bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P3-07-31: Immune activation signatures identify a subset of ER+ breast cancers with increased pathologic complete response to neoadjuvant chemotherapy
Authors: Stover DG , Waks AG , Erica ML , Brugge JS , Winer EP , Selfors LM
Published: May 2016

Background: Proliferation is the strongest predictor of response to neoadjuvant chemotherapy in estrogen receptor-positive (ER+) breast cancer. Evidence of immune activation has also been associated with improved response to neoadjuvant chemotherapy in breast cancer. We hypothesized that immune signatures may be associated with response independent of proliferation in ER+ breast cancers.

Approach: We compiled microarray expression data from breast cancer biopsies obtained prior to neoadjuvant chemotherapy on 465 ER-positive/HER2-negative patients by reported pathologic receptor status. We evaluated the association of 118 published gene expression signatures with response to neoadjuvant chemotherapy, based on study-defined pathologic complete response (pCR) versus residual disease (RD).

Results: Overall, 42 of 118 signatures were significantly associated with response to neoadjuvant chemotherapy in ER+ breast cancer (FDR-corrected p<0.05, simple logistic regression). Of those signatures that achieved significance, 52% (22/42) of signatures were proliferation-associated based on correlation to the 11-gene PAM50 proliferation index (Pearson's R2>0.30, p<1e-10). Among signatures that were NOT proliferation-associated, 50% (10/20) were immune-related. Using unsupervised hierarchical clustering of all 118 signatures, these ten immune signatures formed a distinct cluster. Of the 10 signatures, nine were designed to reflect "immune activation" and were highly correlated with each other in ER+ tumors (R2>0.4, p<0.001). The mean of each of these nine signatures was significantly higher in patients with pCR versus RD (FDR-corrected p<0.05, t-test). Patients with higher "immune activation" signatures had increased likelihood of pCR within multiple subgroups of ER+ breast cancer, including luminal B and non-luminal PAM50 subgroups, as well as intermediate- and high-proliferation ER+ breast cancers. For luminal A or low-proliferation breast cancers, "immune activation" signatures were not significantly associated with response, though very few patients achieved pCR in these two subgroups.

Conclusions: Gene expression signatures associated with "immune activation" identify a subset of ER+ breast cancers with higher rates of pCR to neoadjuvant chemotherapy. These "immune activation" signatures appear to be proliferation-independent and may provide additional predictive information to existing gene expression-based approaches for ER+ breast cancer.

Citation Format: Stover DG, Waks AG, Erica ML, Brugge JS, Winer EP, Selfors LM. Immune activation signatures identify a subset of ER+ breast cancers with increased pathologic complete response to neoadjuvant chemotherapy. [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 P3-07-31.
Abstract P3-07-47: Comprehensive profiling of metaplastic breast carcinoma reveals frequent over-expression of PD-L1
Authors: Gatalica Z , Joneja U , Ghazalpour A , Swensen J , Feldman R , Cai F , Chen W , Xiao N , Reddy S , Palazzo J
Published: May 2016

Background: Metaplastic breast carcinoma (MBC) is a rare subtype of breast carcinoma less responsive to conventional chemotherapy relative to usual breast carcinomas (UBCs) such as ductal and lobular subtype. In molecular terms MBC usually clusters with triple negative breast cancer (TNBC), but MBCs portray a worse prognosis in comparison to TNBCs. Published studies investigating MBCs for specific biomarkers of therapy response are rare and limited by the methodological approaches.

Methods: This study included 132 patients with 38 histologically confirmed MBCs and 94 UBCs. Amongst the 94 UBCSs, 44 were estrogen receptor positive, 33 were triple negative and 17 were HER2 positive. Direct sequencing analysis was performed on genomic DNA isolated from formalin-fixed paraffin-embedded tissue (FFPE) using the Illumina MiSeq Next Generation Sequencing platform (NGS). Immunohistochemistry for PD-L1 (SP142, Spring Bioscience), PD-1 (NAT105, Ventana) and EGFR (31G7, Life Technologies) was performed using automated procedures. Additionally, mutation analysis for EGFRvIII was performed on RNA extracted from FFPE tissue.

Results: At the genomic level, numerous cases of MBC had multiple genomic alterations with the most frequent genetic mutation in TP53 gene (14/24), similar to the TNBC controls (17/33). BRCA1 mutations were detected in 2/10 cases. Potentially actionable mutations were rare and included PIK3CA gene. Importantly, PD-L1 expression on cancer cells was detected in significantly higher proportion of MBCs (37%) than in the UBC cohort (6%) or TNBC control (14%) (p=3.7x10-5 and p=0.03, respectively). PD-1 positive tumors infiltrating lymphocytes (TILS) varied greatly in MBC (0 to >50/mm2). Over-expression of EGFR was frequent in MBCs (62%); however no mutations in the gene including EGFRvIII were detected.

Conclusion: Comprehensive profiling of a large cohort of this rare carcinoma highlighted predominance of TP53 mutations, wild type EGFR gene expression, a distinct increase in proportion of PD-L1 expression in carcinoma cells, and PD-1 expression in TILS. The latter properties can be exploited in clinical trials utilizing immune check point inhibitors.

Citation Format: Gatalica Z, Joneja U, Ghazalpour A, Swensen J, Feldman R, Cai F, Chen W, Xiao N, Reddy S, Palazzo J. Comprehensive profiling of metaplastic breast carcinoma reveals frequent over-expression of PD-L1. [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 P3-07-47.
Abstract P3-07-48: Prediction of complete pathologic response to veliparib/carboplatin plus standard neoadjuvant therapy in HER2 negative breast cancer: Exploratory protein pathway marker results from the I-SPY 2 trial
Authors: Wulfkuhle JD , Yau C , Wolf DM , Gallagher RI , Deng J , Brown-Swigart L , Hirst G , I-SPY 2 Trial Investigators , Rugo H , Olopade OI , Esserman L , Berry D , van't Veer L , Petricoin EF
Published: May 2016

Background: In the I-SPY 2 TRIAL, HER2- patients were randomized to receive standard chemotherapy or chemotherapy plus the oral PARP inhibitor veliparib in combination with carboplatin (V+C), which graduated in the HR-/HER2- arm. Exploratory analysis of protein signaling was performed to identify biomarker candidates that correlated with pCR in the HER2- population. We evaluated 110 key signaling proteins using reverse phase protein microarray (RPPA) data from pre-treatment LCM purified tumor epithelium.

Methods: Of 115 patients, 97 (V+C: 61 controls: 36) had RPPA and pCR data. RPPA data was correlated to pCR in both the treated and control patients using parametric (t-test) or non-parametric (Wilcoxon) statistical analysis, depending on data distribution. Only analytes whose pre-treatment levels were associated with response in the V+C but not the control arm were identified (P<0.05). Markers are analyzed individually; p-values are descriptive and were not corrected for multiple comparisons.

Results: 11 protein/phosphoprotein markers were significantly associated with pCR in the V+C arm but not in controls. Two were positive predictors of response: YAP S127 p= 0.03 and LC3B p=0.04. Negative predictors of response included Cyclin D1 p=0.001, and a number of phosphorylated RTKs: ROS Y2274 p=0.03, IGF1R Y1135/Y1136-IR Y1150/Y1151 p=0.03, ERBB4 Y1284 p=0.002, total HER2 p=0.04, and total IGF1R p=0.01. Moreover, a number of AKT-mTOR pathway proteins were found to be negative predictors of V+C response: ACC S79 p=0.005, p70S6K S371 p=0.01, and B-RAF S445 p=0.01.

Conclusion: Our sample size is too small to draw definitive conclusions and the results are exploratory. Coordinated RTK-mTOR pathway activation appears to be a hallmark signature of lack of response to veliparib in HER2- tumors. We also found that HER2 levels were correlated paradoxically with lack of response in this HER2- population, suggesting potential added clinical value of quantitative HER2 measurement techniques. Such exploratory results merit evaluation in larger trials with HER2- breast cancer patients.

Citation Format: Wulfkuhle JD, Yau C, Wolf DM, Gallagher RI, Deng J, Brown-Swigart L, Hirst G, I-SPY 2 Trial Investigators, Rugo H, Olopade OI, Esserman L, Berry D, van't Veer L, Petricoin EF. Prediction of complete pathologic response to veliparib/carboplatin plus standard neoadjuvant therapy in HER2 negative breast cancer: Exploratory protein pathway marker results from the I-SPY 2 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 P3-07-48.
Abstract P3-07-63: Ki67 cut-off point to predict the benefit of adjuvant chemotherapy in ER+ HER2- breast cancer patients
Authors: Rouzier R , Rossi L , Lerebours F , Savignoni A , Falcou M-C , Bonneau C , Pierga J-Y , Guinebretière J-M
Published: May 2016

BACKGROUND: The benefit of chemotherapy for patients with estrogen receptor(ER)+ HER2- breast cancers is an ongoing question. The evaluation of the tumor's proliferation by Ki67 may guide the indication but no consensual predictive cut-off has been set yet.

MATERIALS AND METHODS: This study included women with a first ER+HER2- invasive breast cancer treated by primary surgery between 2003 and 2008. Data was collected prospectively. Ki67 cut-off was sequentially set each 1% from 5% to 30%. For each threshold, the interaction between Ki67 and adjuvant chemotherapy was integrated in a multivariate Cox model to determine when it became statistically significant in predicting distant-disease free survival (DDFS). Using different Ki67 cut-offs, we also compared DDFS of patients who had or not an indication of chemotherapy, depending on whether they actually received it or not.

RESULTS: Among the 3221 breast cancers, median Ki67 was 10%, with a mean of 15% (S.D = 14). Ki67 was an independent prognosis factor whether the cut-off was set to 14% or to 20% (p<0.001). The interaction between Ki67 and chemotherapy became significant only for a threshold of Ki67 above 20% (p<0.05). When adjuvant chemotherapy was not indicated according to St Gallen guidelines 2013 with a Ki67 cut-off set to 20%, DDFS was not significantly improved if patients did receive chemotherapy.

CONCLUSION: In ER+ HER2- breast cancers, a Ki67 level of expression >= 20% was predictive of benefit from adjuvant chemotherapy. A threshold at 14% was not as discriminant. Based on this large cohort study, we recommend the use of a cut-off at 20% to decide whether patientes with ER positive tumors should receive chemotherapy or not/.

Citation Format: Rouzier R, Rossi L, Lerebours F, Savignoni A, Falcou M-C, Bonneau C, Pierga J-Y, Guinebretière J-M. Ki67 cut-off point to predict the benefit of adjuvant chemotherapy in ER+ HER2- breast cancer patients. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P3-07-63.
Abstract P3-12-20: The impact of everolimus on radiation-induced pulmonary fibrosis in wistar albino rats: Results of an experimental study
Authors: Eren MF , Ay Eren A , Yücel B , Elagöz S , Özgüven Y , Altun A , Kiliçkap S , Matsuno R , Bese N
Published: May 2016

Objectives:

This study was performed to evaluate the effects of everolimus on pulmonary fibrosis, given concurrently or sequential with irradiation in rats.

Materials and Methods:

Forty female Wistar albino rats were randomized into five groups. The first group (Group 1: sham) was observed with identical conditions of the animals in other groups. The second group (Group 2) had only everolimus application. The third group (Group 3) had thoracic irradiation first and they had everolimus injection 22 hours after radiotherapy. The fourth group (Group 4) had irradiation to whole thoracic region in 2 hours of following everolimus administration. The last group (Group 5) had irradiation to whole thoracic region. As an end point the percentage of lung with fibrosis for each rat was quantified with image analysis of histological sections of the lung. Groups were compared using the linear regression method, Fisher's exact and chi-square tests were used to assess the association between treatment groups.

Results:

There was a statistically significant difference in the distribution of fibrosis scores among all groups (p=0.0022). Linear regression analyses revealed that all treatment groups showed an increased risk of lung fibrosis compared to control group. Group 2 (p=0.0020), group 3 (p<0.0001), group 4 (p<0.0001), and group 5 (p=0.0002). Chi-square test demonstrated that there was no statistically significant difference between the sequentially and concurrent treatment groups.

Conclusion:

This experimental study showed that of everolimus by itself may lead pulmonary fibrosis and addition to radiotherapy either sequentially or concomitantly increased the severity of the lung damage in rats.

Keywords: Radiotherapy, everolimus, rats, pulmonary fibrosis.

Citation Format: Eren MF, Ay Eren A, Yücel B, Elagöz S, Özgüven Y, Altun A, Kiliçkap S, Matsuno R, Bese N. The impact of everolimus on radiation-induced pulmonary fibrosis in wistar albino rats: Results of an experimental 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 P3-12-20.
Abstract P3-14-12: Phase 0 study evaluating COX2 inhibition on circulating PGE2 levels from obese subjects
Authors: Lengfelder L , Brenner A , Bowers L , Apte S , Galván G , Kist K , deGraffenried L
Published: May 2016

Introduction: Obesity is associated with poor breast cancer outcomes in postmenopausal women in response to aromatase inhibitor therapy. Our prior studies have shown an association between reduced recurrence rate and use of cyclooxygenase-2 (COX-2) inhibiting non-steroidal anti-inflammatory drugs (NSAIDs) in obese breast cancer patients. The mechanism proposed was a decrease in prostaglandin E2 (PGE2) and reduced activation of the aromatase promote locally in the breast.

Methods: Postmenopausal women of varying body habitus were recruited at the CTRC in San Antonio and underwent randomized assignment to 1 of 3 arms: ASA 81mg daily, 1500mg of docosahexaenoic acid (DHA) and 2500mg eicosapentaenoic acid (EPA) given daily, or combined ASA and DHA/EPA. Sera were collected prior to and following 28 days of exposure, and cytokines including prostaglandin E2 were assessed via enzyme-linked immunosorbant assay (ELISA). Conditioned media was generated by exposing macrophages to patient sera in order to see if the patient sera induced PGE2 concentration in vitro.

Results: Thirty of the planned 120 subjects have completed assessment. No toxicity has been noted. In 71% of the patients, serum PGE2 levels decreased, but only 60% demonstrated concurrent decrease in serum PGE2 levels as well as macrophage PGE2 production, while almost all (88%) of the patients whose serum did not demonstrate a decrease in PGE2 levels also demonstrated no decrease in induced levels.

Conclusion: NSAIDs appear to effectively decrease circulating levels of PGE2 in most obese women. However, one third of the subjects did not demonstrate concurrent suppression of induced PGE2 from macrophages. These data suggest that circulating levels of PGE2 may not be reflective of local tumor microenvironment levels, and other pro-inflammatory circulating factors may be responsible for regulating local inflammatory responses. Final analysis will be completed and presented at the SABCS meeting.

Citation Format: Lengfelder L, Brenner A, Bowers L, Apte S, Galván G, Kist K, deGraffenried L. Phase 0 study evaluating COX2 inhibition on circulating PGE2 levels from obese subjects. [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 P3-14-12.
Abstract P4-01-03: Digital measurement of mammographic density and the risk of overlooking cancer in Japanese women
Authors: Sawada T , Ikeda M , Kuwayama T , Akashi ST , Nakamura S
Published: May 2016

Introduction: mammographic density is a strong risk factor for breast cancer and an essential determinant of screening sensitivity, but the breast density of Japanese women has yet to be objectively examined. Single mammographic examination fails to detect approximately half of breast cancers in women with dense breasts. We objectively evaluated mammographic density to clarify the relation between breast density and incidence of breast cancer in Japanese women.

Method: We enrolled 269 patients diagnosed with breast cancer and 397 healthy controls, who participated in screening mammography at Showa University Hospital. Breast density was measured using Volpara (a software application which analyses breast composition volumetrically).

Results:

The average age of both control and breast cancer groups was 55. Of the control group, 308 (78%) were characterized as having dense breasts, being defined as volumetric density grade (VDG) 3 or 4. More than 50% of the total population (all ages) had dense breasts (VDG 3 or 4) and more than 20% were extremely dense (VDG:4). Of the breast cancer patients, 232 (87%) were VDG 3 or 4. 52 (23%) of those with dense breasts showed negative results in single mammography examination. In primary breast cancer patients with dense breasts, 30% of those under age 70, but only 13% of those over age 70, showed no abnormal findings from mammographic examination.

Conclusion and Discussion: In comparison of our present results and other papers, In the Japanese control group, 72% of patients had dense breasts (VDG 3 or 4) at age 50 or greater, but in Holland, for example, only 36% of women had breasts classed as dense. 87% of Japanese women with breast cancer had dense breasts, approximately the same ratio (88%) as found in Korea. For Japanese and other Asians, women under 70 years old with dense breasts may warrant additional screening.

Citation Format: Sawada T, Ikeda M, Kuwayama T, Akashi ST, Nakamura S. Digital measurement of mammographic density and the risk of overlooking cancer in Japanese women. [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 P4-01-03.
Abstract P4-02-13: The pattern of tumor shrinkage is associated with prognosis in low grade luminal early breast cancer during neoadjuvant chemotherapy
Authors: Fukada I , Araki K , Kobayashi K , Gomi N , Horii R , Akiyama F , Takahashi S , Iiwase T , Ohno S , Ito Y
Published: May 2016

BACKGROUND: In neoadjuvant chemotherapy (NAC) for early breast cancer, the pathological response rate in estrogen receptor (ER)-positive tumors has been low in comparison with those of ER-negative tumors. Therefore, surrogate makers other than the pCR rate are needed during NAC for luminal breast cancer. Using MRI, we analyzed the patterns of tumor shrinkage after NAC as a surrogate prognostic factor in low grade luminal breast cancer. METHODS: Of 854 patients who had received NAC in a single institute from Jan. 2000 to Dec. 2009, 183 patients with low grade luminal breast cancer were retrospectively evaluated for this study. They were defined as ER and/or PgR positive in more than 10% of cancer cells and HER2 negative (IHC 0, 1+ or FISH <2.0) with nuclear grade 1 and 2. RESULTS: The median observation period was 67.9 months following surgery, and recurrence was observed in 31 patients (16.9%). The median age was 49 (22-76) years. One hundred eighty patients received anthracycline-containing chemotherapy, and 158 received taxane. There were 16 deaths (8.7%) related to breast cancer. We categorized the patterns of tumor shrinkage by MRI into 6 types: concentric shrinkage (CS), diffuse decrease (DD), reduction to small foci (RSF), decrease of intensity only (DIO), no change (NC), and enlargement (EL). According to our categorization, CS occurred in 97 (53.0%), RSF in 7 (3.8%), DD in 62 (33.9%), DIO in 7 (3.8%), NC in 5 (2.7%), and EL in 5 (2.7%). As expected, there were statistically significant differences in both the median DFS and OS in each pattern of tumor shrinkage (p <0.001 and p=0.001, respectively); in particular, the CS pattern had excellent prognosis. Multivariate analysis demonstrated that concentric shrinkage was the only significant good prognostic factor for OS (p=0.015). CONCLUSIONS: Tumor shrinkage patterns as revealed by MRI could be important surrogate prognostic factors for NAC in early low grade luminal breast cancer.

Citation Format: Fukada I, Araki K, Kobayashi K, Gomi N, Horii R, Akiyama F, Takahashi S, Iiwase T, Ohno S, Ito Y. The pattern of tumor shrinkage is associated with prognosis in low grade luminal early breast cancer during neoadjuvant chemotherapy. [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 P4-02-13.
Abstract P4-03-06: Near-infrared diffuse optical imaging for early prediction to neoadjuvant chemotherapy in patients with primary breast cancer
Authors: Ogura H , Yoshizawa N , Ueda S , Hosokawa Y , Matsunuma R , Tochikubo J , Nasu H , Shigekawa T , Takeuchi H , Osaki A , Saeki T , Yoshimoto K , Ohmae E , Suzuki T , Ueda Y , Yamashita Y , Sakahara H
Published: May 2016

Background: Diffuse optical spectroscopic imaging (DOSI) can be exploited as a marker of tumor blood volume quantified by tissue hemoglobin (tHb) concentration. In DOSI, frequent measurement is possible for breast cancer patients because of its non-invasiveness. The tHb concentration determined by DOSI is expected to be a new biomarker for prediction of breast cancer response to neoadjuvant chemotherapy (NAC).

Purpose: Our objective is to determine whether early change of tumor tHb concentration predicts pathological complete response (pCR) to NAC in patients with operable breast cancer.

Methods: In a prospective study, one hundred patients with primary breast cancer were enrolled for primary objective analysis. The regimens of NAC were according to the standard of care. Patients underwent sequential scans using DOSI at baseline, after 1st course and 2nd course of chemotherapy. The mean value of tHb (tHbmean) concentration of the targeted lesion was measured and the percentage change in tHbmean (ΔtHbmean) concentration was calculated. Receiver operating curve analysis demonstrated diagnostic performance of DOSI for predicting a pCR.

Results: In interim analysis, it was regarded as a good outcome that area under the curve (AUC) for ΔtHbmean after 1nd course was 0.797 (SE 0.104, 95%CI 0.633-0.911), and after 2st course was 0.867 (SE 0.06, 95%CI 0.715-0.956).

Conclusion: DOSI could predict accurately a pCR to neoadjuvant chemotherapy in patients with primary breast cancer.

Citation Format: Ogura H, Yoshizawa N, Ueda S, Hosokawa Y, Matsunuma R, Tochikubo J, Nasu H, Shigekawa T, Takeuchi H, Osaki A, Saeki T, Yoshimoto K, Ohmae E, Suzuki T, Ueda Y, Yamashita Y, Sakahara H. Near-infrared diffuse optical imaging for early prediction to neoadjuvant chemotherapy in patients with primary 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 P4-03-06.
Abstract P4-04-02: Amplification of chromosome 9p24 targeting PD-L1 and JAK2 correlates with altered tumor microenvironment expression profiles in triple negative breast cancer
Authors: Anderson KS , Andreozzi M , Pockaj BA , Hopper M , McCullough AE , Barrett MT
Published: May 2016

Introduction: In a pilot study, we recently described amplification of chromosome 9p24 encoding PD-L1, PD-L2, and JAK2 (the PDJ amplicon) in 29% of early stage triple negative breast cancers (TNBC). Amplification was associated with poor DFS and OS. The impact of this amplicon on immune function is not known.

Methods: Fresh frozen tumor samples from 36 subjects with newly-diagnosed TNBC (stages I-III) were evaluated for copy number aberrations using DNA-based flow cytometry to enrich nuclei based on ploidy status (tetraploidy or aneuploidy), followed by oligonucleotide CGH arrays. Targeted immune expression profiling was performed using the Nanostring PanCancer Immune Profiling Panel (n=770 genes).

Results: 23 samples had measurable abnormal ploidy (evidence of sufficient tumor content) for CGH analysis; the remaining samples were omitted from analysis. High level (log2ratio >1) amplification was detected in 6/23 (26.1%) of TNBCs, and low-level copy number gain (CN log2ratio >0 and<1) was detected in 3/23 (13.0%). The remaining samples had no copy number change (n=9) or low level copy number loss (CN log2ratio>-1 and <0, n=5). Increased RNA expression of STAT1, STAT2, STAT3, and STAT6 was associated with PDJ amplification, as well as upregulation of the immune regulatory genes CD47 and IDO1.

Conclusion: Amplification of chromosome 9p24.1 involving PD-L1, PD-L2, and JAK2 is present in a significant proportion of TNBCs. This amplicon is associated with altered gene expression of the tumor microenvironment.

Citation Format: Anderson KS, Andreozzi M, Pockaj BA, Hopper M, McCullough AE, Barrett MT. Amplification of chromosome 9p24 targeting PD-L1 and JAK2 correlates with altered tumor microenvironment expression profiles in triple negative breast cancer. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P4-04-02.
Abstract P4-04-13: Comparison of tumor and stroma CD73 expression with TLR9 and survival in breast cancer
Authors: Selander K , Mella M , Kauppila J , Karihtala P , Jukkola-Vuorinen A , Auvinen P , Soini Y , Kauppila S , Haapasaari K-M , Harris K , Vuopala K
Published: May 2016

CD73 is a 5' ectonuclease that catalyzes the conversion of cyclic AMP into the highly immunosuppressive adenosine in extracellular space. In addition to the cells of the immune system, CD73 is highly expressed in various cancer cell lines and clinical cancer tissues. Toll like receptor-9 (TLR9) is a cellular DNA-receptor that is highly expressed in breast cancer. Both CD73 and TLR9 expression have recently been associated with TNBC prognosis but the mechanisms how these proteins possibly contribute to TNBC pathophysiology remains poorly understood. TLR9 and CD73 expression has been shown to be mutually regulated in various cell types. Whether this is the case in cancer is unknown. The aim of this study was to investigate the mutual role of TLR9 and CD73 in breast cancer (BC). Specifically our hypothesis was that TLR9 and CD73 expression correlate in TNBC. We compared immunohistological tumor TLR9 and CD73 expression scores using a previously characterized breast cancer (BC) cohort (n=184) with follow-up time of > 10 years. We did not discover a connection between TLR9 and CD73 expression in tumor cells in BC. There was a trend for increased survival among patients that had high tumor cell CD73 expression, as compared with the lower tumor cell CD73 expression groups. There was a trend for a better survival among TNBC patients that had lower stromal CD73 expression, as compared with those TNBC patients that had higher stromal CD73 expression. No such difference was detected among patients with non-TNBC tumors. Our results suggest that stromal vs. tumor cell CD73 expression have opposite effects on survival in TNBC, but there is no connection between CD73 and TLR9 expression. Our conclusions are limited by low sample numbers.

Citation Format: Selander K, Mella M, Kauppila J, Karihtala P, Jukkola-Vuorinen A, Auvinen P, Soini Y, Kauppila S, Haapasaari K-M, Harris K, Vuopala K. Comparison of tumor and stroma CD73 expression with TLR9 and survival in 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 P4-04-13.
Abstract P4-06-01: Human mammary tumor virus, HMTV, is an MMTV-adapted human pathogen
Authors: Holland JF , Jaffer S , Melana S , Nartey T , Gnjatic S , Pogo BG-T
Published: May 2016

Forty percent of American women's breast cancers contain HMTV, a betaretrovirus 90-98% homologous to MMTV, the causative agent for breast cancer in mice. HMTV is reverse-transcribed and integrated into chromosomal DNA of human breast cells. MMTV contamination of our analyses has been definitively excluded. In 8% of unselected American mothers HMTV is found in cells in milk, but in 21% of milks from women previously biopsied for unconfirmed cancer suspicion. In breasts biopsied for suspicion of cancer before a later diagnosis of breast cancer, we see suggestive morphologic changes if the subsequent breast cancer is HMTV+. HMTV is infectious in vitro for B and T lymphocytes, dendritic cells, and human mammary epithelial cells. Infecting MCF-10 breast cell lines with HMTV produces molecular changes of epithelial-mesenchymal transition with upregulation of vimentin ,and downregualtion of E-cadherin. The excess of breast cancer in western European countries and their former colonies (age standardized rates of 47 to 92 per 100,000 per year) compared to Asian incidence (29 to 43 ASR/y) can be explained by excess HMTV-related breast cancer incidence. In 7 West European, American and Oceania countries, 30 to 60% contain HMTV, while in 5 Asian nations HMTV in breast cancers ranges from 0 to 22% Different indigenous murine species with disparate MMTV burdens parallel these findings: mus domesticus in the West with much MMTV in its genome, and m. castaneus or m.musculus in the East with less. Ancient contamination of the food chain, and current lactational transfer perpetuate infection. HMTV poses a new and challenging dimension to breast cancer research involving diagnosis, molecular mechanisms, epidemiology, therapy and prevention.

Citation Format: Holland JF, Jaffer S, Melana S, Nartey T, Gnjatic S, Pogo BG-T. Human mammary tumor virus, HMTV, is an MMTV-adapted human pathogen. [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 P4-06-01.
Abstract P4-07-06: Imipramine Blue - A safe and potent therapeutic regimen that suppresses breast cancer growth and progression by targeting DNA damage surveillance pathway
Authors: Rajamanickam S , Subbarayalu P , Timilsina S , Gorthi A , Drake MT , Chen Y , Vadlamudi R , Bishop AJR , Arbiser JL , Rao MK
Published: May 2016

Despite improvement in overall survival of breast cancer patients, many women don't survive this disease. Moreover, the quality of life for patients who do survive is often substantially reduced due to the toxicity associated with the chemotherapy. Here, we report that imipramine blue (IB), a novel analogue of anti-depressant imipramine that we recently synthesized, may serve as a safe and potent therapeutic agent for treating breast cancers. We show that IB reduced cell growth, migration and invasion of breast cancer cells. Systemic delivery of IB using nanoparticle-based drug delivery approach suppressed breast cancer growth and metastasis without inducing any toxicity in pre-clinical orthotropic mouse models. Notably, using ex-vivo model of tumor explants from breast cancer patients, we demonstrated that IB inhibited breast cancer growth without affecting normal mammary epithelial cell proliferation. Furthermore, IB improved the sensitivity of breast cancer cells to chemotherapy drugs paclitaxel and doxorubicin. Our results revealed that IB mediated its anti-tumor effect by targeting genes involved in cell cycle progression, microtubule dynamics and DNA damage surveillance pathway including Forkhead Box M1 (FOXM1), stathmin1, S-phase kinase-associated protein 2 (Skp2) and XRCC3, which we show to be highly expressed in breast cancer patients. Importantly, we demonstrated that IB inhibited breast cancer cell's ability to repair DNA strand breaks by impairing homologous recombination events. These findings highlight the potential of IB to be used as a potent therapeutic regimen for treating breast cancer patients. Since IB-1 is derived from a FDA approved drug it has potential to be rapidly translated to the clinic.

Citation Format: Rajamanickam S, Subbarayalu P, Timilsina S, Gorthi A, Drake MT, Chen Y, Vadlamudi R, Bishop AJR, Arbiser JL, Rao MK. Imipramine Blue - A safe and potent therapeutic regimen that suppresses breast cancer growth and progression by targeting DNA damage surveillance pathway. [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 P4-07-06.
Abstract P4-08-03: DEAD-box RNA helicase DP103 as a novel regulator of Wnt/β-catenin signaling pathway and promotes cancer stem cell-like behavior in triple negative breast cancers
Authors: Cai W , Cheong JK , Edison E , Banerjee A , Tan TZ , Gaboury L , Yousef EM , Thiery JP , Lobie PE , Virshup DM , Yap CT , Kumar AP
Published: May 2016

Despite recent advances in breast cancer therapeutics, mortality of metastatic triple negative breast cancer (TNBC) subtype remains high; due to their lack of hormone receptors expression for targeted therapy. Aberrant activation of Wnt/β-catenin signaling has been associated with breast cancers; where 40% of total breast cancers have elevated β-catenin levels with increased Wnt activity. Recently, we identified DEAD-box RNA helicase DP103 as a novel prognostic biomarker and metastasis-driving oncogene; highly expressed in TNBC subtype. Interestingly, we found high DP103 expression to be positively correlated with high β-catenin expression in clinical specimens (n=400). This led us to hypothesize a possible role of DP103 in modulating the Wnt/β-catenin pathway in TNBCs. Depletion of DP103 in metastatic TNBC cells decreases Wnt/β-catenin activity and expression of downstream Wnt target genes, while overexpression of DP103 increases Wnt activity. Depletion of DP103 also decreases phosphorylation of LRP6 and several important Wnt modulators required for downstream Wnt activation. Moreover, induction of Wnt/β-catenin signaling in Wnt responsive TNBC cells also significantly increased DP103 expression, indicating a possible positive feedback loop. Both canonical and non-canonical Wnt signaling is known to independently promote stem cell growth in mammospheres. Herein, we will also provide evidence on the role of DP103 in promoting breast cancer stem cell-like properties. Collectively, our data show a novel regulatory role of DP103 in the Wnt/β-catenin signaling pathway and in promoting breast cancer stem cell-like behavior, presenting itself as a potential drug target in TNBC patients.

Citation Format: Cai W, Cheong JK, Edison E, Banerjee A, Tan TZ, Gaboury L, Yousef EM, Thiery JP, Lobie PE, Virshup DM, Yap CT, Kumar AP. DEAD-box RNA helicase DP103 as a novel regulator of Wnt/β-catenin signaling pathway and promotes cancer stem cell-like behavior in triple negative breast cancers. [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 P4-08-03.
Abstract P4-09-06: The prognostic role of RANK single nucleotide polymorphisms in breast cancer patients with bone metastases
Authors: Casimiro S , Ferreira A , Alho I , Melo M , Brás R , Costa AL , Sousa AR , Mansinho A , Abreu C , Pulido C , Macedo D , Pacheco TR , Costa L
Published: May 2016

Background: Receptor activator of NF-kB (RANK) pathway regulates bone remodeling and is involved in breast cancer (BC) progression. RANK over-expression in primary BC associates with poor prognosis and metastasis development. In patients with BC, RANK and RANK-ligand (RANKL) single nucleotide polymorphisms (SNPs) have also been associated with BC risk and bone metastasis (BM)-free survival , respectively. Here we analyze the association of five RANK missense SNPs with prognosis of patients with BM from BC.

Methods: Missense RANK SNPs (rs34945627, rs12721431, rs35184120, rs35993683, rs61751992) were genotyped in germline DNA from a retrospective cohort of 76 patients with BM from BC, under bisphosphonates; and a cohort of 80 healthy volunteers (samples from Biobanco-IMM, Lisbon, Portugal). Genotypic allelic frequencies were assessed using TaqMan assays (Applied Biosystems). SNP rs34945627 was analyzed with regard to clinicopathological characteristics, skeletal-related events (SRE), bone progression-free interval (BPFi), and overall survival (OS). Univariate association with clinicopathological characteristics was performed using Fisher's exact test and Wilcoxon rank-sum test; univariate differences between survival rates were tested for significance using the log-rank test, while multivariate analysis for survival was tested using Cox proportional hazards models.

Results: SNP rs34945627 had a minor allele frequency of 11.84% (n=9) in BC patients versus 1.25% (n=1) in healthy individuals, whereas the remaining SNPs had a minor allele frequency of 2.60% (n=2) in BC patients. Therefore, for inferential analysis only SNP rs34945627 was considered. No differences between alleles were observed regarding clinicopathological characteristics, including treatment. Homozygous patients (CC) had increased OS when compared to heterozygous patients (CT) controlling for age at diagnosis, visceral involvement, radiographic pattern of BM and baseline urine NTX (adjusted HR 5.76, 95% CI 2.10-15.81; p<0.001). No association could be seen with regard to SREs (adjusted HR 1.43, 95% CI 0.46-4.46; p=0.54) or BPFi (adjusted HR 0.26, 95% CI 0.06-1.17; p=0.08).

Conclusions: RANK SNP rs34945627 seems to be a marker of poor prognosis in patients with BC and BM. Further studies are required to characterize the biological and clinical significance of this finding.

Citation Format: Casimiro S, Ferreira A, Alho I, Melo M, Brás R, Costa AL, Sousa AR, Mansinho A, Abreu C, Pulido C, Macedo D, Pacheco TR, Costa L. The prognostic role of RANK single nucleotide polymorphisms in breast cancer patients with bone metastases. [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 P4-09-06.
Abstract P4-09-11: microRNA markers and risk of recurrence in women with ductal carcinoma in situ
Authors: Liu Y , Baker C , Wang X , Colditz G
Published: May 2016

Background: Women with DCIS have increased risk for invasive breast cancer. Limited abilities to accurately identify aggressive DCIS have resulted in inappropriate treatment for many women with mammographically-detected DCIS. miRNAs play a critical role in tumorigenesis. It remains unknown if miRNAs are associated with recurrence and refine risk assessment for DCIS patients.

Methods: The study included 16 DCIS cases that developed ipsilateral recurrence (cases) and 32 DCIS cases that did not develop any type of recurrence (controls). Cases and controls were treated with breast conserving surgery and matched on age, race, and year of diagnosis. Expression of 96 cancer-related miRNAs was quantified using RT-PCR.

Results: Two miRNAs were differentially expressed between cases and controls (p<0.05). A combination of the two miRNA markers was significantly associated with recurrence risk (odd ratio=2.60 per 1-unit increase, 95% confidence interval=1.14-8.13), which was independent of family history of breast cancer, radiation and endocrine therapy. The miRNA-based score (c-statistic=0.73) performed better than individual miRNAs (c-statistic=0.64-0.69) in discriminating DCIS cases who did and did not develop recurrence.

Conclusion: miRNA expression in DCIS was associated with recurrence risk, which warrants further investigations to identify new miRNA markers by including more DCIS cases and analyzing all breast cancer-related miRNAs.

Citation Format: Liu Y, Baker C, Wang X, Colditz G. microRNA markers and risk of recurrence in women with ductal carcinoma in situ. [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 P4-09-11.
Abstract P4-09-15: BRCAness and PD-L1 expression of basal-like and not basal-like triple negative breast cancer
Authors: Mori H , Kubo M , Yamada M , Kai M , Osako T , Nishimura R , Arima N , Okido M , Kuroki S , Oda Y , Nakamura M
Published: May 2016

Background: Triple Negative Breast Cancer (TNBC) subtype occurs in approximately 20% of all patients with breast cancer and is associated with rapid growth, early metastasis and poor prognosis compared with other subtypes. TNBCs are a heterogeneous disease entity and further subclassification is needed, but still ongoing. In this study, we assessed BRCAness, defined as shared characteristics between sporadic and BRCA1-mutated tumors, in a cohort of basal-like and non-basal-like TNBCs.

Patients and Methods: DNA was isolated from formalin-fixed paraffin-embedded tumor tissues and BRCAness status was analyzed in 262 patients with primary TNBCs resected at our three hospitals between 2004 and 2014. Classification of BRCAness was performed by using Multiple Ligation-dependent Probe Amplification (MLPA) with the probemix P376 BRCA1ness by MRC (Amsterdam, Holland). The tumor subtypes were routinely determined immunohistochemically by using resected specimens. Basal-like phenotype was defined as being positive for Epidermal Growth Factor Receptor (EGFR) and/or Cytokeratin 5/6 (CK5/6). Moreover, TNBCs were stained and analyzed for programmed cell death ligand-1 (PD-L1) expression as a target of new immune therapies.

Results: Of 262 TNBCs, 232 tumors (88.5%) was a basal-like phenotype. The results of MLPA assay showed that 159 (68.5%) of 232 tumors had a BRCAness profile. Patients with basal-like BRCAness tumors were younger than patients with basal-like non-BRCAness tumors (p<0.0001). There was no significant difference between the two groups regarding pathological stage. The basal-like BRCAness group had shorter relapse-free survival (RFS) and overall survival (OS) than the basal-like non-BRCAness group (p=0.028 and p=0.13, respectively), and anthracycline-based regimens provided greater benefit to the basal-like BRCAness group significantly (p=0.01 in RFS and p=0.007 in OS). PD-L1 was expressed in 71 (44.7%) of 159 basal-like TNBCs with BRCAness.

Conclusion: We reported the majority of basal-like TNBCs showed a BRCAness profile and PD-L1 expressed in approximately 50% of BRCAness tumors. It is known that about 30% of BRCAness tumors are BRCA1-mutated tumors. Those biomarkers are essential for subclassification of TNBCs and may offer not only platinum-based chemotherapy but also novel therapies, such as immune-targeted therapies of PD-1/PD-L1 inhibitors and PARP inhibitors, to patients with basal-like TNBCs with BRCAness.

Citation Format: Mori H, Kubo M, Yamada M, Kai M, Osako T, Nishimura R, Arima N, Okido M, Kuroki S, Oda Y, Nakamura M. BRCAness and PD-L1 expression of basal-like and not basal-like triple negative breast cancer. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P4-09-15.
Abstract P4-09-27: ABC transporter expression: Clues into chemoresistance of triple negative breast cancers
Authors: Feldman R , Abbott B , Reddy S , Gatalica Z , Castro M
Published: May 2016

Background: Triple-negative breast cancer (TNBC) is an aggressive histological subtype with high rates of recurrence and metastatic disease. Intrinsic or acquired multidrug resistance facilitated by the over-expression of drug efflux pumps (ABC transporters: BCRP [ABCG2], MRP1 [ABCC1] and PGP [ABCB1]), may contribute to the aggressive nature of this disease. We examined the expression patterns of drug efflux pumps for insight on their potential role in chemoresistance of TNBC.

Methods: 1393 TNBC patients molecularly profiled with a commercial assay (Caris Life Sciences) were evaluated retrospectively for expression of BCRP, MRP1 and PGP by immunohistochemistry. Antibodies used: PGP (C494), BCRP (6D171) and MRP1 (33A6). IHC threshold: positive = ≥1+ and ≥10%). This data set also included metachronous paired samples from 71 TNBC patients. JMP was used to ascertain distributional differences.

Results: PGP and MRP1 positive expression rates were 8.4% (117/1393) compared to 78.2% (248/317), respectively, displaying an inverse association (p=0.0001). BCRP was over-expressed in 52% (90/173). Co-expression data for all three transporters was available for 153 patients. BCRP/MRP1 co-expression was most abundant, 39% (59/153), followed by MRP1/PGP at 4% (6/153) and BCRP/PGP at 2% (3/153). Furthermore, 12% (18/153) of TNBC exhibited positive expression for all three drug pumps (16/18 or 89% were from patients with metastatic disease) and 14% (22/153) exhibited negative expression for all three drug pumps (15/22 or 68% were from patients with metastatic disease). Interestingly, positive MRP1 status, but not BCRP or PGP, correlated with metastatic disease (p=0.0061). In the paired data analysis, PGP expression was absent in 77% (55/71) and retained lack of expression in subsequently profiled specimens, 3% (2/71) retained positive PGP status, 13% (9/71) lost expression of PGP and 7% (5/71) gained expression of PGP. 36/40 patients that lacked PGP expression and 8/9 patients that "lost" PGP expression, exhibited and retained expression of MRP1 in the initial and subsequently-profiled specimens.

Conclusion: Biomarker expression patterns of drug efflux pumps may provide insight to the chemoresistance phenotype observed in TNBC. Expression of MRP1 is favored in TNBC and is correlated with metastatic disease status. Although PGP expression may be absent or lost during TNBC progression, MRP1 (and BCRP) expression are almost always retained. Utilization of chemotherapies that are substrates of PGP, but not MRP1, (based on expression status) may be worthy of future investigation.

Citation Format: Feldman R, Abbott B, Reddy S, Gatalica Z, Castro M. ABC transporter expression: Clues into chemoresistance of triple negative breast cancers. [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 P4-09-27.