bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P2-02-17: T-DM1 in HER2-negative metastatic breast cancer patients with HER2-amplified circulating tumor cells: Current status of the CirCe T-DM1 phase II trial
Authors: Bidard F-C , Romieu G , Jacot W , Cottu P , Dieras V , Lerebours F , Servent V , Luporsi E , Lortholary A , Tubiana-Mathieu N , Espie M , Bollet M , Bourgeois H , Renaud N , Pelissier S , Armanet S , Baeten K , Pierga J-Y
Published: May 2016

Background: Liquid biopsy can reassess key therapeutic targets in metastatic breast cancer. Several studies showed that a low albeit significant rate of metastatic breast cancer initially considered as HER2-negative can be reclassified as HER2-positive by systematic biopsy procedures. We report here the current status of the CirCe T-DM1 trial [NCT01975142] which aims to demonstrate the clinical utility of HER2 status reassessment on circulating tumor cells (CTCs).

Methods: The first step of the trial consists in CTC count and HER2/CEP17 FISH on detected CTCs (CellSearch, Janssen Diagnostics) in patients (pts) with measurable disease progressing after the second line of chemotherapy. Pts with amplified CTCs (HER2/CEP17 ratio equal or higher than 2.2) are eligible to the treatment step of the study in two distinct cohorts: low CTC count (1 or 2 HER2-amplified CTCs) and high CTC count (3 and more HER2-amplified CTCs). In the treatment step has a Simon's two stage design, the anti-HER2 antibody-drug conjugate T-DM1 being administered until tumor progression. The primary objective of the trial is the confirmed response rate (RECIST). This trial is supported by Roche.

Results: CirCe T-DM1 has been initiated in 10 centers in France. As of June 2015, 105 metastatic breast cancers pts considered as HER2-negative were screened. 29 pts (27%) had no CTC detected, 68 pts (65%) had at least 1 CTC detected with no HER2 amplification, and 8 pts (8%) exhibited HER2-amplified CTCs. Among the 8 pts, 1 pt had 5 HER2-amplified CTC, 2 pts had 2 HER2-amplified CTC and 5 pts had 1 HER2-amplified CTC. HER2/CEP17 ratios among HER2-amplified CTCs ranged from 2.5 to 7. Five of the 8 pts were treated by T-DM1. One objective confirmed partial tumor response has been observed (20%).

Conclusion: The accrual is ongoing; the first efficacy assessment will occur after having treated 14 pts. This innovative trial highlights the promise and the complexity of liquid biopsy-based programs in the era of precision medicine: scarcity of the target, reliability and reproducibility of the target assessment, major efficacy when the target is matched to the appropriate drug.

Citation Format: Bidard F-C, Romieu G, Jacot W, Cottu P, Dieras V, Lerebours F, Servent V, Luporsi E, Lortholary A, Tubiana-Mathieu N, Espie M, Bollet M, Bourgeois H, Renaud N, Pelissier S, Armanet S, Baeten K, Pierga J-Y. T-DM1 in HER2-negative metastatic breast cancer patients with HER2-amplified circulating tumor cells: Current status of the CirCe T-DM1 phase II trial. [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 P2-02-17.
Abstract P2-03-04: Down-regulation of Bcl2-related ovarian killer (BOK) by miR-296-5p protects breast cancer cells from paclitaxel-induced apoptosis
Authors: Onyeagucha BC , Rajamanickam S , Subbarayalu P , Bansal B , Bansal H , Chang Y-F , Timilsina S , Abdelfaltah N , Eedunuri VK , Rao MK
Published: May 2016

Accumulating evidence shows that miRNAs play a role in drug resistance. Despite these observations, little is known about the identities of the miRNAs involved in drug resistance and their downstream targets. In the present study, we identified miR-296-5p for which a tumor suppressive role has been previously described, as a miRNA that is involved in paclitaxel drug resistance in triple negative breast cancer (TNBC) cells. Enforced expression of miR-296-5p suppressed cell growth, migration, and invasion in MDA-MB-231 breast cancer cells. Using a microarray approach, we identified BCL2-related Ovarian Killer (BOK), a pro-apoptotic gene as a target of miR-296-5p. BOK levels were validated BOK levels in miR-296-5p transfected MDA-MB-231 and MDA-MB-468 cells using real-time PCR and Western blot. Our results demonstrated that over-expression of miR-296-5p down-regulated BOK expression in TNBC cells. Transfection of miR-296-5p significantly suppressed luciferase reporters containing wild-type BOK-3'-UTR constructs. In contrast, mutant BOK-3'-UTR constructs were unaffected by ectopic miR-296-5p. Furthermore, BOK expression was induced in the presence of paclitaxel, but ectopic miR-296-5p significantly suppressed BOK induction by paclitaxel treatment compared to the control cells. These data provide new insights on the role of miRNAs in drug resistance and suggests that therapeutic strategies against miR-296-5p may be warranted.

Citation Format: Onyeagucha BC, Rajamanickam S, Subbarayalu P, Bansal B, Bansal H, Chang Y-F, Timilsina S, Abdelfaltah N, Eedunuri VK, Rao MK. Down-regulation of Bcl2-related ovarian killer (BOK) by miR-296-5p protects breast cancer cells from paclitaxel-induced apoptosis. [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 P2-03-04.
Abstract P2-03-07: Rhythmic time oscillations of microRNAs in human breast epithelial normal and cancer cell lines
Authors: Chacolla RJ , Trevino VM , Scott SP , Moreno JE
Published: May 2016

BACKGROUND: Diverse molecular mechanisms are being reported in human breast cancer (BC), which can affect the biochemical functions throughout malignant cells development. The microRNAs (miRNAs) are an emerging class of modulators of gene expression with relevant roles in several biological processes, as oncogenic, tumor-suppressive, and metastatic-influencing in BC cells. Recently, a few reports have implied the possible pattern of expression (time oscillation) of miRNAs in time that may be related to molecular changes in mammalian cells. These findings suggest a biological connection among normal and cancer cells, and rhythmic regulation of some miRNAs, but such connection has not yet been studied. In this study, we aimed to identify and compare the rhythmic expression of miRNAs in human breast epithelial normal and cancer cell lines.

METHODS: We used cell culture to explore three cell lines, one breast epithelial normal (MCF10A) and two cancer (MCF-7 and MDA-MB-231) cell lines under standard growth conditions in vitro. The cells were synchronized by serum shock (50% horse serum for 2 h), and we collected sample cells (triplicate) for intervals of 4 hours during 48 hours. Collected cells at 12h to 40h (8 time-points) were genome-wide analyzed of miRNA expression using high-throughput Agilent Human miRNA microarray of 2006 human miRNAs. Analysis for identification of rhythmic miRNAs was developed by cosine analysis in R software.

RESULTS: We observed diverse oscillation patterns (minimum 6 patterns, i.e. cosine or sine oscillation) of miRNAs in cell lines. Each cell line shows approximately 85 miRNAs with rhythmic oscillation. These also showed distinct phases between cell lines, which could suggest as part of molecular changes in breast normal and cancer cell lines.

CONCLUSIONS: Our results suggest that miRNAs may present rhythmic oscillation in the regulation of molecular changes of human breast normal and cancer cells.

Citation Format: Chacolla RJ, Trevino VM, Scott SP, Moreno JE. Rhythmic time oscillations of microRNAs in human breast epithelial normal and cancer cell lines. [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 P2-03-07.
Abstract P2-05-12: Epithelial paradox; clinical significance of co-expression of E-cadherin and vimentin in invasive breast cancer
Authors: Yamashita N , Tokunaga E , Inoue Y , Tanaka K , Ueo H , Saeki H , Oki E , Maehara Y
Published: May 2016

Background: E-cadherin and vimentin are now regarded as major and conventional canonical markers of epithelial-mesenchymal transition (EMT). It is commonly assumed E-cadherin is uniformly lost during the process of EMT. We previously reported that the elevated expression of vimentin contributes to the aggressive phenotype in invasive breast cancer. On the other hand, the role of E-cadherin in breast cancer biology might be unclear and more complex. Although, cell cohesion during breast cancer invasion is often overlooked, accumulating evidences indicate breast tumor cells are typically cohesive and often display membrane-localized E-cadherin in both the primary tumor and distant metastases, termed collective invasion. Multiple mechanisms have emerged to address how epithelial breast tumors invade.

Aims: The aim of this study is to reveal the clinical importance of the expression of E-cadherin and vimentin in breast cancer.

Methods: The E-cadherin and vimentin protein expression were evaluated by immunohistochemistry (IHC) in 177 invasive breast cancer samples. Among these, E-cadherin and vimentin expression were evaluated in the set of primary breast cancer and metastatic lymph nodes in 65 cases.

Results: The positive vimentin expression was highly correlated with poor disease-free survival (DFS) and overall survival (OS) (p=0.019 and p=0.0044), however, the E-cadherin expression alone did not correlate with prognosis. Interestingly, Both E-cadherin and vimentin positive tumor had the worst DFS and OS among all breast cancer (p=0.03 and p=0.0089). Vimentin expression was highly correlated between primary tumors and metastatic lymph nodes. However, E-cadherin expression levels were significantly elevated in metastatic lymph nodes (p=0.0017), Co-expression of E-cadherin and vimentin in the metastatic lymph nodes also showed worst DFS and OS (p=0.12 and p=0.027). Conclusions: Co-expression of E-cadherin and vimentin seems to be associated with the most aggressive phenotype and poorest prognosis in breast cancer, and positive E-cadherin expression may not always play roles for tumor suppression.

Citation Format: Yamashita N, Tokunaga E, Inoue Y, Tanaka K, Ueo H, Saeki H, Oki E, Maehara Y. Epithelial paradox; clinical significance of co-expression of E-cadherin and vimentin in invasive 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 P2-05-12.
Abstract P2-05-14: The long noncoding RNA BHLHE40-AS1 as a functional biomarker of invasive breast cancer
Authors: DeVaux RKS , Davis S , Sheftel J , Coarfa C , Behbod F , Herschkowitz JI
Published: May 2016

Increased emphasis on breast cancer screening has led to a dramatic surge in diagnosis of pre-cancerous ductal carcinoma in situ (DCIS) over the past 30 years. Unfortunately, diagnosis of late stage invasive and metastatic disease has not proportionally declined, suggesting significant over diagnosis and over treatment of innocuous DCIS. Due to a lack of biomarkers, the heterogeneity of DCIS lesions, and an insufficient understanding of mechanisms of invasive progression, there is currently no clinically relevant method of predicting which DCIS lesions will advance to invasive disease. As a result, all DCIS patients undergo an aggressive treatment regimen to prevent disease progression. Therefore, there is a critical need to determine the underlying mechanisms driving breast cancer progression to better inform patient treatment options and nominate novel therapeutic entry points for treatment of invasive disease.

Recently, long noncoding RNAs (lncRNAs) have gained attention as critical regulators of epigenetic states and gene expression. Although the study of lncRNAs is in its infancy, they are being uncovered as pivotal regulators of development and tumorigenesis, thus they are a rich source of potential drivers of breast cancer progression. We propose that lncRNAs functionally drive breast cancer invasion and their expression can discriminate between innocuous and potentially invasive DCIS.

Using biopsies from women that exhibit tandem DCIS and invasive breast cancer lesions, we have identified a cohort of long noncoding RNAs (lncRNAs) that are enriched in the invasive biopsy. From this cohort we have identified the lncRNA BHLHE40-AS1 as increasing in a step-wise manner in a breast cancer progression series that escalates from normal, non-transformed cells to highly invasive disease. Preliminary evidence suggests that BHLHE40-AS1 expression regulates invasive potential in vitro. Using a GeneChip® Human Transcriptome 2.0 Array (HTA) we have identified several targets previously associated with driving breast cancer invasion as being potentially regulated by BHLHE40-AS1. Future directions will focus on determining the effect of BHLHE40-AS1 expression on tumor cell invasion in an orthotopic xenograft model, mechanistically elaborating its function, and determining the utility of BHLHE40-AS1 as a clinically relevant biomarker of invasive breast cancer.

Citation Format: DeVaux RKS, Davis S, Sheftel J, Coarfa C, Behbod F, Herschkowitz JI. The long noncoding RNA BHLHE40-AS1 as a functional biomarker of invasive 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 P2-05-14.
Abstract P2-05-24: OPG+ bone marrow B cells induced by non-metastatic tumors inhibit the pre-metastatic bone niche induced by T cells
Authors: Bonomo A , Monteiro AC , Leal AC , Fontão AP , Spinetti E , Balduino A
Published: May 2016

Using the 4T1 model of experimental breast cancer we had recently shown that cancer induced bone disease starts before metastatic colonization and is mediated by RANKL expressed by tumor specific T cells. The role of anti-tumor B cell immune response in the context of cancer induced bone disease has never been investigated. There is evidence in the literature that B cells are good prognostic markers for metastatic breast cancer. B cells have an intimate relationship with bone cells as they differentiate from HSC present on endosteal surfaces; cross-talk with skeletal system through the RANK-RANKL-OPG signaling axis; and produce OPG, a decoy receptor of RANKL.

Here we used the BALB/c derived 4T1 (metastatic) and 67NR (non-metastatic) sibling cell lines of mammary mouse carcinomas. By day 7, 14 and 21 after tumor injection, B220+ BM B cells from 67NR+ animals produce high amounts of OPG in vitro in contrast to B220+ BM B cells of 4T1+ mice. In vitro, BM B cells from 67NR+ mice, but not from 4T1+, could inhibit the RANKL dependent, anti-4T1 T cell mediated-OC differentiation ascertained by TRAP enzymatic activity, morphology and osteolytic disk assay.

Transference of BM B cells from 67NR+ mice together with 4T1 tumor cells to BALB/c mice led to inhibition of osteoclastogenesis, increased numbers of bone lining cells and mesenchyme stem cell. Besides acting directly on the bone remodeling system, these B cells also modulated T cell activity evidenced by diminished RANKL and IL-17F production.

All the anti-osteolytic and pro-osteogenic activity of B cells modulate and inhibits the pre-metastatic niche formation. Indeed transference of such cells to 4T1 animals inhibited LN and BM metastatic colonization.

We conclude that 67NR induced OPG+ B cells can inhibit pro-osteoclastic / pre-metastatic activity of tumor induced T cells, favoring a bone metastasis free-phenotype. These findings have implications, not only for the understanding of the direct contribution of B cells in the control of bone metastasis but also it might be a promising prognostic tool for predicting cancer-induced bone metastasis.

Citation Format: Bonomo A, Monteiro AC, Leal AC, Fontão AP, Spinetti E, Balduino A. OPG+ bone marrow B cells induced by non-metastatic tumors inhibit the pre-metastatic bone niche induced by T cells. [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 P2-05-24.
Abstract P2-05-26: Focal adhesion kinase is required for efficient tumor-induced osteoclastogenesis via control of macrophage colony stimulating factor expression
Authors: Landon K , Howe GA , Zhao H , Addison CL
Published: May 2016

Breast cancer most commonly metastasizes to bone where metastases cause the potentiation of the vicious cycle. During this process, breast cancer bone metastases inhibit osteoblasts from forming new bone, while also activating osteoclasts to degrade bone. This in turn causes release of bone-matrix bound growth factors which propagate tumor growth. The overall net bone destruction can lead to significant adverse clinical consequences for patients including fractures or substantial pain. While agents such as bisphosphonates or the monoclonal antibody to RANKL denosumab, both of which inhibit the osteolytic activity of osteoclasts, are currently used in breast cancer bone metastatic patients to alleviate these adverse clinical outcomes, they have not been associated with increased patient survival. Given the lack of successful treatments for bone metastases providing survival advantages, we sought to evaluate the role of focal adhesion kinase (FAK), a novel therapeutic target, in breast cancer mediated osteolysis. FAK is a non-receptor tyrosine kinase that regulates many pathways that contribute to enhanced tumor progression and metastasis. FAK is also expressed in all the cell types involved in the vicious cycle. We thus hypothesized that FAK plays an important role in breast tumor-induced osteolysis and that its inhibition would lead to restoration of bone homeostasis, in addition to inhibition of tumor progression. Using in vitro siRNA-targeted depletion of FAK in breast cancer tumor cell lines, we found that production of numerous soluble growth factors, many of which are known contributors to osteoclastogenesis, was inhibited. We confirmed that FAK regulates the expression of the osteoclastogenic factor macrophage colony stimulating factor in breast cancer cell lines. Further, using conditioned media from FAK expressing versus depleted breast cancer tumor cells in osteoclastogenesis assays, we show that FAK-depletion results in impaired osteoclastogenesis. These data suggest that in addition to its proven direct anti-tumor effects, inhibition of FAK may also result in therapeutic blockade of bone degradation.

Citation Format: Landon K, Howe GA, Zhao H, Addison CL. Focal adhesion kinase is required for efficient tumor-induced osteoclastogenesis via control of macrophage colony stimulating factor expression. [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 P2-05-26.
Abstract P2-06-02: BIN3 is an 8p21 tumor suppressor regulating the epithelial attachment checkpoint
Authors: Silva J , Marshall N , Sanchez-Garcia F , Parsons R , Pe'er D
Published: May 2016

An important characteristic of multicellular organisms is the control that tissue architecture exerts on the growth and differentiation of individual cells. Epithelial cells sense their location through interactions with the extracellular matrix (ECM) as well as neighboring cells. These interactions generate input signals, including survival, that are critical to maintain tissue and cellular homeostasis. When attachment is compromised, epithelial cells undergo an intrinsically programmed cell death (apoptosis) that is termed anoikis (from the Greek "loss of home"). Importantly, failure to execute the anoikis program could result in adherent epithelial cells surviving in suspension or being able to proliiferate at sites different from their original environment. Thus, anoikis is a line of defense that must be circumvented by cancerous epithelial cells for them to leave their home niche, thrive in inappropriate ECM environments, and establish long distance metastases. Thus, elucidating how epithelial cancer cells escape anoikis is critical to understanding cancer progression.

In order to uncover genes that modulate the anoikis response and are altered in human cancers, we performed a functional genomics study. We couple genome wide RNAi to identify gene functions that, when silenced, induce resistance to anoikis with a novel computational method, ISAR-DEL, specifically aimed at pinpointing candidate tumor suppressor genes based on recurrent loss of copy number. Our studies identified Bridging Integrator 3 (BIN3) as a novel tumor suppressor located on the chromosomal region 8p21.3, one of the most frequently lost regions in epithelial cancers.

Mechanistically, we link BIN3 tumor suppression function to its ability to sense changes in the curvature of the cell membrane and relocate to the cell membrane after cell detachment to induce a proapoptotic cascade. Once BIN3 has translocated to the cell membrane it modulates the relocation and function of CDC42. In these conditions, CDC42 transmits the signal that leads to the activation of the stress protein P38-α and programmed cell death mediated by accumulation of the apoptotic facilitator BimEL. Overall, our results explain how changes in cell geometry are integrated in the cellular signaling network and present, for the first time, BIN3 as a novel breast cancer tumors suppressor.

Citation Format: Silva J, Marshall N, Sanchez-Garcia F, Parsons R, Pe'er D. BIN3 is an 8p21 tumor suppressor regulating the epithelial attachment checkpoint. [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 P2-06-02.
Abstract P2-06-11: Wild type N-Ras, overexpressed in basal-like breast cancer, promotes tumor formation by inducing IL8 secretion via JAK2 activation
Authors: Zheng Z-Y , Bu W , Tian L , Fan C , Gao X , Zhang X , Yu C , Wang H , Liao Y-H , Li Y , Lewis MT , Edwards D , Zwaka TP , Hilsenbeck SG , Medina D , Perou CM , Creighton CJ , Zhang XH , Chang EC
Published: May 2016

"Basal-like" breast cancer (BLBC) is a very aggressive subtype of breast cancer. BLBC has very poor prognosis — median time to distant recurrence is just 2.6 years vs. 5 years overall, and survival time from diagnosis of distant metastatic disease is 9 months vs. 22 months. BLBC tumors usually do not express ER, Her2, or progesterone receptor. As such, they cannot be treated by the current targeted therapies, which target these molecules. What drive the formation and progression of BLBCs is largely unclear.

Ras GTPases are best known for mediating growth factor signaling. Oncogenic mutations in the RAS genes, K-RAS in particular, are found in more than 30% of human tumors. Surprisingly, oncogenic RAS mutations are rare in breast cancer. However, we found that wild-type N-RAS is overexpressed in BLBCs, possibly partly via promoter demethylation, but not in other breast cancer subtypes. Repressing N-RAS inhibits transformation and tumor growth, while overexpressing it enhances these processes even in preinvasive BLBC cells. In contrast, in breast cancer cells of other subtypes, repressing N-RAS expression does not affect growth and transforming activities. We identified N-Ras-responsive genes, most of which encode chemokines and cytokines, e.g., IL8. High expression levels of these N-Ras-responsive genes as well as of N-RAS itself in tumors correlate with poor patient outcome. N-Ras, but not K-Ras, induces IL8 by binding and activating the cytoplasmic pool of JAK2; IL8 then acts on both the cancer cells and stromal fibroblasts.

In conclusion, N-Ras drives BLBC by promoting transformation in epithelial cells, which may in turn remodel the tumor microenvironment to create a proinvasive state. Although oncogenic mutations affecting RAS are common in many other human cancers, tumorigenesis in an important subset of breast cancers is driven instead by increasing activity of wild-type N-Ras. Thus, to fully assess the impact of Ras on tumorigenesis, the role of wild-type as well as mutant Ras proteins must be carefully examined.

Citation Format: Zheng Z-Y, Bu W, Tian L, Fan C, Gao X, Zhang X, Yu C, Wang H, Liao Y-H, Li Y, Lewis MT, Edwards D, Zwaka TP, Hilsenbeck SG, Medina D, Perou CM, Creighton CJ, Zhang XH, Chang EC. Wild type N-Ras, overexpressed in basal-like breast cancer, promotes tumor formation by inducing IL8 secretion via JAK2 activation. [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 P2-06-11.
Abstract P2-08-05: Association between the neutrophil-to-lymphocyte ratio (NLR) and the 21-gene recurrence score
Authors: Srikanthan A , Bedard PL , Goldstein S , Templeton A , Amir E
Published: May 2016

Introduction: A high neutrophil-to-lymphocyte ratio (NLR) has been reported to be a poor prognostic indicator in several malignancies including breast cancer. It is unknown whether the prognosis associated with high NLR can be explained by other prognostic factors such as proliferation or estrogen receptor signalling. Here we explore the association between NLR and the 21-gene recurrence score (RS).

Methods: The associations between RS, NLR, tumor size, histologic grade, and estrogen receptor (ER) and progesterone receptor (PgR) expression (assessed by immunohistochemistry) were explored in sequential women with early-stage, lymph node-negative (or with lymph node micrometastases), ER-positive and HER2-negative breast cancer treated at Princess Margaret Cancer Centre in Toronto, Canada and in whom results of the RS were available. NLR was measured prior to surgery. Patients with a documented history of pre-existing infectious/inflammatory condition were excluded. Associations were explored using simple linear regression and statistical significance was defined as p<0.05.

Results: A total of 130 women diagnosed between January 2006 and April 2015 were included in the analysis. Median age was 55 (range 32-79), 87% were lymph node negative and 13% had nodal micrometastases. The median NLR was 2.2 (range 0.9-9.1) and was collected at a median of 12 days prior to surgery (range 0-60). The median RS was 18 (range 0-41). There was no association between RS and NLR (R=-0.10, p=0.31), grade (R=0.13, p=0.15), age (R=-0.05, p=0.58) or tumor size (R=0.06, p=0.48). RS was negatively associated with the magnitude of expression of both ER (R=-0.22, p=0.01) and PgR (R=-0.44, p<0.001). There was no association between NLR and grade (R=0.20, p=0.15), age (R=-0.13, p=0.17), tumor size (R=0.14, p=0.93), ER (R=0.01, p=0.94) or PgR (R=0.13, p=0.23)

Conclusion: The poor outcomes associated with high NLR are unlikely explained by proliferation of estrogen receptor signalling.

Citation Format: Srikanthan A, Bedard PL, Goldstein S, Templeton A, Amir E. Association between the neutrophil-to-lymphocyte ratio (NLR) and the 21-gene recurrence score. [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 P2-08-05.
Abstract P2-08-14: Validation of gene expression profiling in prediction of locoregional recurrence for breast cancer patients after surgery
Authors: Cheng H-CS , Horng C-F , Huang T-T , Tsou M-H , Shih L-S , Yu B-L , Tsai Y-C , Chen C-M
Published: May 2016

Purpose

We have identified 34 genes of interest that can distinguish post-mastectomy cancer patients into high- and low-risk of locoregional recurrence (LRR) in 2006. In this study, we validate the performance of 18 of 34 genes in prediction of LRR after mastectomy and breast conserving surgery (BCS).

Materials and methods

A total of 124 breast cancer patients who underwent mastectomy from 2005 to 2012 and had DNA microarray study on the primary tumor tissues were chosen for this study. Eligible patients should have no post-mastectomy radiotherapy (PMRT) with a minimum of 2-year follow-up. Among the 34 genes in previous report, only 30 genes could be found in Affymetrix U95 to U133 Plus2.0 array. Eighteen of 30 genes (18-Gene classifier) were used to distinguish low-risk and high-risk patients. Patients with 18-Gene score 31 or more were defined as high-risk and score less than 31 defined as low-risk group. We then examined the performance of the 18-Gene classifier in breast conserving patients (n=87) who usually had adjuvant radiotherapy (94%) and were treated in the same study periods.

Results

The overall accuracy of the prediction of LRR is 87% (sensitivity 91% and specificity 87%). By the 18-Gene classifier, the 5-year LRR rate in high-risk group was 42%, and in low-risk group <1% (p < 0.0001).

LRR and 18-gene score in non-PMRT patients18-Gene scoreNo LRR #LRR #5-year LR control probabilityScore <31971100%Score >= 31151157.7% 11212 LRR: Locoregional recurrence

Multivariate analysis of mastectomy patients revealed that 18-Gene score is the independent prognostic factor that predicts the likelihood of LRR regardless of nodal status and breast cancer subtype. The predictive ability of LRR by the 18-Gene classifier in BCS patients is independent from adjuvant radiotherapy, and is consistent with the performance in mastectomy patients.

Multivariate analysis in BCS patientsRisk factorPatient #LRR #LRR rateUnivariate analysisMultivariate analysisLVI     Absent/focal7345.5%  Prominent14321.4%4.3(1.0,19.3)5.1*(1.1,23.1)18-Gene score     Low risk6623.0%  High risk21523.8%9.4*(1.8,48.4)10.4*(2.0,54.2)Adjuvant R/T     No5240%10.3*(2.0,53.9) Yes8256.1%  LVI: lymphovascular invasion; R/T: radiotherapy; *p<0.05

Discussion

This 18-Gene classifier is capable of identifying high-risk patients of LRR after mastectomy and breast conserving surgery.

Citation Format: Cheng H-CS, Horng C-F, Huang T-T, Tsou M-H, Shih L-S, Yu B-L, Tsai Y-C, Chen C-M. Validation of gene expression profiling in prediction of locoregional recurrence for breast cancer patients 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 P2-08-14.
Abstract P2-09-03: Breast cancer prevention: Is it time for population-based mutation screening of high risk genes?
Authors: Campbell IG , Trainer AH , Devereux L , Young M-A , Mitchell G , Rowley S , Li N , James PA , Thompson E
Published: May 2016

The traditional model of familial breast cancer practice involves ascertaining high-risk individuals based on family history. However, most individuals who carry a BRCA1 or BRCA2 mutation will not have a family history of breast or other cancer in a close relative. Moving germline testing from the familial cancer center to the population will result in many essentially unanticipated findings of great significance in regards to the risk of breast cancer in healthy relatives. To date there have been no large studies that have directly measured the frequency of BRCA1 and BRCA2 mutations in a cancer-free general population. As a first step toward population based BRCA1 or BRCA2 screening, we sequenced the entire coding region of these genes in 1,997 cancer-free Australian women recruited from the lifepool study (www.lifepool.org) which is a cohort of women attending the Australian population based mammographic screening program. Sequencing data were filtered for known pathogenic or novel loss of function mutations. Twelve individuals were identified with pathogenic mutations in either BRCA1 (4 mutations) or BRCA2 (8 mutations), which is consistent with previous indirect estimates for Caucasian populations but to our knowledge this is the largest direct assessment of their prevalence.

Interestingly, the self-reported cancer family history of the majority of the 12 mutation positive women was unremarkable. All 12 women subsequently accepted an invitation to attend a Familial Cancer Centre for advice on whether to proceed with formal clinical genetic testing.

A population carrier frequency of 0.6% for mutations in BRCA1 and BRCA2, coupled with the rapidly declining costs of gene panel sequencing, suggests that population-based screening for these genes will be a highly cost effective way of reducing the incidence of breast and ovarian cancer through preventative strategies.

Citation Format: Campbell IG, Trainer AH, Devereux L, Young M-A, Mitchell G, Rowley S, Li N, James PA, Thompson E. Breast cancer prevention: Is it time for population-based mutation screening of high risk genes?. [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 P2-09-03.
Abstract P2-09-06: Multi-gene panel testing for hereditary cancer risk
Authors: Yadav S , Ladkany R , Fulbright J , Dreyfuss H , Reeves A , Campian S , Thomas V , Zakalik D
Published: May 2016

Background: Multi-gene panels are widely available for assessing hereditary cancer risk in high risk individuals. Due to the use of these panels, many genetic mutations other than BRCA 1 or 2 can be detected which can potentially affect management. This study presents the results of multi-gene panel testing performed at Beaumont Health System.

Methods:All patients who underwent multi-gene panel testing at Beaumont Health System between November 1, 2012 and January 15, 2015 were included in this study. This cohort consisted of patients who met criteria for genetic testing due to personal or family history. All patients received comprehensive pre and post-test genetic counseling. The panels ranged from 5 to 43 genes associated with risk for breast and other cancers.

Results: 653 multi-gene panel tests were performed. The majority of these consisted of either a 5 gene high risk breast panel (25%), an 18 gene moderate to high risk breast panel (21%), or a 9 gene high risk breast and gynecologic panel (17%). 184 variants of undetermined significance (VUS) were identified with a pooled VUS rate of 28%. Among the commonly used panels, there was a positive correlation between VUS rate and the number of genes included in the panel (r = 0.86, p = 0.01, Range 6% to 70%). A pathogenic mutation was identified in one or more genes in 65 (10%) panels for a total of 67 mutations. Of these, 17 mutations were in BRCA1 or BRCA2 gene. Fifty non-BRCA deleterious mutations were identified with the following frequencies: CHEK2(12), MUTYH(7 monoallelic, 1 biallelic), TP53(4), PTEN(4), ATM(4), MSH6(3), PALB2(3), MSH2(2), CDH1(2), APC(2), NF1(2), BARD1(2), MLH1(1) and PMS2(1). Of these non-BRCA mutations, 41(82%) had a significant impact on management.

Conclusions: Our study demonstrates that multi-gene panel testing identifies several genes that can impact management and would likely not have been discovered by pedigree analysis alone. However, this added detection is associated with a higher VUS rate, especially using larger panels. Further research is needed to better define the role of multi-gene panel testing in high risk patients, with a focus on choosing appropriate genes, understanding the magnitude of cancer risk and delineating impact on management.

Citation Format: Yadav S, Ladkany R, Fulbright J, Dreyfuss H, Reeves A, Campian S, Thomas V, Zakalik D. Multi-gene panel testing for hereditary cancer risk. [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 P2-09-06.
Abstract P2-09-14: Surveillance program for women carrying BRCA1/BRCA2 genetic predisposition
Authors: Sidoni T , Cocciolone V , Cannita K , Di Giacomo D , Ciccozzi A , Bafile A , Pizzorno L , Resta V , Marsecano C , Ferrari F , Di Cesare E , Ficorella C , Ricevuto E
Published: May 2016

Between January 2003 and January 2015, women at genetic risk of developing breast and/or ovarian cancer were selected for a surveillance program. The monitoring strategy consisted of the association of breast ultrasound (US), every six months, and Rx-mammography (XM) and breast MRI (MRI) to be performed annually. To date, 29 women have been included in the surveillance program: BRCA1 mutation carriers, 18; BRCA2 mutation carriers, 11.

Eight women (28%) had already developed breast and/or ovarian cancer, while 21 (72%) were unaffected carriers.

At a median surveillance time of 33 months (range 0-144), 4 incidental breast cancers were diagnosed in 4 BRCA1/2 mutation carriers (4/29, 14%), 2 BRCA1+ and 2 BRCA2+; 3/21 (14%) unaffected carriers and 1/8 (13%) previously affected carriers. Two cancers (50%) were detected by all three diagnostic tools; 2 (50%) were identified only by US and MRI in patients aging 33 and 43, respectively.

Characteristics of patients with breast cancer diagnosed during surveillanceagemutated geneprevious cancerUSXMMRIhistologysurveillance time (months)33BRCA2-+-+ductal74+43BRCA2-+-+ductal40+45BRCA1-+++ductal60+40BRCA1yes+++ductal74+

One of these four patients, affected by breast cancer, died at 31 months of overall survival; three underwent surgery, chemotherapy and radiotherapy and are still disease free, in follow-up at 59, 28 and 26 months, respectively.

In conclusion, 29 BRCA1/2 carriers have been included in the surveillance program, the median age was 42.5 months (range 27-68) and the median surveillance time was 33 months (range 0-144). Our preliminary data confirm the 3% expected rate of diagnosed cancers and the effectiveness of performing the triple diagnostic surveillance with US, XM and MRI.

Citation Format: Sidoni T, Cocciolone V, Cannita K, Di Giacomo D, Ciccozzi A, Bafile A, Pizzorno L, Resta V, Marsecano C, Ferrari F, Di Cesare E, Ficorella C, Ricevuto E. Surveillance program for women carrying BRCA1/BRCA2 genetic predisposition. [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 P2-09-14.
Abstract P2-12-06: Prevention of seroma formation after mastectomy by local methylprednisolone injection - A randomized controlled clinical trial
Authors: Qvamme G , Axelsson CK , Lanng C , Wegeberg B , Mortensen MB , Okholm M , Arpi MR , Szecsi PB
Published: May 2016

Objective: To investigate the effect of local steroid administration on seroma formation until 14 days of dryness.

Background: Seroma formation, the most prevalent postoperative complication after mastectomy, is an inflammatory process that is potentially preventable via local steroid administration.

Methods: This double-blind, randomized, placebo-controlled intervention study included 212 women who were scheduled for mastectomy for primary breast cancer. The patients were classified according to the surgical axillary procedure: mastectomy with sentinel lymph node biopsy (M+SLNB) or mastectomy with level I-II axillary lymph node dissection (M+ALND). The participants received either 80 mg of methylprednisolone or saline intracavitary via the drain orifice upon drain removal.

Results: After M+SLNB, 46% (32 of 69) of the patients developed seromas in the steroid group vs. 78% (52 of 67) in the saline group (p<0.0001). The mean cumulative seroma volume in the intention-to-treat material for the first 10 and 30 days was significantly lower in the steroid group than in the saline group (24 vs.127 mL and 177 vs. 328 mL, respectively) (p<0.0001). After M+ALND, 94% of the patients developed seromas in both the steroid (35 of 37) and saline (34 of 36) groups, and steroid administration displayed no significant effect on seroma formation. Additionally, no difference in the infection rate was observed.

Conclusion: Methylprednisolone administered intracavitary on the first postoperative day after M+SLNB exerted a highly significant preventive effect against seroma formation during the first 30 days. Future studies may clarify whether higher or repeated steroid doses enhance these effects.

Citation Format: Qvamme G, Axelsson CK, Lanng C, Wegeberg B, Mortensen MB, Okholm M, Arpi MR, Szecsi PB. Prevention of seroma formation after mastectomy by local methylprednisolone injection - A randomized controlled clinical trial. [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 P2-12-06.
Abstract P2-12-07: Treatment of the breast in occult breast cancer: Results of a prospective Dutch national cohort study with 5 years follow up
Authors: de Maat M , Bretveld R , Steevens J , Vissers Y , Hulsewé K
Published: May 2016

Introduction

Occult breast cancer (cT0N+M0) is a rare diagnosis and there is insufficient evidence on which to base treatment of the ipsilateral breast.

Methods

From 2006 till 2008 the Dutch National Cancer Registry identified patients nationwide with possible occult breast cancer. These patients were diagnosed with axillary nodal metastases with no evidence of the primary tumor on physical examination, mammography and ultrasound. Data on histopathology, imaging diagnostics and treatment were collected. After a minimum of 5 years follow up, data on recurrence and survival were gathered and analyzed.

Results

A group of 71 patients with occult breast cancer was identified. Of these, 7 patients received no local therapy. The remaining 64 patients all underwent surgery with axillary lymph node dissection. Ninety-two percent had MRI of the breast. Three groups were formed: Group A (n=25) had surgery of the breast, group B (n=28) received breast irradiation and group C (n=11) consisted of patients who had no treatment of the breast. Recurrence in the ipsilateral breast was seen in 4 patients (14%) in group B and in 3 patients (30%) in group C. There was 1 local recurrence in group A after breast conserving treatment and no significant differences between groups. After a median of 85 months of follow-up, overall survival (72%, 82% and 82% for group A, B and C, respectively) and distant recurrence rates (12%, 11% and 10% for group A, B and C, respectively) were not significantly different between groups assessed by log-rank test.

Conclusion

In occult cT0N+M0 breast cancer, conservative treatment of the breast is an acceptable modality which results in similar distant recurrence and overall survival rates. Surgery and breast irradiation appear to reduce the risk of ipsilateral breast recurrence. This is the first prospective study that provides evidence in the current clinical setting to guide treatment for patients with occult breast cancer.

Citation Format: de Maat M, Bretveld R, Steevens J, Vissers Y, Hulsewé K. Treatment of the breast in occult breast cancer: Results of a prospective Dutch national cohort study with 5 years follow up. [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 P2-12-07.
Abstract P2-12-11: Does postmastectomy radiotherapy improve survival in patients with 1-3 positive axillary lymph nodes? A systematic review and meta-analysis of the current literature
Authors: Kasem A , Headon H , Mokbel K
Published: May 2016

In breast cancer with more than four positive axillary lymph nodes, it is common practice to deliver radiotherapy to the affected site following a mastectomy. However, less is known about the benefit s this might offer in women with 1-3 positive lymph nodes. In order to assess whether postmastectomy radiotherapy has any benefit in these women, a meta-analysis was performed to assess whether postmastectomy radiotherapy improved overall survival or reduced locoregional recurrence in this group of women. It was found that postmastectomy radiotherapy significantly reduced the risk of locoregional recurrence, with a relative risk ratio of 0.3 (95% confidence interval 0.23-0.38), and resulted in a small benefit in overall survival, with a relative risk ratio of 1.03 (95% confidence interval 1.00-1.07). Therefore, in women with 1-3 positive lymph nodes, postmastectomy radiotherapy reduces the risk of locoregional recurrence and is associated with a small benefit in overall survival, so should be recommended within this group after careful multidisciplinary discussion.

Citation Format: Kasem A, Headon H, Mokbel K. Does postmastectomy radiotherapy improve survival in patients with 1-3 positive axillary lymph nodes? A systematic review and meta-analysis of the current literature. [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 P2-12-11.
Abstract P2-12-13: Modified pectoral nerves block for postoperative analgesia after modified radical mastectomy: A comparative study
Authors: Kanitkar R , Mane A , Agashe A , Kulkarni M , Deshmukh S
Published: May 2016

Background: The modified pectoral nerves block (Pecs II) as a method of analgesia for breast cancer surgery has shown excellent results in recent publications. This technique blocks the long thoracic nerve, thoracic intercostal nerves from T2-T6 & thoracodorsal nerve.

Aims: To evaluate the effectiveness of the Pecs II block for pain relief in the postoperative period of patients undergoing modified radical mastectomy (MRM).

Methods: A prospective comparative study was conducted at our instituition between November 2014 and March 2015. Patients scheduled to undergo MRM were randomly assigned to the test group (endotracheal anaesthesia along with Pecs II) and the control group (endotracheal anaesthesia only). In the postoperative period the patients were evaluated using a visual analogue scale to determine pain scores at 6, 12 & 24 hours.

Results: Fifty patients (25 in each group), between the ages of 24 to 76 years (54.76 ± 10) were included in the study. There was no significant difference in ages between test & control groups. In the postoperative period, the test group had significantly lower median pain scores at 6, 12 & 24 hours as compared to the control group.

Table 1: Comparative analysis for Pecs II block TestControlp valueeffect size (r)Average age (years)54.44 ± 10.7855.08 ± 9.370.824-Median postoperative pain scores  6 hours2 (1)6 (2)<0.001*- 0.8312 hours3 (1)5 (2)<0.001*- 0.6224 hours3 (1)0.034*- 0.30* statistically significant

Conclusion: The modified pectoral nerves block (Pecs II) is a novel & effective technique for postoperative analgesia in patients undergoing modified radical mastectomy.

Citation Format: Kanitkar R, Mane A, Agashe A, Kulkarni M, Deshmukh S. Modified pectoral nerves block for postoperative analgesia after modified radical mastectomy: A comparative study. [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 P2-12-13.