bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract P4-01-17: Expression of circulating tumor cells (CTC) and CK-19 mRNA (CK19) as prognostic factors in heavily pretreated metastatic breast cancer
Authors: Morales Serafin , Velasco Ana , Vidal Jose , Serrate Anna , Valls Joan , Samame Juan Carlos , Gisbert Rafael , Moral Desamparados , Llombart Antonio , Salud Antonieta , Matias-Guiu Xavier
Published: May 2015

Introduction: The expression of CTC and CK19 holds prognostic value for patients with breast cancer but their clinical significance remains still controversial.

Methods: This clinical observational study included 58 preteated metastatic breast cancer patients who started a standard new treatment line. CTC and CK19 was measured with CellSearch® and RT-PCR respectively at inclusion time. Progression-free survival (PFS) was defined as the time elapsed between the initiation of the treatment and either the date of clinical or radiological progression or death or the last follow-up. Cox proportional hazards regression model was used to assess the univariate prognose value of CTC and CK19 on PFS, and Kaplan-Meier estimates. A multivariate Cox model was also performed to additionaly account for ER and visceral disease. CTC and CK19 positivity was considered when a value of 1 or more was observed.

Results: Mean age was 59.68 (range 35-86), the average number of previous treatments was 2.98 (range 1-10), 38 patients (65.52%) were ER+ and 22 (37.93%) had visceral disease. Median PFS was 7 months (CI 95% 4-9). Univariate analyses showed a significant effect of the positivity of CK19 (HR=2.22, CI 95% 1.15-4.29, p=0.01) but did not reach statistical significance for CTC (HR=1.86, CI 95% 0.93-3.72, p=0.07). The estimate disease-free survival rate at 6 and 12 months were 63.6% and 43.4% for patients with CK19<1 and 47.1% and 10.6% for patients with CK19>=1, respectively. The estimate disease-free survival rate at 6 and 12 months were 77.8% and 28.8% for patients with CTC=0 and 43.6% and 21.5% for patients with CTC>=1, respectively. In the multivariate analysis the effect of CK19 and CTC were similar (HR=2.12 and 1.86, p=0.03 and 0.08 respectively), ER was statistically significant (HR=0.48, p=0.05, + vs -) but visceral diseases appeared not significant (HR=1.57, p=0.17, yes vs no).

Conclusions: The expression of CK19 appeared clinically meaningful in pretreated metastatic breast cancer patients, even when adjusting by ER and visceral diseases. CTC showed a similar but minor effect and not statistically significant. These results support CK19 as an interesting biomarker for predicting clinical response in metastatic breast cancer.

Citation Format: Serafin Morales, Ana Velasco, Jose Vidal, Anna Serrate, Joan Valls, Juan Carlos Samame, Rafael Gisbert, Desamparados Moral, Antonio Llombart, Antonieta Salud, Xavier Matias-Guiu. Expression of circulating tumor cells (CTC) and CK-19 mRNA (CK19) as prognostic factors in heavily pretreated metastatic breast cancer [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P4-01-17.
Abstract P4-04-20: Tumor infiltrating lymphocytes in inflammatory breast cancer
Authors: Colpaert Cecile , Marsan Melike , Vermeulen Peter , Dirix Luc , Van Laere Steven ,   Inflammatory Breast Cancer International Consortium
Published: May 2015

Inflammatory breast cancer (IBC) is a rare and aggressive form of breast cancer with poor prognosis. Multimodal treatment including neo-adjuvant chemotherapy is the treatment of choice for IBC patients, with pathological complete response (pCR) being one of the most important prognostic factors.

Tumor infiltrating lymphocytes (TILs) are an independent predictor of response to neo-adjuvant chemotherapy in breast cancer (Denkert C. et al., 2010). In a search for multigene predictors of pCR in IBC using DNA micro-arrays, a 107 gene signature was found that distinguished between responders and non-responders. This signature was enriched for immunity-related genes showing that in IBC, as in non-IBC, response to neo-adjuvant chemotherapy is associated with immunity related processes (Bertucci F. et al., 2014).

Standard H&E stained sections of formalin-fixed paraffin-embedded pre-treatment tumor tissue of 77 IBC patients were used to evaluate the presence of TILs according to the Denkert method: mononuclear cell infiltrates were considered to be intratumoral TILs when making direct contact with the tumor cells, while stromal TILs are mononuclear cells being present in the tumor stroma without making direct contact with tumor cells.

Intratumoral TILs were observed in 10,4% (8/77) of the patients. Stromal TILs (strTILs ) were present in all tumors: in 35% (27/77) strTILs occupied less than 5% of the tumor stroma, in 62 % (48/77) strTILs occupied between 5 and 50% of the tumor stroma and 2,6% (2/77) were "lymphocyte predominant breast cancers (LPBC)" with strTILS occupying more than 50% of the tumor stroma. One LPBC was HER2 positive, the other one was triple negative. There was a significant association of strTILs with response to neo-adjuvant chemotherapy: pCR was obtained in 13% of the patients with strTILs <5%, in 20% of the patients with strTILs between 5 and 50% and in both patients with LPBC (Chi 2 p=0,018).

Tertiary lymphoid structures - lymphoid follicles with germinal centres, known to be associated with better clinical outcomes (Gu-Trantien C. et al., 2013) - were detected in 2.6% of the patients (2/77); both patients had a disease free survival of more than 3 years.

This study shows that IBC tumors do not contain more TILs than non-IBC tumors and that, as in non-IBC, there is a positive association of stromal TILs with pCR in IBC. Further validation of these results and further study of lymphocyte subsets in IBC is warranted.

Citation Format: Cecile Colpaert, Melike Marsan, Peter Vermeulen, Luc Dirix, Steven Van Laere, Inflammatory Breast Cancer International Consortium. Tumor infiltrating lymphocytes in inflammatory breast cancer [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P4-04-20.
Abstract P4-04-28: Weight loss reversed the carcinogenic effect of obesity on basal-like breast cancer
Authors: Qin Yuanyuan , Sundaram Sneha , Zhao Liyang , Essaid Luma , McNaughton Kirk K , Bendt Katharine M , Darr David B , Troester Melissa A , Makowski Liza
Published: May 2015

Epidemiologic studies demonstrate that obesity is associated with a subtype of breast cancer called basal-like breast cancer (BBC), which is fast proliferating and aggressive, with no targeted therapies. Using the C3(1)-TAg murine model of BBC, we have previously shown that obese adult mice displayed significantly decreased tumor latency compared to lean mice with induction of the oncogenic HGF/c-Met pathway. Reducing adiposity is predicted to lower incidence of BBC in human population studies. Thus, the objective of this study was to investigate how a diet-switch to reduce obesity affected BBC onset and early progression. C3(1)-TAg mice were placed on a control low fat diet (LFD) and remained lean, high-fat diet (HFD) to induce obesity, and a group made obese by HFD that were then switched to LFD to induce weight loss. Just two weeks after weight loss, mice in the diet switch group had weights and adiposity similar to lean mice. Weight loss caused a 51.4% decrease in adiposity (P<0.0001) compared to obese mice. Importantly, mice that lost weight displayed significantly delayed tumor latency (17 week) compared to obese mice on HFD (15 week), with no differences detected in tumor burden or growth. Therefore the obese microenvironment that promotes early tumor onset can be reprogrammed with weight loss and restoration of a lean phenotype. Ongoing studies are examining HGF/cMet expression by immunohistochemistry in normal mammary glands and tumors to determine the contribution of this obesity-responsive pathway. Metabolomics and RNAseq analysis will identify novel obesity-dependent pathways relevant to BBC. In conclusion, we demonstrated that loss of adiposity protected against early BBC onset. Further research will identify important biomarkers associated with obesity and weight loss that can be compared through conserved biology approaches.

Citation Format: Yuanyuan Qin, Sneha Sundaram, Liyang Zhao, Luma Essaid, Kirk K McNaughton, Katharine M Bendt, David B Darr, Melissa A Troester, Liza Makowski. Weight loss reversed the carcinogenic effect of obesity on basal-like breast cancer [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P4-04-28.
Abstract P4-05-01: Genetic analysis of the role of Brca1 in suppression of basal-like breast cancer
Authors: Pei Xin-Hai , Bai Feng , Chan Ho Lam , Scott Alexandria , Smith Matthew D , Fan Cheng , Herschkowitz Jason I , Perou Charles M , Livingstone Alan S , Robbins David J , Capobianco Anthony J
Published: May 2015

BRCA1 mutation carriers are predisposed to developing basal-like breast cancers with high metastasis and poor prognosis. However, the mechanism underlying the function of BRCA1 in suppressing basal-like breast cancer remains unclear.

We previously reported that deletion of p18Ink4c (p18), an inhibitor of CDK4 and CDK6, functionally inactivates the RB pathway, stimulates mammary luminal stem cell proliferation, and leads to spontaneous luminal tumor development whereas germline mutation of Brca1 alters the fate of luminal cells and causes luminal-to-basal mammary tumor transformation. We report here that disruption of Brca1 by both germline and epithelium-specific mutation of Brca1 in p18 deficient mice activates epithelial-to-mesenchymal transition (EMT) and induces basal-like breast tumors. Introduction of BRCA1 suppresses expression of EMT-inducing transcription factors (EMT-TFs) and inhibits EMT in mammary tumor cells. Knockdown of BRCA1 in human luminal cancer cells activates EMT and increases tumor formation potential. Consistently, EMT features are inversely correlated with BRCA1 in human breast cancers. These findings provide genetic evidence suggesting that BRCA1 suppresses EMT and inhibits basal-like tumor development.

Citation Format: Xin-Hai Pei, Feng Bai, Ho Lam Chan, Alexandria Scott, Matthew D Smith, Cheng Fan, Jason I Herschkowitz, Charles M Perou, Alan S Livingstone, David J Robbins, Anthony J Capobianco. Genetic analysis of the role of Brca1 in suppression of basal-like breast cancer [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P4-05-01.
Abstract P4-05-05: A novel TP53-dependent role for TLR4 in driving breast cancer
Authors: Haricharan Svasti , Brown Powel H
Published: May 2015

Breast cancer is a leading cause of cancer-related death. Toll-like receptor (TLR)-4 is an important mediator of cytokine secretion that is expressed in breast cancer cells, and is frequently mutated in a subset of breast tumors associated with high mutation load and poor patient survival. We show here that TLR4 plays an important role in driving breast cancer growth in a TP53 context-dependent manner. Using siRNA mediated knockdown, pharmacological inhibition and hyperactivation of TLR4 we demonstrate that TLR4 inhibits growth in breast cancer cells with wildtype TP53 by inducing G0/G1 arrest and decreasing mitotic entry. Conversely, in breast cancer cells with mutant TP53, TLR4 acts as an oncogene and drives cell growth by inducing mitosis. Furthermore, we identify the underlying mechanism for this dual TP53-dependent effect of TLR4 on breast cancer cell growth: differential interferon gamma (IFN-γ) secretion by tumor cells into their microenvironment. Hyperactivated TLR4 signaling in TP53 wildtype breast cancer cells results in increased IFN-γ secretion, identified by an unbiased cytokine array and validated by ELISA, thus inhibiting growth in an autocrine/paracrine fashion. Moreover, secreted IFN-γ is both necessary and sufficient for TLR4-induced growth inhibition in TP53 wildtype breast cancer cells. Finally, the dual TP53-dependent effect of TLR4 extrapolates to several other cancer types in silico, attaching potentially global significance to these results. Taken together, the data presented in this paper strongly suggest a novel role for TLR4 as a suppressor of cell growth in TP53 wildtype tumors, and identify differential IFN-γ secretion as the underlying mechanism. The results of this study are also translationally relevant, delineating the TP53 mutant breast cancer subset as a group that can benefit from pharmacological TLR4 inhibition. Most importantly, these results indicate a need for studying the effect of drivers of cancer growth pleiotropically rather than as isolated events.

Citation Format: Svasti Haricharan, Powel H Brown. A novel TP53-dependent role for TLR4 in driving breast cancer [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P4-05-05.
Abstract P4-05-10: PRKCQ, a novel protein kinase C preferentially expressed in triple negative breast cancer, drives oncogenic growth, survival and migration
Authors: Irie Hanna Y , Halstead-Nussloch Gwyneth , Ito Koichi
Published: May 2015

Background/Rationale: PRKCQ, a member of the novel protein kinase C family, was identified in a functional genomic screen for regulators of anoikis resistance/anchorage-independent survival (Irie et al., 2010). Interestingly, it is preferentially expressed in the triple negative/basal subtype of breast tumors compared to Luminal or Her2+ tumors. We sought to determine the functional role of PRKCQ in triple negative breast cancer and evaluate PRKCQ as a candidate therapeutic target for this subtype. Materials and Methods. Consistent with its expression in triple negative patient breast tumors, PRKCQ is also highly expressed in several triple negative breast cancer cell lines. We utilized both gain- and loss-of-function approaches in these cell lines to determine the requirement for PRKCQ in oncogenic growth and survival using in vitro and in vivo models. Results. Isoform-specific downregulation of PRKCQ using shRNA vectors severely impaired growth of triple negative breast cancer cells in 2D monolayer and 3D Matrigel cultures. In 3D cultures, PRKCQ downregulation not only inhibited growth, but impaired the formation of invasive branching. PRKCQ downregulation also inhibited the growth of MDA231 primary tumor xenografts. All of these data support a requirement for PRKCQ expression in triple negative breast cancer cell growth. To determine if PRKCQ is sufficient to drive oncogenic phenotypes, we overexpressed PRKCQ in a non-transformed immortalized breast epithelial cell line, MCF-10A. PRKCQ expression conferred EGF-independent growth, migration and anoikis resistance. In PRKCQ-expressing MCF-10A cells, EGFR phosphorylation and activation were preserved in the absence of exogenous EGF ligand addition. We are currently elucidating the mechanisms responsible for PRKCQ-mediated, sustained activation of EGFR. Conclusions. PRKCQ is critically required for growth of triple negative breast cancer cells. Increased expression of PRKCQ is sufficient to drive oncogenic, growth factor-independent growth, survival and migration. Interestingly, PRKCQ could play a dual role in the development of triple negative breast cancer, as it may also function in the immune microenvironment to support the growth of these tumors. Therefore, PRKCQ is an attractive candidate therapeutic target for patients with triple negative breast cancer.

Citation Format: Hanna Y Irie, Gwyneth Halstead-Nussloch, Koichi Ito. PRKCQ, a novel protein kinase C preferentially expressed in triple negative breast cancer, drives oncogenic growth, survival and migration [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P4-05-10.
Abstract P4-05-17: Mammaglobin is a potent regulator of breast cancer processes leading to disease progression
Authors: Guerrette Roxann , Picot Nadia , Robichaud Gilles
Published: May 2015

Metastasis is the major cause of death in women suffering from breast cancer. To provide a better understand breast cancer progression, we have studied the role of mammaglobine-1 (MGB1) gene in breast cancer pathogenesis. MGB1 has been extensively studied as a diagnostic biomarker due to its abundant expression in mammary cancer cells. Yet, MGB1's role in disease progression is still unknown. Our experimental results demonstrate for the first time that MGB1 in a pivotal regulator on breast cancer malignancy. More precisely, loss of MGB1 expression correlates with a decrease in proliferation, spheroid formation, migration, and invasion capacities of breast cancer cells. Concomitantly, we also observe that MGB1 expression activates pro-malignant signaling cascades such as MAPKs, focal adhesion kinase (FAK) and NFkB pathways. Moreover, MGB1 promote epithelial to mesenchymal (EMT) features which coincide with our findings. Our study provides the first evidence for MGB1 as regulator of breast cancer malignancy and disease progression.

Citation Format: Roxann Guerrette, Nadia Picot, Gilles Robichaud. Mammaglobin is a potent regulator of breast cancer processes leading to disease progression [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P4-05-17.
Abstract P4-07-01: Calorie restriction normalizes global microRNA expression by preventing the loss of dicer expression during mammary tumorigenesis
Authors: Devlin Kaylyn L , Sanford Tiffany , Mambo Elizabeth , Hursting Stephen D
Published: May 2015

MicroRNA (miRNAs) are small, post-transcriptional regulators that play an integral role in maintenance of cellular functions and whose dyregulation has been shown to promote many types of cancer, including breast cancer. Global downregulation of miRNAs has emerged as a common theme in human breast tumors and has been shown to contribute to oncogenesis. One of the primary mechanisms through which miRNAs are globally dysregulated is downregulation of enzymes involved in miRNA biogenesis. We aim to establish how calorie restriction (CR), which potently inhibits breast cancer progression, regulates global miRNA expression and miRNA biogenesis enzyme functionality in mammary tumors. To address these questions, 100 female Sprague Dawley rats were administered either dimethylbenz(a)anthracene (DMBA) or vehicle control at 50 days of age, then randomized to receive either control (AIN-76A) diet ad-libitum (n=40) or a 30% CR diet regiment (n=60). Resultant mammary tumors were allowed to develop for 12 weeks. Calorie restriction significantly increased survival to study endpoint relative to control diet (75% vs 35%, respectively) (p=0.0047). Furthermore, of the animals that developed tumors, CR significantly decreased median tumor area by 56% compared to control diet (109.4 mm2 vs 250.9 mm2, respectively) (p=0.0286). Global miRNA expression was analyzed through miRNA-specific sequencing. Calorie restriction had a broad effect on miRNA expression, illustrated by the fact that of all the miRNAs with a greater than two-fold expression difference between CR and control, 80% are overexpressed in CR tumors compared to control tumors. These results can be explained by the additional finding that CR was able to prevent the loss of Dicer expression, a key miRNA biogenesis enzyme, observed in control-fed mammary tumor tissue compared to normal tissue. This important finding suggests that global miRNA normalization through the retention of Dicer expression during cancer progression could be a contributing mechanism to CR’s anticancer effects. We plan to further these investigations by exogenously manipulating Dicer expression in Rama25 cells, which were originally derived from a DMBA-induced mammary tumor from a Sprague Dawley rat, and analyzing the resultant tumorigenic potential in vitro and in vivo. The results obtained will provide insights into the mechanisms of breast cancer progression and how CR inhibits progression through microRNA modulation.

Citation Format: Kaylyn L Devlin, Tiffany Sanford, Elizabeth Mambo, Stephen D Hursting. Calorie restriction normalizes global microRNA expression by preventing the loss of dicer expression during mammary tumorigenesis [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P4-07-01.
Abstract P4-11-02: Worse breast cancer prognosis in insulin treated diabetic patients - A population based registry study in Sweden
Authors: Olsson Håkan , Einefors Rickard , Broberg Per , Landin Olsson Mona
Published: May 2015

Background. Diabetes may be linked to incidence of different tumor diseases and prognosis through various mechanisms such as the disease itself, hyperglycemia, obesity and anti-diabetes therapy.

Material and methods. The study includes all women with BC diagnosed in Sweden between 2000 through 2008 (n=54406). The women had no previous cancer diagnosis during the period of 1958-1999. Dates of birth, BC diagnosis and TNM-stage where directly extracted from the cancer registry. The women’s anti-diabetes therapy was gathered from the Swedish Prescribed Drug Registry. Information regarding the cause of death and date of death was obtained from the Cause of Death Registry and tbe Swedish Population Register up until the 31st of December 2012 and 31st of December 2013 respectively. Analyses have been restricted to patients receiving insulin therapy (n=2463) and their breast cancer prognosis has been calculated in comparison with breast cancer patients without diabetes. All analyses were adjusted for TNM-stage and age at diagnosis.

Results. Patients with insulin treated diabetes had a worse prognosis compared with other women with breast cancer (HR 1.7, 95%CI 1.5-2.0). The worse prognosis could be seen both for patients with ER+ and ER- tumors. The worst prognosis was seen for patients treated with NPH insulins (HR 2.8, 95% CI 2.4-3.3) while patients treated with long-acting insulin analogs had an intermediate prognosis (HR 1.6, 95% CI 1.2-2.2). Those women treated with NPH insulins and metformin had a slightly worse prognosis (HR 1.4, 95% CI 1.0-1.8). The results for breast cancer specific survival and total survival were similar.

Conclusion. Our results imply that insulin treated breast cancer patients have a worse survival compared with other women with breast cancer regardless of tumor stage. Metformin therapy may partially counteract the association.

Citation Format: Håkan Olsson, Rickard Einefors, Per Broberg, Mona Landin Olsson. Worse breast cancer prognosis in insulin treated diabetic patients - A population based registry study in Sweden [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P4-11-02.
Abstract P4-12-06: Results of next-generation sequencing panels in a large community-based hereditary cancer risk program
Authors: Blum J L , Garby C A , Simmons A E , Walker S A , Panos L E
Published: May 2015

Background: Next-generation sequencing (NGS) allows for broader germline genetic testing for hereditary cancers. Since the Supreme Court decision of AMP v. Myriad on June 13, 2013, hereditary cancer multi-gene panels can now include BRCA1 and BRCA2, making these panels an option for first-tier testing. However, questions have been raised about the clinical utility and implications of extended panels for medical management given the inclusion of unknown to moderate penetrant genes.

Methods: We reviewed all patients who underwent multi-gene panel testing from July 1, 2013 through May 23, 2014. The indications for testing included personal and/or family history of breast or ovarian cancer. The panels were ordered in a single genetic counseling clinic within a large community-based cancer center.

Results: A total of 136 patients underwent panel testing via a single commercial laboratory. We identified 12 (8.8%) patients who were positive for a pathogenic or likely pathogenic mutation in a cancer susceptibility gene; 4 had prior negative BRCA1 and BRCA2 sequencing and deletion/duplication testing. These positive results included 4 BRCA2 mutations, 2 TP53 mutations, 1 CDH1 mutation, 2 ATM mutations, and 1 patient each with a CHEK2, NBN, or PALB2 mutation. Of the patients found to have a positive test result, 100% met the National Comprehensive Cancer Network (NCCN) guidelines for Hereditary Breast and Ovarian Cancer (HBOC) genetic testing. The CDH1 mutation carrier did not meet NCCN guidelines for hereditary diffuse gastric cancer testing and only one of the TP53 mutation carriers met NCCN guidelines for Li-Fraumeni syndrome. Within our cohort (136), 21 (15.4%) patients had a total of 25 variants of uncertain significance (VUS) and 103 (75.7%) patients had negative test results.

Conclusion: Testing through NGS panels identified 7/12 (58%) patients with a mutation which led to changes in current medical management and 3/7 (43%) had a mutation in a gene other than BRCA1 or BRCA2. Our findings suggest that there is clinical utility of NGS panels for use in this patient population despite the inclusion of unknown to moderate penetrant genes and a higher rate of VUS than single gene testing.

Citation Format: J L Blum, C A Garby, A E Simmons, S A Walker, L E Panos. Results of next-generation sequencing panels in a large community-based hereditary cancer risk program [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P4-12-06.
Abstract P4-12-12: Next generation sequencing-based analysis of BRCA1 and BRCA2 genes: Applicability for fast diagnostics of large samples
Authors: Kong Sun-Young , Cho Eunhae , Lee Junnam , Lim Myong Cheol , Jang Jahyun , Park Boyoung , Yoon Kyong-Ah , Kim Young-Ho , Lee Eun Sook
Published: May 2015

Purpose

BRCA1&2 gene mutation test for hereditary breast ovarian syndrome (HBOC) has been conducted mostly by Sanger sequencing. Currently, the next generation sequencing (NGS) is rapidly incorporated to the fields of cancer research and clinical diagnostics. Here we evaluated NGS-based results of BRCA gene analysis and compared with the results of Sanger sequencing for future diagnostic applications.

Methods

The patients (n=100) who have genetic counseling and decided to perform BRCA test were included. All coding regions of BRCA1 and BRCA2 analyzed by both of Sanger sequencing and NGS using Access Array BRCA1, BRCA2 and TP53 kit (Fluidigm, USA). Access array kit was designed to multiplex 48 samples simultaneously for 184 amplicons and the average sequencing depth per base was 6,500X using MiSeq sequencer (Illumina, USA). We developed analysis pipelines to avoid false negative results and detect all pathogenic mutations. The reads were aligned to a reference genome (NCBI human genome assembly build 37) using the BWA-MEM, then all candidate variants called with minimum filtering parameter.

Results

Total of 765 variants were detected by NGS and among them 616 variants (frameshift:22, nonsense:8, splicing:3, missense:290, and synonymous variants:293) were identical with the results from Sanger sequencing. When we evaluated the results of Sanger sequencing as standard methods, the mean allele frequency showed difference as 41.7% and 12.0% for true positive heterozygous variants (616) and false positive variants (149), respectively.

Conclusions

There was no false negative of pathogenic mutations from NGS. The BRCA mutation detection using NGS represented potential applicability in clinical diagnosis.

Citation Format: Sun-Young Kong, Eunhae Cho, Junnam Lee, Myong Cheol Lim, Jahyun Jang, Boyoung Park, Kyong-Ah Yoon, Young-Ho Kim, Eun Sook Lee. Next generation sequencing-based analysis of BRCA1 and BRCA2 genes: Applicability for fast diagnostics of large samples [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P4-12-12.
Abstract P5-01-03: Opto-acoustic nomograms for improving breast cancer diagnosis
Authors: Lavin Philip , Stavros Thomas
Published: May 2015

Background/Objective:

Diagnostic specificity remains disappointingly low for ultrasound-based methodologies optimized to achieve near 100% sensitivity. Opto-acoustic (OA) imaging is a fusion of real time co-registered, interleaved laser optic and ultrasound imaging that shows fused functional findings (relative de-oxygenation of hemoglobin) and morphologic information (tumor angiogenesis) within and around breast masses using a hand-held duplex OA probe. We assess the improvement in sensitivities and specificities of nomograms (N) based on regression models to classify (CL) benign (B) vs. malignant (M) and to project probability of malignancy (POM) based on five feature-based metrics scored by 21 independent readers (IRs) and an expert reader. We examine OA feature separation and nomogram performance.

Methods:

Masses from a series of 100 subjects with 80 biopsies (42 B, 38 M) were blindly evaluated prior to both core biopsy and excision. The OA algorithm was locked. Three internal OA findings (density of vascularity [V], relative blood oxygen saturation [O], and hemoglobin [H]) and two external OA findings (boundary zone [Z] and peri-tumoral radiating vessels [R]) were assigned 0-5/6 ordinal scores. Feature distributions were compared using a two-sided Kruskal-Wallis test.

Results:

There were consistently significant differences between the feature distributions for benign vs. malignant: density of vascularity (14/21 IRs), relative blood oxygen saturation (11/21 IRs), hemoglobin (16/21 IRs), boundary zone (all IRs), and peri-tumoral radiating vessels (all IRs) always with lower scores for benign vs. malignant. The mean IR sensitivities were near 100% for all IRs. The mean specificities from the nomograms were 49% (POM), 43% (CL), and 48% (averaged).

Conclusions:

The study results indicate that OA findings can be independently and quickly mastered by practicing IRs to consistently differentiate masses. Nomograms offer further confidence to enhance decision making to differentiate. If confirmed in a larger series, OA findings might be useful in differentiation and thus sparing biopsies in addition to more customized surgeries.

Citation Format: Philip Lavin, Thomas Stavros. Opto-acoustic nomograms for improving breast cancer diagnosis [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 P5-01-03.
Abstract P5-01-10: In vitro and in vivo imaging of Her2 with cyclic peptides derived from directed evolution
Authors: Kelderhouse Lindsay E , Hardy Amanda , Pan Yong , Rhea Patrea , Ornelas Argentina , Gammon Seth , Yang Peiying , Fiacco Stephen , Millward Steven W
Published: May 2015

The implementation of personalized medicine for Her2-positive breast cancer treatment (Trastuzumab and Pertuzumab) has made a dramatic effect on overall patient outcome. Unfortunately, there are currently no FDA-approved imaging agents to monitor Her2-positive breast cancer leaving physicians to rely on invasive biopsies and anatomical imaging to monitor treatment with Her2-targeted therapeutics. There is also increasing evidence that women initially diagnosed with Her2-negative breast can present with Her2-positive disease upon recurrence. Noninvasive, whole-body visualization of Her2 would identify Her2-recurrent disease and serve as a powerful tool to monitor the effectiveness of new and emerging Her2-targeted therapeutics (e.g. TDM-1). While many Her2-antibody based imaging agents have been used in preclinical applications, their long circulating half-lives, high liver uptake, and poor synthetic accessibility pose a challenges for clinical translation. Here we describe the use of highly stable cyclic peptides derived from biological display as imaging agents for Her2-positive breast cancer. These peptides have antibody-like high affinity (<10 nM) for Her2-positive tumors and their size (<2 kDa) allows for higher rates of clearance, lower background signal, and consequently higher image sensitivity. Anti-Her2 peptides were pre-optimized for affinity, protease resistance and bio-stability prior to labeling with the Cy7.5 near-infrared dye. Her2-specificity was determined by in vitro assays with Her2-postive (BT474 and SKOV3), HER2-normal (MCF7) and Her2-negative (MDA-MB-231) cell lines. Sequences that showed superior affinity and selectivity were used to acquire fluorescent images of ectopic and orthotopic murine models of Her2-positive breast cancer. Our preliminary imaging data suggests that this scaffold has excellent translational potential for targeted molecular imaging of Her2-postive breast cancer.

Citation Format: Lindsay E Kelderhouse, Amanda Hardy, Yong Pan, Patrea Rhea, Argentina Ornelas, Seth Gammon, Peiying Yang, Stephen Fiacco, Steven W Millward. In vitro and in vivo imaging of Her2 with cyclic peptides derived from directed evolution [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 P5-01-10.
Abstract P5-01-13: The spectroscopic feature of breast cancer
Authors: Ogura Hiroyuki , Yoshizawa Nobuko , Yoshimoto Kenji , Nasu Hatsuko , Taki Yumiko , Hosokawa Youko , Matsunuma Ryouichi , Ide Yoshimi , Yamaki Etsuko , Suzuki Toshihiko , Oda Motoki , Ueda Yukio , Yamashita Yutaka , Sakahara Harumi
Published: May 2015

Objectives: To examine optical properties of breast cancer by time-resolved spectroscopy.

Materials and Methods: We irradiated a pulsed laser of 760, 800, and 830 nm wave-length lights at multiple sites of both breasts including the site just above the cancer and detected the light transmitted through the breast with TRS-20SH (Hamamatsu Photonics K.K.). Absorption coefficient (μa), reduced scattering coefficient (μs’), total hemoglobin (tHb), and oxygen saturation (SO2) of the breast were calculated by photon diffusion equation. The clinical trial started in January 2007. A total of one hundred fifty-two breast cancer patients participated in the trial and written informed consent were obtained from all of the patients.

Results: The absorption coefficient (μa) in 760, 800 and 830 nm wave-length of breast cancer tissue was significantly high compared with contra-lateral normal breast (760nm:cancer;0.078 normal;0.063 p<0.001. 800nm cancer:0.071, normal;0.050, p<0.001. 830nm: cancer;0.084, normal;0.063, p<0.001).

The absorption coefficient and reduced scattering coefficient between breast cancer tissue and contra-lateral normal breast μa (/cm)μs'(/cm) cancernormal breastcancernormal breast760nm0.0780.0639.589.71800nm0.0710.059.239.4830nm0.0840.0639.079.22

There was no difference in reduced scattering coefficient (μs’) between breast cancer tissue and contra-lateral normal breast(760nm: cancer;9.58, normal;9.71, 800nm: cancer:9.23, normal;9.40. 830nm; cancer;9.07, normal;9.22). The tHb of breast cancer tissue was significantly high, compared with normal breast (cancer:32.3±14.6, normal breast;22.0±8.6, p=0.001). There was no difference in oxygen saturation (SO2) between breast cancer tissue and contra-lateral normal breast (cancer:73.2±4.3, normal breast;73.6±5.9, p=0.31).

The tHb and SO2 between breast cancer tissue and contra-lateral normal breast cancernormal breastp-valuetHb(μM)32.3±14.622.0±8.6p<0.001SO2(%)73.2±4.373.6±5.9p=0.32

Conclusion: Absorption coefficient (μa) and tHb increased in breast cancer, whereas reduced scattering coefficient (μs’) and oxygen saturation did not.

Citation Format: Hiroyuki Ogura, Nobuko Yoshizawa, Kenji Yoshimoto, Hatsuko Nasu, Yumiko Taki, Youko Hosokawa, Ryouichi Matsunuma, Yoshimi Ide, Etsuko Yamaki, Toshihiko Suzuki, Motoki Oda, Yukio Ueda, Yutaka Yamashita, Harumi Sakahara. The spectroscopic feature of 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 P5-01-13.
Abstract P5-02-09: Ki67-labelling index of luminal breast cancers: Agreement of core biopsy and surgical specimen results
Authors: Focke Cornelia M , Decker Thomas
Published: May 2015

Objective: To highlight the agreement between Ki67-Labelling indices (LIs) of core biopsies (CB) and related surgical specimens (SSP) in luminal breast cancers (BCs) adopting the recommendations of the St Gallen Consensus Conference 2013 and the International Ki67 In Breast Cancer Working Group.

Methods: We investigated CBs and the related SSPs of 220 luminal Her2 negative BCs. To assess the Ki67-LI a total of 510 tumour cells including hot spot, cold spot and an area of intermediate proliferation was counted by one observer. Every positive nucleus was counted, independent of the staining intensity. A Ki67-LI cut-off <20% as recommended by the St Gallen Consensus was used to define luminal A BCs. Agreement between CB and SSP, rates of under- and overestimation, and positive predictive values (PPV) of CB based subtyping (luminal A vs. B) were calculated.

Results: In SSP 140/220 BCs (64%) were luminal A, 80 (36%) were luminal B. The agreement between CB and OP was 77% (169/220). Overestimation of proliferation in CB occurred in 10 (4%) and underestimation in 41 (19%) cases. The PPVs of CB based Ki67-LI for luminal A and luminal B were 0.76 (130/171) and 0.79 (39/49), respectively.

Conclusion: The agreement of CB and SSP based subtyping of luminal BCs is generally high. However, underestimation of proliferation in CB may result in subtype misclassification in almost one fifth of luminal Her2 negative luminal tumours.

Citation Format: Cornelia M Focke, Thomas Decker. Ki67-labelling index of luminal breast cancers: Agreement of core biopsy and surgical specimen results [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 P5-02-09.
Abstract P5-04-10: Characterization of the immune infiltrate in HER2+ breast cancer
Authors: Ganesan Shridar , Alexe Gabriela , Hirshfield Kim M , Yao Ming , Bhanot Gyan , Deborah Toppmeyer
Published: May 2015

A subset of HER2+ breast cancer is characterized by high expression of lymphocyte-associated genes and the presence of a robust, histologically apparent, lymphocytic infiltrate. The presence of an immune signature and lymphocytic infiltration is associated with improved outcome, and may be a marker of improved response to trastuzumab-based therapy. The relationship of the immune signature in these cancers to the biology and functional status of the lymphocyte subsets present in the tumor microenvironment remain unclear. To better characterize the immune microenvironment of HER2+ cancers, we have analyzed paired RNA sequencing data and histologic image data from The Cancer Genome Atlas. We have confirmed the presence of a strong immune signature in a subset of HER2+ breast cancers that is correlated with the presence of a robust lymphocytic infiltrate. Immuno-phenotyping of the lymphocytic infiltrate was performed on an independent set of HER2+ breast cancers from the Rutgers Cancer Institute of New Jersey. CD8+ T-cells and macrophages are a significant component of the immune infiltrate in these cancers. Interestingly, CD20+ B-cells were also present in the tumor stroma as large aggregates whose organization is suggestive of the presence of ectopic germinal centers. The relationship of lymphocyte subsets present in HER2+ breast cancer, immunoglobulin subtypes expressed and correlation with immune checkpoint protein expression will be presented. The composition of the immune infiltrate and its organization into ectopic germinal centers may give new insight into the role of the immune microenvironment in the development of HER2+ breast cancer and the response to trastuzumab.

Citation Format: Shridar Ganesan, Gabriela Alexe, Kim M Hirshfield, Ming Yao, Gyan Bhanot, Toppmeyer Deborah. Characterization of the immune infiltrate in HER2+ 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 P5-04-10.
Abstract P5-05-10: Leptin peptide receptor antagonist linked to nanoparticles: A novel adjuvant therapy for triple negative breast cancer
Authors: Harmon Tia L , Harbuzariu Adriana , Dill Courtney D , Yang Lily , Gonzalez-Perez Ruben R
Published: May 2015

Background: Obesity and high leptin levels are strongly associated with breast cancer, relapse, drug resistance, and poor patient outcomes. Over expression of leptin and its receptor, Ob-R, induce cell proliferation, angiogenesis, and metastasis in Triple Negative Breast Cancer (TNBC). This aggressive form of the disease has no targeted therapy and chemotherapeutics show several undesirable side effects. We have created a Leptin Peptide Receptor Antagonist, LPrA2, which has been shown to effectively prevent leptin signaling. LPrA2 was coupled to iron oxide nanoparticles (IONPs) and used to determine its potential use as an adjuvant to chemotherapeutics.

Methods: IONPs, bound to LPrA2, were confirmed by Western Blot. TNBC cells were then treated with IONP-LPrA2 plus Cisplatin, Doxorubicin, Paclitaxel, Cyclophosphamide, and Sunitinib. Subsequently, the TNBC cells were analyzed for proliferation, cell cycle progression, and apoptosis with the Cellometer Vision Image Cytometer ®.

Results: IONP-LPrA2 was found to cause a greater decrease in DNA synthesis during the S phase of the cell cycle in TNBC cells than LPrA2 alone. Additionally, IONP-LPrA2 when combined with chemotherapeutics or anti-angiogenic drugs showed synergistic effects on cell proliferation and apoptosis.

Conclusion: These findings indicate that IONP-LPrA2 may be useful in the prevention and treatment of TNBC. Further IONP-LPrA2 treatment may increase the efficiency of chemotherapeutics. These results could be particularly relevant for obese patients, whom show high incidence and the poorest outcome of TNBC.

Acknowledgements: This work was partially supported by the National Institutes of Health and National Cancer Institute Grant 1SC1CA138658-05 and U54 CA118638, and DOD Idea Award BC 123427 to RRGP; and facilities, and support services at Morehouse School of Medicine (NIH RR03034 and 1C06 RR18386) and NIH/NCRR grant 1G12RR026250-03.

Citation Format: Tia L Harmon, Adriana Harbuzariu, Courtney D Dill, Lily Yang, Ruben R Gonzalez-Perez. Leptin peptide receptor antagonist linked to nanoparticles: A novel adjuvant therapy for triple negative breast cancer [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P5-05-10.
Abstract P5-06-05: miRNAs associated with DNA repair capacity in Puerto Rican women with breast cancer
Authors: Matta Jaime , Isaza Clara , Ortiz Carmen , Suarez Erick , Morales Luisa
Published: May 2015

BACKGROUND: MicroRNAs (miRNA) are short non-protein-coding RNAs that regulate gene expression at the post-transcriptional level via binding to 3′-untranslated regions of protein-coding transcripts. Some miRNAs have been used as diagnostic, prognostic and therapeutic markers of breast cancer (BC). It is well established that dysregulation of DNA repair capacity (DRC) is an important risk factor of BC. However, there is little published information as to what specific miRNAs are associated with DRC in women with BC. OBJECTIVE: The main objective of this study was to identify candidate miRNAs associated with dysregulation of DRC in women with BC. METHODS: Plasma samples from 30 BC cases and 30 controls selected based on their DRC levels (low, high) using a proprietary algorithm. Samples were analyzed for miRNA expression utilizing protocols from Applied Biosystems (Life Technologies). The miRNA expression profiling was performed utilizing the RT-PCR TaqMan Array Human MicroRNA A Cards v 2.0 (Applied Biosystems) containing 383 miRNA probes. Single-stranded cDNA was synthesized from 200 ng of total RNA in 8 Multiplex RT primer pool reactions containing stem-looped RT primers that were specific to mature miRNAs. U6 snRNA-001973 was selected for normalization based on our own experimental validations. To quantify the association of the miRNA expression the fold change () was estimated for every detector with the p-values calculated using the t-test. RESULTS: Candidate miRNAs that showed a statistically significant expression were: miR-146, miR-34a, miR-221, Let-7b, miR-193b, miR-132, miR-192, miR-21, miR-197, miR-24, miR-26b, miR-29c were identified based on a false discovery rate of 4%. The results showed that twelve miRNAs differentially expressed in patients with BC. Candidate miRNAs have been reported associated with the expression of twenty seven DNA repair genes. Two of these genes are part of the NER pathway which has been identified by previous studies as important in BC. CONCLUSION: Our preliminary data suggests that differential expression of specific miRNAs might be associated with dysregulation of DRC in BC. The molecular mechanisms by which miRNAs regulate DNA repair genes remain to be elucidated. However, our results lend further promise to the concept of miRNAs as a tool to study the regulation of DRC. We see potentialfuture applications in prognosis and therapy of women with BC. Supported by grants S06 GM008239-20 and 1SCA157250 from the NCI Center to Reduce Health Disparities and NIH-MBRS Program (NIGMS) and NIH-NIGMS #GM082406 (CO).

Citation Format: Jaime Matta, Clara Isaza, Carmen Ortiz, Erick Suarez, Luisa Morales. miRNAs associated with DNA repair capacity in Puerto Rican women 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 P5-06-05.