bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P1-12-14: Retinoic acid sensitizes triple-negative breast cancer cells to tamoxifen treatment
Authors: Coyle Krysta M , Dean Cheryl A , Jo Diana B , Thomas Margaret , Sultan Mohammad , Marcato Paola
Published: May 2015

Tamoxifen, an estrogen receptor (ER) antagonist, is often used as an adjuvant endocrine therapy in the successful treatment of ER+ breast tumors. Tumors that lack ER, progesterone receptor (PR) and HER2 expression (i.e. triple-negative breast cancers) cannot be treated with adjuvant endocrine therapies, like tamoxifen, and are often more aggressive. Inducing ER expression is a potential strategy for sensitization of triple-negative breast cancers to adjuvant endocrine therapies. Given recent evidence suggesting cross-talk between the retinoic acid (RA) and estrogen signaling pathways, we investigated if RA induces expression of ER in triple-negative breast cancer cells. We hypothesize that this would lead to sensitization of the cells to tamoxifen treatment. Quantitative PCR of mRNA isolated from triple-negative MDA-MB-231 cells treated with RA and estradiol had increased ER transcript levels. Furthermore, treatment with estradiol and RA synergistically induced increased expression of RA-inducible genes. In cell proliferation studies, neither RA nor estradiol treatment alone significantly altered the growth of MDA-MB-231 cells; however, when treated with both estradiol and RA together, the growth of the cells increased significantly. This suggests that the RA-mediated increase in ER expression sensitizes MDA-MB-231 cells to estradiol-induced cell growth. Next, we investigated whether the increased ER expression sensitized MDA-MB-231 cells to tamoxifen treatment. Tamoxifen did not decrease the growth of MDA-MB-231 cells; however, when applied in combination with both estradiol and RA, tamoxifen significantly reduced MDA-MB-231 proliferation. Furthermore, tamoxifen treatment reduced the synergistic effects of estradiol/RA on RA-inducible gene expression. Together, these results suggest that the use of RA in combination with tamoxifen warrants further investigation as a potential treatment for triple-negative breast cancers. The success of the combination treatment of tamoxifen and RA in the reduction of triple-negative breast cancer cell tumor xenografts will provide further justification for this strategy in the treatment of triple-negative breast cancers.

Citation Format: Krysta M Coyle, Cheryl A Dean, Diana B Jo, Margaret Thomas, Mohammad Sultan, Paola Marcato. Retinoic acid sensitizes triple-negative breast cancer cells to tamoxifen treatment [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P1-12-14.
Abstract P1-16-06: Margin Probe device is able to reduce re-excision rate of breast conserving surgery in invasive and pre-invasive breast cancer independent from any patient or tumor related factors
Authors: Blohmer Jens Uwe , Tanko Julia , Voelker Ragna , de Grahl Julia , Gross Jessica
Published: May 2015

Background: A positive margin status after breast conserving surgery (BCS) is one of the strongest predictors of local recurrence of intraductal and invasive carcinoma. As much as 20-50% of all patients with BCS need to undergo a second operation in order to receive free margins. In this study we tested the clinical performance of Margin Probe (Dune Medical Devices), a novel device for intraoperative margin evaluation.

Methods: A prospective clinical trial was performed: The device was applied to 150 lumpectomy specimen from consecutive patients with BCS treated during the first three months in 2013. The re-excision rate was compared to the re-excision rate of a historical group of 156 patients treated with BCS during the first three months in 2012, without the application of the device. We analyzed whether Margin Probe is affected by tumor morphology, grading, size of the tumor, breast density, age, body-mass-index or the use of wire-marker.

Results: Due to the application of Margin Probe the re-excision rate decreased significantly by 51% from 39.7% to 14.6%. In the subgroup of intraductal carcinoma (DCIS) the re-excision rate was reduced about two thirds from 66.7% to 23.1%. In the subgroup of invasive lobular carcinomas the re-excision rate decreased from 37.0% to 19.0%. Margin Probe results are not affected by grading, tumor size, breast-density, age, body-mass-index or wire-marker application.

Conclusion: Margin Probe is an effective tool for detection of positive margins during BCS and significantly decreases the re-excision rate. It is not limited to invasive carcinoma but also detects involved margins in DCIS as well as in invasive lobular carcinoma. It does not interfere with any of the factors we examined.

Citation Format: Jens Uwe Blohmer, Julia Tanko, Ragna Voelker, Julia de Grahl, Jessica Gross. Margin Probe device is able to reduce re-excision rate of breast conserving surgery in invasive and pre-invasive breast cancer independent from any patient or tumor related factors [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P1-16-06.
Abstract P1-16-07: Translating the concept of intrinsic subtypes into an oncoplastic cohort of more than 1000 patients-predictors of recurrence and survival
Authors: Rezai Mahdi , Kellersmann Stephanie , Knispel Sarah , Kimmig Rainer , Kern Peter
Published: May 2015

Introduction:

The concept of breast cancer experienced a paradigm shift by Sørlie T. et al. with intrinsic subtypes as prognostic classification of breast cancer [1]. We validated this concept in a large cohort study of oncoplastic surgery.

Patients and methods:

We analysed 1035 patients with oncoplastic surgery (2004-2009) and survival parameters related to histopathological approximated intrinsic subtypes.

Data were retrieved from customized questionnaires and patients charts. Survival data were determined from cancer registries.

Results:

A total of 944 patients with primary unilateral breast cancer, median age 58 years, were eligible for analysis. At a median follow-up of 5.3 years, LRR was 4.1%, with 5-year-OS of 94.5% and DFS of 90.9%. Stage distribution was as follows: T1a 3%, T1b 12 %, T1c 44,2 % and T2 was 22,1%. 70,4 % of patients were nodal-negative and nodal involvement was predominantly low.

Intrinsic subtypes, not T-size, nodal-status, resection margin width nor histopathology, governed the prognosis of this cohort. Triple-negative and Her2 non luminal breast cancer had the highest recurrence and the lowest survival rates compared to Luminal A: Recurrence TNBC 11,3 %, Her2pos non luminal 9,3 %, Luminal A 2,5 %; Overall survival: TNBC 91,3 %, Her2 non luminal 93,7 %; Luminal A: 96,3 %. Our data confirmed the intrinsic subtype concept on a large basis in oncoplastic surgery.

(1) Sørlie T, Perou CM, Tibshirani R, Aas T, Geisler S, Johnsen H, Hastie T, Eisen MB, van de Rijn M, Jeffrey SS, Thorsen T, Quist H, Matese JC, Brown PO, Botstein D, Lønning PE, Børresen-Dale AL: Gene expression patterns of breast carcinomas distinguish tumor subclasses with clinical implications. Proc Natl Acad Sci USA 2001; 98: 10869–10874.

Citation Format: Mahdi Rezai, Stephanie Kellersmann, Sarah Knispel, Rainer Kimmig, Peter Kern. Translating the concept of intrinsic subtypes into an oncoplastic cohort of more than 1000 patients-predictors of recurrence and survival [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P1-16-07.
Abstract P2-01-19: Long-term follow-up of the node-negative breast cancer patients before treatment evaluated with sentinel node biopsy alone following neoadjuvant chemotherapy
Authors: Nogi Hiroko , Uchida Ken , Kamio Makiko , Kato Kumiko , Toriumi Yasuo , Takeyama Hiroshi
Published: May 2015

BACKGROUND Sentinel node biopsy (SNB) for the node negative breast cancer is standard treatment as an accurate assessment of axillary lymph node status. The use of neoadjuvant chemotherapy (NAC) has increased during the past several years. At present, the use of SNB following NAC is controversial. Proponents of SNB after NAC prefer a single surgical procedure with potential for fewer axillary dissections.

OBJECTIVE Our objective was to examine SNB evaluation alone following NAC in patients with clinically node-negative breast cancer before treatment and to evaluate the axillary lymph node recurrence for patients undergoing NAC versus patients undergoing surgery first.

METHODS A total of 1176 patients with clinically node negative breast cancer underwent SNB from 2007 to 2013. Clinicopathologic and survival data were reviewed and comparisons made between patients receiving NAC and those undergoing surgery first. Lymphatic mapping was performed with both radioactive colloid and blue dye. Patients with negative SN for metastasis were followed without axillary lymph node dissection (ALND). Patients with metastases to a SN underwent ALND.

RESULTS Of the patients, 180 (15.3%) underwent SNB following NAC and 996 (84.7%) underwent surgery first. SN identification rates were 98.3% in the NAC group and 98.9% in the surgery first group. 152 (84.4%) patients in the NAC group and 877 patients (88.1%) had negative SNs. At median follow-up of 49.2 months, 1 patient (0.007%) in the NAC group and 3 patients (0.003%) in surgery first group had axillary lymph node recurrences. There were not any significant differences between the NAC group and the surgery first group.

CONCLUSION The SNB following NAC in patients with node-negative breast cancer has a low axillary recurrence rate and could be acceptable.

Citation Format: Hiroko Nogi, Ken Uchida, Makiko Kamio, Kumiko Kato, Yasuo Toriumi, Hiroshi Takeyama. Long-term follow-up of the node-negative breast cancer patients before treatment evaluated with sentinel node biopsy alone following neoadjuvant chemotherapy [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P2-01-19.
Abstract P2-01-27: In patients with micrometastatic in sentinel lymph node biopsies, involvement of the non-sentinel lymph nodes cannot be predicted by clinicopathological variables
Authors: Chauhan Muhammad N , Dave Rajiv V , Ghaus Mauria , Ahmad Sana , Sayers Craig , Kryjak Zbigniew , Ali Deedar
Published: May 2015

Background: The Sentinel Lymph Node Biopsy (SLNB) procedure is recognised to be an accurate method of staging the axilla in patients with early stage breast cancer. There remains a debate as to whether patients with micrometastases should undergo completion axillary lymph node dissection (CALND). We aimed to assess the indicators for positive non-sentinel lymph nodes (n-SLN) following CALND.

Methods: We retrospectively analysed our experience of SNLB between July 2008 to July 2013. A total of 1152 breast cancer patients underwent SLNB based on lymphoscintigraphy, intra-operative gamma probe detection, and blue dye mapping using 99mTc-nanocolloid and Patent Blue V injected peri-areola. Statistical analysis was performed using Fisher’s exact and c2 for categorical data.

Results: Out of 1152 SLNB biopsies performed, 224 (19.5%) were positive for metastatic disease; macrometastases in 150 (67%), micrometastases in 72 (32%) and ITC in 2. CALND was not performed in 20 cases (9 macrometastases, 10 micrometastases, and 1 ITC), largely due to concerns regarding fitness for anesthesia. Macrometastases on SNLB were more likely to predict positive n-SLN on ANC {macrometastases; 39/141(27.7%) vs micrometastases; 9/62 (14.5%), p=0.029}. On univariate analysis, positive n-SLN in CALND for patients with micrometastases on SLNB was not predicted by grade (G0-G2; 6/43, and G3; 3/19, p=0.565), size of primary breast tumour (<40mm; 8/58, 340mm; 1/4, p=0.475), lymphovascular invasion (5/30 vs 4/31, p=0.503), age (<50 years; 3/24 vs 350 years; 6/38, p=0.496), or number of positive SLNB (all patients had <2 positive nodes on SLNB). Recurrences were detected in 4 patients, of which 1 was in a patient with micrometastases on SLNB. Out of the 4 recurrences, 3 were distant (liver and bone) and one was locoregional, with new disease in the contralateral breast).

Conclusion

In our series, 14.5% (9/62) of patients with micrometastases had positive n-SLB on CALND, which was not predicted by any clinicopathological characteristics. We have recently changed our practice toward not routinely offering CALND in patients with micrometastases, in keeping with current vogue. However, it is important to inform our patients that 14.5% of patients with micrometastases on SLNB may have positive n-SLN.

Citation Format: Muhammad N Chauhan, Rajiv V Dave, Mauria Ghaus, Sana Ahmad, Craig Sayers, Zbigniew Kryjak, Deedar Ali. In patients with micrometastatic in sentinel lymph node biopsies, involvement of the non-sentinel lymph nodes cannot be predicted by clinicopathological variables [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P2-01-27.
Abstract P2-01-28: Prognosis of metastatic internal mammary sentinel nodes (IMSN) in breast cancer
Authors: Avisar Eli , Libson Shai , Perez Eduardo
Published: May 2015

Introduction:

The presence of positive internal mammary nodes has historically been associated with a significant worse prognosis. Recent studies have also demonstrated a worse prognosis associated with drainage to the internal mammary nodes. Intense search for those nodes, biopsy, appropriate staging and modern treatment for positive IMSN might improve outcome. We sought to study the prognosis of pathologically positive IMSN at our institution.

Methods:

A retrospective analysis of a prospectively collected database including all breast cancer patients identified with an IMSN that was biopsied between 2005 and 2012 was performed. Demographics, histologic markers, patterns of recurrence as well as survival data were collected. Univariate and multivariate analysis were performed.

Results:

Thirteen of 82 patients with a biopsied IMSN were metastatic (16%). In 8 of those (62%), the biopsy resulted in a change in staging. In all cases of positive IMSN additional radiation to the internal mammary chain was added. Eighteen percent (18%) of positive nodes did not show drainage to the internal mammary basin during Lymphoscintigraphy. There was no statistical difference in regional and distant recurrences between the patients with positive IMSN and those with negative IMSN. Furthermore there was no difference in disease specific survival.

Conclusions:

Intense search for IMSN presence and biopsy of those nodes is associated with changes in staging and treatment for metastatic IMSN. In our study, pathologically positive IMSN were associated with a non- inferior prognosis than negative IMSN.

Citation Format: Eli Avisar, Shai Libson, Eduardo Perez. Prognosis of metastatic internal mammary sentinel nodes (IMSN) in breast cancer [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P2-01-28.
Abstract P2-01-30: A new nomogram to predict axillary metastasis in breast cancer patients without axillary surgery
Authors: Rodionov Valery , Cometova Vlada , Panchenko Sergey , Idrisova Sereda , Savinov Yurij , Rodionova Maria
Published: May 2015

Background: Several studies have concentrated on finding a combination of predictive parameters to establish a mathematical model that can identify patients with no axillary metastasis for whom routine lymph node dissection could be safely avoided. We developed a new model of nomogram (the Ulyanovsk Cancer Center axillary lymph node metastasis nomogram, UCC-ALNM nomogram); it employs clinically and pathologically relevant variables and offers possible advantages over the others nomograms.

The purpose of the study: To assess the predictive power of UCC-ALNM nomogram.

Methods: A total of 530 breast cancer patients treated between 2008 and 2010 were used as the modeling group for validating the UCC-ALNM nomogram. Clinical and pathologic features of patients were assessed by multivariable logistic regression to predict the presence of axillary metastasis in breast cancer patients. The predictive accuracy of our nomogram was measured by calculating the area under the receiver-operating characteristic (ROC) curve (AUC). Clinical factors included into analysis were: patient’s age and localization of the primary tumor. Pathological factors evaluated were: traditional pathological criteria (primary tumor size, histological type, tumor grade, HR- and Her-2 status) and new total pathological index (Ulyanovsk prognostic index - UPI), introduced by pathologists of the Ulyanovsk Regional Cancer Center. UPI is total score of six main pathological criteria that characterize the malignancy of epithelial tumors: degree of cellular differentiation, cellular polymorphism, mitotic activity, growth pattern, lymphovascular invasion, stromal reaction.

Results: By the multivariate analysis, patient’s age (p=0.04), tumor size (p<0.001), UPI (p<0.001), PR (p<0.001) and Her2 status (p=0.02) were identified as independent predictors of axillary metastasis. The nomogram was then developed using the six variables associated with axillary metastasis: age, tumor size, PR, Her2, UPI.

The new model was accurate and discriminating with an AUC of 0.7510 when applied to the modeling group.

Conclusions: UPI is a new predictive factor of axillary metastasis in breast cancer patients. UCC-ALNM nomogram.

Citation Format: Valery Rodionov, Vlada Cometova, Sergey Panchenko, Sereda Idrisova, Yurij Savinov, Maria Rodionova. A new nomogram to predict axillary metastasis in breast cancer patients without axillary surgery [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P2-01-30.
Abstract P2-02-02: A review of the accuracy of wire localisation using digital breast tomosynthesis (DBT) in a prospective series of 81 patients
Authors: Holt Simon DH , Nadi Khaldoun MY , Gurung Amrita , Williams Helen R , Huws Anita M , Sharaiha Yousef M
Published: May 2015

Introduction: Digital breast tomosynthesis allows identification of impalpable lesions in three-dimensions. We have devised a technique for wire placement using a gridded plate that allows localisation of such lesions for subsequent surgical excision.

Method: 81 consecutive cases were studied prospectively between Jan 2012 and May 2014. The localisation is usually performed by the operating surgeon under local anaesthetic shortly before the patient is transferred to the operating theatres. Our surgeons consider the optimal position for the tip of the wire to be 1 cm beyond and 1cm to the side of the lesion placed using only the CC or lateral projection. The patient is positioned with the aperture of a gridded plate over the lesion needing to be surgically removed. A single DBT is acquired and the volumes reviewed. The lesion is identified and the depth in the breast recorded (Z coordinate). The ideal position for the wire is then marked in that plane and the volume scrolled through to bring the calibrated plate into focus. This identifies the X and Y coordinates, which are then recorded. The wire is then placed freehand in the line of the beam and a further DBT performed to check the position of the wire without needing to reposition the patient. A specimen x-ray is performed immediately following excision to confirm the target lesion is included. The results are verified at the subsequent multidisciplinary team (MDT) meeting.

Results: Once the target is identified, the average time for the localisation procedure and check DBT was 4 minutes. This series included both diagnostic (N=24, 30%) and therapeutic excisions (N=57, 70%). Patients presented both from screening (N=64, 79%) and symptomatically (N=17, 21%). The age range of the patients was 42-79y (mean 59y). 54 (67%) cases were undergoing excision for calcifications, 15 (19%) for impalpable masses, 11 (13%) for radial scars or distortions and 1 for glandular asymmetry. The final diagnoses following excision were IDC 34 (42%), DCIS 29 (36%), benign 13 (16%) and 1 case each of ILC, spindle cell tumour, mucinous carcinoma, phyllodes and pleomorphic LCIS.

In all cases the target lesion was shown to be present in the specimen x-ray, confirmed on histology and ratified by the MDT.

Conclusions: Wire localisation using DBT is a quick and accurate method of localising impalpable lesions for surgical excision.

Citation Format: Simon DH Holt, Khaldoun MY Nadi, Amrita Gurung, Helen R Williams, Anita M Huws, Yousef M Sharaiha. A review of the accuracy of wire localisation using digital breast tomosynthesis (DBT) in a prospective series of 81 patients [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P2-02-02.
Abstract P2-04-08: Accuracy of estrogen receptor, progesterone receptor, HER2 Status and Ki67 labeling index between core needle and surgical excisional tumour in 910 patients with breast cancer
Authors: Nakamura Rikiya , Yamamoto Naohito , Itookubo Yoshiyuki , Nakagawa Ayako , Itami Makiko
Published: May 2015

Purpose

Accurate determination of ER, PgR, HER2 status and Ki67 labeling index was very important in making decision for adjuvant and neoadjuvant treatment for patients with breast cancer. However discordance of ER, PgR, HER2 status and Ki67 labeling index (Ki67-LI) between core needle biopsy (CNB) and surgical specimens (SSp) varied among reported studies.

The aim of the present study was to compare the accuracy of CNB with that of SSp for ER, PgR,HER-2 status, Ki67-LI detection in breast cancer.

Patients and methods

All patients diagnosed with an early breast cancer in our Cancer Center Hospital between January 2005 and May 2014 was included but exclusion criteria of patients with large tumour requiring neoadjuvant chemotherapy. All CNB were performed under ultrasound guidance, with at least four 14-gauce core biopsies being obtained for pathological examination. ER, PgR, HER-2 status and Ki67-LI were assessed in CNB and in SSp. ER and PgR were determined by

Immunohistochemistry (IHC). The cut points for ER and PgR positive was10%. The cut point for Ki67-LI high expression was 20%. HER2 was determined by IHC and scored from 0 to 3+. FISH analysis was carried out in HER2 2+ cases and in discordant cases.

Results

A total of 910 patients were assessed. CNB can be used with confidence for ER and HER2 determination. Rates for sensitivity, specificity, negative predictive value and positive predictive value for CNB compared with SSp are 99.3%,95.6%,97.2%,98.5% for ER, 94.5%,81.8%,88.2%,90.7% for PgR ,93.0%,99.1%,99.6%,93.9% for HER2, and 87.7%,69.8%,81.1%,79.4% for Ki67-LI, respectively. Specially, the most impact factor of discordance for Ki67-LI and HER2 is Tumour size and that for ER and PgR is lobular carcinoma.

Conclusion

CNB can be used with confidence for ER and HER2 determination. For PgR and Ki67-LI due to substantial discordance results from CNB should be used with caution.

Citation Format: Rikiya Nakamura, Naohito Yamamoto, Yoshiyuki Itookubo, Ayako Nakagawa, Makiko Itami. Accuracy of estrogen receptor, progesterone receptor, HER2 Status and Ki67 labeling index between core needle and surgical excisional tumour in 910 patients with breast cancer [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P2-04-08.
Abstract P2-05-01: The non-coding transcriptome of hypoxic breast cancer: Novel insights of clinical relevant long non-coding RNA in hypoxia signalling
Authors: Choudhry Hani , Schodel Johannes , Albukhari Ashwag , Haider Syed , Buffa Francesca , Ratcliffe Peter J , Mole David R , Ragousis Ioannis , Harris Adrian L
Published: May 2015

Hypoxia is associated with aggressive and poor prognosis of breast cancer. Generally, pan-genome analyses of hypoxia have focussed on protein-coding genes, however, the role of non-coding RNAs, in particular long non-coding RNAs (lncRNA) in hypoxia is not well characterised. We undertook an integrated genomic analysis of the hypoxic transcriptome in MCF7 breast cancer cells, employing total RNA-seq together with ChIP-seq for the hypoxia-inducible transcription factor (HIF) and for epigenetic marks of transcriptional activation (RNApol2 and histone H3K4me3).

Analyses revealed that all classes of RNA are significantly regulated by hypoxia including piwiRNA, miRNA, tRNA, and sn/snoRNA. Significant numbers of lncRNAs were upregulated in hypoxia and were associated with increased RNApol2 and H3K4me3 markers and with HIF binding, indicating direct transcriptional activation of lncRNAs by HIF. The most hypoxiclly upregulated lncRNA was NEAT1, which is a direct transcriptional target of HIF-2a but not HIF-1a. The role of NEAT1 in cancer has not been previously studied.

We demonstrated that hypoxic NEAT1 induction is common in breast cancer cell lines and xenografts models treated with bevacizumab. NEAT1 directly induces the formation of nuclear paraspeckle bodies in hypoxia. Moreover, it contributes to tumourigenicity by increasing cell proliferation, colony formation, and reducing apoptosis. In addition, we report that NEAT1 is required to retain hypoxia induced hyper edited Junctional Adhesion Molecule A (JAM-A) mRNA in the nucleus, thus preventing export into the cytoplasm for translation. Finally, in a large cohort of 2000 breast cancers, high levels of NEAT1 were associated with poor clinical outcomes and clinicopathological features.

Our results extend knowledge of the hypoxic transcriptional response into the spectrum of non-coding transcripts. These findings provide novel mechanisms of transcriptional regulation in hypoxia and open new avenues to find novel pathways and targets to develop therapies for breast cancer.

Citation Format: Hani Choudhry, Johannes Schodel, Ashwag Albukhari, Syed Haider, Francesca Buffa, Peter J Ratcliffe, David R Mole, Ioannis Ragousis, Adrian L Harris. The non-coding transcriptome of hypoxic breast cancer: Novel insights of clinical relevant long non-coding RNA in hypoxia signalling [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P2-05-01.
Abstract P2-05-04: Gene expression associated with poor prognosis of young TNBC patients
Authors: Karn Thomas , Pusztai Lajos , Liedtke Cornelia , Bianchini Giampaolo , Györffy Balazs , Hatzis Christos , Rody Achim , Müller Volkmar , Schmidt Marcus , Holtrich Uwe , Becker Sven
Published: May 2015

Background:

Among TNBC patients those of very young age (<40 years) display a significantly worse prognosis (Liedtke et al. 2013 Breast Cancer Res Treat). We verified this result in 1161 TNBC samples with full Affymetrix gene expression data of which 845 patients had both detailed age and follow up information.

Materials and Methods:

We split the full sample set into a finding cohort of 394 TNBC and a validation cohort of 767 TNBC encompassing 309 and 536 samples, respectively, with both age and follow up data. We then used significance analysis of microarrays (SAM) in the finding cohort to look for genes whose expression is associated with young age (<40 years). Identified genes were analyzed for their correlations to known metagenes characteristic for different TNBC subtypes. Subsequently the whole analysis was repeated in the validation cohort.

Results:

We identified 98 and 222 probesets by SAM in the finding and validation cohort, respectively. Only a subset of identical probesets (19.4%) was re-identified in the validation. However, the gene lists were similarily enriched for correlations to specific metagenes and the respective TNBC subtypes. We found that young age among TNBC is positively correlated with increased proliferation and a basal-like subtype, but negatively correlated with the molecular apocrine phenotype and the claudin-low subtype. We also observed a negative correlation of young age with stromal enrichment and EMT but no difference in lymphocyte infiltration as judged by specific metagenes. However, despite that TNBC of patients <40 years have a poor prognosis (P=0.006) and are clearly enriched in basal-like subtype (70.7% vs 55.3%; P=0.001), still basal-like TNBC do not differ from non-basal-like TNBC in prognosis. Moreover, even within the group of basal-like TNBC the prognostic effect of age<40 was still observed (P=0.008). Interestingly, the changes in expression of single genes and metagenes were observed over a wide age range of 35-85 years. In contrast the difference in prognosis was markedly pronounced at the 40 year age limit. Thus, potential critical differences in gene expression associated with poor prognosis of TNBC <40 years seem to be concealed by many age-associated changes without prognostic value.

Citation Format: Thomas Karn, Lajos Pusztai, Cornelia Liedtke, Giampaolo Bianchini, Balazs Györffy, Christos Hatzis, Achim Rody, Volkmar Müller, Marcus Schmidt, Uwe Holtrich, Sven Becker. Gene expression associated with poor prognosis of young TNBC patients [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P2-05-04.
Abstract P2-05-16: Gene expression in synchronous primary, axillary nodal and disseminated breast cancer cells
Authors: Broom Reuben J , Blenkiron Cherie , Harman Richard , Whineray Kelly Erica , Allpress Stephen , Gale Katherine , Rice Sam , Shin Peter , Brown Anna , Lasham Annette , Print Cristin
Published: May 2015

Disseminated Tumour Cells (DTC) have been found in the bone marrow (BM) of up to 70% of patients with breast cancer metastatic to axillary lymph nodes. They are a biologically and therapeutically interesting tumour cell population. Although there is uncertainty about their relationship to cancer prognosis, DTC provide a valuable window into the processes by which primary tumour (PT) cells disseminate. DTC can be enriched from BM by immunoaffinity using antibodies directed against epithelial cell markers such as EpCAM. To better understand the changing gene expression patterns that may accompany breast cancer metastasis, we have compared the whole genome RNA expression profiles of matched PT, axillary lymph node metastases (LNM), and EpCAM-enriched cells from the BM of seven patients. Compared to PT, axillary LNM had consistently altered expression of RNAs encoding matrix metalloproteinases (MMP), growth factors, transcription factors, as well as downstream targets of Catenin-a, Tumour Necrosis Factor (TNF)-a and miR-22. Once the gene expression patterns of potentially contaminating BM cells were subtracted, compared to PT and LNM EpCAM-enriched MB cells had consistently elevated expression of RNAs encoding metabolic enzymes, ribosomal proteins, and DNA-binding factors such as YB-1. The gene expression changes we identify are candidate mediators of breast cancer metastases and represent attractive targets for further study on a cell by cell basis.

Citation Format: Reuben J Broom, Cherie Blenkiron, Richard Harman, Erica Whineray Kelly, Stephen Allpress, Katherine Gale, Sam Rice, Peter Shin, Anna Brown, Annette Lasham, Cristin Print. Gene expression in synchronous primary, axillary nodal and disseminated breast cancer cells [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P2-05-16.
Abstract P2-06-07: StemScreen®, an innovative platform technology for the identification of novel cancer stem cell-directed compounds
Authors: Chen Janice , Li Hai , Glicksman Marcie A , Karan Charles , Brooks Christopher , Rowinsky Eric K
Published: May 2015

Cancer stem cells (CSCs) are thought to play significant roles in breast cancer initiation and progression. Conventional therapeutic agents target tumor bulk but spare CSCs, leading to tumor recurrence and relapse. Therefore, drugs that eliminate both tumor bulk and CSCs may represent the most effective treatment strategy for breast cancer. We have developed a proprietary cell-based screen called StemScreen® that permits the rapid testing and identification of novel CSC-directed compounds in a high-throughput manner. StemScreen® takes advantage of a landmark discovery that many cancer cell lines harbor stable populations of CSCs. Our initial studies utilized a breast cancer cell line transfected with a unique expression vector that is only active in the 1-5% putative CSC population. The StemScreen® platform is unique because it allows for screening of compounds within the context of the CSCs’ natural microniche environment and has advantages over traditional drug discovery methods that have been designed to identify compounds that only target tumor bulk, but not CSCs.

In an effort to increase the throughput and make the assay compatible with existing compound libraries, we miniaturized the platform to a 384-well format and optimized the line for high-content screening. A series of compound hits identified from screening these cells against smaller libraries of known active compounds as well as larger and more diverse compound libraries will be presented. We are also currently expanding this technology into other tumor types. We believe that this approach represents a major technological advance in oncology drug discovery and that this platform will be instrumental in the discovery of unique new therapies that have a dual effect on CSCs and non-CSC tumor bulk.

Citation Format: Janice Chen, Hai Li, Marcie A Glicksman, Charles Karan, Christopher Brooks, Eric K Rowinsky. StemScreen®, an innovative platform technology for the identification of novel cancer stem cell-directed compounds [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P2-06-07.
Abstract P2-07-01: EMT activates PERK-eIF2α signaling and sensitizes cells to perturbations in endoplasmic reticulum function
Authors: Feng Yuxiong , Sokol Ethan S , Del Vecchio Catherine A , Sanduja Sandhya , Claessen Jasper HL , Proia Theresa A , Jin Dexter X , Reinhardt Ferenc , Ploegh Hidde L , Wang Qiu , Gupta Piyush B
Published: May 2015

Epithelial-to-mesenchymal transition (EMT) plays an important role in cancer progression. By undergoing an EMT, cancer cells acquire a spectrum of malignant properties, including invasiveness, multi-drug resistance and stem-like traits. Although they play an important role in tumor progression and resistance, few vulnerabilities of EMT cancer cells have been reported to date. To identify specific vulnerabilities of EMT cells, Using small molecule and RNAi screens, we have discovered that induction of EMT greatly sensitizes cells to agents that perturb endoplasmic reticulum (ER) function. This unexpected sensitivity to ER stress is mainly due to the expression and secretion of large amount of extracellular matrix (ECM) proteins by cells that have undergone an EMT. In line with their increased secretory load, EMT cells display a branched ER morphology and constitutively activate the PERK-eIF2α branch of the unfolded protein response (UPR). Using a PERK-specific inhibitor, we found that PERK activation is also required for EMT cells to invade and metastasize. In human tumor tissues, EMT gene expression correlates strongly with both ECM and PERK-eIF2α genes. In summary, our findings identify a novel vulnerability of cells that have undergone an EMT, and demonstrate that the PERK branch of the UPR is required for their malignancy.

Citation Format: Yuxiong Feng, Ethan S Sokol, Catherine A Del Vecchio, Sandhya Sanduja, Jasper HL Claessen, Theresa A Proia, Dexter X Jin, Ferenc Reinhardt, Hidde L Ploegh, Qiu Wang, Piyush B Gupta. EMT activates PERK-eIF2α signaling and sensitizes cells to perturbations in endoplasmic reticulum function [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P2-07-01.
Abstract P2-07-07: Prostaglandin E receptor 2 (EP2) regulates breast cancer stem-cell like property and promotes epithelial-mesenchymal transition
Authors: Shin Vivian Y , Siu Man T , Ho John C , Cheuk Isabella , Chen Jiawei , Kwong Ava
Published: May 2015

Background: Presence of cancer stem-like cells (CSCs) is the main obstacle for poor treatment response and mortality in breast cancer patients. Prostaglandin E receptors have been reported to play a role in epithelial-mesenchymal transition (EMT) and metastasis, however, the contribution on cancer stem cell compartment remains unstudied.

Methods: Human xenograft breast cancer model was used to study the expression of EP receptors during cancer development. Construction of stable EP2-expression cells was used to study tumorigenesis and characterization of EP2 receptor. Functional role of EP2 receptor on cell proliferation, flow cyometry, invasion and EMT gene expression array were performed in transfected cells. Expression of EP2 receptor was compared in primary tumor tissues by immunostaining and real-time PCR.

Results: EP2 receptor was predominantly expressed in animal tissues during cancer development, as well as in human primary tumor tissues. In mouse xenograft model, MB-231-EP2 clone developed a more aggressive tumor with a larger tumor size and showed a significant increase in cancer stem cell marker aldehyde hydrogenase (ALDH1) expression. In vitro study, MB-231-EP2 clone increased colony formation capacity and S-phase entry by the regulation of E-cadherin, TWIST1 and ALDH1. Importantly, we found that Twist1 expression level was higher in breast cancer patients than healthy controls and was associated with ALDH1 expression.

Conclusions: These findings implicated that EP2 receptor was crucial to nurture CSC phenotype and promote tumorigenesis in breast cancer. Blocking of EP2 might be a potential therapeutic strategy to improve treatment response for breast cancer patients.

Citation Format: Vivian Y Shin, Man T Siu, John C Ho, Isabella Cheuk, Jiawei Chen, Ava Kwong. Prostaglandin E receptor 2 (EP2) regulates breast cancer stem-cell like property and promotes epithelial-mesenchymal transition [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P2-07-07.
Abstract P2-08-01: Autoimmunity in breast carcinogenesis. A new paradigm
Authors: Fernández Madrid Félix , Maroun Marie-Claire , Lipovich Leonard , Tait Larry , Stark Azadeh , Zarbo Richard , Polin Lisa , Chitale Dhananjay
Published: May 2015

The B cell response in breast cancer [BC] may have a tumor-promoting effect. The objective of this work was to demonstrate that TAAs and cytokines inflict autoimmune damage to the breast creating a chronic inflammatory milieu that promotes BC progression

Methods: The DCIS.com nude mouse xenograft model of BC was treated with rituximab and dexamethasone. Immunoscreening of a cDNA library, PCR, and sequence determination. Immunohistochemistry [IHC] for inflammatory cytokines, immunoglobulins and complement and the TUNEL assay.

Results: Deposition of immunoglobulins and complement was shown on breast epithelial and stromal cells and the involvement of IL-4, IL-6, IL-17, TNF-alpha and NFkB was demonstrated by IHC staining. We identified autoantibodies recognizing multiple human breast epithelial and stromal antigens and the Fc and V regions of immunoglobulins. Rituximab and dexamethasone treatment markedly modified the macroscopic and microscopic appearance of BC and the pattern of cytokine deposition, induced increase in vascularity, marked stromal hypertrophy and massive debulking of the tumor by drastically reducing the tumor mass through apoptosis.

Conclusions: These results suggest that autoantibody- and cytokine-mediated autoimmune damage, triggered by TAAs creates a chronic inflammatory milieu with generation of pro-inflammatory and tumor promoting signals supporting BC progression. The dramatic results of the treatment of the xenograft model with rituximab and dexamethasone illustrate the potential of treating BC as a chronic rheumatic autoimmune disease. The demonstration of a major role of autoimmunity in BC progression may have a transformative impact on prevention, diagnosis and treatment of BC.

Citation Format: Félix Fernández Madrid, Marie-Claire Maroun, Leonard Lipovich, Larry Tait, Azadeh Stark, Richard Zarbo, Lisa Polin, Dhananjay Chitale. Autoimmunity in breast carcinogenesis. A new paradigm [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P2-08-01.
Abstract P2-08-02: Autoimmunity to centrosomal and proteasome proteins in breast cancer. A link to chromosome instability?
Authors: Maroun Marie-Claire , Olivero Ofelia , Abrams Judith , Chen Wei , Stark Azadeh , Peebles Carol , Tait Larry , Chitale Dhanajay , Fernández Madrid Félix
Published: May 2015

BACKGROUND. Perturbation of centrosomal or centrosome-associated proteins has been observed in nearly all human solid tumors and has been implicated in the origin of chromosomal instability. Ubiquitin-proteasome degradation is highly dependent on the organizing capabilities of the centrosomes. Here we report that autoantibodies in breast cancer [BC] sera target centrosomal proteins as well as important proteins involved in proteasome protein degradation.

METHODS. We immunoscreened a T7 cDNA library of BC proteins and the association of the cloned autoantigens with BC was studied by autoantigen microarray analysis. We used immunohistochemistry [IHC] to investigate the expression of the centrosome and centrosome-associated autoantigens identified.

RESULTS. Immunoscreening with BC sera led to the identification of autoantibodies recognizing epitopes developing in a family of proteins located on the centrosomes such as NIMA-related kinase 7, dynein heavy chain domain 3, peri-centriolar material-1, isomorph CRA, and stathmin-1, the ubiquitin-conjugating enzyme E2, the proteasome 26S subunit and the SUMO/sentrin peptidase. Antibody reactivity to these proteins which are associated with centrosome assembly, microtubule function and protein degradation were highly associated with the diagnosis of BC. IHC staining of paraffin-embedded BC sections with specific antibodies showed that aurora and stathmin-1 and other centrosome antigens are expressed in BC.

CONCLUSIONS. The discovery of autoantibodies targeting important centrosome and proteasome proteins associated with BC suggests that this immune reactivity could be related to autoimmunity developing in BC. Our findings indicate that these autoantibodies might be biomarkers of early BC and suggest the possibility of a link with chromosomal instability in BC.

Citation Format: Marie-Claire Maroun, Ofelia Olivero, Judith Abrams, Wei Chen, Azadeh Stark, Carol Peebles, Larry Tait, Dhanajay Chitale, Félix Fernández Madrid. Autoimmunity to centrosomal and proteasome proteins in breast cancer. A link to chromosome instability? [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P2-08-02.
Abstract P2-09-04: Measurements of advocate participation in and contributions to the Mayo Clinic breast cancer SPORE
Authors: Chauhan Cynthia , Eppard Wayland , Denison Lori , Gearhart Debra , Holmes Lawanna , Kraft Ruth , Miller Linda , Sitta Mary A , Smith Mary L , Ingle James , Goetz Matthew
Published: May 2015

[Background] Patient advocates bring a unique, valuable perspective and presence to SPOREs. However, although patient advocates are recommended members of SPORE grants, guidelines are not given on the role of advocates or on measurement of the performance of advocates.

[Methods] The Mayo Clinic Breast Cancer SPORE Patient Advocate Advisory Committee has developed a participation measurement tool. This tool will underpin and augment evaluations of the individual advocate contributions by the program PI and the SPORE project leaders. The Advocate Advisory Committee Chair will review the evaluations with the individual advocates. Annually, the advocate committee will review the performance of the group by the standard of the metrics.

[Results] Advocates kept a log of their SPORE activities for four months then worked with a statistician to discern patterns of measurable, evaluable behaviors. The determined evaluable areas are training, scientific activity, meeting attendance, communication. Currently, those metrics are being tracked. Advocates developed the metrics together and report activities at the monthly advocate meetings. Currently, activities are simply tracked and categorized. No minimum standards are set. This will be the third step in the development of the metrics.

[Conclusions] This approach is the first step in the development of meaningful evaluation of advocate SPORE activities. It quantifies the participation of advocates in the SPORE and provides the basis for developing a qualitative performance evaluation. The SPORE PI and the Committee Chair will use the evaluations to interpret the involvement of advocates in SPORE research and activities and to plan interventions to strengthen the advocate contributions to the SPORE.

Citation Format: Cynthia Chauhan, Wayland Eppard, Lori Denison, Debra Gearhart, Lawanna Holmes, Ruth Kraft, Linda Miller, Mary A Sitta, Mary L Smith, James Ingle, Matthew Goetz. Measurements of advocate participation in and contributions to the Mayo Clinic breast cancer SPORE [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P2-09-04.