bluuebunny/crossref_metadata_2025 huggingface dataset:
P2-03-03: An Insulin-Like Growth Factor I (IGF-I)-Induced Gene, Solute Carrier Family 7 Member 11 (SLC7A11)/xCT, Mediates IGF-I-Induced Biological Behaviors in Breast Cancer Cells.
Authors: Yang Y , Becker MA , Yee D
Published: June 2012

Our laboratory studied the gene expression profiles of a series of T47D variant cell lines with differential insulin receptor substrate (IRS) adaptor protein expression to develop predictive IGF-I pathway biomarkers. We identified an IGF-I-induced gene, SLC7A11 (or xCT), which is specifically regulated through IRS-1. xCT encodes the cystine/glutamate transporter subunit of the heterodimeric amino acid transport system xc- which is a major plasma membrane transporter for the cellular uptake of cystine in exchange for intracellular glutamate. xCT is involved in the regulations of proliferation, metastasis, and drug resistance in various cancers. However, to date, the linkage between xCT and the IGF-I signaling pathway has not been described. To study the role for xCT in mediating IGF-I-induced biology in breast cancer cell lines, we examined xCT mRNA expression upon IGF-I stimulation in two breast cancer cell lines; the MCF-7 (IRS-1 activated) and MDA-MB-231 (IRS-2 activated) cells. Significant increased xCT expression was observed only in MCF-7 cells after IGF-I treatment. Immunoblots showed that xCT protein expression was elevated after IGF-I treatment and induced glutamate/cystine exchange in MCF-7 cells. shRNA was used to downregulate xCT in MCF-7 and MDA-MB-231 cells. In MCF-7, IGF-I-stimulated cell monolayer growth was suppressed by xCT shRNA or by the xCT inhibitor sulfasalazine (SASP). In MDA-MB-231 cells, xCT downregulation did not affect IGF-mediated Boyden chamber migration. Thus, IGF-I induction of cellular xCT levels is associated with cell growth in the IRS-1 activated MCF-7 cells, while MDA-MB-231 cells were not affected by downregulation of this gene. Therefore, our data imply that xCT may mediate IGF-I induced biological functions in breast cancer cell lines through an IRS-1 dependent pathway.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-03-03.
P2-03-04: Novel Pathways Underlying the Initiation and Transition of DCIS to IDC of HER2−Overexpressing Breast Cancer Model.
Authors: Pradeep CR , Koestler W , Lauriola M , Nair H , Rao R , Mills GB , Yarden Y
Published: June 2012

Background: The HER2/Neu -oncogene is amplified in 20–25% percent of human primary breast cancers and this alteration is associated with disease behaviour. The HER2/neu oncogene encodes a receptor-like tyrosine kinase, whose overexpression in breast cancer predicts poor prognosis and resistance to conventional therapies. Whereas signalling pathways emanating from HER2 have been characterized, much less is known about the transcriptionally regulated genes controlled by HER2 that contribute to the initiation of Ductal Carcinoma Insitu (DCIS) and their transition to an invasive ductal carcinoma (IDC).

Materials and Methods: Normal and HER2 overexpressing mammary epithelial cells (MCF10A) were grown in extracellular matrix to form 3D structures, When grown in extracellular matrix as 3-dimensional spheroids, control cells developed a hollow lumen, but HER2−overexpressing cells populated the lumen by evading apoptosis. On the next step, uopon the growth factror stimulation HER2−overexpressing cells grown in extracellular matrix, which protruded invasive outgrowths. Highly variable genes were selected from the described phenotypes using RNA isolated from the 3D structures and hybridized to an Affymetrix HuGene 1.0 ST oligonucleotide array. Results: Using microarrays we analysed transcriptional events responsible for the morphological changes and found several novel sets of genes such as integral proteins, transcription factors, matrix proteases, Notch genes and chemokines were highly altered in the HER2 overexpressing group which were not appreciated before. Using gene annotation we defined molecular-pathways responsible for the phenotypical changes. More specifically, our study proposes a two hit model describes the pathways involved in the initiation of the Ductal Carcinoma Insitu and their transition to Invasive Cancer underlying the HER2 transcriptional network.

Discussion: According to the proposed model, expansion of ductal hyperplasia is limited by intraluminal apoptosis, unless they overexpress HER2, which drives proliferation and forms DCIS due to HER2 induced Notch pathways genes. Secondly, results obtained with the 3D system and their reflection in clinical outcome, we propose that neither HER2 amplification, nor the presence of GFs, is sufficient for development of IDC, but their co-occurrence can instigate metastasis by the activation of genes of integrin-adhesion signaling. The more virulent scenario combines HER2 amplification with GFs, thereby switching a robust, auto-stimulatory program.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-03-04.
P2-03-06: Endothelin-1/Endothelin A Receptor Signalling in Breast Cancer.
Authors: Tamkus D , Leece C , Gallo K , Madhukar BV , Dimitrov N
Published: June 2012

Background: Endothelin-1 (ET-1) and endothelin A receptor (ETAR) are implicated in breast cancer growth and progression. ET-1 is secreted by both tumor cells and stroma (macrophages, endothelial cells). The purpose of this study is to evaluate ET-1/ETAR role on cell proliferation and its effects on EGFR signaling pathway. Materials and Methods: Two breast cancer cell lines: MCF-7 and MDA-MB-231 were stimulated with ET-1. Proliferation of breast cancer cells was analyzed using MTT assay. Protein expression (EGFR, pEGFR, AKT, pAKT, ERK, pERK) was evaluated by Western blot. siRNA knockdown of endothelin A receptor (ETAR) was performed in MCF-7 and MDA-MB-231.

Results: ET-1 stimulated proliferation in both cell lines. Interestingly, at higher ET-1 concentrations cell proliferation was more pronounced in MCF-7 when compared with MDA-MB-231. Stimulation with ET-1 activated EGFR and downstream signaling proteins (pAKT and pERK) in both cell lines. Incubation of breast cancer cells with ET-1 for 48 hours had different effect on total EGFR level: 3 fold increase of total EGFR was seen in MCF-7, while no change was detected in MDA-MB-231. siRNA against ETAR decreased ET-1 induced cell proliferation and reduced total EGFR level in MCF-7.

Conclusions: These observations suggest that ET-1/ETAR plays an important role in survival and proliferation of breast cancer cells. However, those effects are diverse in different subtypes of breast cancer. Our results may represent an improved selective targeted treatment strategy, especially for estrogen receptor positive breast cancer.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-03-06.
P2-04-02: Identification of a Novel Glycosyltransferase-Like Gene as an Autophagic Inducer in Human Mammary Carcinoma Cells Via Down Regulation of BCL-2.
Authors: Shastri BG , Jung Y , Steiner M , Perry JK , Lobie PE , Liu D-X
Published: June 2012

Here we report the identification of a previously undescribed glycosyltransferase-like gene, GLTx, and its cellular effect on mammary carcinoma cells. GLTx codes for a novel human protein which is highly conserved in primates. Data obtained from the mRNA expression database Oncomine, demonstrate that GLTx is differentially expressed in a variety of cancers including breast cancer. Western blot analysis of a multiple human tissue blot using a rabbit polyclonal antibody against GLTx demonstrated that GLTx was highly expressed in liver, moderately in kidney, intestine and stomach and undetectable in any other tissues. To functionally characterize GLTx, we attempted to establish stable cell lines with forced expression. However, forced expression of GLTx was completely lethal and resulted in severe cell death in the mammary carcinoma cell lines, MCF-7 and BT549, as well as in any other cell lines tested. Staining of MCF-7 cells transiently transfected with a GLTx expression plasmid by either Hoechst 33258 or fluorescein isothiocyanate-conjugated annexin V and propidium iodide confirmed that the observed cell death was not caused by apoptosis. Further investigation confirmed that GLTx expressing cells exhibited cytoplasmic accumulation of autophagosomes, consistent with the induction of autophagy, as measured by the fluorescently labelled autophagosome marker LC3. In addition we have shown that GLTx downregulated BCL-2, an inhibitor of autophagy, as measured by BCL-2 gene promoter luciferase activity assay and western blot assay following transient transfection of a GLTx expressing plasmid. Thus, we have demonstrated that GLTx is a novel autophagic inducer that causes cell death by down regulating BCL-2.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-04-02.
P2-05-07: Combined Antiangiogenic and Anti-Estrogen Therapy in Breast Cancer. Molecular Mechanisms Involved.
Authors: de la Haba J , Berciano M , Cañabate M , Porras I , Valverde A , Cañas A , Rodriguez A , Aranda E
Published: June 2012

Background: Much is known about the mechanisms involved in the response to anti-hormonal treatment or those involved in the response to antiangiogenic therapy. It is also known the association between angiogenesis and hormonal status, both in physiological and pathological settings. However, the molecular and cellular mechanisms contributing to the efficacy of combined antiangiogenic-antihormonal therapy in breast cancer are still unknown. This combination is currently in clinical trials, but unfortunately there are scarce preclinical studies contributing to the rationale of combining antiangiogenic and antihormonal therapies.

Aims: To define the mechanisms involved in the response to combined antiangiogenic-antihormonal treatments in breast cancer cells. Methods: Breast cancer cell lines with different estrogen dependence (MCF-7, BT-474, MDA-MB-231) were subjected to an estrogen gradient (estradiol), and treated with fulvestrant (antiestrogen), plus bevacizumab (anti-angiogenic). Cellular proliferation and apoptosis were determined using the corresponding kits. Proliferation and survival intracellular signaling pathways, estrogen alpha and VEGF receptors activation and COX-2 expression were analyzed by western-blot using specific antibodies.

Results: In estrogen-dependent breast cancer cells (MCF-7 and BT-474) the pro-proliferative effect of estradiol decreased after bevacizumab treatment.

Furthermore, the combination of the antiangiogenic with an antiestrogen enhanced this antiproliferative effect, that was also related to the reduction in the levels of VEGF-A in the culture medium and to diminished ER alpha phosphorylation. The combined treatment also altered the phosphorylation of Akt and Erk1/2 signaling kinases. Interestingly, bevacizumab treatment augmented COX-2 activity in MCF-7.

Conclusions: Our results suggest that in estrogen dependent breast cancer cells the anti-proliferative effect of bevacizumab depends on estradiol concentration, that in turn affects VEGF production levels, using a different mechanism to apoptosis. The combination of bevacizumab with antiestrogens enhances this antitumoral effect, altering intracellular signaling pathways of proliferation and survival.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-05-07.
P2-08-13: Does Routine Use of Preoperative Magnetic Resonance Imaging (MRI) in Breast Cancer Influence the Outcome?
Authors: Große R , Kantelhardt EJ , Steer S , Spinda AK , Vetter M , Ruschke K , Thomssen C
Published: June 2012

Introduction: The role of routine preoperative breast MRI in patients diagnosed with breast cancer is still under discussion. In a milestone review, Houssami et al. (CA Cancer J Clin 2009;59;290) did not demonstrate an advantage with regard to DFS or OS. We reviewed our case series (n=793) in order to add evidence to the current discussion of whether or not preoperative MRI would influence the outcome.

Material and Methods: In our database we identified four years of diagnosis (2004-07) in which a high percentage (72%) of patients (349 of 483) with histologically confirmed invasive breast cancer received preoperative MRI. In the following years (2008/09), MRI was still done in 31% of the cases (97 of 309). All pts were treated according to national guidelines; however, patients with additional lesions in MRI were subjected to an additional MRI-guided needle biopsy or additional wire-guided excision. The median follow-up time was 31.5 months (0-81). The patients were followed clinically. In most cases local surveillance was done by mammography.

Results: In the total cohort, 20% of the patients had additional needle biopsies and/or wire-localisation due to MRI-findings. By MRI, lesions that subsequently led to an additional preoperative needle biopsy were found in 82 pts. Interestingly, only ten contralateral second cancers were found. In 357 pts, MRI was used as guidance during the operation with (n=118) or without MRI-supported wire localization (n=238). The percentage of additional MRI-needle biopsy or MRI-supported localisation did not differ between invasive ductal (n=655) and lobular cancer (n=91): 10% versus 13% needle biopsies and 15% versus 20% wire localisations. The number of surgical procedures to achieve tumor-free margins did not differ between patients with or without preoperative MRI. In this series, 69 of 100 patients who received neoadjuvant chemotherapy, also had MRI-imaging. Considering the patient cohort with follow-up available (n=737), no significant difference of DFS probability was observed between pts with and without preoperative MRI.

Discussion: This retrospective analysis did not demonstrate any advantage for routine preoperative MRI with regard to local treatment and DFS probability. The observed rate of 20% additional operative procedures corresponds to similar published data (16% additional multicentric or multifocal lesions found by MRI, Solin, Breast. 2010;19:7).

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-08-13.
P2-09-03: Breast Cancer Detection Using a Novel Functional Imaging Device Equipped with Multiparametric Computer Analysis.
Authors: Sella T , Sklair-Levy M , Maya C , Rozin M , Tanir A , Maira S , Libson E , Izhaky D
Published: June 2012

Introduction: A novel prototype system performing functional three-dimensional (3D) infrared imaging (RUTH), coupled with multi-parametric computer analysis (MIRA), was evaluated for non-invasive breast cancer detection. The technique utilizes non-ionizing radiation and automatically provides risk assessment based on parameters derived from a clinically known training set.

Materials and methods: Following IRB approval, prospective multi-center, two-armed blinded evaluation, was performed on 102 women, which included 43 patients with biopsy proven breast cancer and 59 women with a normal (BI-RADS 1) mammogram. RUTH imaging (Real Imaging Ltd, Israel) was acquired on a continuous temporal timeline for 5 minutes during which a cold-stress test was applied for one minute. Acquired data were analyzed using multiple computed algorithms (MIRA) which were combined to develop a master algorithm for risk assessment of breast cancer. Analysis was blinded to the clinical status. Results were displayed as risk assessment scale with 100 being suspicious for breast cancer and −100 inferring normal healthy breasts.

Results Thirty-nine of 43 cancers (91.7%) were identified using this novel technology. The sensitivity and specificity of RUTH coupled with MIRA for detecting breast cancer were 90.7% and 67.8% respectively. The AUC defined by the ROC curve was 0.832. Twenty-seven of the 39 (70%) detected cancers were smaller than 20mm in size.

Conclusions: This preliminary study shows adequate performance of the novel technology examined for detecting breast cancer. Performance is comparable to the reported performance of new adjunctive breast imaging technologies such as tomosynthesis and contrast-enhanced mammography, and superior to the published sensitivity of mammography. Our results warrant further evaluation of the RUTH and MIRA technologies in detecting and characterizing breast cancer.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-09-03.
P2-09-12: High Resolution Diffusion MRI Characterizes Tumor Stromal Boundaries.
Authors: McLaughlin RL , Newitt DC , Wilmes LJ , Sinha S , Wisner DJ , Hylton NM
Published: June 2012

Background: Studies of 3D cell cultures have shown that mammary epithelial cell growth and morphogenesis is regulated by extracellular matrix (ECM) stiffness, linking ECM stiffening to malignant transformation. Tumors are consistently stiffer than normal adjacent tissue, and matrix stiffening is caused by ECM cross-linking and increased deposition of collagen. Some evidence suggests that collagen orientation at tumor boundaries can promote tumor metastasis. Measuring the stiffness of the tumor boundaries and adjacent stromal tissue may give additional information 1) about tumor microenvironment and 2) to guide treatment. Diffusion-weighted imaging (DWI) MRI measures the mobility of water in tissue and may be sensitive to this phenomenon.

Material and Methods: MRI data was collected on patients with locally advance breast cancer enrolled in an IRB-approved study at UCSF and signed informed consent. In addition to a standard dynamic contrast enhanced (DCE) MRI, a high-resolution diffusion-weighted image (HR DWI) was acquired with an echo planar imaging sequence and the following parameters: TR/TE=4000/64.8 ms, b=0,600, FOV=70×140mm, matrix=28×64, and voxel size=0.55×0.55×4.0mm. Apparent diffusion coefficient (ADC) maps were created. HR DWI images were segmented into tumor and non-enhancing, surrounding stromal tissue. A proximity mapping method was used to measure ADC values at the inner edge of tumor and at increasing distances from the tumor boundary on HR DWI. The mean was calculated for the voxels in 1 mm increments, starting at 5 mm into the tumor (−5 mm) and ending at 2.5 cm away from the tumor (25 mm).

Results: The average of the changes per 1 mm shell was largest for the transition of the tumor boundary (Table 1). In Table 1, the −5 to 0 mm, 0 to 5 mm, and 5 to 25 mm columns represent inside the tumor, tumor boundary, and outside the tumor, respectively. In general, ADC values were consistently lower inside the tumor than outside. The greatest changes per 1 mm shell was seen in the transition from inside to outside tumor, although the values varied among tumor types. Each of the three cases analyzed had different patterns of ADC values.

Discussion: These preliminary studies show that water mobility measurements change at the tumor boundary, with different patterns observed among individual patients. We are further investigating the influence of density and tumor margin morphology on these ADC measurements.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-09-12.
P2-09-13: The Value of FDG PET/CT in Screening Detected Breast Cancer Patients.
Authors: Koo HR , Moon WK , Cho N , Chang JM , Kang KW , Yi A
Published: June 2012

Background. To evaluate the diagnostic value of FDG PET/CT for initial staging of screening detected breast cancer.

Methods. Between January 2008 and June 2010, a total of 77 women (mean age 54 years, range 31–77 years) with screening detected primary breast cancer (mean invasive tumor size 1.65cm, range 1–70mm) underwent whole body fluorine-18 fluorodeoxyglucose (FDG) PET/CT for initial staging and were included in this retrospective study. Two patients had bilateral breast cancer. The sensitivity of FDG PET/CT for the detection of primary tumor and the sensitivity, specificity, PPV and NPV for the detection of axillary lymph node metastases were determined. Systemic staging with whole body FDG PET/CT was also performed. For analysis of diagnostic performance of FDG PET/CT, quantitative measurement of the maximum standardized uptake value (SUVmax) criteria 1.0 was used. The final histopathology following surgery served as the gold standard.

Results. The primary tumor was FDG PET/CT positive in 65 of 79 lesions (82%). Depending on the tumor size, there was a variation in diagnostic sensitivity (63% in ≤ 1cm tumor, n=19 vs. 88% in > 1cm tumor, n=60) and the uptake of FDG was significantly higher in > 1cm tumor than in ≤ 1cm tumor (mean SUVmax 2.85 vs. 1.11, p<0.05). The uptake of FDG was significantly higher in ductal carcinomas compared to lobular carcinomas (median SUVmax 2.0, n=72 vs. 1.3, n=7, p<0.05). Of the 77 patients included in this study, 16 patients were found to have axillary node metastasis. The sensitivity, specificity, PPV and NPV of FDG PET/CT for detection of LN metastasis were 63% (10/16), 89% (54/61), 59% (10/17) and 90% (54/60), respectively. FDG PET/CT showed distant uptake in 9 patients and 8 of 9 were false positive results. 4 lesions were confirmed histopathologically as benign and 4 lesions were evaluated with radiologic methods. One of nine was true positive result. Distant involvement was skeletal and visible on the conventional bone scintigraphy. The patient staged as cT1N3M1.

Conclusion. FDG PET/CT has limited value for the initial staging of screening detected breast cancer patients. Considering high costs, radiation exposure and false positivity, FDG PET-CT is not recommended for the preoperative evaluation of screening detected breast cancer patients.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-09-13.
P2-10-04: 3D Mapping of Total Choline in Human Breast Cancer Using High-Speed MR Spectroscopic Imaging at 3T: A Feasibility Study.
Authors: Posse S , Royce M , Dayao ZR , Zhang T , Zhao C , Sillerud L , Lopez S , Casey L , Eberhardt SC , Lomo L , Lee S-J , Rosenberg R , Rajput A , Bolan P
Published: June 2012

PURPOSE: To assess the feasibility of quantitative high-speed MR spectroscopic imaging (MRSI) of total Choline (tCho) as an adjunct to dynamic-contrast enhanced (DCE) MRI to improve lesion characterization and monitor treatment response in patients undergoing neoadjuvant chemotherapy.

METHOD AND MATERIALS: Seven patients with biopsy-confirmed, infiltrating ductal carcinoma were studied using a clinical 3T MR scanner (Siemens Medical Solutions, Erlangen, Germany) equipped with 8- and 16-channel breast array (Hologic Inc., Bedford, MA). Measurements were performed using PRESS prelocalized 3D Proton-Echo-Planar-Spectroscopic-Imaging (PEPSI) using TR/TE=2000ms/135ms, matrix size up to 32×16x8, voxel size=1cc, and total acquisition time of 10 minutes (including water reference scan). Additional data were collected at TE 60 ms to enhance sensitivity for detecting tCho and J-coupled resonances. TE-averaging (8 steps, ΔTE: 2.5 ms) was employed to minimize gradient sideband artifacts. Quantification of tCho in reference to tissue water was performed using spectral fitting and relaxation correction.

RESULTS: Strongly elevated tCho with maximum concentration ranging from 0.3 to 4.1 mmol/kg was measured in five patients with single and multi-centric enhancing lesions larger than 2 cc volume (Table 1). The measured tCho concentration in Grade 3 tumors was higher than in lower grade untreated and treated tumors. Strong decreases in tCho concentration were measured in 2 patients undergoing neoadjuvant therapy in a follow-up scan. At short TE an additional resonance was detected that was elevated in enhancing lesions and tentatively assigned to Taurine. Two patients had lesions smaller than 2 cc with surgical clips in which tCho was not detectable due to line broadening. MRSI data sets were preferentially collected before contrast injection, since it increased spectral line width by up to 50%.

CONCLUSION: This study demonstrates feasibility of quantitatively mapping tCho in invasive breast carcinoma using high-speed MRSI. The long-term goals are to utilize high-speed MRSI as an early predictor of treatment failure in women undergoing neoadjuvant therapy (i.e. chemotherapy, endocrine therapy or biologic therapy) for breast cancer and to develop an improved screening protocol for high-risk patients. Grant support: 1RC1EB010617-01

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-10-04.
P2-12-09: Prediction of Residual Risk of Recurrence after 5 Years of Follow-Up by Clinicopathologic Variables and 4 IHC Markers: A TransATAC Study.
Authors: Sestak I , Cuzick J , Dowsett M , Salter J , Quinn E , Zabaglo L , Howell A , Buzdar A , Forbes J
Published: June 2012

Background

Adjuvant endocrine therapy beyond 5 years is known to be of benefit to some ER+ patients but it is unclear which patients have sufficient residual risk (RR) to merit this. We have previously shown that 4 immunohistochemical markers (ER, PgR, Ki67, HER-2), both alone and combined into the IHC4 score (Cuzick et al, JCO 2011) are significantly correlated with time to recurrence (TTR) in the overall follow up of a cohort of 1125 patients from the monotherapy arms of the Arimidex, Tamoxifen, Alone or in combination (ATAC) trial. We have now assessed the relationship of each of these parameters and common clinical variables (nodal status, grade, tumour size, age and treatment option) for predicting outcome beyond 5 years.

Material and Methods: We determined the univariate and multivariate prognostic value of clinical variables and the 4 IHC variables separately and as the IHC4 score for TTR separately in years 0–5 and 5–10 of follow up for all patients, separately for anastrozole and tamoxifen and only in the node-negative patients.

Results: Results in years 5–10 are summarized in the Table. Nodal status, tumour size and grade were at least as strong in years 5–10 as in years 0–5. Ki67 and the overall IHC4 score were the only significant IHC biomarkers related to TTR univariately in this period, but both lost significance in a multivariate model including clinical variables. There were no significant interactions with treatment. Similar results were seen for the node-negative population.

Conclusions: None of the IHC4 markers provided significant additional prognostic information in the 5–10 year period, but nodal status, tumour size and grade continued to be strong prognostic factors.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-12-09.
P2-12-10: Low TCR Diversity (Divpenia) Is a Prognosis Factor of Overall Survival in Metastatic Breast Cancer.
Authors: Manuel M , Tredan O , Bachelot T , Parmentier G , Courtier A , Rabeony T , Chabaud S , Mouret J-F , Grives A , Perez S , Clapisson G , Blay J-Y , Caux C , Pasqual N , Menetrier-Caux C
Published: June 2012

Background: We already showed that lymphopenia (<1000 lymphocytes/μl) or CD4+ T cell lymphopenia (<450/μl) detected before initiation of chemotherapy are prognostic factors for toxicity and death for metastatic breast cancer (MBC) patients. The goal of the present study was to identify the characteristics of the T cells in these lymphopenic patients. TCR diversity was investigated and tested as a prognostic factor for overall survival (OS).

Patients and methods: The ImmunTraCkeR® assay (ImmunID, Grenoble, France), which analyzes through semi quantitative multiplex-PCR the V-D-J combinatorial diversity of TCR-beta chain (TRB), was used to investigate diversity of T cell repertoire on cryopreserved blood samples from a retrospective cohort of MBC patients before chemotherapy administration (n=66). Univariate and multivariate analysis were performed. We then validated our score on a prospective cohort (n=67) using the same eligibility criteria (MBC patients before first line chemotherapy administration).

Results: Using a 33% cutoff for divpenia in our retrospective cohort (T cell diversity below 33%) (average diversity for healthy people is 70%), divpenia was associated with a median OS of 10 months vs 22 months for patients with diversity >33% (logrank p value=0.04). The NDL® score (Numeration Diversity Lymphocytes representation) that combines lymphocyte numeration with TRB diversity, demonstrated that lympho-divpenia (T cell diversity below 33% and lymphopenia below lGiga/L) was associated with a poor OS compared to patients with either lymphocyte <1000/μL & diversity >33% or lymphocyte >1000/μL & diversity <33% or both lymphocyte >1000/μL and diversity >33% (p=0.015). In multivariate analysis, including performance status (PS), hemoglobin level, polynuclear neutrophil count (PNN), age, and liver metastasis, NDL® score was identified as an independent prognostic factor for OS. In our prospective validation cohort, NDL® score was also identified as a prognostic factors for OS (p=0,007), as well as lymphopenia (<1000/μL) (p=0,0003), CD4+ lymphopenia (<450/μL) (p=0,04), menopausal status (p=0,02), hormonal receptor status (estrogen receptor p=0.02; progesterone receptor p=0.002) and lung metastasis (p=0,009). In multivariate analysis, hemoglobin level was the only independent prognostic factor in this cohort.

Conclusion: We showed that Divpenia and NDL® score are prognostic factors for OS in MBC patients. In order to confirm these results, a prospective clinical trial is ongoing on a larger cohort of MBC and lung cancer patients.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-12-10.
P2-12-17: Prospective Evaluation of the Conversion Rate of HER2, ER and PR between Primary Tumors and Corresponding Metastases. CONVERTHER/GEICAM 2009–03 Study.
Authors: Martinez de Dueñas E , Lluch A , Guerrero A , Chacon JI , Perez R , Antolin S , Blancas I , Ferrer-Lozano J , Burgues O , Lopez A , Gonzalez-Angulo AM
Published: June 2012

Introduction: A substantial number of small studies, most retrospective, have shown variable degrees of discordance in HER2, ER and PR expression between primary and metastatic breast cancer. This may have prognostic and therapeutic implications. These discordances could be triggered by tumor somatic mutations, intratumoral heterogeneity, or technical issues with receptor assayes. In this prospective study, we centrally analyzed and determined the changes in receptor status between primary tumor and corresponding metastases.

Methods: In 32 Spanish hospitals, patients where primary tumor sample was available, were prospectively included prior to the biopsy of the presumably metastatic lesion. Determinations of ER and PR status (assessed by immuhistochemistry [IHC]) and HER2 status (IHC and FISH) were centrally and blindly performed in both primary tumor and metastases. Concordance rate between local laboratories and central laboratory receptor determinations were analyzed. We also determined whether the biopsy results modified the oncologist initial treatment strategy.

Results: A total of 236 patients were included. To today, 183 paired cases were centrally evaluated. Patients in first recurrence were 65% and the mean age was 57.2 years. Biopsy samples were taken from locoregional recurrence (25%), liver metastases (28%), bone metastases (11%), lung metastases (7%) and other locations (29%). In 6.6% of the biopsies normal tissue, benign lesion or secondary malignant neoplasm were seen. The changes in HER2, ER and PR were 17%, 26% and 35% (local lab) and 4%, 12% and 26% (central lab), respectively; and were independent of treatment given or biopsy localization. Concordance rate in HER2, ER and PR between local and central determination were 83%, 92% and 78% (kappa index 0.58, 0.80 and 0.55), respectively. Changes in receptor status lead to a modification in medical management in 16% of patients.

Conclusions: A remarkable discordance in receptor status, especially in the hormonal receptors, was found between primary tumor and metastases that implies a change in medical management in 16% of patients. These discordances were reduced but remained after performing in a high-quality central laboratory, so that they could only be partly explained by technical discrepancies in receptor assessments.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-12-17.
P2-12-27: Simply Adding Together the Diameters of Tumor Foci in Patients with Multicentric or Multifocal Disease Does Not Add Any Additional Prognostic Information: An Analysis from NCIC CTG MA.12 Randomized Placebo-Controlled Trial of Tamoxifen after Adjuvant Chemotherapy in Pre-Menopausal Women with Early Breast Cancer.
Authors: Hilton JF , Dong B , Bouganim N , Chapman J-AW , Arnaout A , O'Malley F , Nielsen T , Gelmon K , Yerushalmi R , Levine M , Bramwell V , Whelan T , Pritchard KI , Shepherd L , Clemons M
Published: June 2012

Background: A common clinical conundrum in breast cancer management is whether pathologic T stage in women with multicentric or multifocal disease should be taken as the diameter of the largest focus or as the sum of all foci in the breast. Most staging systems, such as the American Joint Committee on Cancer (AJCC), simply use the largest tumor focus for staging. We examine here the impact of alternate methods of estimating tumour size including measures of total tumor size, volume and surface area.

Materials & Methods: NCIC CTG MA.12 is a randomized placebo-controlled trial of tamoxifen after adjuvant chemotherapy for pre-menopausal women with early breast cancer. Median follow up is 9.7 years. Pathologically reported patient tumor dimensions for up to 3 foci were utilized to examine the effects of tumor size on Breast-Cancer-Free-Interval (BCFI), defined as the time from randomization until recurrence (defined as first local, regional, distant, or contralateral invasive tumor or DCIS). Tumor size was estimated as 1) pathologic T stage as per AJCC criteria; 2) largest dimension of largest tumor focus (cm); 3) sum of largest dimension(s) of tumor foci (cm); 4) sum of surface area(s) of tumor foci (cm2), and 5) sum of volume of tumor foci (cm3). Step-wise forward unstratified Cox regression was used to assess the different effects of tumor size. Results: This study accrued 672 patients, 43% with T1 tumors, 51% with T2 tumors, and 6% with T3/T4 tumors; 25% were node negative and 56% had 1–3 positive lymph nodes. 75% were locally determined to have hormone receptor positive tumors. A higher number of involved lymph nodes was associated with significantly shorter BCFI (p<0.0001). None of pathologic T stage (p=0.14), largest dimension of largest tumor size (p=0.14), sum of largest dimensions of tumor foci (p=0.24), sum of surface area (p=0.38), and sum of volume of foci (p=0.51) were significantly associated with BCFI. Likewise, lymphovascular invasion (p=0.08), grade (p=0.14), nor administration of anthracycline therapy (p=0.08) were associated with BCFI.

Discussion: In the MA.12 population of pre-menopausal women randomized to either tamoxifen or placebo, the sole factor significantly associated with BCFI was nodal status. No measure of tumor size in unifocal or multicentric/multifocal tumors impacted BCFI. The findings of this mature data set suggest that simply adding together the diameters of tumors in patients with multicentric or multifocal disease did not add any additional prognostic information.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-12-27.
P2-12-30: Pre-Operative Haematological Markers and Prognosis in Early Breast Cancer.
Authors: Cordiner RL , Mansell J , Obondo C , Angerson WJ , Lannigan A , McMillan D , Wilson CR , Doughty JC
Published: June 2012

Background

Systemic inflammation may influence survival in women with breast cancer. Both the platelet lymphocyte ratio (PLR) and the neutrophil lymphocyte ratio (NLR) have been shown to be associated with survival in a number of solid tumours. The aim of this study was to assess the role of PLR and NLR in survival of women with early breast cancer.

Methods: Women diagnosed with early breast cancer at two centres between 2003 and 2006 were included. All women had measured white cells, neutrophils, lymphocytes and platelets. NLR and PLR were calculated. Cox regression survival analysis was performed with breast cancer specific survival used as the primary end point. Significant associations of NLR and PLR with pathological variables was assessed using the Chi squared test.

Results: 707 women were included with a median follow up of 3.7yrs. On univariate analysis neither NLR (HR 1.08, 95% CI 0.66−1.79) or PLR (HR 0.96, 95% CI 0.58−1.59) were significantly associated with breast cancer specific survival. An increased PLR was found to be associated with significantly fewer grade 1 tumours (P<0.05) but no other significant associations with pathology was demonstrated for either PLR or NLR.

Conclusions: This study would suggest that the NLR and PLR are not prognostic of survival in women with early breast cancer.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-12-30.
P2-13-03: Comparative Assessment of 636 Women at Risk for Hereditary Breast Cancer within 3 Public Hospitals: The Consortium of Underserved BRCA Testers.
Authors: Beattie MS , Ganschow P , Gabram-Mendola S , Wilson A , Joseph G , Lee R , Loranger K , Stanislaw C , Seelaus C , Farrell R , Trim L , DelPozo S , Luce J
Published: June 2012

Background: Underserved women at risk for Hereditary Breast and Ovarian Cancer (HBOC) are confronted with many unique challenges, such as barriers to accessing appropriate genetic testing and counseling services and decreased resources, that place them at increased cancer risk. It is unclear whether these high-risk women are appropriately referred for genetic counseling and what their genetic test results demonstrate due to minimal practice-based evidence. To study this population, and to establish an infrastructure to further explore long-term outcomes, we formed the Consortium of Underserved BRCA testers in October 2010 from: San Francisco General Hospital (SF), Stroger Hospital of Cook County (Chicago), and Grady Memorial Hospital (Atlanta).

Methods: Using common clinical and research protocols and mixed methods analysis, we examined and compared referral patterns, demographics, and BRCA test results between sites. We used chart reviews and common data collection instruments to gather and pool data. Using descriptive and comparative statistics, we examined similarities and differences between Consortium sites.

Results: SF's program began 9 years ago, Chicago's began 6 years ago, and Atlanta's began 3 years ago. Medicaid funding for BRCA testing has been available in SF since 2011, in Chicago since 2009, and is not yet available in Atlanta. P values were all <0.05 for comparing referral sources between sites. Each site uses unique referral tools and systems. For example, in SF, a family history screening questionnaire is administered during mammography; in Chicago, an extensive primary care network has been educated to provide appropriate referrals; in Atlanta, a pedigree assessment tool is employed by referring clinics.

Conclusions: Despite differences in referral patterns and Medicaid funding, underserved women at all 3 public hospital sites had similar BRCA positive rates, which are in line with positive rates from University Cancer Centers. We believe this relates to the availability of genetic counseling services and a similar testing threshold at each site. The large and ethnically diverse population of this Consortium can serve as a valuable resource for researchers and clinicians. We plan to follow this cohort prospectively to study clinical outcomes and medical decisions after BRCA testing in underserved families at risk of HBOC.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-13-03.
P2-13-05: Breast Cancer, BRCA Mutations and Attitudes Regarding Pregnancy and Preimplantation Genetic Diagnosis.
Authors: Litton JK , Etzel CJ , Jackson MA , Muse KI , Turco D , Schover LR , Theriault RL , Mattair D , Lu KH , Hortobagyi GN , Arun BK
Published: June 2012

Background: Breast cancer is associated with treatment-related infertility and has been demonstrated to be a major concern for premenopausal survivors. Detection of a BRCA deleterious mutation may also affect attitudes regarding future childbearing. Preimplantation genetic diagnosis (PGD) allows women to use in vitro fertilization (IVF) to implant only those embryos without a BRCA mutation. The ability to test the fetus for BRCA mutations is also available through amniocentesis and chorionic villus sampling (CVS). The objective of this study was to evaluate attitudes about childbearing and fertility in women being evaluated for a BRCA mutation. Methods: Women with childbearing potential who were referred to the Clinical Cancer Genetics Clinic to be evaluated for a BRCA mutation were invited to participate in this survey. The questionnaire was administered prior to genetic counseling. A follow-up was administered after the BRCA results were disclosed. The survey queried participants regarding their attitudes on fertility, pregnancy as it may relate to cancer and the potential of a BRCA mutation. Other questions detailed attitudes regarding IVF, PGD, and CVS in these instances. Descriptive statistics were used. Results: One hundred and twenty-eight women completed pre-questionnaires and to date 76 have completed post results disclosure questionnaires. The mean age was 33 (range 21–44) with 69.5% with a diagnosis of breast cancer, 39.8% received chemotherapy and 60.9% already had at least 1 biological child. A future child was desired by 45.3% although 53.1% worried that their children would have an increased risk of cancer. Regarding PGD, although only 30.9% (38/123) said that they would use PGD, 80.2% felt that the testing should be available to families with inherited cancers. Regarding fetal testing via amniocentesis or CVS, 29.7% would have the fetus tested and 7% would consider termination if a genetic mutation was identified. Additionally, 69.5% felt it was important to receive fertility counseling and treatment at the same place where they receive their cancer care. To date 8 women have been diagnosed with a BRCA1 mutation and 4 with a BRCA2 mutation. When asked similar questions after their genetic results were disclosed, 2 women who had previously stated they would not use PGD changed their mind. Conclusions: Future pregnancies are important to many breast cancer survivors. BRCA mutation carriers have the option to have children without passing on their genetic risk for cancer. Although few would use these interventions, a large majority felt it was important to have information about these choices and to have options for fertility preservation options addressed at the center where cancer care is delivered.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-13-05.
P2-13-08: A Retrospective Study on Surveillance Behaviour and Prophylaxis after Genetic Testing for Hereditary Breast and Ovarian Cancer among High-Risk Chinese Females.
Authors: Kwong A , Chu ATW
Published: June 2012

Background

Research in the West has shown that the uptake rates of surveillance and prophylaxis may be different in BRCA mutation carriers and non carriers and this may vary between different ethnicities. To date there is limited comprehensive data available for the behavioural pattern of Asian cohorts. The present on-going study is the first to investigate the behavioural impact and medical compliance of Chinese high risk females who had undergone BRCA1/2 genetic testing up to 1.5 years after genetic testing.

Methods: 88 females with personal or family breast and/or ovarian cancer history who had BRCA1/2 genetic testing performed by the Hong Kong Hereditary Breast Cancer Family Registry were surveyed by a face-to-face or telephone interview. Sociodemographic information, genetic test results, pre-and post-testing surveillance practice, chemoprevention methods used and attitudes towards clinical management were obtained. Retrieval of medical records was performed using a prospective database.

Results: 39 carriers and 49 noncarriers of a BRCA 1/2 mutations were interviewed. 82.1% of carriers and 79.6% noncarriers had breast and/or ovarian cancer prior to testing. The uptake rate of breast surveillance by eligible carriers (85.2%) and noncarriers (71.1%) remained high after knowing the test results. 12.8% and 15.4% of the eligible carriers had prophylactic contralateral mastectomy and salpingo-oophorectomy respectively, while 5.1% chose to have both procedures within 1.5 years of knowing the genetic test result. Significantly more carriers (85.7%) than noncarriers (49%) underwent ovarian surveillance (p= 0.004). 25.6% of carriers and 26.5% of noncarriers with breast cancer history chose Western chemoprevention methods, while no carriers without breast cancer history chose such methods. 34.7% of carriers (30.8% of noncarriers) used Chinese herbal medicine as complementary prevention treatment after knowing the genetic test result. Clinicians’ advice was perceived as the major reason of choosing a particular surveillance or preventative strategy.

Conclusions: As population-based breast screening is unavailable in Hong Kong, the relatively high uptake rate of breast screening among noncarriers may be the result of enhancement in health-consciousness due to prior personal or family cancer experience. Proper education by clinicians is crucial to increase uptake of high risk screening. Traditional Chinese herbal medicine methods are commonly used as a form of prevention. A longitudinal prospective study will contribute to the understanding of attitude-change towards clinical management of this Chinese population who may have different cultural beliefs.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-13-08.
P2-14-05: Racial Differences in the Use of Adjuvant Chemotherapy for Breast Cancer in a Large Urban Integrated Health System.
Authors: Simon MS , Lamerato L , Krajenta R , Booza J , Ruterbusch J , Kunz S , Schwartz K
Published: June 2012

Background: Despite improvements in breast cancer treatment, there continues to be a gap in survival between African American (AA) and White women with breast cancer, which may be due at least in part to racial differences in the patterns of care. In order to better understand breast cancer survival disparities, we evaluated racial differences in the receipt of adjuvant chemotherapy among women treated at a large integrated health care system in Southeastern Michigan. Materials and Methods: The study population included 2,234 women (33 % AA) with stage I through III breast cancer treated at the Henry Ford Health System (HFHS) from 1996 through 2005. Linked datasets from the HFHS, the Metropolitan Detroit Cancer Surveillance System (MDCSS) and the U.S. Census Bureau were used to obtain sociodemographic and clinical information. Co-morbidity was assessed by the Charlson co-morbidity index (CCI), and economic deprivation was categorized using a neighborhood deprivation index (DI). Results: AA women were significantly more likely than Whites to have larger tumors (40% vs. 31% > 2 cm), as well as more aggressive tumors (29% vs 19% estrogen and progesterone negative). AA women were also significantly more likely than Whites to have other co-morbid conditions (15 % vs. 8% with a CCI of 2+) and to reside in a more economically deprived area (45% vs. 5% reside in the most deprived area). While AA women were more likely to receive adjuvant chemotherapy (47% vs. 43%, p=0.0278), they were also more likely to have a delay in initiation of treatment (57% vs 45% delayed beyond 60 days from diagnosis, p=0.0004). After multivariable adjustment for age, DI, CCI, insurance and surgery, there were no racial differences in treatment delay (Odds Ratio 1.20, 95% confidence interval, 0.82−1.77). Discussion: In a large urban integrated health care system, racial differences in the delay in receipt of adjuvant chemotherapy for breast cancer can be explained by clinical and socioeconomic characteristics.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-14-05.