bluuebunny/crossref_metadata_2025 huggingface dataset:
P1-08-13: Determinants of Risk, Characteristics and Prognosis of Breast Cancer Occurring after Hodgkin Lymphoma.
Authors: Rapiti E , Veit-Rubin N , Usel M , Benhamou S , Vlastos G , Vinh Hung V , Bouchardy C
Published: June 2012

Background Several studies have evaluated the excess of breast cancer (BC) risk after Hodgkin lymphoma (HL) but none has reported of all the aspects of BC occurring after HL including determinants of risk, comparison with other BC profile, treatment and outcome. This is the aim of our study.

Patients and Methods We used data from the Surveillance, Epidemiology and End Results (SEER) Programme of the National Cancer Institute. To assess the risk and determinants of BC after HL we included all 9'620 women with first primary HL diagnosed between 1973 and 2007 and calculated age and period standardized incidence ratio (SIR) of BC as compared with general population. We also compared tumour characteristics and treatment between BC after HL (n= 316) vs. all other first primary BC occurring during the same period (n=450'413) by logistic regression. Finally, we compared breast cancer specific mortality between the two groups by Cox model.

Results Overall, HL patients had an increased risk of developing BC (SIR: 2.4, 95% confidence interval [CI]: 2.2−2.7); the risk decreased with increasing age at HL, and disappeared when HL occurred after 50 years. BC risk was the same among those treated with or without radiotherapy until 15 years after HL and then an excess appeared in the irradiated group and persisted over 30 years after diagnosis of HL. Compared with other BC, BC after HL occurred at younger age (Adjusted odds ratio [OR] for age 40–59 vs. < 40 years 0.25, 95% CI: 0.20−0.33), was diagnosed at an early stage (OR for regional extension vs. localized 0.63, 95% CI: 0.42−0.89), expressed less frequently hormone receptors (OR for negative vs. positive status: 1.34, 95% CI: 0.99−1.81), was located more frequently in external quadrants (OR for internal vs. external: 0.61, 95% CI: 0.42−0.89), and received less frequently radiotherapy (OR for use vs. non use: 0.30, 95%CI: 0.22-042). The last two results were limited to patients who had received radiotherapy for HL. BC after HL presented higher breast cancer mortality (Hazard ratio: 1.36, 95%CI: 1.05−1.76) after adjustment for patients, tumour and treatment characteristics.

Conclusion: This study provides valuable results on various aspects of BC occurring after HL. In particular, we found earlier stage at diagnosis of BC probably linked to higher surveillance, more aggressive tumours probably due to iatrogenic effect of HL treatment, more frequent occurrence in outer quadrants less protected during mantel radiation. Also, this study confirms the poorer prognostic of those BC not explained by differences in patient, tumour or treatment characteristics.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-08-13.
P1-08-25: Breast Cancer Burden, Risks and Outcomes in Latin America.
Authors: Justo NA , Wilking NE , Jönsson B
Published: June 2012

BACKGROUND: Breast cancer (BC) burden and care vary substantially across Latin America (LA). Aim: Provide an overview of the epidemiology, outcomes and countries’ risk profiles in LA to identify opportunities for improvement.

METHODS: Review of literature (PubMed, LILACS, SCIELO), public databases (Globocan 2002 & 2008, CEPALSTAT, DIRAC, PAHO, WHOSYS, etc) and conference presentations (ASCO, ISPOR). Local experts & patient organizations were surveyed. Current BC epidemiology in LAC, future trends, countries’ risk profiles and outcomes (such as mortality-to-incidence ratios — MIRs) were analyzed and compared (Sweden as benchmark).

RESULTS: BC annual incidence (114,900 cases) & mortality (37,000 deaths) are the highest of all cancers in LA and twice as many deaths are expected by 2030. Countries more advanced in the demographic transition, like Uruguay and Argentina, present much higher incidence and mortality but also better outcomes as observed in their MIRs. In LA women are diagnosed younger and at a more advanced stage, especially in countries with poor outcomes, such as Ecuador, Mexico or Peru. The imbalanced progress since 2002 seems to widen the gap in BC outcomes, but countries like Costa Rica are catching up.

Age is the main risk factor; while the association of education and wealth with BC occurrence is less significant. Reproductive and lifestyle-related risk factors, proven at a patient level, were not significant at a country-aggregate level and the specialized literature indicates that the evidence on associations between BC and diet is inconclusive.

DISCUSSION: LA's considerable variation embedded within per-countries’ averages may be masking underlying causality. Though, it can be inferred that there is no one-suit-all prevention campaign and policies aimed at reducing the risk of developing BC will succeed as long as they are properly targeted and tailored for different needs. Early diagnosis and population-wide access to treatment following evidence-based best practices remain unresolved matters that endow many of these countries with the opportunity to substantially improve the care for their women.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-08-25.
P1-09-07: Contribution of TP53 p.R337H Mutation to Breast Cancer Incidence in Brazil.
Authors: Giacomazzi J , Toledo de Bueno Osorio CA , Koehler-Santos P , Graudenz MS , Marthel-Planche G , Achatz MIW , Soares FA , Goldim JR , Caleffi M , Hainaut P , Camey SA , Ashton-Prolla P
Published: June 2012

Background: The exact contribution of TP53 germline mutations, associated with Li Fraumeni Syndrome — LFS, to the overall burden of cancer is still only partially known. Studies in Southern and Southeastern Brazil have shown that a particular mutant, TP53 p.R337H, has incomplete penetrance and may be present in a significant number of subjects (estimated frequency at the populational level of 1:300 individuals). In an exploratory approach, the aim of this study is to assess the prevalence of the TP53 p.R337H mutation in women with breast cancer diagnosed before 46 and after 55 years of age, unselected for family history of cancer and resident in Southern and Southeastern Brazil.

Methods: Formalin-fixed paraffin-embedded (FFPE) non-tumoral tissue of 521 women diagnosed with breast cancer between 2000 and 2010 in two pathology laboratories were obtained retrospectively and consecutively, and analyzed after anonimization. Genomic DNA was isolated with the QIAamp DNA Tissue Kit and genotyping performed by allelic discrimination using a TaqMan assay. Confirmation of all mutation-positive and a sample of mutation-negative cases was done by TP53 exon 10 sequencing.

Results: Analysis of the first 299 cases identified the TP53 p.R337H mutation in the germline of 15 (5,0%) cases: 13/142 (9,2%) before 46 years and 2/157 (1,3%) diagnosed after 55 years. The p53 expression pattern assessed by immunohistochemistry in the breast tumors was not different between p.R337H mutation carriers and non-carriers. Conclusion: Preliminary analysis in a sample of women with breast cancer in Southern Brazil indicates that the germline TP53 p.R337H founder mutation is present in a high proportion of cases, especially those diagnosed at a young age. Germline TP53 mutations are considered rare, occurring in about 1:5000 individuals of the general population. The occurrence of this founder mutation at such a high frequency in a particular geographic region has important implications for disease management and cancer risk counseling for these patients and families. This mutation likely contributes to a significant proportion of the health burden associated with breast cancer in Southern Brazil.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-09-07.
P1-10-03: Colony Stimulating Factor Use with Taxane-Based Therapy for Metastatic Breast Cancer: Claims Analysis of Prophylaxis, Treatment, and Costs.
Authors: Force RW , Pugmire BA , Culbertson VL
Published: June 2012

Background: The taxanes (paclitaxel, docetaxel, and albumin-bound paclitaxel) are used in women with metastatic breast cancer (MBC). Clinical trials indicate that these drugs may differ in their toxicity profiles, including rates of neutropenia and need for colony stimulating factor (CSF) supportive therapy. Utilizing medical claims data, we evaluated prophylactic and treatment-related CSF use among women receive taxane-based chemotherapy regimens.

Objective: To determine if differences exist in rates of CSF use for prophylaxis, treatment and associated costs in women receiving taxane-based chemotherapy for MBC.

Methods: Women with MBC were identified with ICD-9-CM codes and by their prior use of adjuvant chemo regimens. Paid medical claims (source: Ingenix Consulting) from May 1, 2006 to April 30, 2009 were analyzed. Study groups were defined according to the first taxane administered. CSF utilization was classified as prophylaxis (0-5 days post-taxane administration), treatment (6-21 days post-taxane,) or non-taxane associated use (>21 days post-taxane). Comparisons were made between the taxane groups. All CSF costs were captured from date of first taxane to end of taxane therapy +21 days and categorized as above. Patients censored if ≥35 days elapsed with no taxane therapy. Two-stage regression analyses were performed. Control variables included age, Romano comorbidity score, and use of prior and other concurrent chemotherapy. Results: 4,503 women (mean age: 53 +/− 10 years) with MBC were identified: 2,599 in the docetaxel group; 1,643 received paclitaxel; and 261 received nab-paclitaxel. More patients receiving docetaxel received any CSF (75.6%) than either paclitaxel (49.8%) or albumin-bound paclitaxel (36.8% P<0.05 for each comparison). For docetaxel 70.5% of women received prophylactic CSF, whereas the rates were 47.0% for paclitaxel and 33.0 for albumin-bound paclitaxel (P<0.05). The rate of women receiving CSF in the albumin-bound paclitaxel group was not different than the other groups. Daily per-patient CSF expenditures during taxane therapy were $176.81 (95% CI, $168.20-$185.52) for docetaxel, $127.71(95% CI, $113.62-$142.60) for paclitaxel, and 46.24 (95% CI, $29.37-$67.25) for albumin-bound paclitaxel.

Conclusions: Compared with docetaxel and paclitaxel, patients receiving albumin-bound paclitaxel had significantly lower prophylactic CSF use, yet did not experience any difference in treatment-related use. Daily per-patient CSF expenditures for albumin-bound paclitaxel were significantly lower than the other two taxanes.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-10-03.
P1-10-07: Comparing Breast Cancer Screening Guidelines: A Stage-Survival-Cost Model in a Public Hospital.
Authors: Friedman DT , Raskind-Hood C , Adams K , Becker E , Habtes I , D'Orsi C , Gundry K , Birdsong G , Gabram-Mendola S
Published: June 2012

Background: Despite advances in diagnosis and treatment of breast cancer in the United States, racial disparities in survival persist. Female breast cancer represented the costliest cancer site in 2010 with further increases projected to 2020. Mammography screening guidelines reflect the inherent difficulty in reconciling mortality reduction with potential harms to patients and cost-effectiveness concerns. Controversy still surrounds the 2009 updated USPSTF recommendations, as critics and advocates alike evaluate lives saved, costs, and how best to compare screening strategies. This study simulates USPSTF and ACS guidelines’ effects on stage, 5-year survival, and cost of treatment vs. usual practice in women treated in an urban public hospital.

Methods: Charts of 84 patients diagnosed with Stage I-III breast cancer in 2008 were reviewed. Published tumor doubling times guided a retrospective model to predict size at diagnosis by simulated ACS or USPSTF guidelines. AJCC-7 stages were then assigned to produce 3 distributions: 1) actual stage; 2) stage under ACS; and 3) stage under USPSTF. Survival estimates by stage and year from diagnosis were drawn from NCDB statistics and applied to each stage distribution, yielding average predicted survival for the actual and hypothetical scenarios. Finally, treatment costs for women continuously enrolled in Medicaid for 18 months were calculated from merged claims and registry data and similarly applied.

Results: Study patients averaged 55 years; 85% were African American. Forty-nine percent were covered by Medicaid and 23% by Medicare. Comparing actual and predicted stages at diagnosis showed significant shifts in stage distribution between all three scenarios (Table 1).

ACS guidelines predicted higher survival (87.5% at 5 years) than actual (85.5%; p<0.01) or USPSTF (84.6%; p<0.0001), while no significant difference was revealed between actual and USPSTF. ACS guidelines also predicted lower costs of treatment relative to USPSTF and actual, while USPSTF guidelines were more expensive than actual (Table 2).

Discussion: To our knowledge, no studies have compared the impact of alternative screening guidelines on outcomes in a predominantly African-American, public hospital population. These data support continued use of and adherence to ACS screening recommendations for inner city patients to achieve the best survival and the lowest cost. Given the likelihood of patients to be uninsured or covered by federal programs, this study carries implications for public policy and patient education, especially in low-resource programs caring for underserved patients.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-10-07.
P1-11-10: Quality of Breast Cancer Care in a Boston Area Patient Navigator Program.
Authors: Raj A , Ko N , Battaglia T , Moy B
Published: June 2012

Background: Elimination of disparities is critically important for lessening the burden of cancer. Patient navigator programs (PNPs) assist with all aspects of care, including access, cancer prevention, screening, post-diagnosis care, and survivorship care. Little is known about the effect of PNPs on patient care and outcomes following the diagnosis of breast cancer (BC). We examined quality measures (QMs) of breast cancer care among women participating in the Massachusetts General Hospital Avon Breast Care Patient Navigator Program (MABCP), which provides patient navigation services to disadvantaged minority communities in the greater Boston area.

Methods: Women diagnosed with BC who participated in the MABCP from 2001 to 2011 were followed to determine the proportion whose care was concordant with American Society of Clinical Oncology/National Comprehensive Cancer Network (ASCO/NCCN) QMs. QMs included 1) hormonal therapy (HT) within 1 year of diagnosis for HR+ tumors > 1 cm; 2)chemotherapy within 120 days of diagnosis of HR- >1cm tumors for women <70 years; and 3) post-lumpectomy radiation therapy (XRT). Descriptive statistics were used to report characteristics of MABCP patients.

Results: Of the 186 MABCP patients diagnosed with BC, some treatment data was available on 158 (85%) and race/stage information was available on 149 (80%) [Table 1]. Among the MABCP patients, concordant care was received by 70/74 (95%) for the HT QM, 15/17 (88%) for the chemotherapy QM, and 65/71 (92%) for the XRT QM. In comparison, available benchmark concordance rates of BC patients treated at 8 NCCN centers from 2003–6 are: 340/382 (89%) for the HT QM, 156/179 (87%) for the chemotherapy QM, and 141/148 (95%) for the XRT QM.

Conclusions: Overall, breast cancer care in the MABCP PNP is concordant with published ASCO/NCCN quality measures. At present, the sample is insufficient to compare concordance rates with NCCN patients but preliminarily, it appears that the quality of care is comparable. Future research should include prospective analyses of quality metrics to assess the process and outcomes of patient navigation in diverse settings, compared with control populations.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-11-10.
P1-11-11: Wait Times for Breast Cancer Care in Manitoba 2009–2010. Time To Face the Challenge.
Authors: Carpenter-Kellett T , Nashed M
Published: June 2012

Background: Wait times for patients with breast cancer to receive oncological treatment vary significantly. In Manitoba, the radiation treatment wait times improved in the year 2005 compared to 2001. However, other parts of the patient's journey have lengthened and negated the reduction seen in radiation treatment wait times. Aims: To examine the overall time from disease suspicion to treatment of breast cancer patients from June 2009 to June 2010 and to compare to the previously published wait times.

Methods: This population-based retrospective study looked at representative samples of women newly diagnosed with breast cancer. Patients were followed from the time of first presentation, either to their family physician or after a suspicious screening mammogram, to the time adjuvant treatment was started. Each patient's journey was subdivided into different chronological stages. The data was compared to the wait times reported in 2005.

Results: 363 patients’ data was collected and analyzed. The median elapsed time in days for each phase of the journey was calculated. Total wait times from suspicion to diagnosis were also calculated. The wait times of most stages of the patients’ journey have worsened when compared to 2005. However, there was some improvement in the diagnostic part of the journey. Delays to surgery and pathology have contributed to lengthening of the total journey. There was also a systematic and repetitive administration delay at each transition. Screen-detected cases had a shorter journey than those who presented through a family physician.

Conclusions: In spite of improvements achieved in the wait times at some stages of the journey, the total length of the patient journey has not been shortened. This represents not only individual team failures but also a built in complexity in the health system. In order to achieve a meaningful cut in wait times for breast cancer patients, the entire trajectory of care needs to be addressed.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-11-11.
P1-12-01: Pregnancy during and Following Adjuvant Trastuzumab in Patients with HER2−Positive Breast Cancer: An Analysis from the HERA Trial (BIG 01-01).
Authors: Azim HA , Metzger-Filho O , de Azambuja E , Loibl S , Focant F , Gresko E , Procter M , Piccart-Gebhart M
Published: June 2012

Background: 1-year of adjuvant trastuzumab (T) is the standard of care in managing patients (pts) with early HER2−positive breast cancer (BC). As T is not known to alter fertility, pts with childbearing potential could become pregnant during or following treatment with T. Cases of oligohydramnios, some associated with fatal pulmonary hypoplasia of the fetus have been reported in women receiving T during pregnancy (preg). Here we report the outcome of preg in all pts enrolled in the HERA trial.

Methods: Pregnancies in the HERA trial are reported on a distinct “pregnancy form” for up to 10 years following T completion. The form includes information on approximate date of conception, preg course & outcome, fetal measurements at birth, and congenital anomalies. Any missing data were retrieved from the study site, if available. For this analysis, pts were grouped into 3 groups: 1) preg during and up to 3 months after T, 2) preg > 3 months of last T dose, and 3) preg with no prior exposure to T.

Results: By March 2010, 70 preg were reported in 58 out of 5102 pts randomized. Five, 30 and 7 completed preg were reported in groups 1, 2 & 3 respectively.

As per protocol, all pts on T were required to use adequate contraceptive measures, yet 16 pts became pregnant during the course of T and up to 3 months thereafter. The percentage of completed preg was lowest in group 1, with 4 spontaneous and 7 induced abortions. In group 2, preg occurred at a mean of 29 months following completion of T, with 6 spontaneous and 4 induced abortions. In group 3, abortion was induced in 3 pts and no spontaneous abortions reported. Across all 3 groups, all but 1 spontaneous abortion occurred during the 1st trimester. Two congenital anomalies were reported; a Down's syndrome in a 43 year old pt >5 years after completing T for which abortion was induced, and one with partial fusion of the 2nd and 3rd toe born to a pt in group 3.

Conclusions: Unintentional exposure to T during preg may be associated with spontaneous abortion, yet the numbers remain low to draw firm conclusions (spontaneous abortion rate in general population is up to 20%). No oligohydramnios or anomalies were observed in group 1. While an increased risk of oligohydramnios has been reported when T is administered after the 1st trimester, T administered to Cynomolgus monkeys during organogenesis did not cause fetal harm (Pentsuk et al; 2009). Nevertheless, women of childbearing potential should be advised to use effective contraception during and up to 6 months after treatment with T. On the other hand, prior exposure to T did not appear to affect the preg course or outcome. We are planning to collect information from the other T adjuvant trials to confirm our findings.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-12-01.
P1-12-06: The Role of MAPK and PI3K/AKT/mTOR Signaling in Innate Lapatinib Resistance.
Authors: McDermott M , O'Brien N , McDonald K , Crown J , O'Donovan N , Slamon D
Published: June 2012

Background: Lapatinib is a tyrosine kinase inhibitor which blocks downstream MAPK and P13K/AKT/mTOR proliferation and survival signaling pathways in HER2 positive breast cancer cell lines, tumor xenografts and HER2 positive breast cancer patients. However, pre-clinical and clinical studies have shown that not all HER2 positive patients respond to lapatinib and thus innate resistance to lapatinib exists. The identification of biomarkers of lapatinib response is therefore critical and would enable individual therapeutic decisions to be based on tumor biology rather than basic histopathology data alone. The aim of this study was to examine the role of MAPK and PI3K/AKT/mTOR signaling in a panel of lapatinib sensitive and resistant HER2−amplified breast cancer cell lines to identify pharmacodynamic markers of response to lapatinib treatment.

Methods: Dose response curves were generated to determine sensitivity to lapatinib in a panel of 17 HER2−amplified breast cancer cell lines. Total and phosphorylated levels of HER2, HER3, EGFR, AKT, ERK, S6K and eEF2 were determined following 24 hours lapatinib treatment in each of the cell lines.

Results: Twelve of the cell lines were sensitive to lapatinib with IC50g < 1 μM while 5 of the cell lines were innately resistant to lapatinib with IC50g > 1 μM. Levels of pHER2 and pHER3 were decreased in response to lapatinib in all cell lines independent of sensitivity to lapatinib. This suggests that inhibition of HER2/HER3 activation is not indicative of response to lapatinib treatment. There was also no correlation between the levels of HER2, HER3, and EGFR and sensitivity to lapatinib in the cell line panel.

In lapatinib sensitive cell lines, lapatinib decreased PI3K (pAKT), mTOR (pS6K) and MAPK (pERK) signaling and increased peEF2 levels. In contrast the levels of pAKT, pS6K, peEF2 and pERK were maintained following lapatinib treatment in lapatinib resistant cells. The continued activation of these proteins in the presence of lapatinib suggests a possible feedback mechanism that warrants further investigation. These data confirm that maintained signaling through either the P13K/AKT/mTOR pathway or the MAPK pathways in the presence of lapatinib can be an early pharmacodynamic biomarker of response.

Conclusions: Maintenance of pAKT, pS6K, peEF2 and pERK levels, in response to lapatinib treatment correlates with lapatinib resistance. These data suggest that alterations in the P13K/AKT/mTOR and MAPK pathways play an important role in innate lapatinib resistance and pharmacologically targeting these pathways is a rational therapeutic approach for overcoming innate lapatinib resistance.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-12-06.
P1-12-18: Change in HER2 Status in HER2 Positive Operable Breast Cancer Patients Treated with Neoadjuvant Chemotherapy with or without Anti-HER2 Therapy: Analysis of Two Consecutive Cohorts.
Authors: Barbieri E , Piacentini F , Dieci MV , Ficarra G , Bettelli S , Conte P , Guarneri V
Published: June 2012

Introduction: emerging literature data have shown a change of HER2 expression from primary tumors to metastatic deposits. Tumor heterogeneity, genetic drift as well as selective pressure of adjuvant therapy have been suggested to explain this phenomenon. Aim of the present analysis is to evaluate the change in HER2 expression after neoadjuvant chemotherapy with or without anti-HER2 agents.

Methods: two consecutive cohorts of HER2+ breast cancer patients treated with neoadjuvant therapy were identified from a prospectively maintained database including 310 patients. The first cohort (A) includes 38 patients enrolled before 2005, treated with chemotherapy alone. The second cohort (B) includes 48 patients treated with neoadjuvant chemotherapy in combination with antiHER2 agents (trastuzumab or lapatinib).

HER2 expression was evaluated by IHC on pre-treatment core biopsy (tru-cut with 14 gauge needle) and on surgical specimen after neoadjuvant therapy. FISH analysis was performed on IHC 2+ samples.

Results: The two cohorts were balanced in respect of tumor stage, patient age, and HR expression. In particular, a co-expression of HER2 and HR was observed in 60% of the patients in cohort A and in 70% of the patients in cohort B (p=0.2).

Patients in cohort B have a significantly higher rate of pathologic complete response (pCR) in comparison to cohort A (45% vs 11%, p=0.001). A change in HER2 expression from biopsy to post-therapy samples was observed in 39% of the patients in cohort A vs 12% of the patients in cohort B (p=0.02). No patients with pCR have recurred so far vs 25% of the patients with less than pCR (p=0.005). The rate of recurrence was significantly higher for patients experiencing a change in HER2 expression (50% vs 19%, p=0.018).

Conclusion: contrary to our expectations, patients not receiving anti-HER2 therapy as part of neoadjuvant therapy were more likely to have a change in HER2 status vs patients receiving anti-HER2 neoadjuvant therapy. The change in HER2 status has a negative prognostic impact.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-12-18.
P1-12-25: Evaluation of PTEN, EGFR and Ki67 Expression as Predictors of Response to a Trastuzumab-Containing Neoadjuvant Chemotherapy Regimen in a HER-2 Over-Expressing Locally Advanced Breast Cancer (LABC) Trial.
Authors: Ho J , Turashvili G , Hayes M , Gelmon KA , Ellard SL , Macpherson N , Pansegrau G , Chia SK
Published: June 2012

Background: In patients with HER-2 over-expressing breast cancer, predictors of trastuzumab response and resistance remain unclear and unvalidated. In an exploratory analysis within a phase II trial, we evaluated various biologic markers as predictors of response or resistance to a trastuzumab containing neoadjuvant chemotherapy regimen.

Methods: A tissue microarray (TMA) was constructed from formalin-fixed paraffin-embedded tumor tissue samples obtained prior to neoadjuvant chemotherapy in 21 (from a total of 30) HER-2 positive stage II-III patients. Protocol treatment consisted of 4 cycles of 5-Fluorouracil, Epirubicin and Cyclophosphamide (FEC100) followed by 4 cycles of Docetaxel, Carboplatin (AUC 6) and Trastuzumab (TCH). Immunohistochemical (IHC) analyses of estrogen receptor (ER), progesterone receptor (PR), HER-2, Ki67, EGFR and PTEN were performed utilizing previously published protocols and cut-offs. HER-2 equivocal (2+) by IHC cases was confirmed by prior FISH analysis. Fisher's exact test was used for statistical analysis.

Results: 21 patients had sufficient tumour for analysis. Median age at diagnosis was 49 years old, 67% were pre-menopausal, and biomarker expression was as follows: 43% ER positive, 29% PR positive, and 100% HER-2 positive. 14 patients (67%) had pathologic complete response (pCR) following the completion of neoadjuvant chemotherapy and trastuzumab. In general, tumour samples showed high Ki67 (>14%) staining (62%), low EGFR positivity (14%), and high PTEN expression (71%). Neither high Ki67 expression (p=0.65), any EGFR expression (p=0.26), nor PTEN loss (p=0.60) was associated with higher or lower rates of pCR respectively.

Conclusion: In our small cohort of locally advanced HER-2 over-expressing tumours, baseline expression of Ki67, EGFR and PTEN were not associated with pCR to a neoadjuvant sequential anthracycline and taxane/trastuzumab combination regimen. Additional biomarkers for an activated PI3K pathway and for p95 will be attempted.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-12-25.
P1-14-05: Surgical Complications from the GeparQuinto Trial of Patients Receiving Preoperative Bevacizumab.
Authors: Eidtmann H , Kittel K , Rezai M , Tesch H , Ulmer HU , Stirnberg S , Belau A , Mau C , Stickeler E , Schrader I , Fasching P , Salat CT , Abdallah A , Holms F , Schwedler K , Nekljudova V , Loibl S , von Minckwitz G
Published: June 2012

Background: Bevacizumab has been reported to increase the risk of surgical complications, especially in pts treated with neoadjuvant chemotherapy.

Methods: In January 2010 it was decided to collect surgical complications prospectively on a specifically developed from. All patients received chemotherapy with epirubicin/cyclophosphamide (EC) followed by docetaxel (T) with or w/o bevacizumab. Patients not responding to the first 4 cycles of EC+/−B were enrolled in another setting. All patients receiving bevacizumab were supposed to be operated at 5 weeks after the last infusion of bevacizumab.

Results: Data from 699 patients have been prospectively collected, 329 received EC-T and 370 ECB-TB. Median age was 48 years in both arms. Median tumor size was 30mm in the ET-C arm vs 35mm in the ECB-TB arm (p=0.13). Multifocal or multicentric disease was present in 21% in the EC-T arm vs 22.4% (p=0.74). 36% of the patients received a sentinel node biopsy prior to start of neoadjuvant chemotherapy in both groups. 68.7% of patients received breast conserving therapy in the EC-T arm compared to 71.1% in the ECB-TB arm (p=0.54). The rate of pathological complete response was 19.5% with EC-T compared to 23.2% with ECB-TB (p=0.26). The median time from last CHT+/− bevacizumab to surgery was 29 days in EC-T and 34 days in ECB-TB. Bleeding or hematoma were reported in 6.7% of the patients treated by EC-T and 7.1% with ECB-TB (p=0.88). Wound infections or abscess were reported in 2.5% with EC-T and 3.9% with ECB-TB (p=0.39). Delayed wound healing was reported in 3.4% with EC-T and 6.3% with ECB-TB (p=0.11). Necrosis was reported in 1.9% with EC-T and 2.5% with ECB-TB (p=0.80). Any complication was reported in 11% with EC-T compared to 15.3% with ECB-TB (p=0.12).

Conclusion: In general the rate of surgical complications was low. Patients treated with bevacizumab had a numerical increase of surgical complications. However, none of the differences were statistically significant. The intervals from last infusion to surgery as defined by the study protocol appear to be safe.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-14-05.
P1-17-09: A Phase 1/2 Dose-Escalation Study of SAR245408 (S08) or SAR245409 (S09) in Combination with Letrozole (L) in Subjects with Hormone Receptor-Positive and HER2−Negative (HR+/HER2−) Breast Cancer (BC) Refractory to a Nonsteroidal Aromatase Inhibitor (AI).
Authors: Baselga J , Tolaney S , Hart L , Gomez P , Gartner E , DeCillis A , Ruiz-Soto R , Lager J , Burris H
Published: June 2012

Background: Upregulation of PI3K activity is a common molecular mechanism involved in resistance to AIs. S08 is a potent, orally bioavailable, pan-PI3K inhibitor. S09 is a potent, orally bioavailable inhibitor of PI3K which also possesses mTOR inhibitory activity. Both compounds exhibit robust PI3K and ERK pathway inhibition in paired human tumor biopsy samples from phase 1 studies (Edelman G, et al. ASCO 2010; Brana I, et al. ASCO 2010).

Methods: This ongoing, open-label, multicenter, phase 1/2 study (NCT01082068) was designed to evaluate the safety and tolerability of L in combination with either S08 (Arm 1) or S09 (Arm 2). Eligible female patients (pts) were ≥18 yrs, ECOG PS 0–1, with advanced or recurrent HR+/HER2− BC whose disease is refractory to nonsteroidal AIs. Phase 1 used an ascending 3+3 dose-escalation design and pts were accrued to each arm until no more than 1/3 pts or ≥33% of 3–6 pts at a given dose level experienced a dose-limiting toxicity (DLT) during the first 28-day cycle. Pts were alternately assigned to Arm 1 or Arm 2 and received 2.5 mg L PO (qd) in combination with different dose levels of either S08 (Arm 1; starting dose 200 mg tablets, PO, qd) or S09 (Arm 2; starting dose of 30 mg capsules, PO, bid). After reaching a preliminary maximum tolerated dose (MTD) for each combination, pts will accrue to the phase 2 portion of the study. A two-stage design will be used evaluate the phase 2 co-primary endpoints of ORR and PFS. Each arm will be evaluated independently and no formal comparisons between arms are planned.

Results: As of June 1st 2011, 17 pts were enrolled to Arm 1 (8 pts) or Arm 2 (9 pts). Median age was 54 yrs. Based on preliminary data, the adverse event profile in both arms were similar to those reported in the single agent phase I studies for S08 and S09 respectively. SAEs reported in Arm 1 (2 subjects) included 2 cases of Gr1 pneumothorax and 1 Gr4 pneumonitis; in Arm 2 (2 subjects) included Gr4 lumbar pain and Gr4 elevation of ALT and AST. No DLTs were reported in Arm 1 while one DLT (Gr3 skin rash) was reported in Arm 2 at a dose level of S09 50 mg bid (table 1). Arm 1 dosing data from S08 400 mg qd + L 2.5 mg qd dose level (maximum dose level allowed per protocol) will be completed by August 2011. For Arm 2, the MTD was S09 50 mg bid + L 2.5 mg, qd.

Discussion. Both S08 and S09 can be combined with L, and maximal doses for both combinations were established. Additional safety, PK and efficacy data will be presented from phase 1.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-17-09.
P2-01-11: Fish Oil Targets miR-21 To Increase PTEN Expression for Inhibition of Bone Metastatic CSF-1 in Breast Cancer Cells.
Authors: Mandal CC , Ghosh-Choudhury T , Ghosh Choudhury G , Ghosh-Choudhury N
Published: June 2012

Current therapy for the breast cancer patients is aimed at treating the cancer mainly at the primary site with limited ability to block metastasis. We have recently shown that fish oil (FO) diet significantly prevents MDA MB-231 breast cancer cell (MDA) metastasis to bone in a mouse model. Colony stimulating factor-1 (CSF-1) is one of major osteoclastogenic cytokines responsible for osteolytic metastasis of breast cancer. Quantitative RT-PCR (qRT-PCR) analysis showed significant inhibition of CSF-1 mRNA expression in fish oil fed mice xenograft tumor samples and docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), two ω-3 fatty acids in fish oil, inhibited CSF-1 mRNA and protein expression in MDA cells. A positive correlation was observed between expression of CSF-1 and the bone metastatic potential of different breast cancer cells. We have demonstrated recently upregulation in PTEN expression by FO diet as a mechanism of breast tumor growth arrest in mice. Over expression of PTEN inhibited migration of MDA cells. PTEN is a negative regulator of PI 3 kinase/Akt signaling. We investigated the mechanism of CSF-1 expression by breast cancer cells. Cotransfection of the CSF-1 promoter-driven luciferase plasmid (CSF-1-Luc) in MDA with PTEN or dominant negative (DN) PI 3 kinase or DN Akt, inhibited the transcription of CSF-1, indicating requirement of PI 3 kinase/Akt signaling. Several oncogenic microRNAs including miR-21 have been shown to induce metastasis of cancer cells. Fish oil fed mice tumors and DHA- and EPA-treated MDA cells significantly blocked miR-21 expression in MDA. PTEN mRNA 3'UTR contains miR-21 recognition element. Over expression of miR-21 inhibited PTEN 3'UTR-Luc reporter activity but it was increased by downregulation of miR-21, indicating functional targeting of PTEN mRNA by miR-21 in MDA. Similarly, expression of either miR-21 or miR-21 Sponge (plasmid-coded anti-miR-21 sequences) in MDA significantly suppressed or increased PTEN protein, respectively. Moreover, DHA-increased PTEN expression is reversed by over expression of miR-21 indicating a critical regulation of PTEN by miR-21. miR-21 over expression increased migration of MDA cells and partially reversed DHA- or EPA-mediated inhibition of migration. Since PTEN regulates expression of CSF-1, we examined the involvement of miR-21. Expression of miR-21 enhanced while miR-21 Sponge reduced expression of CSF-1 mRNA and its transcription. Furthermore, DHA-inhibited CSF-1 mRNA expression is reversed by over expression of miR-21. Together these results uncover a novel function of fish oil, which beheads miR-21 signaling to dampen expression of CSF-1 thus preventing osteoclastogenesis and osteolytic lesions in breast cancer.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-01-11.
P2-01-15: RT-PCR Gene Profiling for the Prediction of Bone Metastases in Primary Breast Cancer.
Authors: Ibrahim T , Mercatali L , Ricci R , Scarpi E , Liverani C , Serra L , Guerra D , Ciani S , Amadori D
Published: June 2012

Background: Patients with breast cancer frequently develop bone metastases, which are responsible for high morbidity and mortality. It is known that numerous molecules are involved in the bone metastasization process and that only cancer cells with a specific molecular profile are capable of reaching and colonizing bone tissue. The early detection of patients with a high probability of relapsing to bone could be used to select candidates for tailored therapy with bone-specific drugs such as bisphosphonates or RANK-L inhibitors. The aim of the present study was to identify a pattern of tissue markers from primary breast cancer capable of predicting bone metastatization.

Material and methods: The expression of different markers was retrospectively analyzed in frozen breast cancer tissue samples from 40 patients comprising 15 cases with no evidence of disease (NEDP), 15 with bone metastases (BMP), and 10 with visceral metastases (VMP). Twelve transcripts were analyzed by Quantitative Real time PCR: trefoil factor 1(TFF1), DKK1, bone sialoprotein (IBSP), heparanase (HPSE), osteopontin (SPP1), Agr2, SPARC, CTGF, COMP, follistatin (FST), osteoprotegerin and RANK. The predictive accuracy of each marker was calculated using receiver operating characteristic (ROC) curves.

Results: Analysis of marker expression in the 3 different subgroups showed twofold higher median values of COMP and HPSE in BMP with respect to either NEDP or VMP (no significant difference). Furthermore, TFF1 median value was about sevenfold higher in BMP than in the other 2 subgroups (p=0.05). The area under the curve (AUC) was 0.73 for TFF1 and 0.62 for COMP and HPSE. Considering markers as dichotomous variables, TFF1 expression in BMP reached 67% compared to 21% and 20% in NEDP and VMP, respectively. In dichotomous variable analysis, the most important result was observed for TFF1; in particular, TFF1 positivity was observed in 67% of cases in the BMP subgroup compared to only 21% and 20% in NEDP and VMP, respectively. The combination analysis of TFF1 and other markers showed that positivity to at least one of the three markers, TFF1, DKK1, or IBSP, was observed in 80% of cases for the BMP group, and in only in 21% and 20% of cases in NEDP and VMP subgroups (p=0.041). Another interesting marker was RANK: although sensitivity was fairly low (12%), specificity reached 100% in both control groups.

Conclusions: Our preliminary study identified a RT-PCR gene expression pattern in primary breast cancer that could predict patients destined to bone relapse. Such patients could consequently benefit from adjuvant treatment with bone-targeted therapy.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-01-15.
P2-01-16: Dissecting the Cellular Mechanism of Induction and Sustenance of Dormancy by Rho GTPases in Breast Cancer.
Authors: Chatterjee M , van Golen KL
Published: June 2012

Tumor dormancy has been a very real problem in breast cancer survivors where recurrences are common and usually fatal. Molecular mechanisms involved in this process are often poorly understood and are a hot spotin breast cancer research today.

We have found that Rho GTPases, specifically RhoC, is hyperactivated in low metastatic dormant breast tumor cells while RhoA is downregulated. We also found RhoC causes phosphorylation of c-jun NH2 terminal kinase (JNK) in these cells which in turn activates the process of autophagy, a possible mode of cell survival in dormant tumor cells. Modulating RhoC affected phosphorylation of JNK. Also, abrogating JNK by a pharmacological inhibitor SP600125 inhibited expression of the autophagy marker LC3B-II. Autophagy genes like Atg 5 and Atg 12 were expressed along with the autophagy marker LC3B-II. On inhibiting autophagy by a specific inhibitor 3-Methyladenine (3-MA), cells did not readily undergo apoptosis but on stressing them doubly with LY294002, an inhibitor of Class I PI3K, along with 3-MA (inhibitor of Class III PI3K), significant number of dormant cells underwent apoptosis as observed by TUNEL staining. We also found that p190RhoGAP was activated in these dormant cells which may lead to the observed downregulation of RhoA activation levels. Involvement of beta-1 integrin along with c-src in activation of RhoC was observed in these dormant cells. We are in the process of further elucidating these signal transduction processes that might be involved in cellular dormancy of breast cancer.

A better understanding of some of the cellular mechanisms of induction and sustenance of dormancy may aid in formulating therapies for annihilating these dormant tumor cells thus preventing the often fatal relapses in breast cancer survivors.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-01-16.
P2-01-21: The Biological Influence of Brain Derived Neurotrophic Factor (BDNF) on the Aggressiveness of Human Breast Cancer Cells.
Authors: Yang MX , Martin TA , Jiang WG
Published: June 2012

Introduction: Brain derived neurotrophic factor (BDNF) is a member of the neurotrophin superfamily and has been indicated in the pathophysiology of the nervous system and is important in a number of neurological and psychological conditions. Recently, BDNF has also been shown to play a role in the development and progression of solid tumours myeloma, lung cancer and prostate cancer. It has been recently reported that BDNF is also aberrantly expressed in human breast cancer and that a raised level of BDNF is associated with poor clinical outcome and reduced survival. The present study aimed to investigate the biological role of BDNF in human breast cancer. Materials and methods: A panel of human breast cancer cells were used in the present study. The expression profile of BDNF was evaluated using RT-PCR. We constructed a set of anti-BDNF transgenes which were used to transfect breast cancer cells in order to generate BDNF knockdown cells (MCFΔBDNF). The impact of BDNF knockdown on growth and cellular migration were evaluated using standard growth assays and ECIS (electronic cell impedance sensing) technology.

Results: BDNF gene transcripts were successfully detected in the breast cancer cell lines, MCF-7, MDA MB-231 and ZR 75-1. MCF-7 wild type cells were subject to transfection of anti-BDNF transgenes, followed by the establishment of BDNF knockdown sub-lines and empty vector control cells (MCFpef). Loss of BDNF in breast cancer cells resulted in reduction of cell growth rate (growth rate in MCFΔBDNF by day 3 1.60+/−0.11, compared with 2.06+/−0.04 in MCFpef, p=0.006). Using electric cell-substrate impedance sensing, it was found that loss of BDNF in breast cancer cells resulted in a significant increase in the rate of cellular attachment and migration (migration rate over 14 h in ohms: MCFΔBDNF 6310.125+/−595.183 compared with MCFpef 5115.5 +/−422.394, p=0.0003).

Conclusion: When BDNF was stably knocked down in MCF-7 cell lines, the growth and proliferation were decreased. The ECIS results also showed that cell migration was decreased in MCF-7 cells stably transfected with ribozymes for BDNF compared with the vector control. It is concluded that BDNF, a neurotrophic growth factor aberrantly expressed in cancers such as breast cancer, has a profound impact on the cellular behaviour of breast cancer cells and that loss of BDNF is associated with a reduction of the aggressiveness of breast cancer. BDNF is therefore a potential therapeutic target in breast cancer and its effect in human breast cancer requires further investigation.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-01-21.
P2-02-05: Molecular Mechanism for Src Homology Phosphotyrosyl Phosphatase 2 Regulation of Cell Motility and Migration.
Authors: Hartman ZR , Agazie YM
Published: June 2012

Background: The Src homology phosphotyrosyl phosphatase 2 (SHP2) acts as a transducer of mitogenic and cell survival signaling downstream of receptor tyrosine kinases, cytokine receptors and integrins. As such, SHP2 promotes cell growth, transformation, survival, and motility, among other processes. Recent studies suggest that SHP2 is overexpressed in breast cancer and is essential for the maintenance of the transformed phenotype both in the HER2−positive and the basal-type/triple-negative breast cancer (BTBC) cells.

Materials and Methods: The expression of SHP2 in BTBC cells was suppressed constitutively by lenti virus-mediated transduction of specific shRNA. Impact of SHP2 inhibition on cell migration was determined by the monolayer wound healing assay. The SHP2 substrate-trapping mutant was used to identify FAK as an SHP2 substrate. Site-directed mutagenesis, binding studies and phosphatase assays were employed to further characterize FAK as a bona fide SHP2 substrate. Immunofluorescence microscopy was conducted to visualize the amounts of phosphorylated FAK and its location in the cell after adhesion.

Results: We demonstrate that SHP2 is essential for the migration of BTBC cells as evidenced by loss of the enhanced migratory behavior upon its inhibition. Using the SHP2 substrate-trapping mutant, FAK was identified as a biological substrate of SHP2. Site-directed mutagenesis, binding studies and phosphatase assays confirmed that SHP2 indeed associates with and dephosphorylates FAK at the pY397 site, a site known to promote the focal adhesion activity of FAK. Immunofluorescence of breast cancer cells with SHP2 inhibition were found to have higher levels of FAK pY397 further confirming that FAK is an SHP2 substrate. The increase in pY397 level correlated with loss of cell polarity, enhanced focal adhesion formation, and acquisition of non-transformed morphology.

Discussion: These results suggest that dephosphorylation of pY397 of FAK by SHP2 is the major mechanism by which it is able to modulate cancer cell migration. Together with previous findings, the current studies show that SHP2 is a suitable target for therapeutic intervention to combat breast cancer metastasis.

Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P2-02-05.