bluuebunny/crossref_metadata_2025 huggingface dataset:
Differential Gene Expression Profiles in Breast Cancer Tumors from Young and Post-Menopausal Patients.
Authors: Moreira-Filho C. , Ramos P. , Ramos P. , Barreta L. , Brentani H. , Corradini B. , Maciel M. , Carraro D. , Achatz M. , Michelli R. , Lyra E. , Grosso S. , Folgueira M. , Katayama M. , Brentani M.
Published: December 2010

Breast cancer in young patients (<35 yrs) has been hypothesized to be more histologically aggressive than those occurring in older cohorts, based mainly in their histopahtological features and worse prognosis. Our current knowledge of the genes involved in such differences is limited. To obtain insight into the molecular processes underlying tumorigenesis in young women we have used 44K cDNA microarrays (Agilent) to compare the gene expression profiles of 30 samples obtained from patients younger than 35 years (median 32,5, range 25-35) and 19 post-menopausal patients (median 66, range 55-88 ), all of them with ductal invasive carcinoma. Both age groups showed similar incidence of ER and PR-positive cases (70 and 60%, respectively). We considered as differentially expressed genes those presenting a p value below 0.01(t test) and fold variation of+/- 2.5.Gene ontology was performed using the GOTree machine tool (GOTM). By such criteria, 121 genes (86% down-regulated) were identified between the two groups here analyzed. In tumors arising in younger women we found an overrepresentation of genes involved with G-protein receptor signaling,cytoskeleton and muscle development, including ankyrin 3,filamin B, gfsarcoglycan and small proline- rich protein, one of the cornified-envelope structural precursor protein. In addition, solely in the younger women set we compared the gene profiles between women with (n =19) or without (n = 10) familial history of breast carcinoma. Using the same criteria, 59 transcripts distinguished the two subsets: GO categories over represented in tumors from patients with familial history included cell-cell adhesion, cytoskeleton organization,cell motility and protein binding related genes. These results indicate that breast cancer tumors arising in young women, besides their uniqueness regarding tumorigenesis, also differ among familial and non-familial cases as revealed by gene expression profiles.Support: FAPESP 04/14029-1

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 6138.
Gene Transcripts in the Tumor Suppressor Region of Chromosome 3 (3p21).
Authors: Al Sarakbi W. , Sasi W. , Jiang W. , Roberts T. , Newbold R. , Mokbel K.
Published: December 2010

Background:There is increasing evidence that the short arm of chromosome 3 has a tumour suppressor region that has been associated with permanent growth arrest of tumour cells. We have identified 9 closely related genes in this region (CCDC12, KLH9, KLH18, MYL3, NBEAL2, PTPN23, SCAP, SETD2, and TESSP2) located which could be possible candidates for tumour suppressor genes (TSG).Our objective was to determine, using quantitative PCR, whether the mRNA expression levels of these genes were consistent with a tumour suppressive function in human breast cancer.Methods:A total of 153 samples were analysed. The levels of transcription of all 9 genes were determined using quantitative PCR and normalized against (CK19).Transcript levels within breast cancer specimens were compared with non-cancerous breast tissue and analyzed against conventional pathological parameters and clinical outcome over a 10 year follow-up period.Results:Quantification of CCDC12, KLH9, KLH18, MYL3, NBEAL2, PTPN23, and SCAP mRNA expression after CK19 normalisation showed no statistically significant difference between malignant and normal breast tissue (p=0.23, 0.46, 0.3, 0.141, 0.8, 0.696, and 0.11 respectively). The mRNA expression of TESSP2 was higher in normal samples compared with malignant lesions (347 vs. 35, p=0.0061) but showed no significant associations with clinical outcome.The levels of SETD2 mRNA were significantly lower in malignant samples (406 vs. 17, p=0.0345) and decreased with increasing tumour stage. SETD2 expression levels were significantly lower in samples from patients who developed metastasis, local recurrence, or died from breast cancer when compared to those who were disease free for > 10 years (p=0.041).Conclusion:These findings are consistent with a possible tumour suppressor function of SETD2 in human breast cancer.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 6139.
Genes Regulating the Progression of Human Ductal Carcinoma In Situ to Invasive Breast Cancer.
Authors: Lee S. , Wu Y. , Dao B. , Lim A. , Medina D. , Stewart S. , Allred D.
Published: December 2010

Rationale: The progression of human ductal carcinoma in situ (DCIS) to invasive breast cancer (IBC) represents a major potentially life-threatening event. Our understanding of this process is rudimentary. Improved understanding could lead to better ability to predict and new therapeutic strategies to prevent progression, which would be major contributions to health care. Objectives: To identify genes involved in regulating the progression of human DCIS to IBC, and to confirm their involvement utilizing relevant in vitro and in vivo model systems. Methods: Gene expression profiles were evaluated in five complimentary cohorts of human DCIS and IBC (total n=59 in each category) using cDNA microarray technology. Supervised statistical comparisons were used to identify differentially expressed genes in each cohort. The genes were distilled to top priority candidates by the following vetting strategy: (1) expression changed in same direction in 2 or more cohorts; and (2) they possessed known biological functions which might reasonably be involved in regulating tumor invasion; and (3) they had not been studied in the context of breast cancer invasion. Initial studies have focused on candidate invasion-suppressor genes (down-regulated in IBC vs. DCIS). A human DCIS-like cell line (DCIS.COM) was stably transduced with shRNAi against the genes (3-5 hairpins each) and evaluated in vitro and in vivo for changes in invasion-related behaviors vs. controls (growth and migration in 2D and 3D cell cultures, invasion in 3D culture, and a human-in-mouse [HIM} DCIS xenograft model). The latter is a novel method we developed where cells are injected into the nipple duct of intact mammary glands of immune-compromised mice. They initially grow as DCIS which progress to IBC at varying rates depending on the cells. Results: Two novel invasion-suppressor genes (CSTA and FAT1) were identified based on demonstrating significantly increased growth, migration, and invasion in the assays described. In particular, shCSTA (n=22) and shFAT1 (n=22) HIM DCIS xenografts progressed to large IBCs by 10 weeks or less, significantly (P<.0001) greater than observed in controls (n=26) (67%, 64%, vs. 26%, respectively). Conclusions: We have identified two genes (CSTA and FAT1) whose normal functions appear to suppress the progression of human DCIS to IBC. These results could lead to new therapeutic strategies to prevent or suppress progression.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 6141.
SRC-1 Promotes Breast Cancer Metastasis by Activating MAPK Pathway.
Authors: Qin L.
Published: December 2010

Background: Steroid receptor coactivator (SRC-1) was defined as a nuclear receptor coactivator. It interacts with nuclear receptor and other transcriptional factors and promotes target genes expression. In breast cancer, SRC-1 expression correlates positively with HER2 expression, disease recurrence and resistance to endocrine therapy. Our previous work demonstrated that knockout of SRC-1 in mice suppressed breast cancer metastasis without affecting primary tumor initiation and growth in the MMTV-PyMT transgenic mice. However, the molecular mechanism responsible for SRC-1 in breast cancer metastasis is still unclear.Material and Methods: To further study the function of SRC-1 in breast cancer metastasis, we generated transgenic mice with overexpression of human SRC-1 in the mouse mammary epithelial cells. The mammary glands of these mice exhibited normal development. After crossing these mice with MMTV-TVA (avian subgroup A receptor gene, TVA) transgenic mice, we obtained MMTV-TVA/MMTV-hSRC-1 (tester) and MMTV-TVA/WT (control) biogenic mice. RCAS (avian leukosis virus) viruses, which contain MMTV-PyMT expression cassette and can bind to TVA, were injected into mammary gland to induce mammary tumor. Tumor initiation, growth and metastasis were carefully examined with these mice. To investigate mechanism, multiple PyMT tumor cell lines were generated form individual primary tumor with or without SRC-1 and cell migration and invasion were compared with those cells and human breast cancer cells.Results and discuss: No difference in tumor initiation and growth was observed in those mice, but metastasis incidence of breast cancer to lung increased significantly in PyMT/TVA/hSRC-1 mice compared with PyMT/TVA/WT control mice. Tumor metastasis in lung also presented higher index in PyMT/TVA/hSRC-1 mice. In vitro study with PyMT/SRC-1 WT and KO cell lines showed SRC-1 deficiency disturbed cells migration and invasion. WT tumor cells adhered faster to extracellular matrix fibronecetin (FN) and presented high levels of phosphorylated forms of FAK, c-Src, Erk, Jnk and activated Rac1. Western blotting and real-time RT-PCR revealed lower integrin alpha5 and Gem, one of GTP binding protein, levels in SRC-1 deficient tumor cells. In human breast cancer MDA-MB-231 cells, SRC-1 knockdown led to decreased cell migration and invasion. Therefore, SRC-1 may regulate integrin expression and MAPK pathway to promote cancer cell migration and metastasis. Our study provides a possibility to inhibit breast cancer metastasis by targeting SRC-1 function.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 6150.
Outcomes of Stereotactic Radiosurgery in the Management of Brain Metastasis in Patients with HER2 Positive Metastatic Breast Cancer.
Authors: Raza S. , Narayana A. , Morsi A. , Parker E. , Rush S. , Golfinos J. , Novik Y.
Published: December 2010

Background:We present single institution experience on the effect of Her 2 status on results of Gamma knife radiosurgery (SRS) for brain metastases (BM) as to local control, pattern of relapse and survival.Material and Methods:From 2004 to 2008, 65 patients with metastatic breast cancer (MBC) were treated with GK-SRS at our center, 52 had documented Her 2 status. We performed analysis based on Her 2 status of the original tumor defined as positive by 3+ DAKO or FISH. SRS was delivered using cobalt leksell GKS to a median dose of 20Gy (range 16-20 Gy) at 50% isodose line. Responses were evaluated by gadolinium enhanced MRI. We analyzed the data as to: objective response (OR) defined as radiologic response or stable disease, and local progression –free survival (PFS) defined as freedom from progression of treated BM, appearance of new brain metastases and/or leptomeningeal progression, as well as overall survival (OS) measured from the time of SRS .ResultsTwenty-eight patients were identified as HER2 positive (Group A) and 24 patients were HER2 negative (Group B). Median time to develop BM in group A was 60 months (4 -240 months) versus 48 months (4-156) in group B. Median follow-up was 12 months (1-48) from the time of onset of BM. 66% patients in group A and 82% patients in group B have no more than 2 BM at the time of SRS.In group A, OR was 78.6% (22 pts)and 21.4%(6 pts) had progressive disease (PD) where as in group B the OR was 83% (20 pts) and 17%(4 pts) had PD. Median time to local recurrence of the treated lesion in group A was 13 months (95% CI 5.84 to 20.15) compared to 8 months (95%CI 6.75 to 9.25) in group B. Local PFS at 1 year for Group A vs. B was: control of the treated lesions 69% versus 41.2% respectively, p= 0.97 by log rank test; and development of new BM 67% compared to 86% (p=0.2).Out of all CNS relapses, almost a quarter of patients in both groups received second SRS whereas 25% patients in group A and 31% patients in group B received whole brain irradiation respectively. Interestingly, 28% (9 pts) failed as leptomeningeal disease following SRS, 7 of them with Her 2 positive tumors. Leptomeningeal relapses were seen both as only pattern of recurrence (4 pts) or associated with other types of CNS progression: local re-growth or new BM (5pts).ConclusionsSRS provides high rate of objective response and local control for BM from both Her 2 positive and Her 2 negative breast cancer. No statistically significant difference in local PFS was seen. More frequent pattern of relapse as leptomeningeal disease was seen in Her 2 positive after SRS. Evaluation of novel systemic targeted therapies in conjunction with local therapies is needed for effective management of BM from breast cancer.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 6152.
Predominance of Brain and Lung Metastases in Triple-Negative Breast Cancer Patients.
Authors: Gadiyaram V. , Kurian S. , Abraham J. , Hobbs G. , Vona-Davis L. , Vona-Davis L.
Published: December 2010

Background: Patients with triple-negative breast cancer have an increased likelihood of recurrence compared to other types of breast cancer, however, little is known about their pattern of metastatic spread. Our object was to evaluate the metastatic patterns of women diagnosed with triple-negative breast cancer compared to other subtypes. Methods: We studied a cohort of 572 white patients diagnosed with invasive breast cancer at West Virginia University Hospital between 1999 and 2004. Hospital registry, charts, and pathology records provided clinical data including tumor receptor status and biopsy-proven metastatic spread to bone, brain, liver and lung. Breast cancers that were negative for estrogen, progesterone, and HER2neu, otherwise known as triple-negative were compared with HER2neu-postive and HER2neu-negative (endocrine receptor positive) disease. Body mass index was calculated and a value of ≥30 considered indicative of obesity. Specimens of primary carcinoma were available for analysis of Ki67 mitotic index and expression of p53. Results: 134/572 (23.4%) had triple-negative breast cancer, while the frequencies were 108/572 (18.9%) and 330/572 (57.7%) in HER2neu-positive and HER2neu-negative (endocrine receptor positive) groups. Women with triple-negative disease were more likely to have brain-metastasizing breast cancer; 10.5% versus 4.6% for HER2neu-positive and 3.3% for HER2neu-negative (P<0.05). They were also more likely to have metastasis to the lung; 10.5% versus 2.8% for HER2neu-positive and 7.0% for HER2neu-negative (P<0.05). Triple-negative breast cancer patients who developed brain and lung metastases were younger <50 years and significantly more obese (P=0.0236). High Ki67 labeling index and p53 expression were associated with more advanced disease indicating an aggressive phenotype for this group.Patterns of metastasis in breast cancer subtypesSite of metastasisTriple-negativeHER2-positiveHER2-negativeP valueBone20/134 (14.9)7/108 (6.5)38/330 (11.5)0.1037Brain14/134 (10.5)5/108 (4.6)11/330 (3.3)0.0136*Liver11/134 (8.2)4/108 (3.7)22/330 (6.7)0.3261Lung14/134 (10.5)3/108 (2.8)23/330 (6.9)0.0507*Other5/134 (3.7)3/108 (2.8)3/330 (0.91)0.1093 Conclusion: The excess risk of brain and lung metastasis in women with triple-negative breast cancers versus other subtypes needs further validation. The unique biology of triple-negative tumors may explain this pattern of metastatic spread.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 6159.
Fatty Acid Synthase (FASN) as a Therapeutic Target for Breast Cancer Metastasis.
Authors: Gonzalez-Guerrico A. , Lupu R.
Published: December 2010

Numerous solid tumors confirm increased expression of FASN. FASN expression is upregulated in early steps of breast cancer, such as DCIS, and is associated with poor clinical outcome in more advanced disease. The objective of this study is to evaluate FASN inhibition as a prevention of breast cancer progression and as an effective treatment of metastasis. We utilized the model the MCF10CA1 series of cell lines, which complete the spectrum of breast cancer progression ranging from non-transformed breast epithelial cells to metastatic breast cancer cells. These metastatic cell lines are CA1A and CA1D cells (short for MCF10A-CA1A and MCF10A-CA1D), which metastasize to the lung following a tail vein injection. Interestingly, we determine that with the progression of breast cancer in these model series the level of FASN was correlated with the evolution to a more aggressive phenotype. The CA1A and CA1D cells expressed the highest levels of FASN. To determine whether FASN plays a role in the aggressiveness of these cells we introduced a specific FASN silencing RNA into CA1A and CA1D cells and isolated stable cell lines in which FASN expression was significantly reduced. Stable expression of a FASN shRNA completely abrogated the anchorage-independent growth ability of the metastatic cells and furthermore these cells reverted from EMT to MET (Mesenchymal Epithelial Transition) as demonstrated by a Matrigel 3Dimentional (3D) assay and the immunofluorescence staining of E-Cadherin in matrigel. These results emphasize the potential of FASN as a therapeutic target and as a key modulator of breast cancer progression and metastasis.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 6160.
Hammerhead Ribozyme Suppresses the Expression of MLN64 and Affects the Function of Breast Cancer Cells In Vitro.
Authors: Cai W. , Cai W. , Ye L. , Mansel R. , Jiang W.
Published: December 2010

Background: The MLN64 gene, which is localized in q12-q21 of the human chromosome 17, encodes a novel transmembrane protein that shares homology with the cholesterol binding domain (START domain) of the steroidogenic acute regulatory protein. The function of MLN64, initially reported in the metastatic lymph nodes, remains unclear, but has been indicated to probably regulate cholesterol transport. MLN64 is highly expressed in certain breast carcinomas, often co-amplified with erbB-2, and thus may contribute to the development and progression of breast tumours. The present study investigated the molecular and cellular impact of MLN64 on the functions of breast cancer cells.Methods: An anti-human MLN64 hammerhead ribozyme transgene was constructed using the mammalian expression vector pEF6/V5-His TOPO, and transfected into breast cancer cells MDA MB 231 and MCF-7 by electroporation. Expression of MLN64 mRNA and protein was determined using RT-PCR and Western blotting respectively. Cell growth, invasiveness, adhesion, motility and migration assays were used to assess, in vitro, the impact of MLN64 knockdown on the cells. In addition, expression of MLN64 was evaluated in a cohort of breast tumours, at protein and message levels.Result: Loss of MLN64 in MCF-7 cells resulted in a significant reduction of cellular growth (growth index for MCF-7ΔMLN64 being 0.87±0.07, P<0.01 vs wild type control (MCF-7WT 1.13±0.06) and transfection control (MCF-7pEF 1.27±0.05). Interestingly, the same change in cell growth was not seen with MDA-MB-231 cells, in that MDA-MB-231ΔMLN64 didn't exhibit a significant effect on the cell growth comparing with corresponding WT and PEF controls. In matrix adhesion assay, MDA-MB-231ΔMLN64 cells showed a significant increase in adhesion (86±14), compared with MDA-MB-231WT and MDA-MB-231pEF (61 ±20 and 45±27, respectively, P<0.01). In MCF-7 cell line, no significant differences were observed between the knockdown mutants and the wild types and pEF type. Breast cancer tissues showed a higher levels of MLN64 transcript than normal tissues (median (IQR) 1.08 (0.19-4.7) copies vs 0.5 (0.18-1.3) copies. Furthermore, ER positive tumours had significantly higher levels of MLN64 transcript (2.86(0.53-11.9) copies) than ER negative tumours (0.59 (0.15-2.68) copies), p=0.0012.Conclusion: In this first report on the functions of MLN64 in breast cancer cell lines, we have shown that MLN64 levels are correlated with the growth and adhesion of breast cancer cells in vitro. It is suggested that MLN64 has a direct influence on the aggressiveness of breast cancer cells and may play a role in progression and metastasis of breast cancer.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 6163.
Lympho-Vascular Invasion by Detecting D2-40 Was the Potent Prognostic Predictor in Breast Cancer Patients.
Authors: Woo S. , Ryu W. , Lee J. , Kim H. , Kim C. , Son G. , Lee E. , Lee J. , Bae J.
Published: December 2010

(Introduction) Distant metastasis and recurrence are the major factors of prognosis in breast cancer patients. Both of lymphovascular invasion (LVI) and blood vascular invasion (BVI) are known to be the important route for metastasis to regional lymph nodes and systemic metastasis. Despite the importance of vascular invasion as a prognostic factor, the application of vascular invasion as a histopathologic criterion is controversial. The aims of this study were to distinguish between LVI and BVI for the role of prognosis and recurrence in breast cancer patients by using an endothelial subtype specific immunohitochemical stain (podoplanin, D2-40 and CD31)Methods) Sections from 80 paraffin-embedded archival specimens of invasive breast cancer were stained for expression of podoplanin, D2-40, and CD31. The results of immunohistochemical staining were correlated with clinicopathologic features, such as tumor size, the status of lymph node metastases, estrogen receptor status, progesterone receptor status and HER2 expression, and recurrence. The patients with ductal carcinoma in situ and stage 4 breast cancer were excluded.Results) A significant correlation was found in the higher expression of D2-40, and lymph node metastasis. (P=0.02) We found a significant correlation between the expression of D2-40 and the recurrence of breast cancer. (P=0.04) However, there was no significant correlation between BVI and recurrence. A poorer disease free survival was shown in the higher expression of D2-40. (P<0.01) In multivariate analysis, the higher expression of D2-40 revealed a significant association with decreased disease-free survival.Conclusion) The higher expression of D2-40 was the more prognostic predictor than blood vascular invasion in breast cancer patients.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 6171.
A Single Nucleotide Polymorphism in Steroid Receptor Coactivator-1 Diminishes the Coactivation of ERα.
Authors: Hartmaier R. , Hartmaier R. , Hartmaier R. , Richter A. , Richter A. , Richter A. , McGuire S. , McGuire S. , Wang J. , Lee A. , Lee A. , Lee A. , Osborne C. , Osborne C. , O'Malley B. , Brown P. , Brown P. , Brown P. , Xu J. , Skaar T. , Skaar T. , Philips S. , Philips S. , Rae J. , Rae J. , Azzouz F. , Azzouz F. , Li L. , Li L. , Henry N. , Henry N. , Nguyen A. , Nguyen A. , Stearns V. , Stearns V. , Hayes D. , Hayes D. , Flockhart D. , Flockhart D. , Oesterreich S. , Oesterreich S. , Oesterreich S. , Oesterreich S.
Published: December 2010

Estrogen signaling is critical in the progression of a large fraction of breast cancers as well as in the maintenance of bone mineral density. Generally speaking, Tamoxifen, a selective estrogen receptor modulator (SERM), blocks estrogen signaling in the breast by acting as an antagonist at the same time as promoting estrogen signaling in the bone by acting as an agonist. This tissue specific action characteristic of SERMs is thought to be maintained by the balance of levels and/or activities of estrogen receptor (ER) coregulators. Because of this critical role, single nucleotide polymorphisms (SNPs) in ER coregulators could have a dramatic effect on tamoxifen action.Previous studies have implicated steroid receptor coactivator-1 (SRC-1) as a critical coregulator for the mixed antagonist-agonist specificity of tamoxifen. By using an ERE-Tk-Luciferase transient transfection assay, we have shown that a SNP causing the amino acid change P1272S in SRC-1 dramatically reduces its ability to coactivate ERα.To assess the impact of this SNP on tamoxifen action in breast cancer we decided to genotype the SRC-1 P1272S SNP in a breast tumor DNA bank. This bank was generated from archived tumors from individuals either exclusively receiving adjuvant tamoxifen or not receiving any adjuvant therapy following surgical resection. Currently, the bank is composed of 1000 samples divided equally among the two groups. Genotyping for the SRC-1 P1272S SNP is currently underway.Since SRC-1 has also been shown to be important in proper maintenance of bone mineral density (BMD) and in the agonist action of tamoxifen in certain tissues, we also assessed the impact of the P1272S SNP in bone following tamoxifen therapy. We genotyped breast cancer patients exclusively receiving tamoxifen therapy (i.e. no chemotherapy, no radiation) who had lumbar (n=113) and/or hip (n=108) BMD scans before and after 12 months of tamoxifen therapy. We observed a significant association between the SNP and bone loss; women harboring the P1272S SNP lost 6.4% of their lumbar BMD, while women with the WT version lost only 1.3% of their lumbar BMD.These findings illustrate the critical role of SRC-1 in tamoxifen action in bone, and potentially other hormone responsive tissues. Studies are ongoing to decipher the mechanism for decreased coactivation activity of the SNP, and also to extend the clinical association studies.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 66.
Changes in the Dynamic Relationship between ER and HER2 in Paired Breast Cancer (BC) Biopsies from Patients (Pts) with Recurrent Tamoxifen-Resistant Breast Cancer.
Authors: Leary A. , Drury S. , Drury S. , Detre S. , Martin L. , Dowsett M. , Dowsett M. , Johnston S.
Published: December 2010

Background: Paired biopsies at diagnosis and relapse on tamoxifen (TAM) provide information regarding adaptive changes associated with endocrine resistance. We previously reported that conversion to a HER2+ phenotype occurs infrequently1, however modest increases in HER2 may be relevant in the context of TAM resistance. Also while TAM resistance in general is associated with a fall in mean ER and PgR levels, this masks changes in individual pts.Aim: To determine whether a subset of HER2 negative BC demonstrated modest increases in HER2 by IHC and FISH, and if so whether this correlated with changes in ER and PgR expression in individual pts.Methods: Triplicate TMAs were constructed of paired primary and recurrent tumor samples from pts relapsing after adjuvant TAM. TMAs were stained for ER and HER2, whole sections for PgR. HER2 amplification was assessed by FISH.Results: Paired samples from primary and TAM-resistant recurrences were available on 53 ER+ and/or PgR+ pts. 10% had a HER2+ primary (IHC3+ and FISH+). 19% (10/53) showed increasing HER2 in the relapsed tumor (0 to 1+ in 5 pts, 1+ to 2+ in 2 pts and 0/1+ to 3+ in 3 pts). The 3 pts converting to 3+ also became FISH+. 71% (38/53) had stable or decreasing HER2. DFI was 50.3 months in the subset with stable or decreasing HER2, 40.2 in the subset with increasing HER2, and 35.2 in the HER2+ subset (p=NS). We next compared changes in ER and PgR in the 3 subsets. Among pts with stable or decreasing HER2 (N=38), there was a significant decrease in mean H-scores for both ER (p=0.001) and PgR (p=0.002) at relapse (Fig 1); the majority recurred with reduced ER (76%) or PgR levels (84%). In contrast, among pts with increasing HER2, there was no change in mean ER or PgR levels (Fig 2A); however when considering changes in individual pts, there was a balanced split between tumors relapsing with up- vs. down-regulation in ER/PgR (Fig 2B,C). In the HER2 positive subset, 100% of the pts relapsed with suppressed ER and PgR.Conclusion: In an updated series of TAM-resistant pts, we show that while conversions to HER2 3+/FISH+ remain rare (5%), modest increases in HER2 are more frequent (19%). TAM resistance in general is associated with decreasing ER and PgR. However, among pts with adaptive increases in HER2, TAM resistance is associated with either down- or up-regulation in ER expression and function. This is consistent with data from acquired endocrine resistance models we previously reported2,3.1Gutierrez et al, JCO 20052Pancholi et al, ERC 2008,3 Martin et al, JBC 2003

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 704.
A Randomised Controlled Trial of Forward-Planned Intensity Modulated Radiotherapy (IMRT) for Early Breast Cancer: Interim Results at 2 Years Follow-Up.
Authors: Coles C. , Barnett G. , Wilkinson J. , Moody A. , Wilson C. , Twyman N. , Hoole A. , Burnet N. , Wishart G.
Published: December 2010

Background: This large single centre trial was designed to investigate whether correction of dose inhomogeneities using IMRT reduces late toxicity and improves quality of life in patients with early breast cancer. We report interim results at 2 years follow-up.Materials and Methods: Standard tangential breast radiotherapy plans of 1145 trial patients were assessed for dose inhomogeneity as defined by ICRU recommendations. Patients with inhomogeneous plans were randomised to either IMRT or standard radiotherapy. Patients with homogenous plans were treated with standard radiotherapy, but were followed up within the trial as per randomised patients. The primary endpoint of the trial was photographic assessment of breast shrinkage. Secondary endpoints included photographic assessment of final cosmesis, clinical assessment of toxicity, and patient-reported symptoms and quality of life. All endpoints were scored using validated assessment methods. Data were collected on potential confounding factors which included: age at randomisation, smoking history, body mass index, co-morbidity such as diabetes mellitus, cardiovascular and peripheral vascular disease, use of hormones and/or chemotherapy, treated breast volume, and presence or absence of breast boost.Results: 330/1145 (29%) of patients had adequate dosimetry with standard 2D radiotherapy and were not randomised. In the 815 randomised patients, dosimetry was significantly improved in the interventional (IMRT) arm compared to the control. Currently, data from 632 randoimsed patients who have reached 2 years of follow up have been analysed. This interim analysis demonstrates that the incidence of telangiectasia in the control arm was significantly greater than in the interventional group (P = 0.024). Pre-radiotherapy surgical cosmesis significantly affected photographic assessment of final cosmesis, and clinical assessment of breast shrinkage and induration (P < 0.0001). In randomised patients who had good surgical cosmesis, there was a significant difference improvement in final cosmesis with the use of IMRT (P = 0.038). The quality of life data has not been analysed at this stage.Discussion: This trial shows that the use of forward-planned IMRT can significantly improve dose homogeneity, and suggests that this translates into improvements in late toxicity at only 2 years from completion of radiotherapy. Surgical cosmesis appears to be a very important determinant of overall final cosmesis after 2 years of follow-up, and may mask any beneficial effects of IMRT at this early stage. It is highly likely that these trial patients will have not expressed their final level of toxicity at 2 years post-radiotherapy, and re-analysis is planned at 5 years from completion of radiotherapy.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 71.
Testosterone Undecanoate Treatment Reduces Joint Morbidities Induced by Anastrozole Therapy in Postmenopausal Women with Breast Cancer: Results of a Double-Blind, Randomized Phase II Trial.
Authors: Birrell S. , Tilley W.
Published: December 2010

Background: Approximately 50% of women taking 3rd generation aromatase inhibitors (AI) for adjuvant breast cancer therapy develop arthralgia, tenosynovitis or arthritis. In many women, the morbidity is significant and affects therapy compliance. The pathology of AI-induced joint morbidity is poorly understood, but clinical features in severe cases mimic rheumatoid arthritis. Afflicted women are treated with anti-inflammatory medication, but this does not address the etiology, which appears to involve steroid hormone imbalance. Testosterone can be metabolised to estrogen or the more potent androgen, 5alpha-dihydrotestosterone (DHT). Androgen levels decrease with age and this may be exacerbated by chemotherapy and, paradoxically, aromatase inhibition. We hypothesise that AI-associated joint morbidities are the result of testosterone deficiency and that testosterone treatment during AI therapy will be beneficial. Testosterone replacement had been contraindicated in women with breast cancer until the advent of powerful 3rd generation aromatase inhibitors that are highly efficacious in blocking conversion of testosterone to estrogen, thus allowing the examination of combinations of drugs such as anastrozole and testosterone.Materials and Methods: 90 women taking Arimidex® 1mg daily and registering a visual analogue scale (VAS) pain or stiffness score of ≥ 40mm on a 100mm scale, were randomized into 3 equal arms: placebo, 40 mg or 80 mg testosterone undecanoate daily, with follow-up at 1 and 3 months. The primary efficacy endpoint was reduction in pain at 3 months and the primary safety endpoints were signs of androgen excess with secondary safety endpoints being serum hormone levels, serum lipids and bone absorption markers.Results: The VAS scores at 3 months fell for all groups, with a marked placebo effect of 35% compared with 43% and 70% for the 40 and 80mg treatment arms (p=0.06 and p=0.04, respectively). No significant androgenic side-effects (hirsutism, alopecia and acne) were observed and serum lipids remained unaltered. Serum total testosterone and free-testosterone levels stabilized at 3 months in a dose-dependent fashion into a physiological range. There was no elevation in serum estradiol in either treatment arm and the serum sex hormone binding globulin level fell significantly in the 80mg treatment group (p=0.03). There was no significant change in bone absorption markers in any of the patient groups.Discussion: This study has demonstrated for the first time that testosterone replacement may be a viable option in the management of AI-induced arthralgia, tenosynovitis and arthritis with good tolerability and safety, without adversely reducing aromatase inhibitor efficacy (as indicated by serum estradiol levels).This study was supported AstraZeneca (UK).

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 804.
Prospective Evaluation of Neurotoxicity in Breast Cancer Patients Treated with Adjuvant Paclitaxel.
Authors: Crew K. , Weimer L. , Brafman L. , Fuentes D. , Awad D. , Hershman D.
Published: December 2010

Background: Taxanes are increasingly used in the treatment of early stage breast cancer (BC). Up to 30% of patients develop moderate-severe neurotoxicity during treatment. We recently reported that about 60% of BC patients complain of some neuropathy up to 2 years after completing adjuvant paclitaxel treatment. The prevalence, severity, and natural history of this syndrome have not been well described.Methods: Using a prospective cohort study design, women with stage I-III breast cancer undergoing adjuvant paclitaxel were enrolled. At baseline, 4 and 8 weeks during treatment, then 3, 6 and 12 months after finishing treatment, all patients completed the Functional Assessment of Cancer Therapy-Taxane (FACT-Tax) and quantitative neurosensory testing (touch perception and vibration threshold) was conducted. The FACT-Tax includes a 16-item subscale with components for neurotoxicity (11 items) and other taxane-related symptoms (5 questions), where lower scores represent worsening symptoms. Missing data was imputed. We used descriptive statistics, t-test and analysis of variance statistics.Results: A total of 50 patients enrolled, however only 41 are evaluable; median age: 47 (32-78); White/Hispanic/Black: 19/16/6; median BMI (kg/m2): 28 (21-44); stage I/II/III: 5/21/9. Mean self-reported scores on the Neurotoxicity component of the FACT-Tax decreased from 37.5 at baseline to a low of 24.8 at week 8 (p=0.0002). These scores remained lower than baseline at 6 and 12 months after completing paclitaxel treatment (32.0 and 32.4, respectively). Similar trends were seen with the Taxane component (P=0.001).The percentage of patients that reported any hand numbness increased from 14% at baseline to 57% at 8 weeks; while foot numbness increased from 20% to 72% in this timeframe. In both groups, 40% reported persistent symptoms at 12 months. Compared to baseline, there was a significant decrease in vibration sense at 8 weeks during active treatment in the great toe (p<0.01), but only small, clinically insignificant changes in the fingers. No significant change from baseline was seen in touch perception.Discussion: The majority of BC patients receiving adjuvant taxanes develop moderate to severe self-reported numbness and discomfort during active treatment, which improves but remains below baseline up to 12 months later. Similar, but less impressive trends were seen with quantitative neurosensory testing. Taxane-based chemotherapy has been shown to improve survival, however, neurotoxicity is the main long-term side effect. Understanding the appropriate measures to be used in interventions to prevent and treat taxane-induced neuropathy are critical.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 807.
Breast Cancer Risk Single Nucleotide Polymorphism (SNP) Genotypes Do Not Correlate with Risk Estimates from Existing Prediction Models.
Authors: Comen E. , Balistreri L. , Kirchhoff T. , Gaudet M. , Dutra Clark A. , Hudis C. , Offit K. , Robson M.
Published: December 2010

Background:Breast cancer risk assessment professionals employ models to estimate individual probabilities of developing breast cancer. The Gail, Claus and BRCAPro models integrate personal, reproductive and family history variables to generate absolute risk estimates. A number of common DNA variants (single nucleotide polymorphisms, SNPs) are associated with modest elevations (relative risk < 2) of breast cancer risk. We investigated the correlation between genotype at 18 published risk SNPs and model-derived breast cancer risk estimates.Methods:Subjects were 475 unaffected, BRCA mutation-negative women who were evaluated for a family history of breast or ovarian cancer. At the time of testing, subjects provided information that was used to estimate absolute breast cancer risk by Gail, Claus, and BRCAPro models. Subjects were genotyped by Sequenom iPLEX MassArray (Sequenom, Inc., San Diego, CA) at loci reported to be associated with elevations in breast cancer risk (1p11.2, 2q35, 5p12, 6q22.33, 8q24, CASP8, COX11, FGFR2, LSP1, MAP3K1, RAD51L1, SLC4A7, TNRC9). Kruskal-Wallis and Spearman correlation tests were used to compare risk estimates for individuals by genotype and number of risk alleles.Results:The median age at sample donation was 46 years (range 24-83). Most subjects (94.9%) were white, and 65.5% were of Ashkenazi Jewish (AJ) ancestry. Median breast cancer risk scores risk estimates were significantly lower from BRCAPro (5 yr: 1.1%, lifetime (LT): 11.6%) than those derived from Gail (5 yr: 1.8%, LT: 18.5%) or Claus (5 yr: 1.5%, LT: 11.8%) models. No significant differences (p>0.05) in distribution of estimated breast cancer risk scores were found between women carrying 0, 1, or 2 risk alleles at any locus except 6q22.33, where AJ women homozygous for the risk allele rs2180341 were noted to have higher LT Claus (p=0.001) and BRCAPro (p=0.01) estimates. When comparing subjects with no risk alleles to those with 1-2 risk alleles for each SNP, no significant differences in distribution of estimated risk were noted at any loci, including 6q22.33. There was no significant correlation between the total number of risk alleles carried by a subject and her estimated lifetime Gail (ρ=-0.052, P=0.27), Claus (ρ=0.051, P=0.32), or BRCAPro (ρ=0.024, P=0.60) risk.Conclusions:In this analysis, risk allele genotype did not correlate with absolute breast cancer risk estimated by standard models. This suggests that the contribution to risk of genotype at these SNPs is insufficient to impact the modeled estimate. Further studies are necessary to evaluate whether incorporation of genotypic information can improve the calibration or discriminatory accuracy of existing models.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 901.
The Contribution of Founder Mutations to Early-Onset Breast Cancer in French-Canadian Women.
Authors: Ghadirian P. , Robidoux A. , Royer R. , Zhang P. , Akbari M. , Zhang S. , Fafard E. , Costa M. , Martin G. , Potvin C. , Patocskai E. , Larouche N. , Younan R. , Nassif E. , Giroux S. , Narod S. , Foulkes W.
Published: December 2010

Background: In an ethnically-homogeneous population, it is valuable to identify founder mutations in cancer-predisposing genes. Founder mutations have been found in four breast cancer predisposing genes in French-Canadian breast cancer families. The frequencies of the mutant alleles have not been measured in a large series of unselected breast cancer patients from Quebec, nor in healthy controls. These estimates are necessary to measure their contribution to the hereditary burden of breast cancer in Quebec and to help develop genetic screening policies which are appropriate for the province.Methods: We studied 564 French-Canadian women with early-onset invasive breast cancer who were treated at a single Montreal hospital. Patients had been diagnosed at age 50 or less, and were ascertained between 2004 and 2008. We screened all 564 patients for nine founder mutations; four in BRCA1, three in BRCA2 and one each in PALB2 and CHEK2. We also studied 6433 DNA samples from newborn infants from the Quebec City area to estimate the frequency of the nine variant alleles in the French-Canadian population.Results: We identified a mutation in 36 of the 564 breast cancer cases (6.4%) and in 35 of 6443 controls (0.5%). In the breast cancer patients, the majority of mutations were in BRCA2 (54%). However, in the general population (newborn infants), the majority of mutations were in CHEK2 (54%). The odds ratio for breast cancer to age 50, given a BRCA1 mutation was 10.1 (95% CI: 3.7 to 28) and given a BRCA2 mutation was 29.5 (95% CI: 12.9 – 67). The odds ratio for breast cancer to age 50, given a CHEK2 mutation was 3.6 (95% CI: 1.4 – 9.1). One-half of the women with a mutation had a first or second-degree relative diagnosed with breast or ovarian cancer.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 902.
Breast Cancer Events Associated with Germline Mutations of the CDH1 Gene (Hereditary Diffuse Gastric Cancer).
Authors: Hebbard P. , MacMillan A. , McCarthy J. , Laing K. , Wadden N. , Green J. , Fernandez B. , Kwan A. , Wirtzfeld D.
Published: December 2010

Breast cancer events associated with germline mutations of the CDH1 gene (Hereditary Diffuse Gastric Cancer).Background: Mutations of the CDH1 (e-cadherin) gene are known to cause an autosomal dominant familial cancer syndrome, commonly referred to as hereditary diffuse gastric cancer. The lifetime risk of gastric cancer may be up to 80 percent. An associated risk of lobular breast cancer has been identified, but less well studied. Lifetime risk of breast cancer in affected women may be up to 50 percent.Method: We identified 17 women in our centre with known CDH1 mutations that have been offered genetic counseling for breast cancer risk. Counseling included discussion of prophylactic surgery, and breast screening via MRI/mammogram. We performed a chart review to identify breast cancer screening, prophylaxis, and treatment events in these patients.Results: 16 patients had adequate chart data to study. Of these, one patient had a breast cancer (T3N0M0) prior to mutation identification. 13 of 15 remaining patients entered into a high-risk breast-screening program with annual MRI and mammogram. Two breast cancers were identified in the first two years of patient screening; both were T1N0M0. Both women opted for bilateral mastectomy. No patient opted for prophylactic breast surgery prior to cancer detection, although several were in the process of prophylactic gastrectomy.Conclusion: To our knowledge, this is the first in-depth description of breast cancer events in patients with inherited CDH1 mutations. A majority of patients have opted for a breast-screening program, with successful early detection in two patients. Patients to date have not chosen primary prophylactic breast surgery, but two have chosen bilateral mastectomy upon detection of a malignancy. Further epidemiological studies are required to fully understand the breast cancer pattern and natural history that occurs in this genetic syndrome.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 904.
Local Therapy in BRCA1/2 Carriers with Operable Breast Cancer: Comparison of Breast Conservation and Mastectomy.
Authors: Pierce L. , Phillips K. , Griffith K. , Buys S. , Gaffney D. , Moran M. , Haffty B. , Ben-David M. , Garber J. , Merajver S. , Balmanya J. , Meirovitz A. , Domchek S.
Published: December 2010

Background: Women with inherited germline BRCA1/2 mutations have a 55-85% cumulative risk of breast cancer (BC) by age 70. Thus, knowledge of expected outcomes following various treatments is needed to advise these patients should they be diagnosed with BC. It is unclear whether breast conservation (BCT) offers similar rates of tumor control as mastectomy (M) in BRCA1/2 carriers and it is doubtful whether a randomized trial comparing these options could be performed. Thus, this analysis compares the outcomes between similarly staged women with BRCA1/2 mutations treated with BCT vs. M.Methods: Women with deleterious BRCA1/2 mutations diagnosed with operable BC and who consented to longitudinal studies were identified at 10 institutions in the US, Australia, Spain and Israel. Patient, clinical and treatment characteristics were compared between those receiving BCT and those receiving M +/- RT. Time-to-event endpoints included first failure of treatment, diagnosis of contralateral breast cancer (CBC), and overall and BC-specific survival. Cox regression models were constructed to detect significant associations between patient and clinical characteristics and time-to-event endpoints.Results: Clinical characteristics and outcomes for 302 BCT and 353 M patients were compared. With a median F/U of 8.2 years for BCT patients and 8.9 years following M, 15-year local failure as first failure was significantly higher with BCT vs. M (23.5% vs. 5.5%, p<0.0001). Multivariate analysis indicted choice of local therapy as the only factor significantly predicting local recurrence, with a 4.5-fold risk of local failure with BCT compared to M (p<0.0001). Local failure analyses by cohort revealed the presence of a BRCA2 mutation (HR 2.8; p=0.024) and no use of adjuvant chemotherapy (HR 5.4; p=0.0001) as significant predictors within the BCT group; presence of invasive lobular cancer (HR 9.9; p=0.004) was the only significant predictor within the M cohort. No significant differences were seen in distant failure, BC-specific or overall survival by local therapy. 15-year estimates of CBC were 52.1% with BCT; 41.4% with M; and 37.9% with M+RT (p=0.44). Analyses of BCT vs. M +/- RT and surgery +/- RT did not reveal significant differences in CBC by cohort.Conclusions: The higher risk of local failure in BRCA1/2-associated BC treated with BCT compared to M did not translate into an increased risk of distant failure or mortality. RT did not result in a detectable increase in CBC at 15 years above baseline elevated rates. These results at both the involved and contralateral breasts should be discussed when patients with BRCA1/2-associated BC are considering local treatment options.

Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 959.