bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P1-16-07: Pharmacogenomics-pharmacokinetics study of selective estrogen-receptor modulators with intra-patient dose-escalation for Japanese breast cancer patients
Authors: Ohno S , Ishiguro H , Yamamoto Y , Takao S , Sato N , Fujisawa T , Kadoya T , Kuroi K , Bando H , Teramura Y , Iwata H , Tanaka S , Toi M
Published: May 2016

Background

The association between CYP2D6 polymorphisms and efficacy of tamoxifen (TAM) is inconclusive, partly due to inaccurate prediction of active metabolite, endoxifen exposure by solely CYP2D6 genotype. Moreover, TAM dose escalation is not effective for poor metabolizers. Since the contribution of CYP2D6 to toremifen (TOR) activation is small, TOR might be a good alternative to TAM for poor metabolizers.

Methods

Patients who maintained good compliance with TAM or TOR without regular use of strong CYP2D6 inhibitors were enrolled in a screening study. The pharmacokinetics (PK) of TAM or TOR and the pharmacogenomics (PGx) of metabolizing enzymes and transporters were assessed. Associations between TAM and TOR PK and PGx, other CYP inhibitor use, and smoking status were examined by regression analysis. An intra-patient dose escalation study was conducted for patients showing low endoxifen levels during TAM treatment (n = 14). TAM was switched to 40 mg of TOR, and then increased to 120 mg for ≥24 weeks with periodic PK sampling. Total TAM or TOR activity was calculated as the sum of the concentration of each active metabolite adjusted by their respective in vitro activity.

Results

CYP2D6 genotype was the major determinant for TAM activity (p < 0.01) for Japanese breast cancer patients. Current smoking status (p = 0.07) and CYP2C19 (p = 0.07), but not CYP2D6 genotype (p = 0.61), showed marginally significant effects on TOR activity. TOR activity sufficiently increased with dose escalation from 40 mg to 120 mg, even among the poor TAM metabolizers, and was maintained for ≥24 weeks.

Total Activity of TOR (ng/mL) Screening (n=271)Intra-patient dose escalation (n=14)Dose (mg)4040120120*120/40120*/40120*/120Mean (ng/mL)4168.53218.18758.67268.03.12.60.9Median (ng/mL)3865.43020.38924.67579.72.82.50.8SD1446.21207.01707.11793.21.20.80.2p-value0.04  <0.01<0.010.054*(Sample taken at ≥24 weeks)

Conclusion

Since the contribution of CYP2D6 to TOR metabolism is relatively small, TOR is a valid alternative to TAM, especially in patients predicted to be poor TAM metabolizers. Further clinical trials are warranted to validate this concept.

Citation Format: Ohno S, Ishiguro H, Yamamoto Y, Takao S, Sato N, Fujisawa T, Kadoya T, Kuroi K, Bando H, Teramura Y, Iwata H, Tanaka S, Toi M. Pharmacogenomics-pharmacokinetics study of selective estrogen-receptor modulators with intra-patient dose-escalation for Japanese breast cancer patients. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P1-16-07.
Abstract P2-01-07: Neutrophil lymphocyte ratio and breast cancer: A future prognostic indicator for outcome after surgery?
Authors: Williams GJ , Huws AM , Holt SDH , Sharaiha YM
Published: May 2016

There is a growing body of literature highlighting the prognostic use of pre-operatively comparing certain white blood cell differentials, namely neutrophils and lymphocytes, with outcomes in many different types of solid tumour cancers. It has been postulated that a raised neutrophil to lymphocyte ratio (NLR) may reflect a greater tumour burden on the patient, and therefore lead to poorer outcomes. There is some evidence that a raised pre-operative NLR may help to predict poorer outcomes following surgery for breast cancer.

This observational retrospective proof of concept study aims to assess the association between pre-operative NLR and breast cancer outcomes over a three year period of follow-up from a cohort of 147 patients. The results of this study revealed that the cohort of patients with a raised pre-operative NLR was associated with a clinically significant increased rate of local recurrence of their breast cancer following surgery with curative intent (with or without adjuvant chemotherapy and radiotherapy) compared to those that had a low or normal NLR (20.4% ± 4.9% vs 5.1% ± 4.9% respectively, CI 95%; p=0.004). In relation to intra-operative sentinel lymph node biopsy, the analysis also showed that the incidence of a raised pre-operative NLR was not related to whether there was nodal involvement or not at the time of surgery (33% vs 33%, p=1). This suggests that a raised pre-operative NLR may be an independent prognostic indicator for breast cancer outcomes.

This study provides a template for a prospective observational study which is needed to corroborate this result and to account for confounding factors such as concomitant chronic disease. Additionally, a larger cohort sample size with a follow-up of five years is necessary to assess the relationship between pre-operative NLR and mortality due to the latter's relatively low incidence in breast cancer. However, this study further supports that consideration of the pre-operatively calculated NLR may provide an early prognostic indicator for breast cancer outcomes and warrants further investigation.

Citation Format: Williams GJ, Huws AM, Holt SDH, Sharaiha YM. Neutrophil lymphocyte ratio and breast cancer: A future prognostic indicator for outcome after surgery?. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P2-01-07.
Abstract P2-02-06: Impact of apoptotic circulating tumor cells in metastatic breast cancer
Authors: Wallwiener M , Deutsch TM , Riethdorf S , Hartkopf AD , Taran F-A , Trumpp A , Brucker S , Schütz F , Rom J , Pantel K , Schneeweiss A
Published: May 2016

Background

Circulating tumor cells (CTC) are a heterogeneous cell population and an independent negative prognostic factor for progression-free (PFS) and overall survival (OS) in patients with metastatic breast cancer (MBC). The study aimed to prospectively assess CTC status for the subtypes apoptotic CTC (aCTC) and intact CTC (iCTC) at baseline (CTCBL) and after one cycle of a new line of systemic therapy (CTC1C). Changes from CTCBL to CTC1C (CTC kinetics, CTCKIN) were evaluated for their utility in predicting response, progression-free (PFS) and overall survival (OS).

Methods

423 MBC patients were included in a prospective trial prior to a new regimen of treatment. Intact and apoptotic CTC were analyzed at baseline (CTCBL) and after one cycle of systemic therapy (CTC1C) using CellSearch™ (Veridex) and morphologic criteria. Samples with ≥5 CTC/7.5ml blood were regarded as positive. Therapy response was assessed using the RECIST-criteria on three-monthly radiological controls. CTCKIN were characterized by ≥25% change from CTCBL to CTC1C to differentiate stable, increased and decreased CTC kinetic.

Results

35% of patients were iCTCBL-positive and 28% aCTCBL-positive at baseline (CTCBL). PFS and OS differ significantly between the iCTCBL-positive and the iCTCBL-negative group (PFS 4.5 vs. 8.0; OS 12.5 vs. 27.2 (months)). Positive aCTC in conjunction with positive iCTCBL at baseline has worst prognostic impact (PFS 6.3; OS 8.7).

Regarding the CTCKIN (BL to 1C), aCTC-decrease (≥25%) is a positive prognostic factor compared to aCTC-stable and aCTC-increase (PFS 7.6 vs. 3.7; 3.3 and OS 21.0 vs. 4.8; 5.7). Decreasing aCTCKIN shows favorable prognostic impact versus decreasing iCTCKIN (PFS 7.6 vs. 5.9 and OS 21.0 vs 16.4).

Conclusion

Elevated aCTC levels at baseline have an unfavorable prognostic impact on both OS and PFS in conjunction with elevated iCTC. Additionally, the decrease of aCTC is a relevant prognostic value for systemic therapy response. aCTCKIN allows better differentiation for therapy response in patients with positive CTC-status at baseline. Differentiated enumeration of intact and apoptotic CTC should be considered in clinical application.

Citation Format: Wallwiener M, Deutsch TM, Riethdorf S, Hartkopf AD, Taran F-A, Trumpp A, Brucker S, Schütz F, Rom J, Pantel K, Schneeweiss A. Impact of apoptotic circulating tumor cells in metastatic breast cancer. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P2-02-06.
Abstract P2-02-08: Initial circulating tumor cell count and venous thrombosis-free interval in the course of metastatic breast cancer
Authors: Beinse G , Berger F , Cottu PH , Dujaric M-E , Kriegel I , Guilhaume M-N , Dieras V , Beuzeboc P , Pierga J-Y , Bidard F-C
Published: May 2016

Background: Circulating tumor cells (CTC) count is a major prognostic factor in metastatic breast cancer. It has been also reported to be associated with venous thrombosis in short-term studies on advanced metastatic breast cancer patients.

Methods: We assessed whether an early CTC detection (CellSearch®), before the start of the first line chemotherapy, impacts Thrombosis-Free Interval during the whole course of metastatic breast cancer. Electronic medical files of all patients included in the large prospective IC 2006-04 CTC detection study (NCT00898014, Pierga et al, Ann Oncol 2012) and treated at Institut Curie (Paris, France) were manually and automatically searched for thrombosis events in the course of their metastatic breast cancer. Superficial venous thromboses were not taken into account.Thrombosis-Free Interval (TFI) was defined as the first event occuring between venous or arterial thrombosis.

Results: In the 142 patients studied, with a median follow-up of 64 months [25-81 months], venous (deep venous thrombosis, pulmonary embolism, catheter thrombosis and portal vein thrombosis) and arterial thrombosis occurred in 21.8% and 4.2% of patients, respectively. Twenty-four (30%) of the 80 patients with ≥1 CTC/7.5ml of blood before the start of first line chemotherapy experienced at least one thrombotic events in the course of their disease, while only 10 (16%) of the 62 CTC-negative patients experienced a similar event (p=0.06). Baseline positive CTC was associated with a higher risk of experiencing a venous or arterial thrombosis (TFI: HR=2.6 [1.2-5.5], p=0.009). The median TFI was 64 months in CTC-positive patients and not reached in CTC-negative patients. Among the other patients characteristics tested at baseline, Performance Status (PS=1 vs PS=0: HR=0.85 [0.4-1.9], p=NS; PS=2 or 3 vs PS=0: HR=3.7 [1.5-8.7], p=0.003) and LDH levels at baseline (LDH>UNL vs LDH<UNL HR=2.4 [1.2-4.9], p=0.012) were also associated with TFI.

Conclusions: Metastatic breast cancer patients with ≥1 CTC before the start of chemotherapy are at higher risk of venous thrombosis; the clinical relevance of thrombo-prophylaxis should be investigated in this population.

Citation Format: Beinse G, Berger F, Cottu PH, Dujaric M-E, Kriegel I, Guilhaume M-N, Dieras V, Beuzeboc P, Pierga J-Y, Bidard F-C. Initial circulating tumor cell count and venous thrombosis-free interval in the course of metastatic breast cancer. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P2-02-08.
Abstract P2-03-06: Dysregulation of miR-34a-SIRT1 axis reduced breast cancer stemness
Authors: Xiao GG , Ma W , Mao J , Li L
Published: May 2016

Recent studies show that enforced expression of miR-34a results in elimination of cancer stem cells (CSCs) in many malignant tumors. Sirtuin-1 (SIRT1) being confirmed as a direct target of miR-34a was reported to be involved in regulation of growth and survival of leukemia stem cells (LSCs). In this study, we aim to understand regulatory mechanism of miR-34a -SIRT1 axis in breast cancer stem cells (BCSCs). Lower endogenous level of miR-34a and higher level of Sirtuin1 (SIRT1) gene were identified in CD44+/CD24- BCSCs than breast cancer cells. Either ectopic expression of miR-34a or silenced SIRT1 in MCF-7 cells inhibited cellular proliferation, and led to cell apoptosis. Overexpression of miR-34a also suppressed expression of ALDH1, BMI1 and Nanog, and decreased capacity of mammosphere formation significantly.. Studies in vivo showed that stable expression of miR-34a reduced tumor burden significantly in nude mice xenografts. Taken together, our results showed that miR-34a inhibit the proliferative potential of BCSCs in vitro and in vivo, at least partially through downregulating SIRT1. miR-34a-SIRT1 axis may play an important role in self-renewal and stemness maintenance of BCSCs. This study may provide a novel BCSCs specific therapeutic strategy to improve breast cancer treatments.

Citation Format: Xiao GG, Ma W, Mao J, Li L. Dysregulation of miR-34a-SIRT1 axis reduced breast cancer stemness. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P2-03-06.
Abstract P2-04-03: SUMO wrestles breast cancer: SUMO posttranslational modification directs breast cancer cell epigenome
Authors: Bawa-Khalfe T , Kumar S , Lin F-M
Published: May 2016

Epigenetic reprogramming allows breast cancer (BCa) cells to bypass normal growth checkpoints and acquire an aggressive morphology. Function of critical epigenetic proteins is directed by small ubiquitin-like modifiers (SUMO) posttranslational modification (PTM). Components of the SUMO system are dysregulated in BCa. Our objective is to delineate how changes to SUMO-PTM affect epigenetic programs that coordinate BCa pathology. We report that chromatin-bound SUMO protease SENP7L is upregulated in metastatic tamoxifen-resistant (TamR) and triple-negative BCa. SENP7L facilitates chromatin remodeling that potentiates the growth and invasiveness of these BCa subtypes. SENP7L interacts with and deSUMOylates heterochromatin protein 1 alpha (HP1-alpha/CBX5) to mediates these responses. Canonical models suggest that HP1-alpha reads the histone mark to localize predominantly at constitutive heterochromatin. Our studies challenge the old paradigm as we show that SUMOylated HP1-alpha is enriched outside heterochromatin at euchromatin, including E2F-responsive and mesenchymal gene loci. Recruitment of SUMOylated HP1-alpha supports de novo heterochromatin formation at these loci. However, SUMOylated HP1-alpha is lost in aggressive BCa due to SENP7L induction. Consistently, SENP7L upregulation prompts chromatin de-condensation and gene transcription. Nuclei acid and chemical targeting of SENP7L reduces self-renewal/differentiation properties of TamR mammospheres as well as tumor volume and secondary tumor formation of TamR cells. Hence, SENP7L inhibitors can serve as promising tools to modulate epigenetic abnormalities in TamR BCa.

Citation Format: Bawa-Khalfe T, Kumar S, Lin F-M. SUMO wrestles breast cancer: SUMO posttranslational modification directs breast cancer cell epigenome. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P2-04-03.
Abstract P2-05-05: Expression profiling of in vivo DCIS progression models identified BCL9 as a molecular driver of invasive progression
Authors: Behbod F , Elsarraj H , Hong Y , Valdez K , Chien J , Godwin A , Fields T
Published: May 2016

Introduction: There are an estimated 60,000 new cases of ductal carcinoma in situ (DCIS) each year. At present, it is not clear why some DCIS remain non-invasive for decades while others become invasive. A lack of understanding in DCIS pathobiology has led to overtreatment of more than half of DCIS patients. To identify factors that promote DCIS invasion, we have profiled the temporal molecular changes during DCIS transition to invasive ductal carcinoma (IDC) using two in vivo models, MIND (mouse-intraductal) and DCIS/IDC tandem lesions. These studies led to the identification of B cell lymphoma-9 as a potential molecular driver of early invasion. BCL9 is a newly found co-activator of Wnt-stimulated β-catenin-mediated transcription. BCL9 has been shown to promote progression of multiple myeloma and colon carcinoma. However its role in breast cancer progression had not been recognized.

Methods: Microarray and RNA sequencing were utilized to characterize the sequential and temporal changes in mRNA expression during DCIS invasive transition. BCL9 shRNA knockdown was performed to assess the role of BCL9 in in vivo invasion, EMT and canonical Wnt signaling. Immunofluorescence of 28 patient DCIS samples was used to assess a correlation between the expression of BCL9 and biomarkers of high risk DCIS. TCGA data was analyzed to assess the status of BCL9 gene alterations in 959 human breast cancers.

Results: Analysis of BCL9, by RNA and protein showed BCL9 up-regulation to be associated with DCIS transition to IDC. Analysis of patient DCIS revealed a significant correlation between high nuclear BCL9 and pathologic characteristics associated with DCIS recurrence: ER and PR negative, high nuclear grade, and high HER2. In vivo silencing of BCL9 resulted in the inhibition of DCIS invasion and reversal of epithelial-mesenchymal transition (EMT). Analysis of the TCGA data showed BCL9 gene to be altered in 26% of breast cancers. This is a significant alteration when compared to ERBB2 (19%) and ESR1 (8%). A significantly higher proportion of basal like invasive breast cancers showed BCL9 amplification.

Conclusion: BCL9 is a molecular driver of DCIS invasive progression and may predispose to the development of basal like invasive breast cancers. As such, BCL9 has the potential to serve as a biomarker of high risk DCIS and as a therapeutic target for prevention of IDC.

Citation Format: Behbod F, Elsarraj H, Hong Y, Valdez K, Chien J, Godwin A, Fields T. Expression profiling of in vivo DCIS progression models identified BCL9 as a molecular driver of invasive progression. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P2-05-05.
Abstract P2-05-08: p53 deficiency enhances metastatic potential of triple negative breast cancer by promoting growth in primary and metastatic sites
Authors: Powell E , Shao J , Mistry N , Yuan Y , Tieu T , Decker K , Chen H-C , Cai S , Do K-A , Heffernan TP , Edwards J , Piwnica-Worms D , Piwnica-Worms H
Published: May 2016

TP53 is one of the most commonly mutated genes in human cancer. Mutation or loss of TP53 can occur during the early stages of oncogenesis or as a later event as tumors progress to more aggressive forms. We used a patient-derived xenograft (PDX) model of triple negative breast cancer (TNBC) to investigate how p53 deficiency contributes to metastasis. The PDX line was generated by engrafting the primary breast tumor of a patient with TP53 wild-type metastatic TNBC into the humanized mammary fat pad of NOD/SCID mice. Isogenic lines differing only in TP53 status were generated from this model by silencing p53. Here we demonstrate that the p53-proficient and -deficient tumors metastasized from the mouse mammary gland and colonized the lung, liver, bone, brain, and lymph nodes with similar frequency. p53-deficient tumors grew faster in both the primary and metastatic sites as a result of increased mitosis and decreased apoptosis. Circulating tumor cells (CTCs) are currently being evaluated in p53-proficient and –deficient PDX models. Gene expression profiling identified candidate genes with potential clinical relevance that may be responsible for increased tumor growth upon p53 loss in late stage tumors.

This model is also being used to identify genes that promote breast cancer metastasis. To enrich for metastasis, metastatic subpopulations of tumor cells were isolated from lungs and serially passaged in vivo. This serial passaging enriched for metastasis to multiple secondary sites. A transcriptional signature of putative metastasis genes was generated and is being functionalized with high throughput screening in vivo.

Citation Format: Powell E, Shao J, Mistry N, Yuan Y, Tieu T, Decker K, Chen H-C, Cai S, Do K-A, Heffernan TP, Edwards J, Piwnica-Worms D, Piwnica-Worms H. p53 deficiency enhances metastatic potential of triple negative breast cancer by promoting growth in primary and metastatic sites. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P2-05-08.
Abstract P2-05-13: A novel mutagenesis screen identifies SHARPIN as a breast cancer metastasis gene that predicts survival of breast cancer patients
Authors: Bii VM , Rae DT , Trobridge GD
Published: May 2016

Breast cancer (BC) is a heterogeneous disease and the second leading cause of malignancy among women in the U.S.. Metastasis of the primary tumor results in poor prognosis and increased mortality and the molecular mechanisms by which metastatic tumors occur are not well understood. Identifying the genes that drive the metastatic process could provide targets for improved therapy and biomarkers to improve outcomes for BC patients. Here, we utilized a replication-incompetent gammaretroviral vector (γRV) to perform a forward insertional mutagenesis screen to identify genes involved in BC metastasis. In this approach, BC cells mutagenized with a γRV were xenotransplanted into the mammary fat pad of immunodeficient mice, and primary tumors and metastases were allowed to develop. Metastatic lesions were collected and analyzed for proviral integration sites to identify vector integration sites and nearby candidate metastasis genes. The γRV has a bacterial origin of replication and kanamycin resistance gene that allows for rescue in bacteria and rapid identification of vector integration sites. Using this approach, we identified the previously described metastasis gene WWTR1, and several other novel candidate metastasis genes including SHARPIN. SHARPIN was then independently validated as a BC metastasis gene in vivo using RNAi. Analysis of patient data showed that SHARPIN expression predicts metastasis-free survival after adjuvant chemotherapy (p < 0.005, Concordance Index = 55.3, Risk Groups Hazard Ratio = 1.87). Our replication-incompetent γRV approach is efficient and has broad potential to identify genes involved in oncogenic processes for BC and other cancers.

Keywords

Insertional mutagenesis screen, Gammaretroviral vector (γRV), Metastasis, Inducible shRNA, Prognostic biomarker.

Citation Format: Bii VM, Rae DT, Trobridge GD. A novel mutagenesis screen identifies SHARPIN as a breast cancer metastasis gene that predicts survival of breast cancer patients. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P2-05-13.
Abstract P2-05-15: NOS2&COX2 activation of TLR4 & EGFR signalling causes poor outcome in oestrogen receptor-negative breast cancer via pro-survival signals and immune polarisation
Authors: Glynn S , Garrrido-Cuesta P , Wink D , Ridnour L , Ambs S , Keane M , Walsh E , Callagy G
Published: May 2016

Background

We seek to further elucidate mechanisms by which inflammatory mediators promote estrogen receptor (ER)-negative breast cancer progression and poor survival. We previously reported association between inducible nitric oxide synthase (NOS2) and poor outcome in ER-negative tumours. In tumours aberrant NOS2 induction facilitates tissue remodelling and stimulates neovascularisation. Also involved in inflammation and wound healing is cyclooxygenase-2 (COX2). We demonstrated that COX2 is associated with Akt activation and poor outcome in ER-negative tumours. We hypothesise that co-expression COX2 with NOS2 in ER-negative tumours amplifies effects of NOS2 on poor outcome, via EGFR and TLR4 signalling loop activation, and polarization of the tumour immune-compartment to pro-tumorigenic M2 phenotype.

Methodology

We determined the association of NOS2 and COX2 co-expression on breast cancer specific survival in ER-negative and triple negative breast tumours (N=102), via immunohistochemistry and cox regression statistical analysis. To explore the mechanism of NOS2 induction of COX2 through transactivation of EGFR, NO donors in combination with EGFR inhibitors were used to determine if NO exposure results in amplified EGFR and PGE2 pro-survival and pro-metastatic signalling in triple negative breast cancer cell lines. Finally, we explored the ability of NO to modify the ability of triple negative breast cancer secretome to induce polarisation of macrophages to a pro-tumorigenic M2 phenotype.

Results

Co-expression of NOS2 and COX2 in triple negative breast cancer results in poor outcome, via activation of pro-survival signalling and modification of the immune compartment to a pro-tumorigenic M2 associated phenotype. NO induces activation of growth factor signalling pathways and secretion of M2 promoting cytokines that induce THP1 macrophage polarization to an M2 phenotype.

Citation Format: Glynn S, Garrrido-Cuesta P, Wink D, Ridnour L, Ambs S, Keane M, Walsh E, Callagy G. NOS2&COX2 activation of TLR4 & EGFR signalling causes poor outcome in oestrogen receptor-negative breast cancer via pro-survival signals and immune polarisation. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P2-05-15.
Abstract P2-05-21: The AKT-mTOR pathway as a potential organ-specific drug target signature of hepatic metastases from breast cancer
Authors: Pierobon M , Wong S , Reeder A , Anthony SP , Robert NJ , Northfelt DW , Jahanzeb M , Vocila L , Wulfkuhle J , Dunetz B , Aldrich J , Byron S , Craig D , Liotta L , Petricoin EF , Carpten J
Published: May 2016

Background: The identification of organ-specific targetable signatures may help design more effective treatment for patients with metastatic breast cancer (MBC). We took a multi-OMIC approach to assess whether the AKT-mTOR pathway is globally activated during metastatic progression or whether it represents an organ-specific target.

Methods: Snap frozen biopsies from 25 MBC patients enrolled in a prospective phase II trial were used. Sites of metastasis were classified as liver (n=8) and others (n=17), the latter including cutaneous, lung, lymph nodes, and intra-abdominal lesions. Signaling analysis of the 25 cases was performed using Reverse Phase Protein Microarray (RPPA) coupled with Laser Capture Microdissection. Activation of the AKT-mTOR pathway was quantified as phosphorylation of AKT (S473) and the mTOR target p70S6 (T389). Matched exome (WES) and RNASeq data were available for 17 of 25 patients, five with liver metastases. Sequencing data was processed using an in-house developed pipeline to identify somatic events including coding mutations, copy number alterations, gene fusions, and differential expression. Activation of the AKT-mTOR pathway and sequencing data were compared between hepatic and non-hepatic lesions using an integrated RPPA and genomic approach.

Results: Among liver metastases, AKT was activated in 4 of the 8 (50.0%) patients, while 6 of the 8 cases (75.0%) showed activation of p70S6. Sequencing data revealed mutation of PIK3CA in 4 of the 5 liver metastases (80.0%). Three of the PIK3CA mutated specimens with catalytic domain mutations (codons 1023 and 147) demonstrated co-activation of AKT and p70S6, while the fourth case, containing a helical domain mutation (E542K), had activation of p70S6 only. The PIK3CA wild-type liver metastasis demonstrated low activation of AKT and p70S6. For non-hepatic metastases AKT was activated in 2 of the 17 cases (11.8%) and p70S6 in 5 of the 17 patients (29.4%).

Discussion: Although these results need further validation, activation of the AKT-mTOR pathway appears to be a hepatic specific signature in MBC and could be used for the selection of targeted agents for hepatic lesions.

Citation Format: Pierobon M, Wong S, Reeder A, Anthony SP, Robert NJ, Northfelt DW, Jahanzeb M, Vocila L, Wulfkuhle J, Dunetz B, Aldrich J, Byron S, Craig D, Liotta L, Petricoin EF, Carpten J. The AKT-mTOR pathway as a potential organ-specific drug target signature of hepatic metastases from breast cancer. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P2-05-21.
Abstract P2-08-06: Usefulness of the pre-treatment neutrophil-to-lymphocyte ratio in predicting first-line progression free-survival in triple-negative breast cancer patients
Authors: Fontanella C , Fanotto V , Gerratana L , Bonotto M , Cinausero M , Bozza C , Iacono D , Russo S , Andreetta C , Minisini AM , Moroso S , Mansutti M , Fasola G , Puglisi F
Published: May 2016

Background: The neutrophil-to-lymphocyte ratio (NLR) is an independent predictor of poor prognosis inunselected breast cancer patients with NLR >3.3. Moreover, pre-treatment NLR has been associated with disease-free and overal survival (OS) in patients with early triple-negative breast cancer (TNBC). We aimed to determine whether the NLR is predictive of progression-free survival (PFS) in metastatic TNBC.

Methods: We reviewed the records of 48 TNBC patients who received at least one administration of first-line (1°L) chemotherapy for advanced disease from October 2004 to April 2014. The NLR (absolute neutrophil count/absolute lymphocyte count) was calculated from the full blood count routinely performed immediately before the initiation of first-line treatment. The association between categorical variables was calculated by X2 test. PFS (from start of 1°L treatment to disease progression or death) and OS (from start of 1°L treatment to death) were estimated using Kaplan Meier method. Multivariable Cox regression was used to determine the independent prognostic significances of the NLR (co-variables stage at diagnosis, histology, and tumor grade).

Results: NLR was not associated with stage at diagnosis (p=0.214), histology (p=0.597), or tumor grade (p=0.775). After a median follow-up of 10.9 months (range 1.3-54.9), 88.6% of TNBC patients with NLR≤3.3 versus 0.0% of patients with NLR>3.3 had a 1°L PFS>3 months (p<0.001). Similarly, 62.9% of TNBC patients with NLR≤3.3 versus 30.8% of patients with NLR>3.3 had an OS>10 months (p=0.047). Metastatic TNBC patients with NLR≤3.3 had a longer median 1°L PFS (5.2 months) and median OS (13.5 months) compared with patients with NLR>3.3 (1°L PFS 2.1 months, p<0.001; OS 7.7 months, p=0.018). In multivariable analysis, NLR>3.3 is associated with a shorter PFS (hazard ratio [HR] 22.4; 95% confidence interval [CI] 6.7-75.1, p<0.001) and higher risk of death (HR 3.2, 95%CI 1.4-7.4, p=0.005).

Conclusion: Our study showed that pre-treatment NLR is associated with 1°L PFS and OS in patients with metastatic TNBC. However, further investigation in larger series of metastatic TNBC is warranted.

Citation Format: Fontanella C, Fanotto V, Gerratana L, Bonotto M, Cinausero M, Bozza C, Iacono D, Russo S, Andreetta C, Minisini AM, Moroso S, Mansutti M, Fasola G, Puglisi F. Usefulness of the pre-treatment neutrophil-to-lymphocyte ratio in predicting first-line progression free-survival in triple-negative breast cancer patients. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P2-08-06.
Abstract P2-08-07: Prognostic significance of immune checkpoint receptors in node-negative breast cancer
Authors: Schmidt M , van de Sandt L , Heimes A-S , Battista M , Lebrecht A , Almstedt K , Hoffmann G , Rahnenführer J , Hengstler JG
Published: May 2016

Background: Immune checkpoint blockade is increasingly discussed in breast cancer. We examined the prognostic significance of the immune checkpoint receptors cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) and programmed cell death protein 1 (PD-1) in node-negative breast cancer.

Methods: Microarray based gene-expression data for CTLA-4 (221331_x_at) and PD-1 (207634_at) were analysed in four previously published cohorts (Mainz, Rotterdam, Transbig, Yu) of node-negative breast cancer patients not treated with adjuvant therapy (n=824). A meta-analysis of previously published cohorts was performed using a random effects model. Prognostic significance of CTLA-4 as well as PD-1 for metastasis-free survival (MFS) was examined in the whole cohort and in different molecular subtypes: luminal (ER+/HER2-), basal-like (ER-/HER2-) and HER2+. Independent prognostic relevance was analysed using multivariate Cox regression.

Results: Higher RNA expression of CTLA-4 was related to better MFS in a meta-analysis of the whole cohort (HR 0.58, 95% CI 0.38-0.88, P=0.0102). Prognostic significance was most pronounced in the HER2+ positive molecular subtype (HR 0.23, 95% CI 0.08-0.65, P=0.0062) as compared to luminal (HR 0.68, 95% CI 0.39-1.18, P=0.1744) and basal-like (HR 0.53, 95% CI 0.25-1.15), P=0.1087) carcinomas of the breast. PD-1 RNA expression, however, was not associated with outcome in the whole cohort of patients (HR 0.88, 95% CI 0.32-2.43, P=0.1853). A trend for improved survival was noticed in basal-like breast cancer (HR 0.40, 95% CI 0.15-1.08, P=0.0701). Neither luminal (HR 0.81, 95% CI 0.28-2.36, P=0.2122) nor HER2+ (HR 0.85, 95% CI 0.27-2.68, P=0.7759) patients showed an association of PD-1 with MFS. CTLA-4 showed independent prognostic significance (HR 0.393, 95% CI 0.224-0.688, P=0.001) in multivariate analysis. In addition to CTLA-4, only histological grade of differentiation (HR 2.335, 95% CI 1.490-3.660, P<0.0001) and tumor size (HR 1.924, 95% CI 1.260-2.937, P=0.002), but neither PD-1 nor age nor HER2 status nor hormone receptor status retained an independent prognostic association with MFS.

Conclusions: The immune checkpoint receptor CTLA-4 has independent prognostic significance in node-negative breast cancer. Higher expression of CTLA-4 is associated with improved outcome. The prognostic impact of CTLA-4 differs between molecular subtypes and is most pronounced in HER2+ breast cancer.

Citation Format: Schmidt M, van de Sandt L, Heimes A-S, Battista M, Lebrecht A, Almstedt K, Hoffmann G, Rahnenführer J, Hengstler JG. Prognostic significance of immune checkpoint receptors in node-negative breast cancer. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P2-08-07.
Abstract P2-08-17: Prognostic impact of breast cancer subtypes in elderly patients
Authors: Bergen ES , Tichy C , Berghoff AS , Rudas M , Dubsky P , Bago-Horvath Z , Mader RM , Gnant M , Dieckmann K , Zielinski CC , Steger GG , Preusser M , Bartsch R
Published: May 2016

Background

We aimed to analyze the impact of BC subtypes on the clinical course with special emphasis on the occurrence of brain metastases (BM) and outcome in an elderly breast cancer population.

Patients and Methods

571 patients ≥65 years receiving treatment for BC from 2007-2011 were identified from a BC database. BC subtypes and clinical characteristics including overall survival (OS) were obtained by chart review. Statistical analysis was performed using the Chi Square test, the log rank test and time depended covariate cox regression model as appropriate.

Results

Three-hundred-eighty/571 (63%) were grouped among the young-old (65-74 years), 182/571 (31.9%) among the old-old (75-84 years), and 29/571 (5,1%) among the oldest-old (≥85 years). 392/571 (68.8%) patients presented with luminal BC, 119/571 (20.8%) with HER2 positive and 59/571 (10.3%) with triple negative BC. After a median follow up of 38 months (range 0-204), 115/571 (20.1%) patients presented with metastatic recurrence. Highest recurrence rate was observed in HER2 positive BC patients (43/119 (36.1%)), followed by triple negative (15/59 (25.4%) and luminal BC (57/392 (14.5%); p<0.001; Chi Square test). BM occurred significantly more frequently in HER2 positive BC patients (9/119 (7.6%) compared to triple negative (2/59 (3.4%) and luminal BC patients (6/392 (1.5%); p=0.003; Chi Square test). Occurrence of metastases (HR 7.7; 95% CI 5.2-11.4; p<0.001) as well as development of BM (HR 3.5; 95% CI 1.9-6.4; p<0.001) had a significant impact on OS prognosis as entered in a time depended covariate cox regression model.

Conclusions

In contrast to younger BC patients, HER2 positive BC subtype and not triple negative BC subtype was linked to the most aggressive clinical course including the development of metastatic disease and BM in our elderly cohort.

Citation Format: Bergen ES, Tichy C, Berghoff AS, Rudas M, Dubsky P, Bago-Horvath Z, Mader RM, Gnant M, Dieckmann K, Zielinski CC, Steger GG, Preusser M, Bartsch R. Prognostic impact of breast cancer subtypes in elderly patients. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P2-08-17.
Abstract P2-08-21: Stromal co-expression of urokinase-type plasminogen activator (uPA) and plasminogen activator Inhibitor (PAI-1) protein by IHC predicts poor disease outcome in endocrine-treated postmenopausal patients with receptor-positive early breast cancer
Authors: Singer CF , Filipits M , Jahn S , Abete L , Jakesz R , Greil R , Bauernhofer T , Kwasny W , Seifert M , Fitzal F , Schmitt M , Moinfar F , Gnant M
Published: May 2016

Background Elevated levels of intratumoral uPA and PAI-1 in ELISA-based measurements are associated with a high recurrence risk and allow to identify patients who might particularly benefit from adjuvant chemotherapy. The clinical utility of ELISA-based protein analysis is, however, greatly limited by the requirement of fresh tumor tisssue. We have therefore evaluated in a large clinical trial patient cohort with long-follow-up whether immunohistochemical detection of uPA and PAI-1 in FFPE archived tumor samples is also able to identify women with poor prognosis.

Patients and Methods 547 postmenopausal women with hormone receptor–positive, early breast cancer who had received at least 5 years of endocrine therapy in the prospectively designed ABCSG-06 trial, and in whom FFPE tumor tissue was available, were included in this analysis. uPA and PAI-1 protein expression was evaluated by immunohistochemistry, and correlated with distant-disease free (DDFS) and overall survival (OS).

Results Stromal co-expression of uPA and PAI-1 was detected in 166 of 276 (60%) evaluable tumor samples and was weakly associated with tumor size (p=0.012, Chi Square test) but not with age, nodal status, grading, or receptor status. Women with intratumoral uPA/PAI-1 expression were more likely to have a shorter DDFS in multivariate analysis (HR for relapse p=1.870; 95% CI 1.184-2.955; p=0.007 Cox regression analysis) and and OS (adjusted HR for death p=1.291; 95% CI 0.928-1.795; p=0.129) than women without. After a median follow-up of 10 years, women with uPA/PAI-1-positive tumors experienced a significantly shorter DDFS (p<0.0001 log rank test) and OS (p=0.020).

Conclusions Stromal co-expression of uPA and PAI-1 in breast cancer samples predicts poor DDFS and OS in postmenopausal women with hormone-receptor positive early-stage breast cancer who receive endocrine therapy.

Citation Format: Singer CF, Filipits M, Jahn S, Abete L, Jakesz R, Greil R, Bauernhofer T, Kwasny W, Seifert M, Fitzal F, Schmitt M, Moinfar F, Gnant M. Stromal co-expression of urokinase-type plasminogen activator (uPA) and plasminogen activator Inhibitor (PAI-1) protein by IHC predicts poor disease outcome in endocrine-treated postmenopausal patients with receptor-positive early breast cancer. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P2-08-21.
Abstract P2-09-02: Panel testing for familial breast cancer: Tension at the boundary of research and clinical care
Authors: Campbell IG , Thompson ER , Rowely SM , Li N , McInerny S , Devereux L , Wong-Brown MW , Trainer AH , Mitchell G , Scott RJ , James PA
Published: May 2016

Gene panel sequencing is revolutionizing germline risk assessment for hereditary breast cancer. Despite scant evidence supporting the role of many of these genes in breast cancer predisposition, results are often reported to families as the definitive explanation for their family history. We assessed the frequency of mutations in 18 genes commonly included in hereditary breast cancer panels among 2,000 index cases from breast cancer families and 1,997 population controls. Cases were predominantly breast cancer-affected women referred to specialized familial cancer centers (BRCA1 and BRCA2 wild-type). Controls were cancer-free women from the LifePool study (www.lifepool.org). Sequencing data were filtered for known pathogenic or novel loss of function mutations.

The frequency of pathogenic mutations in BRCA1 and BRCA2 in the control group was 0.2% (4 mutations) and 0.4% (8 mutations), respectively, which is consistent with previous indirect estimates for Caucasian populations but to our knowledge this the largest direct assessment of their prevalence.

Excluding 18 mutations identified in BRCA1 and BRCA2 among the cases and controls, a total of 69 cases (3.5%) and 26 controls (1.3%) were found to carry an "actionable mutation". PALB2 was most frequently mutated (22 cases, 3 controls), while no mutations were identified in PTEN or STK11. Among the remaining genes, loss of function mutations were rare with similar frequency between cases and controls.

The frequency of mutations in most breast cancer panel genes among individuals selected for possible hereditary breast cancer is low and in many cases similar or even lower than that observed among cancer-free population controls. While multi-gene panels can significantly aid in cancer risk management, they equally have the potential to provide clinical misinformation and harm at the individual level if the data is not interpreted cautiously.

Citation Format: Campbell IG, Thompson ER, Rowely SM, Li N, McInerny S, Devereux L, Wong-Brown MW, Trainer AH, Mitchell G, Scott RJ, James PA. Panel testing for familial breast cancer: Tension at the boundary of research and clinical care. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P2-09-02.
Abstract P2-09-15: Rapid BRCA analysis: Experience of a single institution
Authors: Mailliez A , Herin H , Revillion F , Adenis C , Peyrat J-P , Bonneterre J
Published: May 2016

The interest of rapid BRCA analysis, even before the initial treatment could modify the surgical indications (ie total mastectomy and immediate reconstruction even in case where tumorectomy would be suitable). The aim of this study was to evaluate the indications and consequences of this rapid determination of BRCA status in our institution.

Patients

Rapid BRCA analysis was conducted in 129 patients (pts) treated for breast cancer and addressed to the genetician at Oscar Lambret Center between 2009 and 2014: in 98 pts to discuss breast and axilla surgical modalities, in 4 cases to discuss oophorectomy, in 3 cases to have access to PARP inhibitors. In the other 24 pts, the reasons were debatable: patient leaving abroad, patient requiring reproductive assisted technology.....

Results

38 deleterious mutations (29,4%) were identified (31 BRCA1, 7 BRCA2) in 38 pts.

Cancer treatment was on going for 107 pts, programmed for 12 pts and finished for 10.

The median time between cancer diagnosis and genetic counseling was 81 days (5-5160), between blood sample collection and BRCAs analysis 37 days (13-132), between availability of the laboratory results and the information to the patient: 39 days (1-234); 7 pts (5.4%) never came to obtain the laboratory results.

Treatment was modified after genetic counselling for 8 pts out of 119 (7%). Six of them had BRCA1 deleterious mutations. Three decided subsequent radical mastectomy and adjuvant radiation was cancelled, another had a radical mastectomy before radiation, another treated for bilateral breast cancer had a bilateral radical mastectomy before radiation and the last treated for unilateral breast cancer had a bilateral radical mastectomy before radiation.

Out of these 8 pts, two did not have a deleterious mutation. One had a bilateral radical mastectomy before laboratory result and one with BRCA variant of unknown significance had a homolateral radical mastectomy.

Conclusion discussion

Our experience shows that rapid BRCA analysis is feasible. In this selected population the mutation rate is very high. However, first, it appears that indications for rapid BRCA analysis were variable and not always relevant. Second, genetic result impacts on treatment only in 7% of pts. Indications could be justified by the use of PARP inhibitors. Up to now, the indications of rapid genetic analysis are rare.

Citation Format: Mailliez A, Herin H, Revillion F, Adenis C, Peyrat J-P, Bonneterre J. Rapid BRCA analysis: Experience of a single institution. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P2-09-15.