bluuebunny/crossref_metadata_2025_split huggingface dataset:
Stromal-epithelial cell cross-talk in the breast tumour microenvironment elicits changes in tumour epithelial cell gene expression.
Authors: Potter SM , Dwyer RM , Kerin MJ 
Published: December 2010

Abstract
Abstract #1053
Introduction Tumour growth & progression in breast cancer is a product of autocrine & paracrine actions of factors secreted by malignant epithelial & stromal populations in the tumour microenvironment. Stromal cells, recognised as prominent modifiers of cancer progression, are thought to play a critical role in controlling neoplastic epithelial cell function. Although chemokines, which are chemotactic cytokines, have been postulated as mediators of stromal-epithelial cell signalling, the precise nature of such intricate interactions remains incompletely understood. The aim of this study was to investigate stromal cell-mediated modification of breast cancer epithelial cells.&#x2028; Methods Two breast cancer cell lines with different hormone receptor profiles (ER+ MDA-MB-231 & ER- T47D) were cultured individually & on confluent monolayers of stromal cells isolated from primary breast tumours. Co-culture supernatants were harvested for chemokine quantitation by ELISA. Following direct co-culture breast cancer epithelial cells were retrieved from mixed populations with EpCAM specific magnetic beads & RNA extracted. Changes in gene expression in epithelial cells resulting from interaction with stromal cells were identified by RQ-PCR using primers for the chemokines Monocyte Chemotactic Protein-1 (MCP-1/CCL2), & Stromal Cell Derived Factor-1α (SDF-1α/CXCL12), as well as their respective receptors, CCR2 & CXCR4. Changes in expression of genes involved in invasion (MMP11 & HER-2) and proliferation (Ki67) were also analysed.&#x2028; Results Expression of CCR2 & CXCR4 was confirmed in epithelial cell lines. Co-cultures of epithelial & stromal cells secreted significantly higher levels of MCP-1 than the cells cultured alone, while the reverse was true of SDF-1α secretion (Table 1). RQ-PCR revealed changes in gene expression of epithelial cells retrieved following co-culture with tumour stromal cells. The highly invasive MDA-MB-231 cells showed upregulation of both CCL2 & CXCL12 genes (2-3 fold increase) following interaction with tumour stromal cells, while the non invasive T47D cells showed downregulation of these genes. Expression of genes specifically associated with invasion, Her2 & MMP11, were elevated (1-2 fold increase) following exposure to tumour stromal cells, with little or no change observed in the proliferative marker Ki67.&#x2028; &#x2028; Conclusion Tumour stromal cells induce altered secretion & expression of tumourigenic chemokines & invasion-associated genes in breast cancer epithelial cells, mediated through direct cell contact. These results further define the tumour promoting role of stromal cells & highlight these interactions as potential therapeutic targets.
Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 1053.
The impact of adipose stromal cells on migration and invasion of breast cancer cells.
Authors: Walter M , Liang S , Ghosh S , Hornsby P , Li R 
Published: December 2010

Abstract
Abstract #1056
Epidemiological studies have identified obesity as a significant risk factor for developing breast cancer among postmenopausal women. Adipose tissue is an important extragonadal source of estrogen. Although circulatory estrogen levels in postmenopausal women decline, tissue-specific estrogen levels remain virtually unchanged and are even found elevated in breast tissue from obese patients and patients with breast cancer. Therefore, excessive estrogen production due to overall adiposity is considered to be a critical contributing factor to obesity-associated breast cancer risk. However, it is not clear whether adipose tissue per se can also affect breast cancer development in a hormone-independent manner. In the current study, we used adipocyte stromal cells (ASCs) isolated from lipoaspirates from cancer-free patients and studied their possible influence on tumor cell behaviors. Using the Boyden-chamber assay, we found that ASCs had a stimulatory effect on the migration of both ER-positive (MCF-7) and ER-negative breast cancer cells (MDA-MB-231), suggesting that ASCs can influence the behaviors of breast cancer cells in an estrogen-independent manner. A similar effect of ASCs on MDA-MB-231 cells was also observed in a wound-healing assay. Furthermore, in a xenograft experiment using co-transplanted MDA-MB-231 and ASCs, we observed a significant role of ASCs in promoting the invasiveness of MDA-MB-231 cells. These data indicate that, in addition to their established role in supplying local estrogen, ASCs may facilitate tumor invasion into surrounding tissue and subsequently metastasis. The possible molecular basis for this effect of ASCs on tumor cell migration and invasion will be discussed.
Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 1056.
Fingerprinting genomic heterogeneity in primary breast carcinomas and among sentinel lymph node metastases: implications for clinical management of breast cancer patients.
Authors: Ellsworth DL , Ellsworth RE , Patney HL , Oviedo A , George A , Croft DT , Love B , Jordan RM , Deyarmin B , Becker TE , Hooke JA , Shriver CD 
Published: December 2010

Abstract
Abstract #1059
Background: Sentinel lymph node (SLN) status is a key prognostic factor for breast cancer patients. The SLN hypothesis postulates that cancer cells initially spread from the primary tumor to the first-draining (sentinel) node(s) before reaching higher echelon nodes. Although SLN biopsy is clinically useful for staging patients, little is known about the genomic heritage of cells that define the genetic makeup and clinical behavior of nodal metastases. We used allelic imbalance (AI) coupled with high-density SNP genotyping to examine the extent of genetic heterogeneity in primary tumors and relationships among metastases in sentinel and non-sentinel axillary lymph nodes in 30 patients with node positive breast cancer.&#x2028; Material and Methods: Pathologically positive nodes were identified by H&E/IHC, while sentinel nodes were localized by standard scintigraphic techniques. For each patient, nodal metastases and 6-15 areas from multiple paraffin blocks of the primary carcinoma were isolated by microdissection. AI was assessed by microsatellite typing at 26 chromosomal regions; genome-wide copy number variation was examined using Affymetrix GeneChip Human Mapping 500K arrays and the Genotyping Consol. The genomic heritage of sentinel and non-sentinel nodal metastases in relation to multiple areas of the primary tumor was assessed by hierarchical clustering and phylogenetic analyses.&#x2028; Results: Extensive genomic heterogeneity was observed in primary tumors and among nodal metastases (0-44% in primaries, 0-36% in metastases). Overall levels of variation were significantly higher (P&lt;0.01) in primary tumors (17%) than metastases (10%), but did not differ between sentinel (10.2%) and non-sentinel (10.0%) metastases. Within patients, many (but not all) metastases appeared to be descended from different areas of the primary tumor, but patterns of descent were complex. As a group, sentinel node metastases were not genomically distinct from other axillary node metastases, and there was no evidence that SLN metastases are seeded by cells that colonize the axilla early in tumorigenesis.&#x2028; Discussion: Although metastases develop in a sequential manner that maintains the known capability of SLN biopsy to accurately stage the axilla, metastases appear to develop from genomically-diverse progenitor cells descended from different areas of the primary tumor. Genomic diversity in primary tumors and metastases may be an important factor in prognosis and treatment resistance in breast cancer patients.
Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 1059.
DNA repair protein biomarkers in triple negative breast cancer.
Authors: Alexander BM , Sprott K , Wang X , D'Andrea AD , Schnitt SJ , Collins LC , Weaver DT , Garber JE 
Published: December 2010

Abstract
Abstract #1064
Introduction: Triple negative breast tumors form a distinct, aggressive, subgroup of breast cancers that exhibit a high degree of genomic instability and have many phenotypic similarities to BRCA1 deficient tumors. These observations suggest that aberrant DNA repair may be involved in triple negative tumor carcinogenesis. We profiled tumor DNA repair capacity using immunohistochemistry in order to develop prognostic biomarkers and suggest targets for novel therapies.&#x2028; Methods: We identified 143 previously treated women with triple negative breast cancers and used their archived, formalin-fixed, paraffin-embedded primary excision biopsies to create a tissue microarray (TMA). The TMA was stained using antibodies against proteins in various DNA repair pathways including XPF, FANCD2, PAR, MLH1, and MK2. Stained tissue was evaluated using machine-based image analysis and scoring that represented both the intensity and quantity of positive tumor nuclei. Biomarker scores and clinical data were assessed for correlations with outcome. Patients were randomized into training and test cohorts for the development of a multiple marker model. A set of critical threshold marker values were determined that maximally separated the training samples into low and high risk recurrence groups. Training set thresholds and marker combinations were applied towards the test set. Kaplan-Meier and Cox proportional hazards were used to test time to recurrence.&#x2028; Results: Clinical data for 115 patients with primary treatment data was available with a median follow up of 58 months. There were 37 recurrences: 18 were distant first, 12 were local first and 7 were simultaneous. Low XPF (p=0.002), pMK2 (p=0.02), MLH (p&lt;0.001) and FANCD2 (p=0.05) were associated with shorter time to recurrence. In the training cohort, the high-risk group defined by a four marker model had a relative risk of recurrence of 3.0 (p&lt;0.00001) and shorter median time to recurrence than the low risk group (13.1 months versus not reached). This was superior to single markers and to other markers such as P53 (p = 0.02) or Ki67 (p = 0.07). In the test set, the model produced similar results with a relative risk of 2.1 (p=0.029) for the high-risk group and shorter median time to recurrence (14.1 months versus not reached).&#x2028; Conclusions: Triple negative breast cancers show variable expression of proteins involved in DNA repair. Levels of four DNA repair proteins correlated significantly with recurrence free survival, and were used to develop a DNA repair profile model in a training set which was prognostic in a test set. DNA repair biomarker panels may be useful as prognostic or predictive indicators as well as suggest possible targets for novel therapies such as PARP inhibition. Further study of the model in another validation set with other clinical variables is warranted.
Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 1064.
Local versus central HER2 immunohistochemistry correlates with kinetic RT-PCR but only central immunohistochemistry and RT-PCR predict pathological complete response: results from the neoadjuvant multicenter GeparTrio trial.
Authors: Loibl S , Müller B , Roller M , Kronenwett R , Darb-Esfahani S , Komor M , von Toerne C , Wirtz R , von Minckwitz G , Denkert C 
Published: December 2010

Abstract
Abstract #1070
Background: There are several methods for determination of HER2 overexpression. Immunohistochemistry (IHC) has been set up as the gold standard in routine diagnostics to identify patients eligible for anti-HER2 therapy. Nonetheless, there is still controversy about the predictive value of HER2 overexpression and pathological response to neoadjuvant anthracycline/taxane-based chemotherapy.&#x2028; Methods: In this study, we investigated the HER2 expression levels in formalin-fixed, paraffin-embedded pre-treatment punch biopsies by kinetic one-step RT-PCR (kRT-PCR); local IHC performed at the pathological institutes of the participating centers; and centrally performed IHC in 150 patients from the neoadjuvant GeparTrio study (NCT00544765), where patients were treated with docetaxel, doxorubicin and cyclophosphamide (TAC) but received no trastuzumab therapy regardless of the individual HER2 status. HER2 was considered positive for IHC if expression was 3+ and for mRNA expression if normalized level was &gt;/= 18.0 arbitrary units (as defined by bimodal expression for this cohort). We evaluated the associations by Spearman's rho correlation between the different methods as well as the best method that might predict a pathological complete response to this neoadjuvant chemotherapy.&#x2028; Results: A randomly selected subset out of 150 of 2,090 patients was included in this analysis. This is a representative collective of TAC-treated patients from the GeparTrio trial with a pCR rate (ypT0; ypTis/ypN0) of 23%. The different methods revealed a HER2 positivity of 31% for the local analyses, 16% for the central HER2 testing and 20% positivity for kRT-PCR analyses for this study collective, compared to 28% for the entire trial population. Local determination of HER2 showed a correlation with central laboratory (RS=.418, P&lt;.001, N=106) and kRT-PCR results (RS=.428, P&lt;.001, N=120), and centrally assessed HER2 expression correlated with the results of kRT-PCR (RS=.923, P&lt;.001, N=115). Binary logistic regression with the locally determined HER2 expression revealed no significant results in prediction of a pathological complete response (P=.802, N=121), whereas the results from central IHC (P=.038, OR 2.84, N=119) as well as the results from kRT-PCR (P=.047, OR 2.47, N=134) were significant.&#x2028; Conclusions: The value of HER2 expression and response to neoadjuvant chemotherapy is still controversial. Our results on this subset of patients show that kRT-PCR of HER2 mRNA expression and centrally performed IHC but not local HER2 assessment predicts pCR for neoadjuvant TAC chemotherapy. Theses results need further confirmation on a larger group of patients.
Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 1070.
Risk assessment of relapse in Swiss participants of BIG 1-98 with grade 2 early breast cancer by gene expression profiling using RNA from formalin-fixed, paraffin embedded (FFPE) tissue.
Authors: Aebi S , Antonov J , Popovici V , Delorenzi M , Giobbie-Hurder A , Viale G , Thürlimann B , Altermatt H , Jaggi R 
Published: December 2010

Abstract
Abstract #1073
BIG 1-98 is a randomized controlled clinical trial for postmenopausal patients with early, estrogen and/or progesterone receptor (ER/PgR) positive breast cancer. It compares adjuvant therapy with 5 years of tamoxifen (T) or letrozole (L), two years of T followed by 3 years of L (T→L) or vice versa (L→T). L improved disease free survival (DFS) over T by 18% (hazard ratio 0.82, JCO 2007;25:486ff). Switzerland contributed 598/8010 patients. The patient and tumor characteristics of Swiss patients was similar to the entire BIG 1-98 population. The purpose of this study was to test a prognostic score in RNA derived from FFPE tissues.&#x2028; Scores representing ER and progesterone receptor (PgR) function, proliferation (PRO14, AURKA/STK15, BIRC5/SURV, CCNB1, CCNB2, CCNE2, CDC2, ENPF, KIF20A, MKI67, MYBL2, ORC6L, PRC1, SPAG5, TOP2A), and HER-2 were built from multiple genes for each score. Genes were selected according to an analysis of publicly available cDNA array databases. In a prior set of 82 pairs of breast cancer samples the Pearson correlation coefficients of the scores between fresh frozen and FFPE tissues were at least 0.94. Cut-offs for ER and HER-2 scores were defined as the antimodes of fitted bimodal Gaussian distributions; the ER and HER-2 scores closely agreed with immunohistochemical (IHC) data (kappa(ER)=0.868, kappa(HER-2)=0.836).&#x2028; FFPE tissues were available from 437 Swiss BIG 1-98 patients. RNA of sufficient quantity and quality was extracted from 388 blocks (89%). Forty-seven genes were quantified by QRTPCR on TaqMan Low Density Arrays (TLDAs) as reported previously (Proc AACR 2008, Abstract 974).&#x2028; The PRO14 score was positively correlated with the histological grade. PRO14 scores greater than the median and PGR scores lower than the median were associated with shorter DFS. In patients with grade 2 tumors, PRO14 scores were significant predictors of DFS in univariate Cox regression analyses. Multivariate analyses will be presented. Our findings are in agreement with an analysis of fresh frozen tissues of Belgian participants in BIG 1-98 (European Breast Cancer Conference 2008, Abstract 481).We show that RNA can be extracted from FFPE in sufficient quantity and quality to be used in QRTPCR using TLDAs. This opens the possibility of using FFPE tumor banks from controlled clinical trials for the investigation of gene expression patterns.
Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 1073.
Quantitative HER2 homodimer levels correlate with time to first recurrence in HER2-positive breast cancer patients who did not receive trastuzumab in the adjuvant setting.
Authors: Bates M , Koestler W , Fuchs E , Sperinde J , Leitzel K , Ali S , Weidler J , Wu Y , Chappey C , Huang W , Lipton A 
Published: December 2010

Abstract
Abstract #1074
BACKGROUND: HER2-positive breast cancer patients are currently treated with trastuzumab in the adjuvant setting, but prior to 2005 these patients were not routinely offered trastuzumab, creating an opportunity to examine the relationship between HER2 expression and disease progression. The VeraTag technology is a proximity-based assay system that permits quantitative measurements of total HER2 protein expression (H2T) as well as HER2 homodimers (H2D) in formalin-fixed paraffin-embedded (FFPE) tissues. We measured H2T and H2D in a cohort of patients who were HER2-positive but did not receive trastuzumab in the adjuvant setting, and correlated those measurements with time to first recurrence of disease.&#x2028; METHODS: Patients were selected for study because they were treated with a trastuzumab-containing regimen for HER2+ metastatic breast cancer. HER2-positivity was defined as IHC (Herceptest) 3+ or 2+, FISH+ (Vysis). 96 HER2+ patients who had been treated (but not with trastuzumab) in the adjuvant setting were identified. The VeraTag assay was used to quantitate H2T and H2D. Kaplan-Meier and Cox proportional hazards analyses were employed.&#x2028; RESULTS: The distribution of H2T ranged over approximately 135-fold, and H2D varied over approximately 100-fold. In univariate Cox proportional hazards regression analysis, H2T trended toward significance (HR=1.44, p= 0.088) while both H2D (HR=1.39, p=0.022) and the ratio H2D/H2T (HR=2.01, p=0.03) were significantly correlated with time to first recurrence. Kaplan-Meier analysis of the H2T distribution divided into tertiles showed no significant difference in time to first recurrence among the tertiles (HR=0.7, p=0.16 for low vs. high comparison) and no trend was observed (Log Rank test for trend p=0.2). However, the same analyses performed using H2D (HR=0.59, p=0.03) or H2D/H2T (HR=0.56, p=0.01) demonstrated significantly longer time to first recurrence for the lowest tertile compared with the highest tertile respectively. A trend was observed in both cases (H2D: Log Rank p=0.048; H2D/H2T: Log Rank p=0.026)&#x2028; CONCLUSION: In a population of HER2+ patients who did not receive trastuzumab in the adjuvant setting, and who subsequently developed metastatic disease, higher levels of HER2 homodimers and the ratio of homodimers to HER2 total expression correlated with time to first recurrence while total HER2 expression levels did not. These data suggest that measures of the activated forms of HER2 (dimers) may be better predictors of disease progression than simple quantitation of HER2 alone.
Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 1074.
Invasive breast carcinoma of 1 cm or smaller: risk of nodal involvement and prognosis.
Authors: Ferro A , Caldara A , Triolo R , Frisinghelli M , Micciolo R , Barbareschi M , Leonardi E , Pellegrini M , Bernardi D , Cazzolli D , Berlarnda G , Mussari S , Galligioni E 
Published: December 2010

Abstract
Abstract #1075
Background: Screening mammography has increased the frequency of diagnosis of small (≤1 cm) invasive breast cancer (BC), for which the prognosis after locoregional therapy remains controversial.&#x2028; Patients and methods: We have performed a retrospective analysis of 882 patients with ≤1 cm BC and positive or negative-nodes, treated in our Institution from 1990 to 2005. Risks of axillary nodes involvement (ANI), relapse-free (RFS) and overall survival (OS) were analyzed according to: age, tumor size, histologic type and grading, ER and PgR status and MIB-1, HER-2 and p53 immuno histochemical expression.&#x2028; Results: Median age at diagnosis was 57 years (range: 20 - 86 years). Tumor size was: 71 Tmic (a single focus of invasive carcinoma ≤ 2 mm or up to 3 foci ≤1 mm in greatest dimensions, according to Silver and Tavassoli), 225 T1a (&gt;2 -5 mm) and 586 T1b (&gt;5–10 mm). ANI was found in 122 cases (13,8%): only 1 inT1mic (1,4%), 21 in T1a (9,3%) and 100 in T1b (17%). Tumor size, young age, poor differentiation and ductal histology were significantly associated with ANI (p=0.01). Local relapses were: 3 in Tmic pts(2 DCIS and 1 invasive), 13 in T1a, 22 in T1b. Distant metastases were: 1 in Tmic pts, 11 in T1a, 17 in T1b. At a median follow up of 92 months, RFS was 86,4% (Tmic: 86,8%, T1a: 86,2%; T1b: 86,2%). BC-specific mortality was 2% for T1mic, 3,3% for T1a, 3,8% for T1b accounting for an OS of 96,6%. Nodal involvement was the more relevant prognostic factor for survival, followed by young age, poor differentiation, high MIB-1 and p53.&#x2028; Conclusions: In pts with 1 cm or smaller Invasive Breast Carcinoma, the risk of ANI is low, but not negligible, particularly for T1b tumors. Furthermore, the prognosis of these pts is influenced by high histological grade and young age.
Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 1075.