bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P3-07-06: Survival benefit increases after breast conserving therapy compared to mastectomy when axillary node status is positive in early stage breast cancer: A registry based follow-up study of Norwegian women with screening and interval detected breast cancer pr
Authors: Hartmann-Johnsen Olaf Johan , Schlichting Ellen , Kaaresen Rolf , Nygaard Jan Franz
Published: May 2015

BACKGROUND: Recent register studies on early stage breast cancer have shown better survival when women underwent breast-conserving treatment (BCT) compared to mastectomy (MTX). It is unclear if this is due to selection effects or to a systemic benefit of BCT compared to MTX. If there is a systemic benefit this could theoretically be more prominent in some subgroups of breast cancer. The aim of this study is to evaluate if there are any subgroups with early stage breast cancer that benefits more of BCT compared to MTX.

METHOD: A cohort of 6.629 women aged 50-69 with primary operable breast cancer detected in the screening programme (on screening or between screening, interval cancer) were selected from January 1998 until December 2009 and followed up until end of 2010. Life tables for overall survival (OS) and breast cancer specific survival (BCSS) were calculated in surgery groups and stages. Corresponding Kaplan-Meier plots were stratified in T1N1M0 and Ductal Carcinoma. OS and BCSS were compared using Cox proportional hazard for estimating hazard ratio between BCT and MTX in crude and adjusted analysis.

RESULTS: 5-years BCSS in T1N0M0 was 99% for both surgical cohorts (BCT and MTX). 5-years BCSS in T1N1M0 was 98% for women undergoing BCT and 91% for women undergoing MTX. Respectively, 10-years BCSS in T1N1M0 was 96% for women undergoing BCT and 83% for women undergoing MTX.

In adjusted analysis, HR for breast cancer death for women with stage T1N1M0 undergoing MTX compared to BCT was 3.12 (95% CI: 1.71-5.72). Respectively, 3.67 (95% CI: 1.20-11.22) stratified on grade III and 4.89 (67 (95% CI: 1.25-19.01) stratified on hormone receptor negative disease.

CONCLUSION: The benefit of BCT compared to MTX seems to increase with increasing severity of the breast cancer disease, positive nodal status (N1), grade III and hormone receptor negative disease. These finding indicate a systemic benefit of BCT compared to MTX.

Citation Format: Olaf Johan Hartmann-Johnsen, Ellen Schlichting, Rolf Kaaresen, Jan Franz Nygaard. Survival benefit increases after breast conserving therapy compared to mastectomy when axillary node status is positive in early stage breast cancer: A registry based follow-up study of Norwegian women with screening and interval detected breast cancer pr [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P3-07-06.
Abstract P3-09-01: A multicenter randomized study comparing the dose dense G-CSF-supported sequential administration of FEC followed by docetaxel versus docetaxel plus cyclophosphamide as adjuvant chemotherapy in women with HER2-negative, axillary lymph node-positive breast
Authors: Mavroudis Dimitrios , Malamos Nikolaos , Papakotoulas Pavlos , Kakolyris Stylianos , Boukovinas Ioannis , Athanasiadis Elias , Kentepozidis Nikolaos , Ziras Nikolaos , Kalbakis Kostas , Christophyllakis Charalambos , Georgoulias Vassilis
Published: May 2015

Background: The dose dense sequential administration of anthracycline and taxane is very effective as adjuvant therapy in node-positive early breast cancer. The non-anthracycline regimen docetaxel plus cyclophosphamide (TC regimen) was better than four cycles of doxorubicin/cyclophosphamide as adjuvant therapy. This study compared the dose dense sequential regimen versus the TC regimen as adjuvant therapy.

Methods: Women with axillary node-positive, HER2-negative early breast cancer were randomized following surgery to receive either dose dense G-CSF-supported FEC (5FU 500mg/m2, epirubicin 75mg/m2, cyclophosphamide 500mg/m2 every 14 days for 4 cycles) followed by Docetaxel (75mg/m2 every 14 days for 4 cycles) (arm A) or 6 cycles of Docetaxel 75mg/m2 plus Cyclophospamide 600mg/m2 every 3 weeks (arm B). The primary endpoint of the study was the 3-year disease-free survival (DFS).

Results: Six hundred fifty patients were randomized; 326 on arm A and 324 on arm B. Of them 109 (33%) and 90 (28%) were premenopausal, 196 (60%) and 218 (67%) had 1-3 positive nodes, 284 (87%) and 288 (89%) were hormone receptor positive in arm A and B, respectively. Chemotherapy was completed in 97% and 93% of patients in arm A and B, respectively. After a median follow up of 46 and 47 months, there were 37 (11.3%) and 33 (10.1%) disease relapses and the median DFS has not yet been reached (p=0.5) while the 3-year DFS rate was 89.5% and 91.1% for arm A and B, respectively. Neutropenia grade III-IV was more common in arm B and anemia, nausea, vomiting and fatigue grade II-III in arm A. No toxic deaths occurred.

Conclusions: The 3-year DFS rate was similar between the dose dense sequential FEC/docetaxel combination and the TC regimen in women with node-positive HER2-negative early breast cancer.

Citation Format: Dimitrios Mavroudis, Nikolaos Malamos, Pavlos Papakotoulas, Stylianos Kakolyris, Ioannis Boukovinas, Elias Athanasiadis, Nikolaos Kentepozidis, Nikolaos Ziras, Kostas Kalbakis, Charalambos Christophyllakis, Vassilis Georgoulias. A multicenter randomized study comparing the dose dense G-CSF-supported sequential administration of FEC followed by docetaxel versus docetaxel plus cyclophosphamide as adjuvant chemotherapy in women with HER2-negative, axillary lymph node-positive breast [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P3-09-01.
Abstract P3-10-07: Phase I trial of eribulin in combination with S-1 for advanced and recurrent breast cancer patients
Authors: Iwasa Tsutomu , Tsurutani Junji , Nishida Satomi , Komoike Yoshifumi , Nakagawa Kazuhiko
Published: May 2015

Introduction: Eribulin is a non-taxane microtubule dynamics inhibitor that has been proved to prolong overall survival in patients with advanced and recurrent breast cancer compared to treatment of physician’s choice. S-1 is an oral fluoropyrimidine anticancer agent that combines tegafur as the effector drug with two modulators, gimeracil, and oteracil potassium, and has shown promising antitumor activity as a single agent for the treatment of advanced breast cancer. In the basis of these results, we aimed to assess the efficacy of the combination of these two drugs in patients with advanced and recurrent breast cancer. In this trial, we determined maximum tolerated dose (MTD) and phase II recommended dose of Eribulin and S-1 combination therapy.

Methods: A traditional 3+3 dose escalation design was implemented. 12 patients pre-treated with anthracycline and taxane were enrolled. As an initiation level, patients received s-1 50 mg/m2 from day1 to day14, and Eribulin 1.1mg/m2 on day1, and day8. In level 2, Eribulin dose was increased to 1.4mg/m2. In level 3, S-1 dose was increased to 80 mg/m2.

Results: In level 1 or 2, no dose limiting toxicity (DLT) was observed. In level 3, grade 3 hypokalemia occurred in 1 case. The level 3 dose level was determined as phase II recommended dose. Neutropenia was observed in all cases, 3 of which was grade 3 febrile neutropenia. However, other non-hematological toxicity was mild, and the toxicity of the both drugs

Safety (N=12)Adverse EventGrade 3/4Leukopenia3 (25%)Neutropenia10 (83%)Febrile Neutropenia3 (25%)Hypokalemia1 (8%)

was not enhanced by the combination. Partial response

Efficacy (N=12)CR0 (0%)PR5 (41%)SD≥24 weeks2 (17%)SD<24 weeks3 (25%)PD2 (17%)Clinical Benefit7 (58%)Disease Control Rate10 (83%)

is confirmed in 5/11 patients with measurable lesions.

Conclusion: We have shown that treatment with Eribulin and S-1 is active and well tolerated in patients with advanced and recurrent breast cancer, suggesting that this combination therapy may have potential as a new treatment option. A phase II study is being conducted to evaluate the efficacy and safety.

Citation Format: Tsutomu Iwasa, Junji Tsurutani, Satomi Nishida, Yoshifumi Komoike, Kazuhiko Nakagawa. Phase I trial of eribulin in combination with S-1 for advanced and recurrent breast cancer patients [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P3-10-07.
Abstract P3-11-07: Changes in PgR and Ki-67 in residual tumour and outcome of breast cancer patients treated with neoadjuvant chemotherapy
Authors: Montagna Emilia , Bagnardi Vincenzo , Viale Giuseppe , Rotmensz Nicole , Sporchia Andrea , Cancello Giuseppe , Balduzzi Alessandra , Galimberti Viviana , Veronesi Paolo , Luini Alberto , Mastropasqua Mauro G , Casadio Chiara , Sangalli Claudia , Goldhirsch Aron , Colleoni Marco
Published: May 2015

PURPOSE We assessed the frequency of the change in biological features between the diagnostic biopsy and final surgery in breast cancer patients with residual disease after neoadjuvant chemotherapy. We also valued the impact of change on outcome in terms of disease free survival (DFS) and overall survival (OS)

PATIENTS AND METHODS We collected information through the institutional clinical database on all consecutive breast cancer patients treated with neoadjuvant chemotherapy at the European Institute of Oncology (EIO), Milan, Italy, between 1999 and 2011. We selected patients who did not achieved pathological complete response (pCR) at final surgery. All patients had pathological evaluation, including ER, PgR, HER2 protein and ki-67 expression performed at EIO both at diagnostic core biopsy and final surgery.

RESULTS We identified a total of 904 patients. The 5% of patients with ER positive at diagnostic biopsy had ER negative residual tumor at final surgery. For PgR expression, 67% of patients, whose tumors had a PgR> 20% at diagnostic biopsy had a PgR<20% at final surgery. The ki-67 expression changes from >20% to <20% in 40% of patients. At the multivariate analysis the decrease of PgR -immunoreactive cells correlated with improved outcome in terms of DFS (HR 0.73; CI 0.54-1.00 p 0.046). In addition, the decrease of ki-67 expression to < 20% of the cells at final surgery was found to be associated with better outcome both in terms of DFS (HR 0.52; CI 0.40-0.68 p <0.0001) and OS (0.45 CI 0.32-0.64 p<0.0001)

CONCLUSION The decrease of PgR and Ki-67 expression after preoperative chemotherapy have a prognostic role in breast cancer patients with residual disease and should be considered in the adjuvant therapeutic algorithm.

Citation Format: Emilia Montagna, Vincenzo Bagnardi, Giuseppe Viale, Nicole Rotmensz, Andrea Sporchia, Giuseppe Cancello, Alessandra Balduzzi, Viviana Galimberti, Paolo Veronesi, Alberto Luini, Mauro G Mastropasqua, Chiara Casadio, Claudia Sangalli, Aron Goldhirsch, Marco Colleoni. Changes in PgR and Ki-67 in residual tumour and outcome of breast cancer patients treated with neoadjuvant chemotherapy [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P3-11-07.
Abstract P3-11-12: Vacuum assisted core-needle biopsy after neoadjuvant therapy in breast cancer to predict pathologic response
Authors: Grosse Regina , Saegenschnitter Jana , Nguyen Thi-Dao , Unverzagt Susanne , Buchmann Joerg , Kantelhardt Eva J , Papendick Nancy , Thomssen Christoph
Published: May 2015

Introduction: Recently published neoadjuvant breast cancer therapy trials have documented impressive pathologic complete response (pCR) rates of up to 50% and above, depending on grade, hormone receptor and HER2 status and type of therapy. The purpose of our prospective study is to examine the predictive value of a standardized vacuum assisted core-needle biopsy (VAB) of the tumor site after neoadjuvant therapy with respect to pCR status. This is a presentation of the first case series of this single institution study.

Methods: As of 2011, all patients who had completed neoadjuvant breast cancer therapy at our institution were informed about this trial regardless of their clinical response status. In case of consent, a mammography-guided VAB of the initially tagged tumor site was performed directly before the operation. In case of breast conserving surgery, mammographic wire localisation was performed immediately thereafter. Final surgery (breast conservation or mastectomy) was performed according to German guidelines. Ethical approval was obtained from the institutional review board. Pathologic complete response was defined as ypT0 ypN0.

Results: Of 70 patients completing neoadjuvant therapy, 44 consented to the trial. The pCR rate was 47% (n=21). Pre-operative VAB correctly predicted residual disease in 19 of 23 cases (82,6%). We observed incorrect prediction of pathologic response if the tagging clip was not placed centrally, or if the radiologist did not perform the VAB in the tagged area. In 3 cases of pathologic partial response (pPR), all residual tumor (DCIS) was removed by VAB. In these cases, no further malignant lesions were detected in the resulting surgical specimens.

Conclusions: Preliminary observations of this ongoing study are encouraging, although the correct prediction rate of residual disease is nowhere near the sensitivity needed to change clinical practice. However, if these results can be improved and confirmed in a larger series, it may be possible to reduce operative procedures after neoadjuvant therapy based on the results of a preoperative standardized VAB.

Citation Format: Regina Grosse, Jana Saegenschnitter, Thi-Dao Nguyen, Susanne Unverzagt, Joerg Buchmann, Eva J Kantelhardt, Nancy Papendick, Christoph Thomssen. Vacuum assisted core-needle biopsy after neoadjuvant therapy in breast cancer to predict pathologic response [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P3-11-12.
Abstract P3-11-13: Change of Ki-67 after long course of neoadjuvant chemotherapy as prognostic factors in patients with stage II, III triple negative breast cancer
Authors: Yang Yaewon , Kim Tae-Yong , Im Seock-Ah , Keam Bhumsuk , Lee Kyung-Hun , Han Sae-Won , Oh Do-Youn , Kim Tae-You , Moon Hyeong-Gon , Han Wonshik , Park In Ae , Noh Dong-Young
Published: May 2015

Background: Triple negative breast cancer (TNBC) shows poor prognosis despite their higher response rate to neoadjuvant chemotherapy (NAC). Response to NAC such as pathologic complete response (pCR) is a well-known useful prognostic marker for relapse and overall survival. Pre-chemotherapy baseline higher Ki-67 is a positive predictive marker for higher pCR rate but a negative prognostic marker for relapse free survival (RFS) and overall survival (OS). After short course of NAC, Ki-67 index changed in response to the treatment. The purpose of this study is to investigate the prognostic role of Ki-67 index change between before and after the long course of NAC in TNBC patients.

Patients and Methods: Patients with clinical stage II or III breast cancer treated with NAC at Seoul National University Hospital were enrolled and their clinicopathological findings including Ki-67 index were reviewed retrospectively. The definition of Ki-67 threshold of high was ≥14% in current study. TNBC definition was adopted from 2011 St Gallen Consensus Panel.

Results: From Jan 2009 to Dec 2010, a total of 183 patients received NAC. Among them, 46 (25.1%) were TNBC. The pCR rate (19.6% vs 9.5%, P=0.069) and the objective response rate (complete response + partial response) (76.1% vs 70.8%, P=0.153) were higher in TNBC group than in non-TNBC group. The RFS was not different between the two groups (3 year 77.3% vs 75.4%, P=0.877). The change of Ki-67 index from high (≥14%) to low (<14%) was significantly associated with longer RFS in only in TNBC group. As for TNBC patients with high Ki-67 index at diagnosis, RFS of those whose Ki-67 index changed to low after NAC were significantly longer than that of whose Ki-67 index remained high (3 year RFS 87.2% vs 54.5%, P=0.036). In non-TNBC group, change of Ki-67 index was not significantly associated with RFS (P=0.245). The initial Ki-67 index itself (high vs. low) was not associated with RFS in both TNBC and non-TNBC group (P=0.726 and P=0.523 respectively).

Conclusion : The change of Ki-67 index from high to low is associated with longer RFS in TNBC. Ki-67 index change between before and after the NAC can be used for further discrimination of TNBC as a prognostic marker.

Citation Format: Yaewon Yang, Tae-Yong Kim, Seock-Ah Im, Bhumsuk Keam, Kyung-Hun Lee, Sae-Won Han, Do-Youn Oh, Tae-You Kim, Hyeong-Gon Moon, Wonshik Han, In Ae Park, Dong-Young Noh. Change of Ki-67 after long course of neoadjuvant chemotherapy as prognostic factors in patients with stage II, III triple negative breast cancer [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P3-11-13.
Abstract P4-01-02: Prospective characterization of HER2-positive circulating tumor cells in patients with HER2-negative metastatic breast cancer
Authors: Krop Ian E , Macrae Erin , Galler Sarah R , Bischoff Farideh , Fauni Romeo , Sales Edgar , Huynh Lan , Mitchell Christine , Clarin-Tamayo Trisky , Anderson Mark , Abad Leslie , Winer Eric
Published: May 2015

Background: We have previously shown feasibility by CLIA validation of HER2 FISH on circulating tumor cells (CTCs). Several small retrospective studies have identified HER2 amplification in CTCs in a subset of patients (pts) with clinically HER2 negative metastatic breast cancer (MBC). While these findings potentially have profound implications for CTCs as a predictive biomarker, prospective validation and functional characterization of this subgroup is necessary.

Methods: We enrolled a prospective cohort of pts with MBC that was HER2 negative by IHC and/or FISH on all available primary and metastatic biopsies. Pts had ≥1 line of prior chemotherapy for MBC. Blood samples were collected at study entry. CTCs were enumerated based on standard criteria (Cytokeratin (CK)+/CD45- staining) as well as by FISH for HER2 using the OncoCEE-BR™ CTC test (Biocept, Incorporated). This test employs a proprietary antibody cocktail for capture followed by CK/CD45 staining, and FISH analysis directly within a microfluidic device. Samples were reported as positive if the HER2/CEP17 ratio was ≥ 2.0. HER2+ CTCs were classified into two categories: CK+/CD45-/HER2+ and CK-/CD45-/HER2-

Results: CTCs were observed in 208 out of 323 pts (64%). Median number of CTCs was 10 (range 1 to > 34195). 75 pts (23%) had HER2+ CTCs, with a median number of 3 HER2+ CTCs (range 1 to 21). 36% (27/75) of these pts had CK+/HER2+ CTCs and 43% (32/75) of pts had only CK-/HER2+ CTCs. The remaining pts (21%) had both CK+/HER2+ and CK-/HER2+ CTCs present.

Conclusion: HER2 amplified CTCs are present in a subset (23%) of pts with clinically HER2-negative breast cancers. Interestingly, we observed a high prevalence of pts with only CK-/HER2 amplified CTCs (32 out of 323; 10%). The unique multi-antibody CTC capture method used here thus allows for detection of a prevalent population of CK-/HER2+ CTCs that may be largely undetected by other current adopted technologies. The functional significance of CK-/HER2+ and CK+/HER2+ CTCs in patients with clinically HER2 negative breast cancer is currently being evaluated in a prospective study with HER2-directed therapy.

Citation Format: Ian E Krop, Erin Macrae, Sarah R Galler, Farideh Bischoff, Romeo Fauni, Edgar Sales, Lan Huynh, Christine Mitchell, Trisky Clarin-Tamayo, Mark Anderson, Leslie Abad, Eric Winer. Prospective characterization of HER2-positive circulating tumor cells in patients with HER2-negative metastatic breast cancer [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P4-01-02.
Abstract P4-02-10: Detection of H1047R and E545K PIK3CA mutations from peripheral blood in ER positive breast cancer patients
Authors: Garcia-Saenz Jose Angel , Acosta Daniel , Moreno Fernando , Ayllon Paticia , Sotelo Miguel , Caldes Trinidad , Diaz-Rubio Eduardo , Romero Atocha
Published: May 2015

Background: Digital polymerase chain reaction (PCR) is a new technology that enables detection and quantification of cancer DNA molecules from peripheral blood. Detecting tumor-specific mutations in circulating plasma DNA may potentially be useful to select systemic therapies for solid tumors. The aim of our study is to evaluate the feasibility of detecting PIK3CA mutations from plasma of breast cancer patients.

Methods: We have designed an allele-specific PCR assay and used a digital PCR system for the detection of PIK3CA mutations H1047R and E545K. Formalin-fixed paraffin-embedded (FFPE) tumor samples were analyzed by COBAS and by digital PCR. The matched plasma samples were then analyzed by digital PCR. Kappa Cohen’s coefficient (κ) was used to test agreement between methods

Results: 37 ER positive breast cancer and matched plasma were evaluated. 31 (84%) patients were stage IV disease. E545K mutation was detected in 4 out of 37 (11%) tumor specimens and H1047R mutation was identified in 8 out of 37 (22%) tumor samples. The proportion of observed agreement between H1047R and E545K detection in tumor samples by COBAS and digital PCR was 100%. Regarding E545K mutation assay we found that the proportion of observed agreement between mutation status in FFPE samples and circulating tumor DNA (ctDNA) was 94.6% (kappa= 0.770; Sensitivity=100%; Specificity=93.9%). In H1047R assay the proportion of observed agreement between FFPE samples and ctDNA was 83.78% (Kappa=0.561; Sensitivity=60%; Specificity=92.59%).

Conclusions: The agreement between methodologies when assessing the PIK3CA status in tumor samples was perfect. However, a moderate-to-fair agreement was found between FFPE samples and ctDNA which might be due to the heterogeneity of the disease. The methodology presented in this study is a feasible approach for PIK3CA mutation detection in blood derived samples.

Citation Format: Jose Angel Garcia-Saenz, Daniel Acosta, Fernando Moreno, Paticia Ayllon, Miguel Sotelo, Trinidad Caldes, Eduardo Diaz-Rubio, Atocha Romero. Detection of H1047R and E545K PIK3CA mutations from peripheral blood in ER positive breast cancer patients [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P4-02-10.
Abstract P4-02-11: Circulating methylated DNA of pregnancy-associated plasma protein A as a clinically useful biomarker in breast cancer
Authors: Sainsbury Richard , Koehler Karsten , Page Catherine , Velamakanni Saroj
Published: May 2015

Background: A recent report suggests that Pregnancy-Associated Plasma Protein A (PAPPA) plays a pivotal role in normal cell division and is targeted early in breast cancer development. Epigenetic silencing of PAPPA is highly prevalent in precursor lesions and invasive breast cancer, correlating with loss of PAPPA protein expression. In normal breast tissue PAPPA promoter methylation was not observed (Journal of Pathology 344-356: 2014). We sought to test the hypothesis that PAPPA promoter hypermethylation can be detected in cell-free tumour specific DNA (ctDNA) in the bloodstream of patients with metastatic breast cancer.

Methods: Plasma derived from 12 patients with metastatic breast cancer was examined for ctDNA. A semi-quantitative real-time PCR based MethyLight assay was used for detection of PAPPA methylation. Spiking experiments with breast cancer cell lines were performed to develop the assay and confirm linearity of sodium bisulfite treatment.

Results: Percentage methylated reference (PMR) values from spiking experiments with PAPPA methylated and non-methylated DNA in plasma (matrix) demonstrate good linearity for the PAPPA gene and the endogenous control Col2A. PAPPA methylation was detected in 10 out of 12 (83%) of the metastatic breast cancer samples. In stark contrast, epigenetic silencing of PAPPA was not detected in any of the plasma samples from non-cancer bearing patients (n=10).

Conclusions: Given the potential clinical impact of our data, this study is being extended to include early stage breast cancer patients with poor prognostic clinic-pathological indicators. Our proof-of-concept findings indicate that testing for PAPPA promoter methylation in ctDNA may be used as a biomarker of disseminated disease, helping clinicians with treatment decisions and adjusting those choices as conditions change. By developing and refining more sensitive techniques, PAPPA methylation testing in ctDNA could also find practical utility in breast cancer screening.

Citation Format: Richard Sainsbury, Karsten Koehler, Catherine Page, Saroj Velamakanni. Circulating methylated DNA of pregnancy-associated plasma protein A as a clinically useful biomarker in breast cancer [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P4-02-11.
Abstract P4-04-01: A new breast cancer classification scheme based on novel classes of tumor stroma
Authors: Thompson Crista , Bertos Nicholas , Gruosso Tina , Finak Greg , Lesurf Robert , Saleh Sadiq M , Zhao Hong , Souleimanova Margarita , Meterissian Sarkis , Omeroglu Atilla , Hallett Michael T , Park Morag
Published: May 2015

A major challenge in cancer treatment is the heterogeneous nature of the disease. This is particularly evident in breast cancer where gene expression profiling of whole tumours has identified multiple intrinsic subtypes of breast cancer. These subtypes are associated with differential prognoses and are correlated with previously identified clinical biomarkers (i.e., ER and HER2 status) used to stratify patients for targeted therapy. Despite recent studies demonstrating that elements within the tumour microenvironment can affect breast cancer progression and outcome, and that information contained within this compartment carries significant prognostic information for patient stratification above and beyond the information supplied by the intrinsic subtypes and existing therapeutic biomarkers, a limited understanding of stromal heterogeneity across the population has hindered the development of effective prognostic tools and targeted therapies directed against these processes. Here we perform expression profiling of the microdissected tumour-associated stromal components of 49 human breast tumours, and demonstrate that stromal heterogeneity can be captured by categorization into six classes which bear distinct molecular phenotypes. These stromal classes exhibit distinct biological functions and carry prognostic information independent of existing tumor-intrinsic biomarkers and molecular breast cancer subtypes. Specific combinations of stromal class and tumour subtype are significantly over- and under-represented; furthermore, simultaneous stratification of tumors in external datasets by both tumour subtype and stroma class identifies good- and poor-outcome cohorts within four of the five molecular breast cancer subtypes. The stroma classes identified here form the basis for an improved breast cancer classification scheme which takes the contribution of the microenvironment into account.

Citation Format: Crista Thompson, Nicholas Bertos, Tina Gruosso, Greg Finak, Robert Lesurf, Sadiq M Saleh, Hong Zhao, Margarita Souleimanova, Sarkis Meterissian, Atilla Omeroglu, Michael T Hallett, Morag Park. A new breast cancer classification scheme based on novel classes of tumor stroma [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P4-04-01.
Abstract P4-04-21: Host response to microbes found in the tumor microenvironment and healthy adjacent tissue
Authors: Navarrete Marian , Sieling Peter A , Lee Delphine J , Chung Andrew W
Published: May 2015

Next generation sequencing methods have defined the bacterial microbiome in human breast milk and breast tissue, but the role of bacteria or their components in breast cancer has not been well defined. There is growing evidence in humans and animals that alterations of the gut microbiome may contribute to host susceptibility to cancer. However, the presence of bacteria at the site of the tumor microenvironment may also contribute to carcinogenesis (e.g. by promoting chronic inflammation) or immune surveillance (e.g. by stimulating antitumor pathways). Previously, we found that bacterial load in breast cancer tissue is inversely correlated to the progression of disease and that antimicrobial gene expression was comparatively higher in breast tissue from healthy subjects. Our investigation of the microbiome of breast tissue showed that overall microbiome signatures in breast cancer patients are similar when comparing their healthy adjacent tissue vs. the tumor tissue. However, our published preliminary studies did identify some differences in the abundance of specific bacteria between the two tissues. Here we investigated how those bacteria in healthy adjacent vs. tumor tissue may differentially stimulate cells found in breast tissue including breast ductal epithelium, adipocytes, and immune cells. Our preliminary data suggest that bacteria found in healthy adjacent vs. tumor tissue stimulate differential cytokine responses that may shape the local tumor microenvironment and beyond and may also contribute to local/regional immune surveillance.

Citation Format: Marian Navarrete, Peter A Sieling, Delphine J Lee, Andrew W Chung. Host response to microbes found in the tumor microenvironment and healthy adjacent tissue [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P4-04-21.
Abstract P4-05-02: Overexpression of the chemokine receptor CCR2 in the breast epithelium is associated with progression of DCIS
Authors: Mafuvadze Benford , Fang Wei Bin , Cheng Nikki
Published: May 2015

Ductal carcinoma in situ (DCIS) is the most common type of non-invasive breast cancer diagnosed in women and an immediate precursor to invasive ductal carcinoma (IDC). It is largely unclear why some cases of DCIS progress to IDC, while others remain non-invasive. Current cyto- and histopathological approaches do not accurately predict disease progression, resulting in patients being under-treated or over-treated for DCIS. Our long-term goals are to identify key factors that lead to IDC that will enable the development of a molecular based approach to predict the risk of IDC, and a more tailored approach to treat DCIS. The CCL2/CCR2 chemokine pathway is best known for regulating macrophage recruitment to late stage breast tumors. In recent studies, we found that CCR2 and its binding ligand CCL2 were overexpressed in breast ductal carcinomas, correlating with tumor grade and poor patient prognosis. To further investigate the significance of CCR2 and CCL2, we performed flow cytometry analysis of breast epithelial cell lines, and found that increased CCR2 overexpression but not CCL2, was associated with invasive potential of the cell lines. CCL2 was found to be increased in stromal cells, particularly in cancer associated fibroblasts. Using a mammary intra-ductal injection model, we further examined for expression patterns of CCR2 and CCL2 in DCIS lesions derived from DCIS.com and Sum225 cells. Increased CCR2 was detected in DCIS.com lesions, which progressed to IDC within 10 weeks of injection, while CCR2 expression was found to be lowly expressed in non-invasive Sum225 lesions. CCL2 was detected in the stroma of both DCIS.com and Sum225 lesions. Overexpression of CCR2 in cultured non-invasive 67NR mammary carcinoma cells enhanced cell survival and increased wound closure. CCL2 treatment further enhanced cell survival and wound closure of CCR2 overexpressing 67NR cells. Knockdown of CCR2 in DCIS.com cells in 3D Matrigel: collagen cultures inhibited cellular invasion into the matrix. These studies indicate that CCR2 overexpression in ductal carcinoma cells, enhances intracellular signaling, and contributes to development of IDC. These data suggest that targeting the CCR2 pathway may be promising strategy for prevention or treatment of IDC.

Citation Format: Benford Mafuvadze, Wei Bin Fang, Nikki Cheng. Overexpression of the chemokine receptor CCR2 in the breast epithelium is associated with progression of DCIS [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P4-05-02.
Abstract P4-05-13: PTEN deletion is linked to adverse phenotype and poor prognosis in breast cancer
Authors: Burandt Eike , Kluth Martina , Kopperschmidt Valerie , Mittenzwei Anja , Lebeau Annette , Müller Volkmar , Witzel Isabell , Jänicke Fritz , Geist Stefan , Paluchowski Peter , Wilke Christian , Heilenkötter Uwe , Krech Rainer , von d Assen Albert , Simon Ronald , Lebok Patrick
Published: May 2015

Deletions of chromosome 10q23, including the PTEN (phosphatase and tensin homolog) locus, are known to occur in breast cancer, but systematic analyses of its clinical relevance are lacking. We thus analyzed a tissue microarray (TMA) with 2,197 breast cancers by fluorescence in-situ hybridization (FISH) using a PTEN-specific probe, and found deletions in 19% of no special type, 9% of lobular, 46% of medullary and 4% of tubular cancers. 98.7% of deletions were heterozygous and 1.3% were homozygous. PTEN deletion was significantly linked to advanced tumor stage (p=0.0054), high tumor grade (p<0.0001), high tumor cell proliferation (Ki67 Labeling Index; p<0.0001), and shortened overall survival (p=0.0090). PTEN deletions were inversely associated with features of luminal type breast cancers (ER/PR positivity, CCND1 amplification). PTEN deletions were strongly linked to amplification of genes involved in the PTEN/AKT pathway such as MYC (p=0.0430) and HER2 (p=0.0065). Remarkably the combined analysis of MYC, HER2, and PTEN aberrations suggested that aberrations of multiple PTEN/AKT pathway genes have a strong additive effect on breast cancer prognosis. Cancers with two or three of these aberrations behaved significantly worse than cancers with none or one of these changes (p<0,0001). The particularly poor prognosis of patients with HER2 amplification and PTEN deletions challenges the concept of PTEN deletions interfering with trastuzumab therapy. In conclusion, PTEN deletion occurs in a relevant fraction of breast cancers, and is linked to aggressive tumors. Reduced PTEN function cooperates with MYC and HER2 activation in conferring aggressive phenotype to cancer cells.

Citation Format: Eike Burandt, Martina Kluth, Valerie Kopperschmidt, Anja Mittenzwei, Annette Lebeau, Volkmar Müller, Isabell Witzel, Fritz Jänicke, Stefan Geist, Peter Paluchowski, Christian Wilke, Uwe Heilenkötter, Rainer Krech, Albert von d Assen, Ronald Simon, Patrick Lebok. PTEN deletion is linked to adverse phenotype and poor prognosis in breast cancer [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P4-05-13.
Abstract P4-07-10: MiRNAs are important regulators of Pax-5 expression and function during breast cancer progression
Authors: Harquail Jason M , Robichaud Gilles A
Published: May 2015

Recent studies have enabled the identification of important factors regulating cancer progression, one of these being the Pax-5 gene. Pax-5, an essential developmental factor of B cells, is aberrantly expressed in various B cell cancer lesions and solid tumors such as breast carcinoma. Although Pax-5 downstream activity is relatively well characterized, the regulation of aberrant Pax-5 expression in a cancer specific context is poorly understood. To investigate the regulation of Pax-5 expression, we turned our attention to micro-RNAs (miRNAs). MiRNAs are highly conserved, small non-coding RNA molecules that regulate key biological processes. Extensive studies also show their deregulation in multiple cancer lesions. In this study, we aim to elucidate a causal link between differentially expressed miRNAs in cancer cells and their putative targeting of Pax-5-dependent cancer processes. With the help of biobank data and bioinformatics analyses, we observe that miRNAs 484 and 210 are aberrantly expressed in breast cancer cells and cross-reference with their predicted capacity to target the Pax-5 mRNA 3’ untranslated region (3’UTR). Using anti- or pre-miRNAs transfected into Pax-5 expressing breast cancer cell lines (MCF-7 and MB231), we demonstrate that miRNAs 484 and 210 are capable of regulating Pax-5 expression. In addition, miRNA-regulated Pax-5 expression resulted in a concomitant alteration in Pax-5-mediated phenotype and cancer processes. This is the first study demonstrating the regulation of Pax-5 expression and function by non-coding RNAs in cancer cells. We believe that the aberrant expression of Pax-5 in cancer cells is in part due to deregulated miRNA expression profiles. This study will bring insight in regards to cancer regulating processes associated with miRNA and Pax-5 deregulations and help us better understand aberrant Pax-5 expression levels within cancerous states. This study can therefore provide the eventual possibility of earlier, more efficient diagnostics as well as more targeted treatments for cancer patients.

Citation Format: Jason M Harquail, Gilles A Robichaud. MiRNAs are important regulators of Pax-5 expression and function during breast cancer progression [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P4-07-10.
Abstract P4-07-12: Re-introduction of tumor suppressor miR-34a harbors therapeutic efficacy in triple negative breast cancer
Authors: Adams Brian D , Pusztai Lajos , Rimm David L , Slack Frank J
Published: May 2015

Place Holder Abstract

Date Expected for Obtaining Final Information for Abstract: 9/14 - 10/14

Triple negative breast cancer accounts for a disproportionate share of the total breast cancer morbidity because of its aggressive behavior, increased incidence in younger women, and lack of effective targeted therapies. MicroRNAs are global regulators of survival and proliferation pathways important in cancer development and tumor maintenance. MicroRNAs function as tumor suppressors and oncogenes, and are highly dysregulated in cancer. Here, we identified miR-34a to be aberrantly lost in triple negative cancer cell lines when compared to both a luminal cancer subtype as well as normal breast cells. Re-introduction of miR-34a in triple negative cancer lines results in inhibition of cell proliferation, reactivation of senescence, and enhances sensitivity to apoptotic-inducing agents. Furthermore, intratumoral delivery of miR-34a into subcutaneously implanted tumors in nude mice delays tumor growth as compared to a scrambled control. In conclusion re-introduction of miR-34a in triple negative breast cancer promotes anti-tumorigenic phenotypes both in vitro and in vivo, and could therefore be used as a therapeutic agent to treat the disease.

Citation Format: Brian D Adams, Lajos Pusztai, David L Rimm, Frank J Slack. Re-introduction of tumor suppressor miR-34a harbors therapeutic efficacy in triple negative breast cancer [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P4-07-12.
Abstract P4-11-09: Comparison of immunohistochemical residual risk panels to predict risk in early breast cancers treated with endocrine therapy
Authors: Stephen Jacqueline , Murray Gordon , Cameron David , Thomas Jeremy , Kunkler Ian , Jack Wilma , Kerr Gill , Piper Tammy , Brookes Cassandra , Rea Daniel , van de Velde Cornelis , Hasenburg Annette , Markopoulos Christos , Dirix Luc , Seynaeve Caroline , Bartlett John
Published: May 2015

Background: We compare two residual risk models combining immunohistochemical (IHC) biomarkers, IHC4 and Mammostrat, in the Edinburgh Breast Conservation Series (BCS) and in the Tamoxifen versus Exemestane Adjuvant Multinational (TEAM) trial.

Materials and Methods: The primary cohorts comprised 831 and 2,513 estrogen receptor (ER)-positive patients who did not receive adjuvant chemotherapy from the Edinburgh BCS and TEAM cohorts respectively. We evaluated prognostic scores for distant recurrence-free survival (DRFS).

Results: Low scores for both IHC4 and Mammostrat are associated with better DRFS. In multivariate Cox regression analyses the addition of both scores to clinical factors provided independent information on residual risk (p<0.05). In the larger TEAM cohort, IHC4 was the stronger predictor of DRFS but additional information was gained from including the Mammostrat score for all ER-positive patients (p<0.001).

Conclusion: The results showed that the scores have different capabilities in predicting DRFS depending on the study and subgroup of patients. However, significant benefit in estimating residual recurrence risk after treatment was observed from a combined use of both marker panels. This provides support for investigating their combined use for risk stratification of ER-positive early breast cancer patients.

Citation Format: Jacqueline Stephen, Gordon Murray, David Cameron, Jeremy Thomas, Ian Kunkler, Wilma Jack, Gill Kerr, Tammy Piper, Cassandra Brookes, Daniel Rea, Cornelis van de Velde, Annette Hasenburg, Christos Markopoulos, Luc Dirix, Caroline Seynaeve, John Bartlett. Comparison of immunohistochemical residual risk panels to predict risk in early breast cancers treated with endocrine therapy [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P4-11-09.
Abstract P4-11-10: Time dependence of biomarkers: Non-proportional effects of immunohistochemical panels predicting relapse risk in early breast cancer
Authors: Stephen Jacqueline , Murray Gordon , Cameron David , Thomas Jeremy , Kunkler Ian , Jack Wilma , Kerr Gill , Piper Tammy , Brookes Cassandra , Rea Daniel , van de Velde Cornelis , Hasenburg Annette , Markopoulos Christos , Dirix Luc , Seynaeve Caroline , Bartlett John
Published: May 2015

Background: We investigate the impact of follow-up duration to determine whether two immunohistochemical prognostic panels, IHC 4 and Mammostrat, provide information on the risk of early or late distant recurrence using the Edinburgh Breast Conservation Series and the Tamoxifen versus Exemestane Adjuvant Multinational (TEAM) trial.

Methods: The multivariable fractional polynomial time (MFPT) algorithm was used to determine which variables had possible non-proportional effects. The performance of the scores was assessed at various lengths of follow-up and Cox regression modelling performed over the intervals 0-5 years and > 5 years.

Results: We observed a strong time-dependence of both the IHC4 and Mammostrat scores with their effects decreasing over time. In the first five years of follow-up only, the addition of both scores to clinical factors provided statistically significant information (p<0.05) with increases in R2 between 5 and 6% and increases in D-statistic between 0.16 and 0.21.

Conclusion: Our analyses confirm that the IHC4 and Mammostrat scores are strong prognostic factors for time to distant recurrence but this is restricted to the first 5 years after diagnosis. This provides evidence for their combined use to predict early recurrence events in order to select those patients who may/will have benefit from adjuvant chemotherapy.

Citation Format: Jacqueline Stephen, Gordon Murray, David Cameron, Jeremy Thomas, Ian Kunkler, Wilma Jack, Gill Kerr, Tammy Piper, Cassandra Brookes, Daniel Rea, Cornelis van de Velde, Annette Hasenburg, Christos Markopoulos, Luc Dirix, Caroline Seynaeve, John Bartlett. Time dependence of biomarkers: Non-proportional effects of immunohistochemical panels predicting relapse risk in early breast cancer [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P4-11-10.
Abstract P4-11-41: Pretreatment neutrophil/lymphocyte ratio and overall survival in African American and white breast cancer patients
Authors: Rimando Joseph C , Campbell Jeff , Kim Jae Hee , Kim Sangmi
Published: May 2015

Previous studies have shown that the pretreatment neutrophil/lymphocyte ratio (NLR) is an independent predictor of mortality in breast cancer patients. Our aim was to further study the relationship between pretreatment NLR and overall survival in African American and white breast cancer patients treated at an academic cancer center. Electronic medical records were reviewed for 589 patients treated between 2002 and 2011, and pretreatment NLR data, determined at an average of 12 days prior to the initiation of cancer treatment, were available from 217 African American and 218 white patients. Other clinical and patient data were obtained from the hospital tumor registry, with annual follow-up for vital status. There were a total of 102 deaths over a mean follow-up of 59 months. For data analysis, patients were divided into quartiles based on their NLR (Q1: <1.54; Q2: 1.54≤2.0; Q3: 2.0≤2.79; and Q4: ≥ 2.79). Patients in the highest quartile of pretreatment NLR showed an increase in overall mortality compared to those in the lowest quartile, with a hazard ratio (HR) of 2.4 (p<0.005). After adjustment for age, tumor stage, and grade, the pretreatment NLR remained significantly associated with overall mortality (HR=2.1, p=0.03). Further adjustment for race, body mass index (BMI) and smoking increased the effect size (HR=3.2, p<0.01), but only half patients remained in the analysis due to incomplete data on BMI and smoking. In our study, race, but neither BMI nor smoking, was associated with pretreatment NLR values: African American patients had higher pretreatment NLR values than white patients regardless of stage. In a stratified analysis by race, HRs for overall mortality associated with the highest quartile of pretreatment NLR versus the remaining three quartiles combined were 2.5 (p<0.01) among African Americans and 1.6 (p=0.24) among white patients, although the interaction between race and pretreatment NLR was not statistically significant. Our data confirm previous findings that pretreatment NLR, a marker of systemic inflammatory response, is unfavorably associated with overall survival in breast cancer patients, and also suggest that the association may be more evident among African American patients whose levels were generally higher than white breast cancer patients. Future studies are warranted to investigate the prognostic utility of pretreatment NLR in different racial groups and whether a heightened systemic inflammatory response is an underlying contributor to racial difference in breast cancer outcomes.

Citation Format: Joseph C Rimando, Jeff Campbell, Jae Hee Kim, Sangmi Kim. Pretreatment neutrophil/lymphocyte ratio and overall survival in African American and white breast cancer patients [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P4-11-41.