Subtypes Of Invasive Breast Cancer Research Articles

Overexpression of the VRK1 kinase, which is associated with breast cancer, induces a mesenchymal to epithelial transition in mammary epithelial cells.

Vaccinia-related kinase 1 (VRK1) is a pro-proliferative nuclear kinase. Mice engrafted with VRK1-depleted MDA-MB-231 breast cancer cells have been shown to develop fewer distal metastases than controls, suggesting VRK1 might play a role in cell migration, invasion, and/or colonization. In work described herein, we investigated the impact of VRK1 overexpression on human mammary epithelial cells. In 2D culture, VRK1 overexpression diminishes cell migration and invasion and impairs the migration-associated processes of cell spreading and cytoskeletal rearrangement. VRK1-overexpressing cells show reduced accumulation of the mesenchymal marker vimentin and increased accumulation of the epithelial markers E-cadherin and claudin-1. VRK1 overexpression also leads to reduced levels of the transcriptional repressors snail, slug, and twist1. Cumulatively, these data indicate that VRK1 overexpression augments the epithelial properties of both MCF10a and MDA-MB-231 cells. We further studied the impact of VRK1 on the epithelial properties of MCF10a cells in 3D matrigel culture, in which cells proliferate and form epithelial sheets that mature into hollow spherical acini. VRK1 overexpression significantly accelerates the initial stages of cell proliferation, leading to larger acini that nevertheless differentiate and mature. Our analysis of human tumor tissue microarrays (TMAs) revealed that VRK1 protein levels are higher in lymph node metastases than in patient-matched mammary tumors. Using public databases, we determined that VRK1 is among the top 10% of overexpressed transcripts in multiple subtypes of invasive breast cancer, and that high levels of VRK1 expression are correlated with decreased relapse-free survival. In sum, overexpression of VRK1, by regulating the transcription repressors snail, slug, and twist1, can promote a mesenchymal-to-epithelial transition (MET) in cell culture. VRK1-mediated MET might facilitate the colonization of distal sites by metastatic breast cancer cells, providing some insight into the frequent association of VRK1 overexpression with breast malignancies and the correlation between VRK1 overexpression and poor clinical outcome.

Exposure to hazardous air pollutants and risk of incident breast cancer in the Nurses\u2019 Health Study II

BackgroundFindings from a recent prospective cohort study in California suggested increased risk of breast cancer associated with higher exposure to certain carcinogenic and estrogen-disrupting hazardous air pollutants (HAPs). However, to date, no nationwide studies have evaluated these possible associations. Our objective was to examine the impacts of mammary carcinogen and estrogen disrupting HAPs on risk of invasive breast cancer in a nationwide cohort.MethodsWe assigned HAPs from the US Environmental Protection Agency’s 2002 National Air Toxics Assessment to 109,239 members of the nationwide, prospective Nurses’ Health Study II (NHSII). Risk of overall invasive, estrogen receptor (ER)-positive (ER+), and ER-negative (ER-) breast cancer with increasing quartiles of exposure were assessed in time-varying multivariable proportional hazards models, adjusted for traditional breast cancer risk factors.ResultsA total of 3321 invasive cases occurred (2160 ER+, 558 ER-) during follow-up 1989–2011. Overall, there was no consistent pattern of elevated risk of the HAPs with risk of breast cancer. Suggestive elevations were only seen with increasing 1,2-dibromo-3-chloropropane exposures (multivariable adjusted HR of overall breast cancer = 1.12, 95% CI: 0.98–1.29; ER+ breast cancer HR = 1.09; 95% CI: 0.92, 1.30; ER- breast cancer HR = 1.14; 95% CI: 0.81, 1.61; each in the top exposure quartile compared to the lowest).ConclusionsExposures to HAPs during adulthood were not consistently associated with an increased risk of overall or estrogen-receptor subtypes of invasive breast cancer in this nationwide cohort of women.

Abstract PD8-02: Mechanisms of recurrence: Paired analysis of primary and metastatic triple negative breast cancer

Abstract Background: Triple negative breast cancer (TNBC) is the most aggressive subtype of invasive breast cancer that lacks ER, PR, and HER2 expression. It is a heterogeneous disease with several molecular subtypes: basal-like1 (BL-1), basal-like 2 (BL-2), mesenchymal (M), and luminal androgen receptor (LAR). Treatment for TNBC is normally limited to chemotherapy, and relapse is common. Here we report molecular alterations associated with TNBC metastasis by analyzing the genomic profiles of paired primary and metastatic TNBCs. Methods:50 paired TNBCs were identified through an IRB-approved protocol via the City of Hope (COH) Biospecimen Repository. DNA mutation and mRNA expression profiles of 10 paired primary and metastatic TNBCs were analyzed. DNA mutations were identified using exome sequencing by FoundationOne®. Affymetrix Human Genechip 2.0 was used for mRNA expression profiling. Raw data were normalized and processed using Expression Console, and linear regression was performed using Limma to identify the differentially expressed genes between primary and metastatic TNBCs. Results: DNA mutation profiling showed that multiple mutations occurred within genes covering pathways of PI3K/AKT/mTOR, DNA repair, RAS/MAPK, cell cycle, and growth factor receptor signaling, reconfirming genomic heterogeneity of TNBCs. Gene expression profiles were analyzed for Lehmann's TNBC molecular subtypes (BL-1, BL-2, M, and LAR). Six of ten TNBCs showed phenotype shift between the primary and metastatic TNBCs. Several unique gene expression patterns were identified by comparing the paired TNBCs. CCNE1 and TPX2 were co-overexpressed in metastatic TNBCs compared to paired primaries. This mirrored prior studies in ovarian cancer, where co-overexpression of CCNE1 and TPX2 were found related to clonal resistance against chemotherapy. Splicing factors TRA2B and SRSF7 were also over-expressed in metastatic TNBCs compared with primaries. The analysis studying the association of CCNE1 and TPX2 with overall survival is ongoing using TCGA. Conclusion: Overall, these results show the comparative changes between primary and relapsed TNBCs and indicate the heterogeneity of molecular mechanisms of recurrence. CNNE1 and TPX2 may represent important genes involved in TNBC metastasis. Further analyses including a total of 50 paired TNBCs are currently underway. Study Contact: Yuan Yuan MD PhD, City of Hope Comprehensive Cancer Center; Duarte, CA 91030; Email: yuyuan@coh.org Citation Format: Zhao Z-M, Yost S, Yuan Y-C, Warden C, Chuang J, Yuan Y. Mechanisms of recurrence: Paired analysis of primary and metastatic triple negative breast cancer [abstract]. In: Proceedings of the 2017 San Antonio Breast Cancer Symposium; 2017 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2018;78(4 Suppl):Abstract nr PD8-02.

Prognostic Value of Tumor-Infiltrating Lymphocyte Density Assessed Using a Standardized Method Based on Molecular Subtypes and Adjuvant Chemotherapy in Invasive Breast Cancer.

This study investigated the prognostic value of tumor-infiltrating lymphocyte (TIL) density as determined by molecular subtype and receipt of adjuvant chemotherapy in invasive breast cancer (IBC). Stromal TIL densities were evaluated in 1489 IBC samples using recommendations proposed by the International TILs Working Group. Cases were allocated to high- and low-TIL density groups using a cutoff of 10%. Of the 1489 IBC patients, 427 (28.7%) were assigned to the high-TIL group and 1062 (71.3%) to the low-TIL group. High TIL density was found to be significantly associated with large tumor size (p=0.001), high histologic grade (p<0.001), and high Ki-67 labeling index (p<0.001). Triple-negative and human epidermal growth factor receptor 2 (HER2)-positive subtypes had significantly higher TIL densities than luminal A or B (HER2-negative) subtypes (p<0.001). High TIL density was significantly associated with prolonged disease-free survival (DFS) by univariate (p<0.001) and multivariate (p<0.001) analyses. In the low-TIL-density group, the patients who did not receive adjuvant chemotherapy showed better DFS (p<0.001), but no such survival difference was observed in the high-TIL group (p=0.222). For the patients who received adjuvant anthracycline, high-TIL density was found to be an independent prognostic factor of favorable DFS in the luminal B (HER2-negative; p=0.003), HER2-positive (p=0.019), and triple-negative (p=0.017) subtypes. Measurements of TIL density in routine clinical practice could give useful prognostic information for the triple-negative, HER2-positive, and luminal B (HER2-negative) IBC subtypes, especially for patients administered adjuvant anthracycline.

Ductal Carcinoma in Situ of the Breast

An adult female underwent a breast-wide local excision for ductal carcinoma in situ. Which of the following statement is false regarding ductal carcinoma in situ (DCIS; intraductal) carcinoma of the breast?a.It may present as an incidental lesion on biopsies performed for other reasons.b.It may be associated with calcifications and appear as linear casts or granular on imaging.c.It is always associated with invasive breast cancer.d.Before widespread imaging mammography, it often was identified as a palpable mass. Answer: c. It is always associated with invasive breast cancer. Ductal carcinoma in situ is not always associated with invasive ductal carcinoma, but they can occur together. Certain subtypes of invasive breast cancer, such as triple negative/basal-like breast cancers, are less often associated with DCIS than are the other subtypes.1Doebar S.C. van den Broek E.C. Koppert L.B. et al.Extent of ductal carcinoma in situ according to breast cancer subtypes: a population-based cohort study.Breast Cancer Res Treat. 2016; 158: 179-187Crossref PubMed Scopus (37) Google Scholar Ductal carcinoma in situ may present as an incidental lesion on biopsies performed for other reasons. Ductal carcinoma in situ may be associated with calcifications and appear as linear casts or granular on imaging. Before widespread imaging mammography, DCIS was often identified as a palpable mass.

Three-dimensional versus two-dimensional shear-wave elastography: Associations of mean elasticity values with prognostic factors and tumor subtypes of breast cancer

PurposeTo explore associations between prognostic factors and subtypes of invasive breast cancer (IBC) and elasticity values using three-dimensional (3D) and two-dimensional (2D) shear-wave elastography (SWE). Materials and methodsMean elasticity values (kPa) of 121 IBCs were measured using both 3D and 2D SWE. Associations between these values and prognostic factors and subtypes were analyzed using linear regression model. ResultsIn both 3D and 2D SWE, larger size and presence of lymphovascular invasion were independent factors influencing higher mean elasticity on multivariate analyses (all p values<0.05). ConclusionsUsing either 3D or 2D SWE, higher mean elasticity values are associated with poor prognostic factors of IBC.

Ethnicity and breast cancer characteristics in Kenya.

There are no published data from specific regions of sub-Saharan Africa describing the clinical and pathological characteristics and molecular subtypes of invasive breast cancer by ethnic group. The purpose of this study was to investigate these characteristics among the three major ethno-cultural groupings in Kenya. The study included women with pathologically confirmed breast cancer diagnosed between March 2012 and May 2015 at 11 hospitals throughout Kenya. Sociodemographic, clinical, and reproductive data were collected by questionnaire, and pathology review and immunohistochemistry were performed centrally. The 846 cases included 661 Bantus (78.1%), 143 Nilotes (16.9%), 19 Cushites (2.3%), and 23 patients of mixed ethnicity (2.7%). In analyses comparing the two major ethnic groups, Bantus were more educated, more overweight, had an older age at first birth, and had a younger age at menopause than Nilotes (p<0.05 for all comparisons). In analyses restricted to definitive surgery specimens, there were no statistically significant differences in tumor characteristics or molecular subtypesby ethnicity, although the Nilote tumors tended to be larger (OR for≥5cm vs.<2cm: 3.86, 95% CI 0.77, 19.30) and were somewhat more likely to be HER2 enriched (OR for HER2 enriched vs. Luminal A/B: 1.41, 95% CI 0.79, 2.49). This case series showed no significant differences in breast cancer tumor characteristics or molecular subtypes, but significant differences in sociodemographic characteristics and reproductive factors, among the three major ethnic groups in Kenya. We suggest further evaluation of ethnic differences in breast cancer throughout the genetically and culturally diverse populations of sub-Saharan Africa.

Biologic Profiles of Invasive Breast Cancers Detected Only With Digital Breast Tomosynthesis.

The purpose of this study was to analyze the clinicopathologic and immunohistochemical features of invasive breast cancers detected only with digital breast tomosynthesis (DBT), compared with those of cancers detected with both DBT and full-field digital mammography (FFDM). The medical records of 261 women (108 without and 153 with symptoms) with invasive breast cancers who underwent FFDM and DBT between April 2015 and June 2016 were retrospectively reviewed. To assess detectability, all DBT and FFDM images were reviewed independently by three radiologists blinded to clinicopathologic information. The reference standard was established by an unblinded consensus review of all images. Clinicopathologic and immunohistochemical features were analyzed according to the detectability status. Of the 261 cancers, 223 (85.4%) were detected with both DBT and FFDM (both-detected group). Twenty-four cancers (9.2%) not detected with FFDM (DBT-only group) were classified by DBT as a mass (58.3%), architectural distortion (33.3%), or asymmetry (8.3%). The remaining 14 cancers (5.4%) were not detected with either DBT or FFDM (both-occult group). On multivariate analysis, a dense breast parenchyma (p = 0.007), small tumor size (≤ 2 cm; p = 0.027), and luminal A-like subtype (estrogen receptor positive or progesterone receptor positive or both, human epidermal growth factor receptor 2 negative, and Ki-67 expression < 14%; p = 0.008) were significantly associated with the DBT-only group. For 108 screening-detected cancers, a dense breast parenchyma (p = 0.007) and luminal A-like subtype (p = 0.008) also maintained significance. The addition of DBT to FFDM in screening would aid in the detection of less-aggressive subtypes of invasive breast cancers in women with dense breasts.

Human Metaplastic Breast Carcinoma and Decorin

Metaplastic breast carcinoma (MBC) is a rare subtype of invasive breast cancer and has poor prognosis. In general, cancers are heterogeneous cellular masses comprised of different cell types and their extracellular matrix (ECM). However, little is known about the composition of the ECM and its constituents in MBC. Decorin is a ubiquitous ECM macromolecule known of its oncosuppressive activity. As such, it provides an intriguing molecule in the development of novel therapeutics for different malignancies such as MBC. In this study, decorin immunoreactivity and the effect of adenoviral decorin cDNA (Ad-DCN) transduction were examined in MBC. Multiple immunohistochemical stainings were used to characterize a massive breast tumour derived from an old woman. Furthermore, three-dimensional (3D) explant cultures derived from the tumour were transduced with Ad-DCN to study the effect of the transduction on the explants. The MBC tumour was shown to be completely negative for decorin immunoreactivity demonstrating that the malignant cells were not able to synthesize decorin. Ad-DCN transduction resulted in a markedly altered cytological phenotype of MBC explants by decreasing the amount of atypical cells and by inhibiting cell proliferation. The results of this study support approaches to develop new, decorin-based adjuvant therapies for MBC.

Likelihood of late relapse assessed by the Breast Cancer Index (BCI) in LN-invasive lobular compared to invasive ductal carcinoma.

e12025 Background: Women with HR+ breast cancer remain at risk for late distant recurrences (DR) despite optimal adjuvant therapy. Invasive lobular carcinoma (ILC) is the 2nd most common subtype of invasive breast cancer. When compared to invasive ductal carcinoma (IDC), ILC has distinguishing clinical and pathologic characteristics that may result in different response to therapy and long term prognosis. BCI is a validated gene expression-based assay for pts with early stage HR+ breast cancer that provides risk of late (5-10 y) DR and predicts likelihood of benefit of extended endocrine therapy (EET). Here, we compared BCI assay results in pts with HR+, LN- ILC vs IDC. Methods: The BCI Clinical Database for Correlative Studies is an IRB-approved de-identified database that contains &gt;50 clinicopathologic and molecular variables from cases submitted for BCI in clinical practice (N=14,463). Molecular variables include BCI Prognostic score, HoxB13/IL17BR ratio (H/I), and molecular grade index (MGI). Clinicopathologic variables were abstracted from pathology reports (available for a subset of cases). LN- pts with available pathologic data were analyzed. Chi-squared tests were used to compare BCI results between IDC and ILC subgroups. Results: Analyses included 2554 LN- pts with available histologic subtype data (80.7% IDC; 13.7% ILC; 2.6% mixed; 2.9% other). Median age was 59.3y (range 27-89y; 74% ≥ 50y). BCI Prognostic had a broad distribution of individual risk assessment in both IDC and ILC. However, BCI Prognostic had a lower median BCI scores (P&lt;0.0001) in ILC, and classified a smaller proportion of pts with ILC as high risk for late DR compared to IDC (36.5% vs 53.1%; P&lt;0.0001). IDC had a higher median molecular proliferative status (MGI) compared to ILC (P&lt;0.0001). BCI Predictive (H/I) identified a slightly decreased proportion of pts with ILC benefiting from extended endocrine therapy compared to IDC (38.2% vs 41.1%). There was no difference in quantitative ER or PR (by PCR) between groups. Conclusions: BCI identified a smaller proportion of pts with ILC at high risk of late DR compared to IDC. Further studies evaluating outcomes are warranted to further validate BCI in pts with ILC.

Association between oral contraceptive use as a risk factor and triple-negative breast cancer: A systematic review and meta-analysis.

Triple-negative breast cancer (TNBC), a unique subtype of breast cancer, which is resistant to endocrine and targeted therapy, usually relapses early, progresses rapidly and is associated with a poor prognosis. Epidemiological investigations focusing on the association between risk factors and the onset of TNBC demonstrated that the incidence of TNBC exhibits a significant correlation with anthropometric, geographical and demographic parameters. The aim of the present systematic review and meta-analysis was to evaluate the strength of the association between the use of oral contraceptives (OCs) and TNBC. Two databases (PubMed Central/PubMed, Web of Science) and secondary references were searched to identify studies meeting the priorly established inclusion criteria. Case-control studies published between January, 2005 and March, 2016 were searched using the key words (triple-negative breast cancer OR basal-like) AND (oral contraceptives). Finally, 9 eligible articles using as control other subtypes of invasive breast cancer and 7 articles using a healthy population as control were incorporated in the meta-analysis. Pooled odds ratio (OR) and 95% confidence interval (CI) were calculated using fixed- or random-effects models according to the heterogeneity between studies. The case-control comparison using other subtypes of breast cancer as the control arm exhibited a significant association between OC use and TNBC (OR = 1.31, 95% CI = 1.18-1.45; Z=5.26, P<0.00001). These results were further confirmed by the case-control comparison using the healthy population as the control arm (OR = 1.21, 95% CI = 1.01-1.46; Z=2.04, P=0.04). The present meta-analysis indicated that women who use OCs have a greater risk of TNBC compared with women who do not. This conclusion prompts that women who used OCs should be examined more closely in population screenings of breast cancer, as they may benefit from prevention and early detection strategies.

Abstract P4-06-09: Synergistic effect of EGFR1 inhibitor and paclitaxel in newly patient derived metaplastic carcinoma cell line which harbored EGFR gene amplification

Abstract Background Metaplastic breast cancer (MBC) is a rare but aggressive subtype of invasive breast cancer, which is generally resistance to conventional chemotherapy. The majority of MBCs do not express the estrogen receptor, progesterone receptor and negative for HER-2/neu over expression. Thus, targeted treatment options to treat MBCs are also limited. Cancer cell lines are important tools used for us to test the therapeutic efficacy of anticancer agents. However, few in vitro systems for studying MBC are developed. Here we report a newly established novel MBC cell line using the recently developed conditionally reprogrammed cell method and search for effective therapeutic regiments to treat MBCs. Methodology The tumor tissue used for cell line establishment was obtained from a 40 year-old Taiwanese female, who was diagnosed with T2N2aM1 metaplastic carcinoma of breast cancer (with squamous metaplasia). The MBC tumor tissue was first digested by collagenase/ dispase mixture. After digestion, the MBC tumor cells were cultured in F medium with irradiated 3T3-J2 fibroblasts supplemented by 5 uM ROCK inhibitor (Liu et al., Am. J. Pathol. 2012). Subsequently, the MBC tumor tissue cells were passaged and propagated in these conditions until the majority of culture is tumor cells. The MBC tumor cell line was further passaged in DMEM/F12 medium without feeder cells and ROCK inhibitor. Finally, the MBC cell line (MBC-1) was characterized using DNA sequencing, immunofluorescence, Western blotting and viability assays. Results The genomic DNA of MBC1 was examined using DNA sequencing, and the MBC-1 maintained known patient's tumor mutation at the TP53 locus (p.V73fs*76). The MBC cell line also harbored EGFR gene amplification as the patient's tumor, according to quantitative PCR assays. To evaluate tumorigenicity, the MBC cell line was injected subcutaneously into immunocompromised mice, and the MBC tumors formed four weeks after injection. The MBC-1 stained positively using pan-cytokeratin antibody and negative of vimentin, confirming the epithelial origin of the cell line. To explore the possible effective treatment options, the cell line was incubated with different targeted therapy and chemotherapy agents, and the MBC cell line was relatively resistant to most of targeted therapy and chemotherapeutic agents. However, EGFR inhibitors (gefitinib and afatinib) could partially inhibit growth of the MBC cell line. We hypothesis that the inhibitory effect could act through inhibiting the amplified EGFR receptors. To further enhance the effect of EGFR inhibitors in the MBC-1, we combine the EGFR inhibitors with chemotherapy agents, and discovered that EGFR inhibitor and paclitaxel were synergistic across the range of concentration tested. The study on downstream mechanism of the synergism is ongoing. Conclusions We have successfully established a novel metaplastic carcinoma cell line with EGFR amplification from a patient using the conditionally reprogrammed cell method. Based on the preclinical study in the cell line, we found that combining EGFR inhibitor and paclitaxel could be a promising strategy to treat MBC with EGFR amplification. Citation Format: Chung P-H, Chen TW-W, Chang D-Y, Lin C-H, Huang S-M, Lu Y-S. Synergistic effect of EGFR1 inhibitor and paclitaxel in newly patient derived metaplastic carcinoma cell line which harbored EGFR gene amplification [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P4-06-09.

Abstract P2-06-08: Race is not a contributing factor to breast cancer-free interval outcome for patients with equal access to healthcare treated at a military medical center

Abstract BACKGROUND Black patients diagnosed with invasive breast cancer (IBC) are more likely to die of the disease than their White counterparts. Differences in IBC subtype, socioeconomic status, and access to healthcare may be contributing factors. We performed this study to determine if there is a racial disparity in breast cancer-free interval (BCFI) for patients treated at a military medical center where equal-access healthcare is provided. METHODS Three datasets were used: 1) TCGA-Breast Cancer cohort with 149 Black and 706 White patients; 2) Clinical Breast Care Project patients from a military medical center (CBCP-WR) with 112 Black and 372 White patients; and 3) CBCP patients from a civilian facility (CBCP-AAMC) without equal-access healthcare (data collection is ongoing and results will be available for the conference). IHC of ER, PR, and HER2 was performed to subtype IBCs as TN (ER-/PR-/HER2-) or Non-TN. Patients with Stage IV IBC at diagnosis were excluded. Kaplan-Meier method was used to generate survival plots. Cox proportional hazards regression model was used for survival analysis between races (Black vs. White). Univariable and multivariable analyses were performed. All analyses were performed using R 3.2.2. A two-tailed p value &amp;lt; 0.05 was considered significant. RESULTS The clinicopathologic properties of Black and White patients in TCGA and CBCP-WR cohorts were analyzed, and the results are consistent with the literature. The median follow-up times were comparable between the two races (TCGA: Black=29.4 M, White=27.5 M; CBCP-WR: Black=61.1 M, White=60.9 M). In TCGA, race significantly affects BCFI in multivariable analysis after adjustment for age, AJCC stage, and PAM50 subtype (HR=1.73, 95%CI=1.02-2.94, p=0.042). Stratifying patients into TN (Black=46.7%; White=53.3%) and Non-TN (Black=14.2%; White=85.8%) subtypes, race trended towards significant in BCFI in TN (HR=2.33, 95%CI=0.98-5.56, p=0.052) but not Non-TN subtypes (HR=1.12, 95%CI=0.55-2.27, p=0.752). Further adjustment for age and AJCC stage did not alter the results. In CBCP-WR, there was no significant racial difference in both univariable (HR=1.52, 95%CI=0.87-2.68, p=0.145) and multivariable analysis of BCFI adjusted for age, subtype and AJCC stage (HR=1.00, 95%CI=0.5-2.01, p=0.989). Stratifying patients into TN (Black=43.5%; White=56.5%) and Non-TN (Black=18.8%; White=91.2%) subtypes, race was not a significant factor in either group (TN: HR=0.920, 95%CI=0.4-2.14, p=0.846; Non-TN: HR=1.17, 95%CI=0.53-2.61, p=0.701). Further adjustment for age and AJCC stage did not alter the results. CONCLUSIONS Our results show that in the CBCP-WR cohort, there is no racial disparity in BCFI at the whole cohort level nor is there any trend in the TN subtype. These results contradict those obtained from the TCGA cohort suggesting unequal healthcare access may be a major contributor to racial disparity in BCFI. Analysis of the CBCP-AAMC cohort may provide further insight into whether treatment at a single medical center needs to be considered when investigating racial disparities in BCFI. The views expressed in this article are those of the author and do not reflect the official policy of the Department of Defense, or U.S. Government. Citation Format: Liu J, Fantacone_Campbell JL, Kovatich AJ, Hooke JA, Kvecher L, Sturtz LA, Shriver CD, Hu H. Race is not a contributing factor to breast cancer-free interval outcome for patients with equal access to healthcare treated at a military medical center [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P2-06-08.

Prognostic significance of centromere 17 copy number gain in breast cancer depends on breast cancer subtype

Increased copy number of chromosome enumeration probe (CEP) targeting centromere 17 is frequently encountered during HER2 in situ hybridization (ISH) in breast cancer. The aim of this study was to clarify the clinicopathologic significance of CEP17 copy number gain in a relatively large series of breast cancer patients. We analyzed 945 cases of invasive breast cancers whose HER2 fluorescence ISH reports were available from 2004 to 2011 at a single institution and evaluated the association of CEP17 copy number gain with clinicopathologic features of tumors and patient survival. We detected 186 (19.7%) cases of CEP17 copy number gain (CEP17≥3.0) among 945 invasive breast cancers. In survival analysis, CEP17 copy number gain was not associated with disease-free survival of the patients in the whole group. Nonetheless, it was found to be an independent adverse prognostic factor in the HER2-negative group but not in the HER2-positive group. In further subgroup analyses, CEP17 copy number gain was revealed as an independent poor prognostic factor in HER2-negative and hormone receptor-positive breast cancers, and it was associated with aggressive histologic variables including high T stage, high histologic grade, lymphovascular invasion, p53 overexpression, and high Ki-67 proliferative index. In conclusion, we found that elevated CEP17 count can serve as a prognostic marker in luminal/HER2-negative subtype of invasive breast cancer. We advocate the use of the dual-colored fluorescence ISH using CEP17 rather than the single-colored one because it gives additional valuable information on CEP17 copy number alterations.

Immunohistochemical Surrogates for Molecular Classification of Breast Carcinoma: A 2015 Update.

-The pioneering works on molecular classification (MC) by Perou and Sorlie et al in the early 2000s using global gene expression profiling identified 5 intrinsic subtypes of invasive breast cancers (IBCs): luminal A, luminal B, normal breast-like, HER2-enriched, and basal-like subtypes, each unique in incidence, survival, and response to therapy. Because the application of gene expression profiling in daily practice is not economical or practical at the present time, many investigators have studied the use of immunohistochemical (IHC) surrogates as a substitute for determining the MC of IBC. -To discuss the continuing efforts that have been made to develop clinically significant and readily available IHC surrogates for the MC of IBC. -Data were obtained from pertinent peer-reviewed English-language literature. -The most commonly used IHC surrogates are estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor-2 (HER2), dividing IBC into luminal, HER2, and triple-negative subtypes. The addition of Ki-67, cytokeratin 5, and epidermal growth factor receptor (EGFR) separates luminal B from luminal A subtypes, and basal-like subtype from triple-negative breast cancer. More recently, biomarkers such as androgen receptor and p53 have been shown to further stratify these molecular subtypes. Although many studies of IHC-based MC have shown clinical significance similar to gene expression profiling-defined MC, its critical limitations are: (1) a lack of standardization in terminology, (2) a lack of standardization in biomarkers used for each subtype, and (3) the lack of a uniform cutoff for each biomarker. A panel of IHC surrogates for each subtype of IBC is proposed.

Abstract 123: Multiplatform molecular profiling of invasive lobular breast cancer

Abstract Introduction: Invasive lobular breast cancer (ILC) is the second most common subtype of invasive breast cancer accounting for 10% of breast cancer diagnosis. ILC has particular histological and clinical characteristics and a distinct response to therapy. Characterizing the molecular alterations in ILC may lead to an improved understanding of its biology and provide new therapeutic options. The purpose of this study is to describe the molecular profile of ILC and compare it to the one of invasive ductal cancer (IDC). Methods: Three-hundred and thirty nine pure ILC specimens profiled from January 2012 - November 2015 were evaluated (Caris Life Sciences, Phoenix, AZ). Multiplatform profiling consisted of gene sequencing (next generation sequencing [NGS]), gene amplification (CISH or FISH), and protein expression (immunohistochemistry [IHC]). Molecular characteristics of estrogen receptor (ER) positive and human epidermal growth receptor factor 2 (HER2) negative pure ILC (n = 236) and IDC (n = 286) were compared. Results: By IHC, ER expression was present in 87.7% (277/316), progesterone receptor in 59.6% (198/313), HER2 in 3.5% (11/313), androgen receptor in 87% (262/301), PD-L1 in 8.1% (12/148) and PTEN in 63.3% (198/313). Amplifications were detected in MYC (7.7%, 2/26), EGFR (8.3%, 2/24), ERBB2 (4.5%, 13/290) and TOP2A (1.3%, 3/236). Mutations were detected in AKT1 (4.7%, 9/191), ATM (3.7%, 7/190), BRCA1 (4.2%, 4/96), BRCA2 (9.5%, 9/95), ERBB2 (7.5%, 14/186), PIK3CA (54.5%, 103/189), PTEN (7.9%, 15/189), and TP53 (13.4%, 25/186). A comparison of ER-positive/HER2-negative invasive lobular and ductal carcinomas revealed significant differences in AR expression (89.7% vs 79.6%, p = 0.0022), and mutations in CDH1 (10.1% vs 0.0%, p = 0.0001), ERBB2 (8.2% vs 2.1%, p = 0.0079), and TP53 (10.3% vs 31.8%, 0.0001). Conclusions: Multiplatform testing of this large series of ILC reveals recurrent alterations and a distinct molecular profile when compared to IDC. These support the definition of ILC as biologically distinct entity. High AR expression and high rates of dysregulation along the PIK3CA/AKT/mTOR pathway are consistent with recent reports in the literature. Citation Format: Raquel A. Nunes, David Arguello, Zoran Gatalica, Sandeep Reddy, Sandra M. Swain. Multiplatform molecular profiling of invasive lobular breast cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 123.

Abstract 496: Androgen receptor expression on circulating tumor cells in metastatic breast cancer

Abstract Background: Androgen receptor (AR) is a nuclear transcription factor and a member of the steroid hormone receptor superfamily that can be detected by immunohistochemical staining in all three subtypes of invasive breast cancer with frequency of &amp;gt;70% of estrogen receptor-positive tumors (ER+), &amp;gt;60% of HER2-positive tumors (HER2+), and 40-50% of triple-negative breast cancers (TNBC) respectively. AR-target therapies are showing early activity in AR+ breast cancer. Given early evidence of activity with agents targeting AR, it is desirable to improve our understanding of the AR biology in breast cancer. Although circulating tumor cells (CTCs) have been proposed as a liquid biopsy for biomarker detection in multiple cancer types, the rate and heterogeneity of AR expression in CTCs in breast cancer is still unclear. In this study, we determine the rate and heterogeneity of AR expression in CTCs of patients with metastatic breast cancer. Methods: Blood samples from patients with metastatic breast cancer were smeared onto glass slides and subjected to nuclear staining with DAPI and reacted with fluorescent-labeled antibodies to detect CD45 on leukocytes and cytokeratin and AR on epithelial cells; the slides were scanned on the Epic Sciences CTC platform and the data analyzed using established algorithms. Results: Among the 59 patients analyzed, 25 (42.4%) had ER+/HER2-, 12 (20.3%) had ER+/HER2+, 6 (10.2%) had ER-/HER2+, and 16 (27.1%) had TNBC based on the most recent staining. CTCs were detected in 50 of 59 (84.7%) patients. Characterization of CTCs identified CK+ CTCs (45 of 59, 76.3%), CK-negative CTCs (16 of 59, 27.1%), CK+ CTC clusters (16 of 59, 27.1%), and apoptotic CTCs (45 of 59, 76.3%), respectively. Number of CTCs per patient ranged from 0 to 282.1 per mL (median = 4.4). Six of the 50 patients (12%) with CTCs had AR+ CTCs. The number of AR+ CTCs per patient ranged from 0.3 to 52.6 per mL (median = 1.15). Among the patients with AR+ CTCs, the percentage of CTCs that were AR+ ranged from 5% to 50% (median = 21%). Among the six patients with AR+ CTCs, five patients had ER+ breast cancer and one had TNBC. Conclusions: We demonstrated that the Epic Sciences non-enriching or selecting platform can detect AR expression in CTCs in breast cancer. These preliminary results suggest the need for clinical validation of CTC AR expression as a potential test to stratify and identify patients who might benefit from AR therapy. The heterogeneity of intra-patient CTC AR expression leads us to another hypothesis that patients with AR+ CTCs might have heterogeneous disease with multiple drivers. Further studies are warranted to allow serial monitoring of changes in AR and to investigate the clinical applicability of AR+ CTCs and their heterogeneity. Citation Format: Takeo Fujii, Yipeng Wang, James M. Reuben, Rachel Krupa, Ryon Graf, Lyndsey Dugan, Jessica Kouw, Dena Marrinucci, Bora Lim, Carlos H. Barcenas, Angela N. Marx, Debu Tripathy, Ryan Dittamore, Naoto T. Ueno. Androgen receptor expression on circulating tumor cells in metastatic breast cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 496.

Abstract 3981: In situ mass spectrometry using desorption electrospray ionization mass spectrometry (DESI-MS) can detect specific lipid profiles across breast cancer molecular subtypes

Abstract INTRODUCTION Lipid profiling as a source for biomarkers identification is an evolving field with some enthusiastic results in cancer. Alterations on lipid metabolism have been increasingly recognized as a hallmark of cancer cells and may help to understand breast cancer carcinogenesis. Analytical chemistry and Pathology have been recently combined in the development of chemical microscopy using in situ mass spectrometry (MS) and may help to decipher breast cancer molecular initiation and progression. OBJECTIVE We aimed to use in situ Desorption Electrospray Ionization Mass Spectrometry (DESI-MS) to investigate lipid profiles across invasive breast cancer (IBC) molecular subtypes and related normal parenchyma (NBP) and ductal carcinoma in situ (DCIS). METHODS: We selected frozen samples from 9 IBC with various ER/PR/HER2 status (ER 0 to 100%, PR 0 to 90%, HER2 0 and 3+, Ki67 10 to 70%). Unstained slides obtained from frozen tissue at 10μm were submitted to DESI-MS analysis with no pretreatment and spatial resolution of 100μm. Molecular profiles (mass to charge [m/z] ratio versus signal intensity) were generated in triplicate directly from tissues to detect changes from areas of interest: IBC (and adjacent normal breast parenchyma and DCIS whenever present) regions were identified by a breast pathologist on HE-stained sequential slides. Experiment was conducted in full-scan mode and m/z range of 100-1200. DESI mass spectra were obtained using a Thermo ScientificTM Q ExactiveTM Hybrid Quadrupole Orbitrap Mass Spectrometer in the positive ion mode. The Biomap software was used to visualize the images and to export the spectrums to metaboanalyst, where the data were statiscally analyzed. RESULTS: The images obtained by lipid profiling presented a direct correlation with HE stained slides, with some ions m/z clearly overlaying areas of normal parenchyma, DCIS and invasive breast cancer. The unsupervised hierarchical clustering analysis showed all technical triplicates remained grouped. All NBP but one clustered together. Triple negative IBC and its related NBP formed a distinct cluster from all other ER+ samples. The ER+/HER2 + tumor segregated from other ER+/HER2- tumors. DCIS and IBC from ER+/PR+/HER2- tumor clustered together. The Ions with m/z 186.9 and m/z 303.2 were consistently related to NBP and IBC respectively. CONCLUSIONS: Chemical microscopy using DESI-MS is a powerful tool to identify specific lipid profiles across molecular subtypes of IBC and can contribute to understand breast cancer carcinogenesis. Citation Format: Victor P. Andrade, Adriana L. Santoro, Sabrina T. Moretti, Nicolas V. Schwab, Marcos N. Eberlin. In situ mass spectrometry using desorption electrospray ionization mass spectrometry (DESI-MS) can detect specific lipid profiles across breast cancer molecular subtypes. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3981.

CLINICOPATHOLOGICAL FEATURES OF NONSPECIFIC INVASIVE BREAST CANCER ACCORDING TO ITS MOLECULAR SUBTYPES

The aim of the present study was to investigate the clinical and morphological features of nonspecific invasive breast cancer according to its molecular subtypes. 163 women with nonspecific invasive breast cancer (T1-4N0-3M0) were included in the present study. Luminal A type of breast cancer was detected in 101 women, luminal B type - in 23 women, overexpression of HER2/neu was identified in 14 women and triple-negative cancer - in 25 women. The study revealed that various molecular subtypes of breast cancer differ in the morphological structure, the expression characteristics of the primary tumor and the rate of lymphogenous and hematogenous metastasis. Lymphogenous metastases were more frequently (in 71%) detected in HER2/neu overexpressing breast cancer than in luminal A (41%), luminal B (39%) and triple-negative tumors (40%). Hematogenous metastasis did not depend on the morphological structure of carcinoma infiltrative component, the state of tumor stroma as well as the proliferative activity in all the investigated groups. The revealed clinicopathological characteristics of different molecular subtypes of invasive breast cancer allow to predict the possible outcome of the disease and select personalized treatment strategy for patients more reasonably.

Tubular Carcinoma of the Breast: A Single Institution's Experience of a Favorable Prognosis

Tubular carcinoma is a subtype of invasive breast cancer that comprises 1 to 4 per cent of invasive breast cancers. Prior studies show nearly 100 per cent 15 year survival rate for tubular carcinoma compared to the 89.2 per cent five year survival of all breast cancers. These encouraging statistics beg the question should tubular cancers be treated as other invasive cancers, or can some patients be spared an invasive procedure or the side effects of adjuvant therapy? Fifty-seven cases of tubular carcinoma over 16 years were analyzed. All relevant aspects of the patient's history, treatment, and outcomes were documented. The aim was compare treatment outcomes of tubular breast cancer outcomes to that of all invasive breast cancers. Of the 57 patients, local recurrence was seen in two patients (3.5%) only one of which recurred as a tubular carcinoma (1.75%). There were no cancer-related mortalities. A look into our institution's data supported the notion that tubular carcinoma of the breast is a less aggressive histological type. Of our 57 cases, only two recurrences (3.5%) were noted and there were no cancer-related mortalities. Interestingly only one patient (1.75%) recurred as tubular carcinoma. Without controlling for adjuvant therapy, recommendations cannot be made for a less aggressive treatment plan at this point. Future randomized controlled trials may lead to a less aggressive treatment plan for this favorable subtype. On the basis of this study and others like it, physicians can give evidenced-based favorable prognosis with a diagnosis of tubular carcinoma of the breast.