step 2, in which tumor response was evaluated if the safety profile was considered acceptable. Irinotecan was given intravenously at a dose of 80 mg/m2 on days 1 and 8 and S-1 orally at 80-120 mg/day depending on body surface area for 2 weeks. The cycle was repeated every 3 weeks.

Results: A total of 21 patients were registered in the study. Median age was 50 years (range, 26 -69 years). Performance status was 0 and 1 in 20 and 1 patients, respectively. Six patients were enrolled in step 1. Grade (G) 3 fatigue and vomiting were observed in 1 patients, G 3-4 neutropenia in 4 and G 3 anemia in 1. The toxicity profile in step 1 was determined acceptable to move on to step 2. Step 2 included 15 patients treated with IRIS. Grade 3-4 neutropenia occurred in 7 out of these 15 patients, fatigue in 1, diarrhea in 3 and anorexia in 1. All patients had previously received anthracyclines and taxanes. Eighteen patients, including 3 patients in step 1, were evaluable for response. A partial response was obtained in 5 patients (27.8%), including in 1 patient who had a history of 4 previous treatments. Progression-free survival in these patients was 137+, 147, 168+, 304+ and 364+ days; no progressive disease was observed in 4 of them. In addition, stable disease was obtained in 5 patients (27.8%) in whom 2 patients experienced SD for longer than 6 months. Conclusion: These results indicate that the IRIS regimen has an acceptable safety profile and modest activity against breast cancer in patients who have been heavily treated with chemotherapy for MBC. We believe the patients responded to the IRIS regimen because irinotecan has a different mechanism of action as compared to other drugs commonly used for breast cancer and complete cross-resistance may not exist between S-1 and capecitabine. This regimen is commonly used for gastric or colorectal cancer in Japan and is considered to be highly tolerable. Therefore, we conclude that this regimen has potential as an option to treat advanced and metastatic breast cancer, although attention must be paid to diarrhea, the dose-limiting toxicity of irinotecan and S-1.

Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P6-11-09.