bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P3-02-03: Increased risk of breast cancer in women with false-positive screening test: can it be entirely explained by misclassification?
Authors: von Euler-Chelpin M , Kuchiki M , Vejborg I
Published: August 2014

Background: Screening for disease in healthy people inevitably leads to some false-positive tests. A recent study (von Euler-Chelpin et al, JNCI 2012) showed that women with a false-positive test from screening with mammography had an increased risk of being diagnosed with breast cancer later in life as compared to women with only negative results (RR = 1.67, 95% CI 1.45 to 1.88). The reasons for this excess risk remain unknown. In the present study we have assessed misclassification and evaluated the impact of misclassification on over all excess risk.

Method: We used data from the population-based screening mammography program in Copenhagen, Denmark, and studied 295 women exposed to a false-positive test that later developed breast cancer. Screening took place between 1991 and 2006, with a follow-up for cancer until 2008. Cancers that developed in the same location as the finding that initially caused the recall were defined as misclassified and excluded in the assessment of the excess risk among the false-positives. Time from diagnosis, breast density, tumor size, receptor and nodal status were also taken into account.

Results: 72 cases were found to be misclassified. When the misclassified tests had been excluded, we still found a significant excess risk for breast cancer of 27% (RR = 1.27, 95% CI (1.11 to 1.46) among the women with a false-positive test compared to women with only negative tests. Women with a false-positive test proven at assessment (Type 1) had an excess risk of 27% (RR = 1.27, 95% CI (1.09 to 1.46), while false-positives proven at surgery (Type 2) had an excess risk of 30 % (RR = 1.30, 95% CI (0.86 to 1.96). In the group of women with misclassified tests there were a significantly higher proportion of breasts with fatty tissue (BI-RADS density classification 1 or 2) than among the non-misclassified. The average time from first false-positive test to time of diagnosis was significantly shorter among women with misclassified tests, 4.0 years versus 7.8 years (p < .0001). There were no significant differences in tumor size, receptor or nodal status.

Conclusion: The results indicate that the increased risk can not be explained only by misclassification. The excess risk remains for both Type1 and Type 2 false-positives, which favors some biological susceptibility, as women with false-positive test Type 2 have undergone surgery, which thus makes residual misclassification unlikely.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P3-02-03.
Abstract P3-02-10: Implementation and uptake of a provincial, population-based, organized breast screening program for high risk women in Ontario: The Ontario breast screening program (OBSP) high risk program.
Authors: Eisen A , Carroll J , Chiarelli AM , Horgan M , Meschino W , Rabeneck L , Shumak R , Warner E
Published: August 2014

Background: Genetic testing for mutations in BRCA1/2 has been clinically available in Ontario since 2000. Over 15000 individuals have been tested. Evidence from clinical trials has consistently shown that women at high risk of breast cancer (BrCa) benefit from BrCa screening that includes both magnetic resonance imaging (MRI) of the breast and mammography, yet access to MRI in Ontario was variable. In 2011, Cancer Care Ontario (CCO) established an expert panel to develop a protocol for expanding the OBSP, initially created for average risk women 50–74, to include MRI and mammography for eligible high risk women.

Methods: The panel's tasks included: 1. determining high risk criteria 2. estimating the prevalence of high risk women 3. selecting a cancer risk model 4. developing a referral and assessment pathway for potentially eligible subjects 5. developing educational resources, training plan and communication strategy for relevant stakeholders 6. developing indicators for program evaluation, 7. providing guidance for post-implementation issues.

Results: The program was initiated July, 2011 and now includes 28 sites. Women aged 30–69 with and without a history of breast cancer are eligible if they 1. are BRCA1/2 mutation carriers or untested first degree relatives of carriers, 2. have a lifetime risk of breast cancer >=25% based on family history according to IBIS or BOADICEA risk models, or 3. received prior radiation therapy to the chest. It is estimated that 34000 high risk women in the target age group live in Ontario. Preliminary volume data for the first 9 mos are shown in Table 1. Of the 5037 women participating, 802 have been referred directly by their physician and 4,235 have been referred for genetic assessment. Of the 2,946 women who received genetic assessment, 31% met the high risk criteria. To date, 729 high risk MRI scans have been performed.

Conclusions: A population based organized screening program for high risk women that includes genetic risk assessment has been implemented in Ontario. Further evaluation of risk assessment and screen results are underway. To our knowledge, this is the first organized screening program for women at high risk of breast cancer.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P3-02-10.
Abstract P3-06-18: Increase of serum androgen and its metabolites in postmenopausal primary breast cancer patients with disease progression during neo-adjuvant exemestane treatment; JFMC 34–0601 TR
Authors: Takada M , Saji S , Honma N , Masuda N , Yamamoto Y , Kuroi K , Yamashita H , Ohno S , Aogi K , Ueno T , Toi M
Published: August 2014

BACKGROUND: We reported a positive correlation between body mass index (BMI) and clinical response to neo-adjuvant hormonal therapy (NAH) with steroidal aromatase inhibitor, exemestane, in post-menopausal breast cancer patients (Takada M et al. Breast 2012). Here, we examined the relationship between serum concentrations of sex steroids and BMI, and explored their predictive value for clinical response.

PATIENTS AND METHODS: Among the 116 post-menopausal patients enrolled in the JFMC 34–0601 clinical trial of 24 weeks (wks) of NAH with exemestane, serums from 60 patients at pre-treatment, at 4wks and end of the treatment (24wks) were subjected to this study. Estradiol (E2), estrone (E1), dehydroepiandrosterone (DHEA), androstenedione (A-dione), 5-androstene-3β, 17β-diol (Aenediol), 5α-androstane-3β, 17β-diol (Aanediol) were measured using LC-MS/MS analysis and E1- sulfate (E1S) was by RIA.

RESULTS: There were no significant correlations between pre-treatment concentrations of sex steroids and BMI, except for moderate correlation of E2 and BMI (p = 0.004). In multivariate analysis, E1 was an independent predictive factor for objective response [odds 6.0, 95% confidence interval (CI) 1.5 — 34.6, p = 0.011], as well as BMI. All of the estrogens decreased to under-detection levels (0.5 for E1 and E2, 5 pg/assay for E1S) at 4 wks of treatment, and maintained through to the end of treatment in almost all patients independently of clinical response. The geometric mean percentage changes in androgens after NAH were: DHEA −0.2% (95%CI −15.3% — +17.6%) for patients without progressive disease (non-PD) and +44.8% (95%CI +22.1% — +71.8%) for patients with progressive disease (PD); A-dione −2.3% (95%CI −17.3% — +15.4%) for non-PD and +45.6% (95%CI +28.3% — +65.3%) for PD; Aenediol −11.5% (95%CI −20.6% — −1.4%) for non-PD and +24.9% (95%CI +1.9% — +53.0%) for PD; Aanediol +21.3% (95%CI −5.5% — +55.8%) for non-PD and +56.3% (95%CI +5.3% — +132.0%) for PD, respectively. The changes in the concentrations of both DHEA and A-dione in patients with PD were statistically significant (p = 0.008 and p = 0.002, respectively). In all of the PD patients, the serum concentrations of DHEA and A-dione were increased after NAH.

CONCLUSION: Pretreatment serum concentration of E1 was an independent predictive factor for clinical response to NAH with exemestane. Measurement of dynamics of the serum androgen concentrations might be helpful for monitoring treatment response, and mechanism of increase of androgens has a value for further investigation. Our results should be validated using a larger dataset. (UMIN ID; C000000345)

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P3-06-18.
Abstract P3-06-25: PTEN mRNA positivity using in situ measurements is associated with better outcome in Tamoxifen treated breast cancer patients.
Authors: Schalper KA , Li K , Rimm DL
Published: August 2014

Background: Aberrant phosphatidylinositol 3-kinase (PI3K) pathway signaling occurs in a high proportion of human breast cancers and is involved in carcinogenesis, tumor progression and resistance to targeted therapies. Reduced levels of the tumor suppressor PTEN has been associated with PI3K pathway activation and is observed in ∼40% of breast carcinomas. PTEN levels have traditionally been determined in breast tumors by qualitative immunohistochemistry (IHC) and its prognostic and/or predictive value has not been clearly established. We used a novel method of quantitative in situ measurements of PTEN mRNA and protein to determine their possible prognostic and predictive value in breast cancer.

Materials and methods: PTEN mRNA and protein were measured on formalin fixed paraffin embedded (FFPE) cell lines and tumor samples on tissue microarrays (TMAs) representing a retrospective cohort of 404 breast cancer cases collected from the Yale Pathology archives between 1975 and 2005. The RNAscope paired-primer method was used for PTEN mRNA in situ hybridization (ISH), with Ubiquitin C and DapB ISH probes used as positive and negative controls for each run. PTEN protein measurements were performed with monoclonal antibody 138G6 (Cell Signaling Technology). Both mRNA and protein were measured using the AQUA method of quantitative immunofluorescence.

Results: Both PTEN mRNA ISH signal and protein were highly reproducible in FFPE tissues and cell lines. PTEN mRNA levels were lower in cell lines with known PTEN downregulation. In breast carcinoma samples, PTEN mRNA scores showed a positive non-linear association with protein levels. Neither PTEN mRNA nor protein showed prognostic value. However, high PTEN mRNA expression using a visually defined cutoff was associated with better outcome in a subset of Tamoxifen treated breast cancer patients.

Conclusion: Initial data suggests that PTEN mRNA expression may be predictive of favorable response to endocrine therapy in breast carcinomas. However, validation in larger independent cohorts is required.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P3-06-25.
Abstract P3-07-01: Endocrine insensitivity defines inheritance of breast cancer prognosis
Authors: Lindström Linda S , Hartman Mikael , Hall Per , Czene Kamila
Published: August 2014

Withdrawn by Author

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P3-07-01.
Abstract P3-07-02: Time-trends in survival in young women with breast cancer in a SEER population-based study
Authors: Ademuyiwa FO , Groman A , Hong C-C , Kumar S , Levine EG , Miller A , Ambrosone CB
Published: August 2014

Background: As mammography is not generally recommended to women under 40, it is reasonable to conclude that documented outcome improvements over time are attributable to treatment advances with screening playing a less important role. In order to determine the contribution of screening and treatment to improvements, we evaluated the odds of presenting with more advanced disease by time-period and examined the time-trends in outcome in a population-based cohort ≤50. We evaluated whether any outcomes differentials existed by ER status.

Methods: Patients in SEER diagnosed with breast cancer (BC) were divided into 4 by year of diagnosis (1990–1994, 1995–1999, 2000–2004, 2005–2008). Patients were also categorized into 2 age-groups: <40 and 40–50 years. Odds ratios for presenting with more advanced disease over the 4 time-periods were calculated for the 2 age-groups. Multivariate analysis was done to investigate the association of survival with time-period for the 2 age-groups by ER status.

Results: 110,629 patient records were included. Multivariate adjusted hazard ratio for mortality in women aged 40–50 with ER positive BC declined over time. Comparing all time periods to 1990–1994, the HR for mortality in 1995–1999 was 0.83 (0.77–0.89), 0.61 (0.57–0.65) in 2000–2004, and 0.35 (0.30–0.40) in 2005–2008 (p <.001). Those <40 years with ER positive BC also had mortality improvements over time (HR 0.86, 0.63, 0.26 for 1995–1999, 2000–2004, and 2005–2008 respectively). On the contrary, no mortality improvements were seen in women with ER negative disease. Trends in progressively worsening mortality over time were observed in this cohort.

A 1.3-fold increase in the odds of presenting with stage I BC was seen in women 40–50 years comparing the 3 latter time periods with 1990–1994. Similarly, in the same age group, there was a 2.12-fold likelihood of presenting with a tumor ≤1cm in 2005–2008 versus 1990–1994. In women <40 years at diagnosis, the probability of presenting with stage I disease was lower in the 3 latter time periods than in 1990–1994; and a 2.84-fold higher probability of presenting with stage III/IV disease was seen in 2005–2008 compared with 1990–1994.

Conclusions: Patients who are ER positive and between 40–50 years have had time-trend changes with improvements in breast cancer outcome and smaller tumors likely attributable to both screening and hormonal therapies. Patients who are <40 years and/or ER negative have not had improvements in breast cancer outcome.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P3-07-02.
Abstract P3-10-01: Epidemiological risk factors and normal breast tissue markers in inflammatory breast cancer
Authors: Atkinson RL , Sexton KR , Ueno NT , El-Zein R , Brewster AM , Krishnamurthy SA , Woodward WA
Published: August 2014

Background: Inflammatory breast cancer (IBC) is a rare form of aggressive breast cancer with no existing identifiers for screening or prevention strategies. Women with triple-negative (TNBC, ER–, PR–, Her2–) non-inflammatory breast cancer are less likely to breastfeed, and we have shown in adjacent normal breast tissue that this tissue has more foci of stem cells compared to non-TNBC cancers. A disproportionately higher percentage of women with IBC have TNBC relative to women with non-IBC. We hypothesized that adjacent normal tissue in TNBC IBC vs. TN non-IBC may also display unique biological features based on epidemiologic characteristics.

Methods: We examined epidemiologic factors by breast cancer receptor subtype in 144 patients diagnosed with IBC in 1991–2011 at MD Anderson Cancer Center. Breast cancer risk factors including parity and breastfeeding were compared between patients with TN and non-TN IBC with chi-square or Wilcoxon rank sum tests. Normal adjacent tissues were stained for stem cell markers CD44+CD49f+CD133/2+ and macrophage marker CD68.

Results: The mean age at diagnosis was 52.3 years (range = 23–80) and 83% of patients were non-Hispanic white, 80% were overweight or obese (BMI >25), and 36% were TN IBC. Patients with TN IBC had significantly lower frequency of breastfeeding compared with non-TN IBC, 28% vs. 55%, (p = 0.01). No differences were found in the frequency of other breast cancer risk factors. All 8 IBC adjacent tissue samples showed a distinct spatial distribution of stem cell staining, not limited to the triple negative patients. Compared with 0/60 non-IBC cases, 0/8 triple negative non-IBC, (p = 0.001 Pearson chi-square). Given the high BMI among IBC patients, we further examined normal tissues for the presence of CD68+ cells distributed individually or as clusters exhibiting a “crown-like” pattern (multiple CD 68+ macrophages found around dead adipocytes), and found that 7 of the 8 IBC adjacent tissues were CD68+. Benign biopsies collected from 2 patients at 10 years before diagnosis displayed similar staining, including both stem cell and CD68 staining. Compared with 12/60 non-IBC adjacent tissues were positive for CD68, with 1/8 TN non-IBC, (p = 0.001 Pearson chi-square).

Conclusion: We describe for the first time a stem-cell staining pattern unique to IBC present in all IBC tissues examined, including pre-cancer biopsies. Tissue samples from additional patients will be examined to further explore the relationship between stem cells and CD68 positivity with IBC subtypes.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P3-10-01.
Abstract P3-14-03: Differences in pathological and biological factors between DCIS, DCIS with microinvasion (DCIS-MI) and DCIS with concomitant invasive ductal carcinoma (DCIS-IDC).
Authors: MacGrogan G , Baranzelli MC , Picquenot JM , Penault-Llorca F , Mathieu MC , Tas P , Fermeaux V , Mery E , Sagan C , Blanc-Fournier C , Brabencova E , Arnould L , Jacquemier J , Ettore F , Velasco V , Gonzalves B , Brouste V , Tunon de Lara C
Published: August 2014

Background: Specific pathological and biological factors differentiating pure DCIS, DCIS with microinvasion (DCIS-MI) and DCIS with concomitant invasive ductal carcinoma (DCIS-IDC) are currently unknown. Identification of such factors would enable us to better understand the transition from an in situ to an invasive carcinoma and improve patient management for patients diagnosed with extensive DCIS.

Methods: We previously performed a prospective surgical study demonstrating the benefit of performing upfront sentinel lymph node (SLN) biopsy in patients with extensive DCIS or DCIS-MI undergoing mastectomy. In that study, 196 DCIS and 31 DCIS-MI diagnosed on vacuum-assisted macrobiopsies underwent mastectomy and SLN biopsy. Final pathological status after mastectomy examination revealed 117 DCIS, 38 DCIS-MI, 69 DCIS-IDC, and in 3 cases no residual disease was found. The mastectomy specimens of 216 cases (111 DCIS, 37 DCIS-MI, 68 DCIS-IDC) were available for central pathological review and a TMA containing DCIS lesions of 214 cases (110 DCIS, 36 DCIS-MI, 68 DCIS-IDC) was performed. DCIS morphological features (histological size, nuclear grade, comedo necrosis, inflammation) and immunohistochemical factors (ER, PR, Ki-67 and Her2) were assessed in each of the three groups and compared using Chi2 or Wilcoxon tests.

Results: Median histological DCIS sizes did not significantly differ between DCIS and DCIS-MI groups (70mm [4–160] and 60mm [4–180], respectively, p = 0.33), nor between DCIS and DCIS-IDC (70mm [4–160] and 65 mm [10–140], respectively, p = 0.98). DCIS nuclear grade was significantly higher in DCIS-MI compared to DCIS (p = 0.01), but there was no difference in nuclear grade between DCIS and DCIS-IDC (p = 0.22). Comedo necrosis was more often present in DCIS-MI compared to DCIS (p = 0.04), and no such difference was found between DCIS and DCIS-IDC (p = 0.81). Stromal inflammation surrounding DCIS was more often found in DCIS-MI and DCIS-IDC compared to DCIS (p = 0.03 and p < 0.001, respectively). DCIS-MI was more often ER-negative than DCIS (p = 0.009), and no such difference was found between DCIS and DCIS-IDC (p = 0.6). No other significant differences concerning PR and Her2 status or Ki-67 index were found between the 3 groups.

Conclusions: In patients with extensive DCIS there are few morphological or current immunohistochemical factors differentiating in situ cases from those that have associated microinvasion or invasive ductal carcinoma. DCIS-MI cases have more aggressive histological features than DCIS. Stromal inflammation surrounding DCIS was the only feature distinguishing DCIS from DCIS-IDC.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P3-14-03.
Abstract P4-03-10: Sonographic-pathological correlation in contrast-enhanced ultrasonography in the diagnosis of breast cancers.
Authors: Kato K , Miyamoto Y , Kamio M , Nogi H , Imawari Y , Mimoto R , Toriumi Y , Nakata N , Takeyama H , Uchida K
Published: August 2014

Background: As contrast-enhanced ultrasonography (CEUS) in the diagnosis of breast masses has been performed less frequently than other imaging modalities, it's efficacy evaluation has not been established. CEUS with Sonazoid, which is microbubble-based contrast agent, allows visualization of the mass bloodstream, therefore the classification of enhancement patterns in CEUS could increase diagnostic yield for breast masses. To achieve this, a clear standard for the CEUS evaluation is needed. The purpose of this study was to identify CEUS findings for malignancies and clarify sonographic-pathological correlation of breast cancers.

Material and Method: Present study included 20 patients with invasive ductal cancer who underwent operation for primary therapy without any chemotherapy or hormonal therapy. We contrasted CEUS features with Sonazoid with surgical specimens retrospectively. As malignant findings, the presence of focal perfusion defects inside masses and enhancement of outside masses were chosen.

Result: All of the 20 cases displayed focal perfusion defects. 9 cases of those could be successfully assessed in the same sections histopathologically and ultrasonographically. Histopathological findings showed those defects to be fibrotic foci inside masses in 7cases (77.7%). While, 6 cases of all (30%) showed stains outside tumor, and only one case was found out to be fibrotic growth with cancer cells.

Conclusion: focal perfusion defects of breast masses in CEUS were considered to be specific for malignancy, and they seemed to be fibrotic foci histopathologically. Desmoplastic reaction has been observed in epithelial solid tumor biology, and if those defects reflect the desmoplasia defects could be the proof of malignancy.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-03-10.
Abstract P4-04-02: Tumor-initiated peripheral myeloid cell expansion is reversed by radiation therapy
Authors: Gough MJ , Savage T , Bahjat KS , Redmond W , Bambina S , Kasiewicz M , Cottam B , Newell P , Crittenden MR
Published: August 2014

Myeloid lineage cells, particularly monocytes and macrophages, have an important role in the development and progression of cancer. Within established tumors, macrophages are critical for angiogenesis, invasion, metastasis, immunosuppression and response to therapy. In addition, both transplantable and transgenic mouse models of mammary cancer are associated with myeloid expansions detectible in the tumor, spleen and peripheral blood, and these myeloid cells are able to suppress T cell activation in vitro. Reducing the tumor burden through chemotherapy and resection has been shown to control the myeloid expansion. However, chemotherapies that reduce tumor burden also have direct effects on myeloid cells, and surgical trauma has been shown to mobilize myeloid cells. Radiation therapy is the third major option to treat cancer, but the consequence of radiation therapy to systemic myeloid populations has not been described. We applied tumor-specific radiation therapy to test the hypothesis that the frequency of myeloid lineage cells correlates with the primary tumor burden. We demonstrate in the MMTV-PyMT model of mammary cancer that myeloid expansion in the blood closely correlates with myeloid involvement in the tumor and with tumor progression. We demonstrate that radiation therapy of 4T1 mammary tumors leads to a decline in myeloid cell numbers in the blood and a decrease in spleen size. The frequency of myeloid cells does not decline to the level seen in tumor-free mice: we demonstrate that metastatic disease can prevent myeloid cell numbers from returning to baseline, and that tumor recurrence from residual disease correlates with re-expansion of myeloid lineage cells. Despite these changes in the frequency of myeloid lineage cells, T cell numbers in the blood and spleen remains constant. However, radiation therapy improves the myeloid cell:CD8+ T cell ratio and results in increased proliferation of T cells in the spleen. Finally, we demonstrate that T cell responses to foreign antigens are not altered by tumor burden or myeloid cell expansion, while T cell responses to tumor-associated antigens are increased after radiation therapy of the tumor. These data demonstrate that myeloid cell numbers are directly linked to primary tumor burden, that this population contracts following radiation therapy, and that radiation therapy may open a therapeutic window for immunotherapy of residual disease.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-04-02.
Abstract P4-06-05: NF-κB Upregulates β1-integrin via Increased Transcriptional Activity in Three-dimensional Culture: a Mechanism by which Malignant Breast Cells Acquire Radioresistance
Authors: Ahmed KM , Zhang H , Park CC
Published: August 2014

The molecular mechanisms giving rise to tumor viability after therapeutic radiation remain elusive. We showed previously that β1-integrin is associated with enhanced cell survival of breast cancer cells after exposure to high dose ionizing radiation (IR). Recently, we discovered that a typical binding site for Nuclear factor-κB (NF-κB), a stress-sensitive heterodimeric transcription factor, was located in human β1-integrin promoter, which indicates that β1-integrin-mediated resistance to radiation may be functionally regulated by NF-κB. The aim of the present study was to investigate the relationship between NF-κB and β1-integrin pathways in radioprotection of malignant T4-2 breast cells in three-dimensional laminin-rich extracellular matrix (3D lrECM). We show that NF-κB is required for radiation-induced β1-integrin transactivation. Inhibition of NF-κB activity significantly reduced clonogenic survival with the inhibition of IR-induced total and phosphorylated (Thr 788/789) β1-integrins, correlated with increased DNA binding activity at the β1- integrin promoter. Treatment of T4-2 colonies with NF-κB activation inhibitor before IR resulted in a reduction of the size of colonies, associated with a decrease in proliferation and increase in apoptosis, indicating an increased susceptibility to IR. Further studies revealed that a direct interaction of β1-integrin with α5-integrin was highly induced, but not with NF-κB p65, in T4-2 cells surviving IR. These interactions were found to be oppositely regulated in radiosensitive S1 cells, suggesting that IR induces the formation of α5β1-integrin complexes leading to increased survival post-IR. Together these results provide the first evidence that NF-κB-induced β1-integrin transactivation is responsible for increased survival post IR in breast cancer. Since β1-integrin is shown to activate NF-κB, our data further suggest a loop-like β1-integrin–NF-κB–β1-integrin pathway in tumor radioresistance. This novel pathway may serve as an efficient drug target to re-sensitize radioresistant tumor cells.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-06-05.
Abstract P4-06-08: Clinical utility of functional analysis of FGFR kinase family for selecting patients who may benefit from FGFR inhibitors
Authors: Hoe N , Zhou J , Kuy C , Jin K , Srikrishnan R , Soundararajan A , Ma Y , Liu X , Singh S
Published: August 2014

Background: Fibroblast growth factor receptor (FGFR) family of receptor tyrosine kinases (RTKs) are currently under investigation as therapeutic targets for the treatment of breast cancer. Members of this kinase family have been associated with breast cancer progression and resistance to current therapeutics. FGFR1 amplifications range from 7.5–17% in breast cancers and are associated with a shorter overall survival. FGFR1 amplified cell lines drive activation of AKT, ERK, and RSK and are highly sensitive to FGFR1 inhibition, suggesting an oncogenic addiction to FGFR1. FGFR2 is activated in lapatinib-resistant, HER2-positive cells. Likewise, FGFR3 expression levels are elevated in tamoxifen-resistant, ER positive patients. FGFR 4 overexpression correlated with poor response to chemotherapy due to activation of MAPK and increase in B-cell lymphoma extra large (BCL-XL) levels. Using an immuno-microarray system, this study seeks to determine FGFR signal transduction pathway modulation post FGFR inhibitor treatment in breast cancer cell lines expressing varying levels of FGFR1, 2, 3 or 4.

Method: FGFR 1, 2, 3, and 4 amplified cell lines (MDA-MB-134VI (KRAS), SNU16, RT112, and MDA-MB-453) were treated with varying dosages (1, 10, 100, and 1000nM) of several FGFR inhibitors (AZD4547 (pan FGFR), PD173074 (FGFR1,3) and Ponatinib-AP24534 (FGFR1)) either in presence or absence of corresponding FGF (FGF1, FGF7, FGF9, and FGF19) stimulation. Expression and activation of FGFR 1, 2, 3, and 4 and downstream signaling proteins FRS2, AKT, ERK, MEK, and RSK were measured utilizing a proximity based immuno-microarray.

Results: FGFR1, 2, 3, 4 protein expression was detected in each of the corresponding four FGFR1, 2, 3, 4 amplified cell lines. Total protein levels remained unchanged with or without FGFs stimulation. Levels of FGFR1 phosphorylation increased 5 folds in MDA-MB-134VI stimulated with FGF1. FGFR2 and FGFR3 activation increased by 2–3 folds in SNU16 and RT112 stimulated with FGF7, and FGF9 respectively. FGFR4 activation levels in MDA-MB-453 remained unchanged due to Y367C mutation. ERK was activated in both FGF1 stimulated and unstimulated group in MDA-MB134 due to the KRAS mutation. RSK activation increased in both SNU16 and RT112 when stimulated with FGF7, and FGF9 respectively. FGFR inhibitors treated data analysis is currently in progress and will be presented.

Conclusion: We have demonstrated utility of specific FGFR analysis using well defined model systems. FGFR- profiling should be considered during the clinical work-up in order to select patients who may benefit from regimen containing specific FGFR inhibitors.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-06-08.
Abstract P4-06-15: Mifepristone modifies the tumor microenvironment increasing the therapeutic efficiency of low doses of Doxorubicin liposomes or paclitaxel-albumin nanoparticles in a murine model of breast cancer.
Authors: Sequeira GR , Vanzulli SI , Lamb CA , Rojas PA , Lanari C
Published: August 2014

Chemotherapy is the standard treatment for metastatic breast cancer today. To overcome undesired side effects and improve drug efficacy in tumor cells, different nanoparticle formulations have been developed. Paclitaxel has been bound with human albumin (Nab-paclitaxel), and pegylated liposomes have been developed for doxorubicin delivery (PEG-LD). The aim of our study was to evaluate the effect of these therapeutic agents in experimental mammary carcinomas expressing different ratios of progesterone receptor (PR) isoforms and to evaluate the effect of combined treatments of chemotherapeutic agents with the antiprogestin mifepristone (MFP). Nab-paclitaxel (Abraxane, Abraxis), at a dosage of 30 and 60 mg/kg body weight administered in three doses every four days, completely inhibited the growth of the antiprogestin-resistant C4-2-HI mouse mammary carcinoma, which is associated with high levels of PR isoform B (PR-B) expression. C4-HI, an antiprogestin-responsive mouse tumor that shows high levels of PR isoform A (PR-A) expression, was not inhibited by the low Nab-paclitaxel dose. However, the combination of MFP (6 mg silastic pellet) with Nab-paclitaxel improved the efficacy of both single treatments. Similar experiments were conducted using PEG-LD (Doxopeg; Laboratorios Raffo). In this case, all tumors tested (C4-HI, CC4-3-HI, 59–2-HI and C4-2-HI) were all highly responsive to high (18 mg/kg/week) or low (9 mg/kg/week) PEG-LD doses. When PEG-LD concentrations were lowered to 4.5 mg/kg/week and they were combined with MFP, an improved response was observed with combined treatments only in tumors with high PR-A levels (p <0.001). Using BALB/c-GFP mice, we demonstrated that MFP treatment induced tissue remodeling in PR-A positive tumors, increasing the stromal/tumor cell ratio and the number of vessels (as shown by CD31 positive cells; p < 0.001). Taking advantage of doxorubicin's red autofluorescence, we showed increased levels of drug accumulation in tumor cells surrounding the stromal tissue. Using a human albumin antibody we demonstrated nab-paclitaxel accumulation in similar locations. However, because increased areas (p < 0.001) of stromal tissue were observed in the antiprogestin-treated mice, the total amount of PEG-LD or Nab-paclitaxel was much higher in the combined-treated tumors. We propose that antiprogestins have the potential to enhance the efficacy of nanotherapeutics in patients with breast tumors with a high PR-A/PR-B ratio.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-06-15.
Abstract P4-09-06: miR-187 is an independent prognostic factor in lymph node-positive breast cancer patients.
Authors: O'Connor DP , Mulrane L , Brennan DJ , Madden S , Gremel G , McGee SF , McNally S , Martin FM , Crown JP , Jirstrom K , Higgins DG , Gallagher W
Published: August 2014

Purpose: MicroRNAs (miRNAs) involved in cancer progression have now become the focus of much attention as they represent a new class of biomarkers and potential drug targets. Here, we describe an integrated bioinformatics, functional analysis and translational pathology approach for the identification of novel miRNAs involved in breast cancer progression.

Experimental Design

Differential gene expression can, in part, be attributed to the activity of specific miRNAs. Given a database of miRNA binding site motifs and gene expression levels determined by transcriptomic profiling, correspondence analysis, between group analysis and co-inertia analysis can be combined to produce a ranked list of miRNAs associated with a specific gene signature and phenotype. Here, using two independent breast cancer cohorts, this approach was employed to produce a ranked list of miRNAs associated with disease progression. Functional studies were subsequently carried out in MCF7 cells assessing for alterations in growth, tumorigenicity and agressiveness and miRNA expression was evaluated in two cohorts of breast cancer patients by locked nucleic acid in situ hybridisation on tissue microarrays.

Results: CIA identified miR-187 as a key miRNA associated with poor outcome in breast cancer.

Ectopic expression of miR-187 in MCF7 cells resulted in a more aggressive phenotype (evidenced by increased anchorage-independent growth, migratory and invasive potential).

In a test cohort (n = 117) breast cancer patients, high expression of miR-187 was associated with a trend towards reduced breast cancer-specific survival (BCSS) (p = 0.058), and a significant association with reduced BCSS in lymph node-positive patients (p = 0.036). In a validation cohort (n = 470), high miR-187 was significantly associated with reduced BCSS in the entire cohort (p = 0.021) and, again, in lymph node-positive patients (p = 0.012).

Multivariate cox regression analysis revealed that miR-187 is an independent prognostic factor in both TMA cohorts (Cohort 1 HR-7.369 (95% CI 2.048–26.509, p = 0.002); Cohort 2 HR-2.798 (95% CI 1.518–5.157, p = 0.001).

Conclusions: miR-187 expression in breast cancer leads to the formation of a more aggressive, invasive phenotype and acts as an independent predictor of outcome.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-09-06.
Abstract P4-13-03: Hormone replacement therapy, is there an increased risk of in situ breast cancer ? data from a french cohort
Authors: Coussy F , Giacchetti S , Hamy A-S , Porcher R , Cuvier C , Lalloum M , de Roquancourt A , Albiter M , Espié M
Published: August 2014

Background: Use of hormone replacement therapy (HRT) has been associated with an increased risk of breast cancer. Few studies focus on correlation between HRT and in situ breast cancer A large cohort study, the Million Women Study (Reeves GK et al. Lancet Oncol. 2006) have shown that the use of HRT has been associated with an increased incidence of in situ breast cancer (RR = 1, 55, IC=1, 4–1,72, p = 0,03).

Purpose: We investigated the association between HRT use and duration and in situ breast cancer incidence from a data base of infraclinic breast lesions.

Materials & Methods: From 2007 to 2011, 2708 patients (pts) with a non palpable breast lesion were referred to our breast disease unit for exploration (Saint Louis Hospital, Paris). Radiological abnormalities were screened by mammography and/or breast ultrasound, and/or MRI. Out of 2708 pts, 1668 had a biopsy. All biopsies were seen by an anatomopathologist dedicated to breast. Here, we focus on the 1017 postmenopausal women (61%).

Results: Biopsies revaled invasive breast cancers in 222 pts (22%), in situ breast cancers in 164 pts (16%) [10 lobular in situ carcinoma and 154 ductal in situ carcinoma], high risk breast lesions in 103 pts (10%) and benign breast lesions in 528 patients (52%). Characteristics of the 164 patients with in situ breast cancer: median age 62 [IQR: 57 to 68], at least one pregnancy (mean number of pregnancies per woman, 1.6) 75,6%, family history of breast cancer:30,2%. Among these 164 patients, 42.6 % had used an oral contraceptive and 53% of them a HRT, with a mean duration of use of 7 years.

The HRT use was not significantly associated with in situ breast cancer (p = 0.66) as well as the duration of HRT.

Discussion: In this study of postmenopausal women, HRT use, whatever is the duration, was not associated with in situ breast cancer. Although a lack of statistical power may be invoked to explain these results, these findings suggest that the type of the HRT could be important to explain the difference between our study and the Million Women Study. In our study, the HRT used is in the majority of the pts, percutaneous oestrogens and natural progesterone as it is the most commun HRT used in France (Fournier A, Int J Cancer. 2005) during this period of time. Use and type of HRT are different in USA where conjugated estrogens and medroxy progestogerone acetate were commonly used.

Conclusion: We therefore encourage further studies to better appreciate the links between type of HRTs and in situ breast cancer.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-13-03.
Abstract P4-13-09: The effect of weight change on breast adipose and dense tissue
Authors: Graffy HJ , Harvie MN , Warren RM , Boggis CR , Astley SM , Evans GD , Adams JE , Howell A
Published: August 2014

Background Observational studies indicate that weight gain or loss alters breast cancer risk. Hormonal prevention with tamoxifen or ovarian suppression reduces mammographic breast density, but has no impact on breast adipose tissue.1 The aim of this study was to assess whether change in weight is associated with change in breast density, breast fat or both.

Methods 74 premenopausal women aged between 35 and 45 years, with a family history of breast cancer, were recruited from our Family History Clinic. 36 were allocated to a 25% calorie-restricted Mediterranean diet and exercise intervention; a further 38 women were separately recruited into a control group and given written diet and exercise advice only. At baseline and 12 months breast dense and non-dense (largely adipose) components of the breast were measured: area using Cumulus2 and volume using the Manchester ‘stepwedge’ method.3 Visual assessment of percentage density was reported to the nearest 5%.4 We used correlations to relate percentage change in absolute dense and non-dense (area and volume) to percentage change in: weight, total body fat (DXA), intra-abdominal and subcutaneous abdominal fat (MRI), as well as serum total IGF-I.

Results At 12 months, 40 (54%) women lost >1.5% body weight, 14 (19%) gained >1.5% and 21 (28%) remained weight stable, <+/−1.5% weight change. There was a range of percentage weight change from −16.3% to +9.8%. There were no significant changes in dense area (r = −0.14 p = 0.29) or volume (r = −0.01 p = 0.97) with weight change. However, both the area and volume of non-dense breast tissue changed significantly in relation to change in body weight (r = 0.52 p < 0.0001 & r = 0.78 p < 0.0001 respectively), total body fat (r = 0.49 p < 0.001 & r = 0.75 p = <0.0001 respectively), intra-abdominal fat (r = 0.45 p < 0.0001 & r = 0.36 p = 0.03 respectively) and subcutaneous abdominal fat (r = 0.55 p < 0.0001 & r = 0.52 p = 0.002 respectively). Visually assessed percentage mammographic density was significantly inversely related to weight (r = −0.27 p = 0.038). Serum total IGF-I was not significantly correlated with breast dense area. (r = −0.21 and p = 0.09).

Conclusions These data indicate that dense area and volume remain stable irrespective of change in weight. Non-dense area changes significantly in response to weight change in parallel with total and abdominal fat stores. These observations were performed on relatively small numbers of women and require confirmation in larger series.

1. Cuzick et al J Natl Cancer Inst. 103(9): 744–52, 2011. 2. Boyd NF et al J Natl Cancer Inst. 87(9) 670–675 1995. 3. Diffy et al IWIDM 2006, LNCS 4046 p1-9. In Astley et al (Eds) Springer-Verlag Berlin Heidelberg 2006 4. Brisson J et al. Cancer Epidemiol Biomarkers Prev 2003;12:7288–32.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-13-09.
Abstract P4-13-10: Do BRCA1 and BRCA2 mutation carriers have an earlier natural menopause than their non-carrier relatives: A study from the Kathleen Cuningham Foundation Consortium For Research Into Familial Breast Cancer (kConFab).
Authors: Collins IM , Milne RL , McLachlan SA , Friedlander M , Birch KE , Weideman PC , Hopper JL , Phillips K-A
Published: August 2014

BACKGROUND: Limited data from the in vitro fertilisation setting suggest that germline BRCA1 mutations are associated with occult primary ovarian insufficiency. We examined whether carriers of a BRCA1 or BRCA2 mutation have earlier natural menopause than their non-carrier relatives.

PATIENTS AND METHODS: Eligible subjects were female mutation carriers and tested non-carriers from families segregating BRCA1 or BRCA2 mutations enrolled in KConFab. Women who had not been tested for the family mutation and those with primary amenorrhoea were excluded. Data were self-reported using uniform questionnaires completed at cohort entry and every 3 years thereafter. Age at natural menopause (NM) was defined as age at cessation of menses for ≥ 12 months in the absence of other causes. Cox proportional hazards analysis was applied, modelling time in years from birth to NM, adjusting for year of birth (continuous), body mass index (continuous), ever regular cigarette smoking (no, yes), and ever regular alcohol use (no, yes). Analysis time was censored at the earliest of: cessation of menses for 12 months in the absence of other reasons (natural menopause), last follow-up, pelvic surgery including oophorectomy or hysterectomy, commencement of hormone replacement therapy or Tamoxifen as a risk reducing medication, or any cancer diagnosis. Hazard ratios (HR) were estimated as a measure of how rapidly mutation carriers reach natural menopause relative to non-carriers.

RESULTS: No difference was observed in the rate at which carriers and non-carriers went through natural menopause, regardless of the gene mutated in the families. The adjusted hazard ratio estimates were 0.97, (95% CI 0.69–1.35, p = 0.8) for BRCA1 mutation carriers versus non-carriers and 1.02, (95%CI 0.72–1.44, p = 0.9) for BRCA2 mutation carriers versus non-carriers.

CONCLUSIONS: This study found no evidence that BRCA1 and BRCA2 mutation carriers do reach natural menopause at an earlier age than their non-carrier relatives.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-13-10.
Abstract P4-14-09: Feasibility of liposuction for treatment of arm lymphedema from breast cancer.
Authors: Doren EL , Smith PD , Sun W , Lacevic M , Fulp W , Reid R , Laronga C
Published: August 2014

Background: Lymphedema is a dreaded complication of breast cancer treatment affecting 20% of women having axillary node dissection. Liposuction minimizes unwanted fat in targeted areas. Our objective was to explore the feasibility of liposuction to reduce fat volume and thus arm lymphedema.

Methods: An IRB-approved prospective trial was conducted of women having unilateral arm lymphedema resulting from breast cancer treatment. At enrollment there was no evidence of cancer recurrence or arm cellulitis. Arm measurements (circumferential), volumes (water displacement and geometric calculation), and muscle strength differences between the affected and unaffected arms and quality of life/functionality were measured pre-operatively and post-operatively at 6 weeks, 6 months and one year(s). Descriptive statistical analysis was performed.

Results: Six breast cancer survivors underwent the liposuction procedure from 12/2008–4/2011. Median age was 54 yrs (range: 43–60) and median volume of fat aspirated was 700mls (range: 350–700). Average volume difference between the affected and unaffected arms at baseline was 522.5 mls (176–867) (geometric) and 589.2mls (280–770) (water displacement). No immediate complications; 1 cellulitis at 4 months post-operative. Average percent volume reductions for 5 of the 6 women at 6 weeks, 6 months and 1 year were 70%, 47%, 71% mls geometrically and 63%, 18%, 54% by water displacement respectively. Quality of life and functionality improved in all patients. Muscle strength remained unchanged. Pain lessened. Average follow-up is 15.49 months (range: 1.8–24.84 months).

Conclusion: Liposuction can safely reduce volume of arm lymphedema and improve functionality/quality of life. Larger studies (longer follow-up) are required to validate the durability of these early results.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-14-09.
Abstract P4-14-13: Therapeutic mammaplasty does not cause a delay in the delivery of chemotherapy in high risk breast cancer patients
Authors: Khan J , Barrett S , Stallard S , Forte C , Weiler-Mithoff E , Reid I , Winter A , Doughty J , Romics L
Published: August 2014

Introduction: oncosurgical safety of therapeutic mammaplasty (TM) is widely investigated. The interval between surgery and delivery of adjuvant chemotherapy is an integral part of overall oncological safety. Therefore, we examined the time between TM and AC, and compared it to wide local excision (WLE) and mastectomy (Mx) with or without immediate breast reconstruction (IBR), respectively.

Methods: data of 174 patients who underwent TM, WLE and Mx±IBR was analyzed retrospectively. All patients were operated within three breast units of Glasgow during a period of 48 months. Time between decision to offer adjuvant chemotherpay and delivery of the first cycle of chemotherapy was analyzed. Significance was calculated with Mann-Whitney and Kruskal-Wallis tests (two and four groups compared, respectively).

Results: median time to adjuvant chemotherapy after TM (n = 36) was 29 [16–58] days, WLE (n = 66) was 29.5 [15–105], Mx only (n = 56) was 29 [15–57], and Mx and IBR (n = 16) was 31 [15–58] days. No significant difference was found in terms of time to adjuvant chemotherpay in patients treated with TM compared to WLE (p = 0.384), Mx only (p = 0.828) or Mx and IBR (p = 0.366). Further, there was no significant difference when a cumulative comparison of the four groups was carried out (p = 0.507).

Conclusions: our data indicate that oncosurgical safety of TM in terms of time to chemotherapy is similar to other high risk breast cancer patients treated WLE and Mx with or without IBR. This also suggests that there is no significant difference in postoperative complication rates after these four ways of surgical treatment of breast cancer, which would possibly be the primary cause for a delay in delivering adjuvant chemotherapy.

Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-14-13.