Resected Breast Cancer Research Articles

A Thrombotic Biomarker-Based Risk Assessment Scoring Model for Disease Recurrence in Breast Cancer Patient Candidate to Systemic Chemotherapy

Background. Hypercoagulability, a common finding in cancer patients, is associated with an increased risk of both thrombosis and tumor development. The HYPERCAN study (ClinicalTrials.gov, ID# NCT02622815) is an ongoing Italian prospective, multicenter, observational study, evaluating the predictive value of thrombotic markers for early cancer diagnosis in healthy subjects and for cancer prognosis and venous thromboembolism in patients with newly diagnosed malignant disease. In this analysis of a large cohort of patients with breast cancer, we evaluated whether pre-chemotherapy thrombotic biomarker levels: 1. are associated with breast cancer-specific clinicopathological features; and 2. may predict for disease recurrence (DR). Patients and Methods. D-dimer, fibrinogen, prothrombin fragment 1+2 (F1+2), and FVIIa/antithrombin complex (FVIIa/AT) levels were measured in 701 early-stage resected breast cancer patients, candidate to adjuvant chemotherapy and prospectively enrolled in the HYPERCAN study. Significant prognostic parameters for DR were identified by Cox-regression multivariate analysis and used for generating a risk assessment model. The study protocol is approved by the local Ethics Committee. Informed written consent is obtained from all study subjects. Results. Increased pre-chemotherapy D-dimer, fibrinogen, and F1+2 levels were significantly associated with a large tumor size (≥ 5 cm) and lymph node positivity. After 3.4 years follow-up, 71 patients experienced a DR. Comparison of the levels of the hypercoagulation variables of patients who experienced relapse versus patients who remained disease-free during follow-up showed that there were no statistical differences for all, but F1+2 biomarker levels, which were significantly higher in the group patients who relapsed [223 (115-618) vs 197 (115-385) pmol/l; p=0.024]. In addition, correlation analyses showed that pre-chemotherapy levels of fibrinogen were significantly and inversely associated with time to relapse (β = -0.317; p=0.012). Cox-multivariate analysis identified F1+2 (HR 2; 95% CI, 1.1-3.6; p=0.019), tumor size ≥ 5cm (HR 2.6; 95% CI, 1.4-4.6; p=0.001), and Luminal B HER2-neg or TN molecular subtypes (HR 3.9; 95% CI, 2.1-7.5; p<0.001) as independent risk factors for DR. Based on these variables, we generated a risk assessment model that significantly identified patients at low- versus high-risk of DR (cumulative incidence: 6.2 vs 20.7%; HR=3.5; p<0.001). Conclusion. Our prospective laboratory data from the HYPERCAN breast cancer subjects were essential for generating a scoring model for DR risk assessment. Future investigations addressing the role of plasma thrombotic biomarkers in breast cancer patients' management are warranted and may provide the rationale for development of new therapeutic strategies. Project funded by AIRC "5xMILLE Multiunit extension program" n. 12237 grant from the "Italian Association for Cancer Research (AIRC)". Disclosures Santoro: Bayer: Consultancy, Speakers Bureau; Amgen: Speakers Bureau; BMS: Consultancy; Sandoz: Speakers Bureau; Eisai: Consultancy, Speakers Bureau; Novartis: Speakers Bureau; Lilly: Speakers Bureau; Celgene: Speakers Bureau; Servier: Consultancy, Speakers Bureau; Gilead: Consultancy, Speakers Bureau; AstraZeneca: Speakers Bureau; Roche: Speakers Bureau; Abb-Vie: Speakers Bureau; Pfizer: Consultancy, Speakers Bureau; Arqule: Consultancy, Speakers Bureau; MSD: Speakers Bureau; Takeda: Speakers Bureau; BMS: Speakers Bureau.

Quantitative Evaluation of Hormesis in Breast Cancer Using Histoculture Drug Response Assay.

Purpose:Hormesis is a phenomenon of growth stimulation at low doses and inhibition at higher doses. In cancer treatment, little is known about how hormesis affects cancer cell proliferation. We evaluated the hormetic dose–response relationship of paclitaxel using surgically resected breast cancer specimens on the basis of histoculture drug response assay (HDRA).Methods:We used surgically resected fresh tumor specimens from 22 patients with breast cancer: 17 invasive ductal, 3 mucinous, and 2 other “special-type” cancers. All patients were female, ranging in age between 40 and 86 (median 60) years. Small pieces of viable cancer tissue were placed on collagen gel and cultured for 7 days with paclitaxel. Inhibition rates of paclitaxel at several concentrations were measured and fitted to a sigmoid dose–response curve.Results:Hormesis was observed in 9 of the 22 cases; ED50 of cytotoxic effect was significantly higher (P = .0036) in hormesis (H) group (44.6 ± 4.2 µg/mL) than in nonhormesis (N) group (26.7 ± 3.5 μg/mL).Conclusion:We evaluated hormesis in breast cancer tissue using HDRA for the first time although previously confirmed in cultured cells. Hormesis seems to occur in patients undergoing treatment with anticancer agents, especially in a metastatic setting. Meanwhile, tumor growth may be stimulated in patients who are resistant to paclitaxel.

Thrombotic biomarkers for risk prediction of malignant disease recurrence in patients with early stage breast cancer.

In cancer patients, hypercoagulability is a common finding. It has been associated with an increased risk of venous thromboembolism, but also to tumor proliferation and progression. In this prospective study of a large cohort of breast cancer patients, we aimed to evaluate whether pre-chemotherapy abnormalities in hemostatic biomarkers levels: (i) are associated with breast cancer-specific clinico-pathological features; and (ii) can predict for disease recurrence. D-dimer, fibrinogen, prothrombin fragment 1+2, and FVIIa/antithrombin levels were measured in 701 early-stage resected breast cancer patients candidate to adjuvant chemotherapy and prospectively enrolled in the HYPERCAN study. Significant prognostic parameters for disease recurrence were identified by Cox regression multivariate analysis and used for generating a risk assessment model. Pre-chemotherapy D-dimer, fibrinogen, and pro-thrombin fragment 1+2 levels were significantly associated with tumor size and lymph node metastasis. After 3.4 years of follow up, 71 patients experienced a recurrence. Cox multivariate analysis identified prothrombin fragment 1+2, tumor size, and Luminal B HER2-negative or triple negative molecular subtypes as independent risk factors for disease recurrence. Based on these variables, we generated a risk assessment model that significantly differentiated patients at low- and high-risk of recurrence (cumulative incidence: 6.2 vs. 20.7%; Hazard Ratio=3.5; P<0.001). Our prospective clinical and laboratory data from the HYPERCAN study were crucial for generating a scoring model for assessing risk of disease recurrence in resected breast cancer patients, candidate to systemic chemotherapy. This finding stimulates future investigations addressing the role of plasma prothrombin fragment 1+2 in the management of breast cancer patients to provide the rationale for new therapeutic strategies. (The HYPERCAN study is registered at clinicaltrials.gov identifier 02622815.)

Cross comparison and prognostic assessment of breast cancer multigene signatures in a large population-based contemporary clinical series

Multigene expression signatures provide a molecular subdivision of early breast cancer associated with patient outcome. A gap remains in the validation of such signatures in clinical treatment groups of patients within population-based cohorts of unselected primary breast cancer representing contemporary disease stages and current treatments. A cohort of 3520 resectable breast cancers with RNA sequencing data included in the population-based SCAN-B initiative (ClinicalTrials.gov ID NCT02306096) were selected from a healthcare background population of 8587 patients diagnosed within the years 2010–2015. RNA profiles were classified according to 19 reported gene signatures including both gene expression subtypes (e.g. PAM50, IC10, CIT) and risk predictors (e.g. Oncotype DX, 70-gene, ROR). Classifications were analyzed in nine adjuvant clinical assessment groups: TNBC-ACT (adjuvant chemotherapy, n = 239), TNBC-untreated (n = 82), HER2+/ER− with anti-HER2+ ACT treatment (n = 110), HER2+/ER+ with anti-HER2 + ACT + endocrine treatment (n = 239), ER+/HER2−/LN− with endocrine treatment (n = 1113), ER+/HER2−/LN− with endocrine + ACT treatment (n = 243), ER+/HER2−/LN+ with endocrine treatment (n = 423), ER+/HER2−/LN+ with endocrine + ACT treatment (n = 433), and ER+/HER2−/LN− untreated (n = 200). Gene signature classification (e.g., proportion low-, high-risk) was generally well aligned with stratification based on current immunohistochemistry-based clinical practice. Most signatures did not provide any further risk stratification in TNBC and HER2+/ER– disease. Risk classifier agreement (low-, medium/intermediate-, high-risk groups) in ER+ assessment groups was on average 50–60% with occasional pair-wise comparisons having <30% agreement. Disregarding the intermediate-risk groups, the exact agreement between low- and high-risk groups was on average ~80–95%, for risk prediction signatures across all assessment groups. Outcome analyses were restricted to assessment groups of TNBC-ACT and endocrine treated ER+/HER2−/LN− and ER+/HER2−/LN+ cases. For ER+/HER2− disease, gene signatures appear to contribute additional prognostic value even at a relatively short follow-up time. Less apparent prognostic value was observed in the other groups for the tested signatures. The current study supports the usage of gene expression signatures in specific clinical treatment groups within population-based breast cancer. It also stresses the need of further development to reach higher consensus in individual patient classifications, especially for intermediate-risk patients, and the targeting of patients where current gene signatures and prognostic variables provide little support in clinical decision-making.

Justification for preservation of the projected skin flap above the tumor during surgical treatment of primary resectable breast cancer

Aim. Improvement of the aesthetic outcomes of reconstructive plastic surgery for nodular breast cancer due to the scientific rationale of conservation of the projected skin flap and moving the postoperative scar to aesthetically acceptable zones.&#x0D; Methods. Data of clinical and instrumental methods of investigation of 240 patients with primary resectable breast cancer were analyzed. The probability of involvement of the front layer of the superficial fascia in the projection of a malignant tumor depending on its clinical and topographic anatomical figures was estimated.&#x0D; Results. The study revealed that the fascia involvement does not depend on the skin flattening. The skin flatte­ning is an indirect sign of superficial tumors having a probability of intimate adherence of the tumor to the fascia of 31.8±0.401%. In turn, the intimate adherence of the tumor to the front layer of the superficial fascia with a pro­bability of its involvement of 39.3±0.41% is a significant risk factor for the projected skin flap involvement. It was also found that involvement of the fascia studied does not depend neither on the size of the tumor, nor on the nodal status, nor on the biological subtype. The key risk factor is the depth of the tumor.&#x0D; Conclusion. The indication for the projection skin flap preservation is the location of a malignant tumor at a depth of more than 1.05 cm, at which the probability of affecting the fascia is statistically insignificant and is 1.7±0.1%, which requires placing the subcutaneous tissue resection line in the tumor projection from the side of preserved skin flap.

Justification for preservation of the projected skin flap above the tumor during surgical treatment of primary resectable breast cancer

Aim. Improvement of the aesthetic outcomes of reconstructive plastic surgery for nodular breast cancer due to the scientific rationale of conservation of the projected skin flap and moving the postoperative scar to aesthetically acceptable zones.&#x0D; Methods. Data of clinical and instrumental methods of investigation of 240 patients with primary resectable breast cancer were analyzed. The probability of involvement of the front layer of the superficial fascia in the projection of a malignant tumor depending on its clinical and topographic anatomical figures was estimated.&#x0D; Results. The study revealed that the fascia involvement does not depend on the skin flattening. The skin flatte­ning is an indirect sign of superficial tumors having a probability of intimate adherence of the tumor to the fascia of 31.8±0.401%. In turn, the intimate adherence of the tumor to the front layer of the superficial fascia with a pro­bability of its involvement of 39.3±0.41% is a significant risk factor for the projected skin flap involvement. It was also found that involvement of the fascia studied does not depend neither on the size of the tumor, nor on the nodal status, nor on the biological subtype. The key risk factor is the depth of the tumor.&#x0D; Conclusion. The indication for the projection skin flap preservation is the location of a malignant tumor at a depth of more than 1.05 cm, at which the probability of affecting the fascia is statistically insignificant and is 1.7±0.1%, which requires placing the subcutaneous tissue resection line in the tumor projection from the side of preserved skin flap.

Combined Fascin-1 and MAP17 Expression in Breast Cancer Identifies Patients with High Risk for Disease Recurrence.

Breast cancer stem cells are considered to be a major cause of disease recurrence in breast cancer as they appear to be chemoresistant. Fascin-1 and MAP17 are stem cell markers whose excessive expression in tumors is associated with aggressive tumor phenotypes. The aim of the present study was to investigate the expression patterns of fascin-1 and MAP17 in breast cancer and to assess their clinical significance. Expression of fascin-1 and MAP17 was assessed via immunohistochemistry in surgical specimens of a cohort comprised of 127 patients with resectable breast cancer. Results were correlated with clinicopathological characteristics and survival data. Progression-free survival (PFS) was defined as the primary outcome of the present study. Fascin-1 and MAP17 expression were strongly associated with the presence of triple-negative cancers (p<0.0001). Tumors displaying high expression of fascin-1 presented correlations with high tumor grade (p=0.002) and high expression of Ki-67 (p=0.004). PFS of patients exhibiting high expression of fascin-1 and MAP17 in cancer cells in the first 5years after surgery was significantly worse than in patients with low expression of the two markers (47.8%, 95% confidence interval [CI] 33-51 vs. 80.5%, 95% CI 47-56; p=0.012) and independent of other clinicopathological characteristics (hazard ratio 0.171, 95% CI 0.034-0.869; p=0.033). Combined expression of fascin-1 and MAP17 in breast cancer cells is associated with a significantly worse 5-year PFS, therefore recognizing a group of patients with high risk for early disease recurrence.

A Retrospective Propensity Score Matched Study of the Preoperative C-Reactive Protein to Albumin Ratio and Prognosis in Patients with Resectable Non-Metastatic Breast Cancer

BackgroundA retrospective study aimed to investigate the association between the CRP to albumin ratio and prognosis in patients with resectable non-metastatic breast cancer in terms of disease-free survival (DFS) and overall survival (OS) using propensity score matching.Material/MethodsPatients with newly diagnosed resectable non-metastatic breast cancer (n=200) who underwent modified radical mastectomy between January 2008 to June 2013 included a group with an increased CRP to albumin ratio ≥0.029 (n=80) and a group with reduced CRP to albumin ratio <0.029 (n=120). Propensity score matching was used to estimate the prognostic role of the CRP to albumin ratio, and a 1: 1 matching using four covariates was performed to overcome selection bias. The prognostic significance of the CRP to albumin ratio was analyzed using receiver operating characteristic (ROC) curves. Kaplan-Meier survival analysis and a Cox proportional hazards model were conducted to identify the impact on DFS and OS.ResultsAn increased CRP to albumin ratio was associated with increased age, post-menopausal status, and a high risk of recurrence or death in patients with breast cancer. An increased preoperative CRP to albumin ratio was significantly associated with reduced disease-free survival (DFS) and overall survival (OS) (all P<0.05). Multivariate analysis showed that an increased CRP to albumin ratio was an independent risk factor for long-term outcome and predicted reduced DFS (HR, 2.225; P=0.024) and OS (HR, 9.189; P=0.003).ConclusionsPreoperative evaluation of the CRP to albumin ratio was an independent prognostic indicator in patients with resectable breast cancer.

A prognostic nomogram to predict the benefit of adjuvant radiotherapy for resected inflammatory breast cancer patients.

e12007 Background: A nomogram is a convenient and reliable tool for individualizing cancer prognosis. Therefore, the purpose of this study was to construct a clinical nomogram for predicting overall mortality (OM) to help clinicians to make individualized estimates of the potential benefit of adjuvant radiotherapy (RT) for resected inflammatory breast cancer (IBC) patients. Methods: We used the Surveillance, Epidemiology, and End Results database to identify 4030 IBS patients treated with surgery between 2004 and 2012, and randomly classified the patients into the discovery (n = 2417) and validation (n = 1613) cohorts. A Cox regression hazards model was used to build a nomogram by determining the prognostic factors for OM in the discovery cohort. The overall accuracy and discriminative ability was measured by Brier score and Harrell's concordance index (c-index). Results: In multivariate analysis, age, race, number of primaries, N stage, estrogen receptor, progesterone receptor status, and marital status, were the independent predictive factors of OM. These variables were selected to generate the nomogram. The calibration curves for 2-, and 5-year OM rate showed excellent agreement between prediction and actual observation. The Brier score and c-index demonstrated the nomogram performed better than the 7th edition TNM staging system for predicting OM possibility. In addition, the nomogram predicted that the magnitude of benefit from adjuvant RT for an individual IBS patient could vary, the certain subgroup of patients (prognostic score: 0-25) would not gain benefit from adjuvant RT, while RT had more pronounced effects in participants with higher prognostic score (prognostic score: ≥25). Conclusions: This nomogram is a reliable prognostic method for predicting OM in IBC patients after surgical resection, which could be used to predict which IBC patients may benefit from adjuvant RT after surgery.

Prognostic value of Tfh-like cells in breast cancer.

e14285 Background: The treatment of breast cancer, one of the most common malignant tumors for female, focuses on the combined treatments based on subtypes including Luminal A, Luminal B, HER2 overexpression and triple negative. Given that none of those subtypes takes into consideration of immunological parameters, it is valuable to explore the biological characteristics and prognostic value of immune cells in breast caner. T follicular helper cells (Tfh cells) have been identified in different types of tumors with rare datas in breast cancer. One of them indicated the positive prognostic value of 8-gene Tfh signature. Exploration of Tfh cells may provide new clues to the potential immunotherapies in breast cancer. Methods: Freshly resected invasive breast cancer tissue from Fudan University Shanghai Cancer Center were collected during 06/2015 to 05/2017. Patients were divided into Luminal A (N = 61), Luminal B (HER2-) (N = 138), Luminal B (HER2+) (N = 72), HER2+ (non-luminal) (N = 57) and triple negative (N = 58) based on St Gallen International Expert Consensus. Results: A subpopulation of PD1hiCD4+ T cells in the tumor tissues of breast cancer was identified as Tfh-like cells, with the specific expression of Bcl-6 and CXCL13, confirmed by IHC. Phenotypes of those cells were checked by flow cytometry. Tfh-like cells were of a high level of ICOS but negative for CXCR5, suggesting that they were atypical Tfh cells. Data showed that high grade (N = 166; mean±SEM, 17.62±0.87) compared with low grade (N = 220; mean±SEM, 12.41±0.46) or high Ki-67 level (N = 300; mean±SEM, 15.38±0.58) with low Ki-67 level (N = 86; mean±SEM, 12.10±0.67), and negative ER expression (N = 115; mean±SEM, 17.47±1.05) compared with positive ER expression (N = 271; mean±SEM, 13.46±0.50) or negative PR expression (N = 177; mean±SEM, 16.87±0.83) compared with positive PR expression (N = 209; mean±SEM, 12.77±0.50) was associated with higher frequency of Tfh-like cells (P &lt; 0.01 to all). Patients with triple negative had higher frequency of Tfh-like cells than those with Luminal A (difference, 6.83; P = 0.0006) and Luminal B (HER2-) (difference, 4.61; P = 0.0124). Patients with HER2+ (non-luminal) had higher frequency of Tfh-like cells than those with Luminal A (difference, 5.34; P = 0.0146). These results indicate that higher frequency of Tfh-like cells could have negative prognostic influence. Conclusions: Our study defined a PD1hiBcl6+CXCL13+CD4+ T cell subpopulation, which specifically secretes cytokine IL-21 as Tfh-like cells. Higher frequency of Tfh-like cells is more likely to be seen in patients with poor prognosis.

63P - Controlling nutritional status (CONUT) score is a prognostic factor in patients with resected breast cancer

63P - Controlling nutritional status (CONUT) score is a prognostic factor in patients with resected breast cancer

Predicting breast cancer response to neoadjuvant chemotherapy based on tumor vascular features in needle biopsies.

In clinical breast cancer intervention, selection of the optimal treatment protocol based on predictive biomarkers remains an elusive goal. Here, we present a modeling tool to predict the likelihood of breast cancer response to neoadjuvant chemotherapy using patient specific tumor vasculature biomarkers. A semi-automated analysis was implemented and performed on 3990 histological images from 48 patients, with 10-208 images analyzed for each patient. We applied a histology-based model to resected primary breast cancer tumors (n = 30), and then evaluated a cohort of patients (n = 18) undergoing neoadjuvant chemotherapy, collecting pre- and post-treatment pathology specimens and MRI data. We found that core biopsy samples can be used with acceptable accuracy (r = 0.76) to determine histological parameters representative of the whole tissue region. Analysis of model histology parameters obtained from tumor vasculature measurements, specifically diffusion distance divided by radius of drug source (L/rb) and blood volume fraction (BVF), provides a statistically significant separation of patients obtaining a pathologic complete response (pCR) from those that do not (Student's t-test; P < 0.05). With this model, it is feasible to evaluate primary breast tumor vasculature biomarkers in a patient specific manner, thereby allowing a precision approach to breast cancer treatment.

Avoiding over- and undertreatment in patients with resected node-positive breast cancer with the use of gene expression signatures: are we there yet?

Prediction of benefit from adjuvant chemotherapy following resection of early breast cancer and, as a result, proper selection of candidates remains an elusive goal since the relative magnitude of benefit is the same regardless of the presence of clinicopathologic factors. Multiple studies, including randomized trials, establish the role of certain gene expression signatures in node-negative disease since they predict the risk of breast cancer relapse being so low that adjuvant chemotherapy can be omitted. In contrast, more limited data are available in higher risk, node-positive breast cancer patients, making the exclusion of adjuvant chemotherapy potentially hazardous. ‘Prospective–retrospective’ studies and limited prospective data show that several signatures, namely Oncotype Dx, MammaPrint, Prosigna, EndoPredict and Breast Cancer Index, select with different levels of success node-positive patients at very low risk for distant recurrence despite not receiving chemotherapy, although the long-term follow-up is still awaited. Pending, however the publication of the results from ongoing randomized studies which enroll patients with node-positive disease, major caution is warranted. Improper use and misinterpretation of these transcriptomic profiles can lead to undertreatment and exposure of patients to unnecessary risks resulting in increased breast cancer mortality for patients with axillary node-positive disease. With this review we critically discuss the available data on gene expression signatures that are used in clinical practice and offer practical recommendations regarding the management of patients with ER-positive, human epidermal growth factor receptor 2 (HER2)-negative, node-positive breast cancer.

Abstract P4-06-12: Biological characteristics and prognostic value of Tfh-like cells in the tumor tissue of breast cancer

Abstract Objective T follicular helper (Tfh) cells formed in germinal center are a distinct subpopulation of CD4+ T cell, with high levels of CXCR5, ICOS, PD-1, Bcl-6 and CXCL13 as their characteristic markers. Secreting IL-21 and helping humoral response are their key features. They have been identified in different types of tumors. However, it still remains unclear in breast cancer. Researches of them may provide new clues to the diagnosis and treatment of breast cancer, especially for the potential immunotherapies. This study was performed to explore the existence and a possible role of Tfh cells in breast cancer. Method Freshly resected invasive breast cancer tissue from Fudan University Shanghai Cancer Center (FUSCC) were collected, with 27 samples for identifying the presence, phenotype and cytokine-producing capacity of Tfh-like cells and 386 for exploring the correlation between Tfh-like cells and clinicopathological characteristics in breast cancer during 03/2015 to 05/2017. Of all those 386 samples, patients were divided into Luminal A (N=61), Luminal B (HER2-) (N=138), Luminal B (HER2+) (N=72), HER2+ (non-luminal) (N=57) and triple negative (N=58) based on St Gallen International Expert Consensus. Data was analyzed based on the result from Multiparameter Flow Cytometry. The correlation between Tfh-like cells and clinicopathological characteristics in breast cancer was examined with t test or one way ANOVA with Tukey's multiple comparisons test. Results A subpopulation of PD1hiCD4+ T cells in tumor tissues of breast cancer was identified as Tfh-like cells, with the specific expression of Bcl-6 and CXCL13. Phenotypes of those cells were checked by flow cytometry. They were of a high level of ICOS but negative for CXCR5, suggesting that they were atypical Tfh cells. They had high levels of activated molecules such as CD38, CD71, CD95 and HLADR, showing that they were highly activated. They had high levels of suppressive markers as Tim3, TIGIT, and LAG3, indicating they could have regulatory functions. In addition, Tfh-like cells specifically secreted cytokine IL-21 with stimulation. Data showed that high grade (N=166; mean±SEM, 17.62±0.87) compared with low grade (N=220; mean±SEM, 12.41±0.46) or high Ki-67 level (N=300; mean±SEM, 15.38±0.58) with low Ki-67 level (N=86; mean±SEM, 12.10±0.67), and negative ER expression (N=115; mean±SEM, 17.47±1.05) compared with positive ER expression (N=271; mean±SEM, 13.46±0.50) or negative PR expression (N=177; mean±SEM, 16.87±0.83) compared with positive PR expression (N=209; mean±SEM, 12.77±0.50) was associated with higher frequency of Tfh-like cells (P&amp;lt;0.01 to all). Patients with triple negative had higher frequency of Tfh-like cells than those with Luminal A (difference, 6.83; P=0.0006) and Luminal B (HER2-) (difference, 4.61; P=0.0124). Patients with HER2+ (non-luminal) had higher frequency of Tfh-like cells than those with Luminal A (difference, 5.34; P=0.0146). These results indicate that higher frequency of Tfh-like cells could have negative prognostic influence. Conclusion Our study defined a PD1hiBcl6+CXCL13+CD4+ T cell subpopulation, which specifically secretes cytokine IL-21 as Tfh-like cells. Higher frequency of Tfh-like cells is more likely to be seen in patients with poor prognosis. Citation Format: Ma L-X, Guo L, Goswami S, Li T, Cao C-M, Zhang X-m, Wu J. Biological characteristics and prognostic value of Tfh-like cells in the tumor tissue of breast cancer [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P4-06-12.

Abstract P1-13-09: Long-term follow-up of two randomized controlled trials (N-SAS-BC01 trial and CUBC trial) comparing oral tegafur-uracil (UFT) versus classical cyclophosphamide, methotrexate, and 5-fluorouracil (CMF) as adjuvant therapy in early breast cancer

Abstract Background: Two randomized controlled trials comparing the efficacy of oral tegafur-uracil (UFT) (2 years) with that of classical cyclophosphamide, methotrexate, and 5-fluorouracil (CMF) (6 courses) were conducted in patients with resected early breast cancer. UFT is an oral fluoropyrimidine that combines tegafur, a prodrug of 5-fluorouracil, with uracil in a 1:4 molar ratio. One study was the N-SAS-BC01 trial (Watanabe et al, J Clin Oncol 2009), which was conducted in patients with high-risk node-negative breast cancer (n=733). The other was the CUBC trial (Park et al, Br J Cancer 2009), which was performed in patients with node-positive breast cancer (n=377). We reported the pooled analysis of these two randomized control trials using individual patient data (Ohashi et al, Breast Cancer Res Treat 2010). This pooled analysis showed that UFT was non-inferior to CMF in terms of inhibiting recurrence of estrogen receptor (ER)-positive early breast cancer. In addition, an exploratory subgroup analysis showed that UFT may be more effective in ER-positive patients who were 50 years or older. The present study was conducted to investigate the long-term efficacy of UFT or CMF in patients with early breast cancer. Methods: Long-term follow-up data for disease recurrence and survival were collected. Hazard ratios (HR) were determined using the Cox model stratified by study and adjusted for clinical characteristics, namely age, tumor size, nodal status, histological type, ER, and progesterone receptor (PgR). Survival curves were estimated by the Kaplan-Meier method. Hochberg multiplicity adjustment was applied in the previous pooled analysis, and non-inferiority of UFT versus CMF was shown in terms of relapse-free survival (RFS) in the ER-positive patient group. We investigated the non-inferiority of UFT in the same patient group with updated data. Restricted mean survival time (RMST) was calculated to consider the relative efficacy of UFT. This study is registered with JapicCTI-163321. Results: In total, 1,057 patients were analyzed (CMF, n=528; UFT n=529). The median follow-up time was 11.1 years (12.1 years in the N-SAS-BC 01 trial and 8.3 years in the CUBC trial). Table 1 shows the 10-year RFS and overall survival (OS) in all patients and ER-positive patients. The difference in RMST between arms at 20 years in terms of RFS was -2.7 months in all patients and 3.4 months in ER-positive patients. Table 2 shows the HR for RFS according to ER status and age. UFT (%)CMF (%)UFT to CMF; HR (95% CI)10-year RFSall patients74.477.11.02 (0.81–1.30)ER-positive patients75.075.10.91 (0.66–1.27)10-year OSall patients86.886.90.98 (0.72–1.34)ER-positive patients89.987.90.86 (0.54–1.37) Age &amp;lt;50 yearsAge ≥50 yearsER negative1.76 (0.95–3.26)0.93 (0.58–1.51)ER positive1.29 (0.74–2.23)0.76 (0.50–1.16) Conclusion: This study shows that UFT was shown to be non-inferior to CMF in terms of RFS in ER-positive early breast cancer. This result is similar to that of the previous pooled analysis. Sponsor: Taiho Pharmaceutical Co., LTD Citation Format: Yonemori K, Ohsumi S, Takao S, Tokuda Y, Ito Y, Nakagami K, Takahashi M, Yoshidome K, Nakayama T, Yamaguchi Y, Ohashi Y, Inaji H, Watanabe T. Long-term follow-up of two randomized controlled trials (N-SAS-BC01 trial and CUBC trial) comparing oral tegafur-uracil (UFT) versus classical cyclophosphamide, methotrexate, and 5-fluorouracil (CMF) as adjuvant therapy in early breast cancer [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr P1-13-09.

Abstract PD9-12: Integrative molecular profiling of breast cancer brain metastasis and patient-derived xenograft organoids from resected breast cancer brain metastases to interrogate and prioritize therapeutic personalized strategies

Abstract Background: Breast cancer brain metastasis (BM) is an area of unmet need in metastatic breast cancer patients. Novel therapeutic interventions to help prevent and treat BM are warranted. We conducted integrative molecular profiling of BM and matched primary tumors (PT) using next-generation DNA and RNA sequencing to examine the molecular landscape. In addition, we established patient-derived xenograft/organoid (PDX/PDO) to examine drug sensitivity according to the molecular and clinical features of the BM. Methods: Archived, formalin fixed paraffin-embedded BM was collected retrospectively. BM were also collected prospectively at the time of clinically indicated surgical resection through the central nervous system tissue banking and the Michigan Oncology Sequencing Center (MI-ONCOSEQ) protocols. Matched archived PT tissues were collected when available. Integrative next-generation sequencing was conducted using the MI-ONCOSEQ platform. The prospectively collected BM were further used to establish PDXs/ PDOs. Successfully established PDXs/PDOs were used for ex vivo drug testing via MiDrugScreen, a novel drug sensitivity testing platform, where testing was performed in a dose-response format with drug selection prioritized by clinical scenario and molecular alterations if known a priori. Results: 12 matched BM-PT pairs were analyzed: 6 triple negative, 5 HER2 positive, and 1 ER positive HER2 negative. All except one (11/12) had TP53 mutations. When present, TP53 mutations in BM were also found in PT (except for 1 unknwon case in PT due to low coverage). ER+HER2- was the only one without TP53 mutation but had hyper-mutation (APOBEC signature). Driver mutations and unique copy number alterations (CDKN2A loss in 1/12, mutations in PIK3CA in 1/12 and ESR1 in 1/12, CCNE1 amplification in 1/12) were noted in BMs. In 75% of cases, mutational burden was higher in BM vs. PT. 2 PDX/PDO were available for drug testing. PDO-BC9 was noted to have RB1 (splice acceptor) and LOH. As predicted by this alteration, PDO-BC9 was insensitive to CDK4/6 inhibitors (palbociclib, abemaciclib) tested on MiDrugScreen panel. PDX-BC4 was established from PIK3CA and ESR1 mutated BM from an ER+HER2- patient who had previously progressed on endocrine therapy with a CDK4/6 inhibitor. As predicted, the PDX-BC4 was resistant to CDK4/6 inhibitor but interestingly sensitive to PIK3CA, ERK, and MEK inhibitors. Conclusions: TP53 mutation was highly prevalent and may be a biomarker for increased risk of BM. Further study is warranted to see if specific TP53 mutations are associated with a risk of BM development and can be used in risk stratification for BM specific intervention. Unique molecular alterations in BM compared to matched PT may have a therapeutic implication as a target or resistance biomarker. Conducting drug testing in addition to molecular profiling has the strong potential of being informative in tailoring or prioritizing therapeutic agents in the era of precision medicine. Additional BM PDXs/PDOs from breast and other solid tumors are being examined using this novel therapeutic tailoring approach with the combination of MIONCOSEQ and MiDrugScreen. Citation Format: Morikawa A, Robinson DR, Soellner M, Wu Y-M, Lonigro R, Gilani R, Cheng X, Lachacz E, Thomas D, McMurray K, Smerage J, Henry NL, Heth J, Chinnaiyan A, Hayes DF, Merajver S. Integrative molecular profiling of breast cancer brain metastasis and patient-derived xenograft organoids from resected breast cancer brain metastases to interrogate and prioritize therapeutic personalized strategies [abstract]. In: Proceedings of the 2018 San Antonio Breast Cancer Symposium; 2018 Dec 4-8; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2019;79(4 Suppl):Abstract nr PD9-12.

Effects of pessimism, depression, fatigue, and pain on functional health-related quality of life in patients with resected non-advanced breast cancer

IntroductionThe aim was to analyze the effects of pessimism, depression, fatigue, and pain on functional health-related quality of life (HRQoL) in patients with resected, non-advanced breast cancer. MethodsA prospective, multicenter study was conducted in 440 breast cancer patients. They completed the Brief Symptom Inventory (BSI), the European Organization for Research and Treatment of Cancer QoL-Questionnaire-Core-30 (EORTC-QLQ-C30), and the Revised Life Orientation Test (LOT-R). ResultsPrevalence rates of pessimism and depression were 23.3% and 40.0%, respectively. Fatigue and pain were the most common symptoms, 8.8% and 4.2%, respectively. Patients without a partner were more pessimistic that partnered ones; those with a lower level of education and subjects without a partner exhibited more depression. Depression was a major factor that proved to have the greatest explanatory power for HRQoL (physical, role, emotional, cognitive, and social functioning) and global health status (R2 range: 0.13 to 0.39). Of the five domains, fatigue had a significant effect on four and pain, on two. ConclusionThis study reveals the impact of depression and pessimism on physical, psychological, social, and quality-of-life aspects and the importance of evaluating them in patients who are going to initiate adjuvant chemotherapy for breast cancer.

Tumor-Infiltrating Lymphocytes in Breast Cancer. Association with Clinical and Pathological Parameters.

In patients with primary resectable breast cancer, a positive correlation between the age and the count of CD16+ lymphocytes and a negative correlation of this parameter with the number of regulatory CD4+CD25+CD127- cells and proliferative activity of Ki-67 tumor cells were revealed. Higher level of Ki-67 was associated with reduced number of effector lymphocytes (CD8+ and CD16+) and elevated content of regulatory CD8+CD11b-CD28- T cells. The absence of expression of estrogen receptors was associated with reduced cytotoxic potential of CD8+ T cell in comparison with ER+ breast cancer. The percentage of CD8+ lymphocytes (CD3+CD8+ and CD8+CD11b+CD28+) among lymphocytes infiltrating the tumor was higher in PR+ breast cancer than in PR- tumors. With increasing the tumor load, the number of lymphocytes expressing CD16 marker and their cytotoxic potential decreased.

Loss of Human Leukocyte Antigen Class I Expression Is Associated with Poor Prognosis in Patients with Advanced Breast Cancer

BackgroundHuman leukocyte antigen class I (HLA-I) molecules play important roles in regulating immune responses. Loss or reduction of HLA-I expression has been shown to be associated with prognosis in several cancers. Regulatory T-cells (Tregs) also play critical functions in immune response regulation. Evaluation of HLA-I expression status by the EMR8-5 antibody and its clinical impact in breast cancer have not been well studied, and its relationship with Tregs remains unclear.MethodsWe evaluated HLA-I expression and Treg infiltration by immunohistochemistry in 465 surgically resected breast cancer samples. We examined the correlation between HLA-I expression and Treg infiltration and clinicopathologic characteristics and survival analyses were performed.ResultsTotal loss of HLA-I expression was found in 84 breast cancer samples (18.1%). Univariate survival analysis revealed that loss of HLA-I expression was significantly associated with worse disease-specific survival (DSS) (p = .029). HLA-I was not an independent prognostic factor in the entire patient group, but it was an adverse independent prognostic factor for DSS in patients with advanced disease (stage II–IV) (p = .031). Treg numbers were significantly higher in the intratumoral stroma of HLA-I–positive tumors than in HLA-I–negative tumors (median 6.3 cells/high power field vs 2.1 cells/high power field, p < .001). However, Tregs were not an independent prognostic factor in our cohort.ConclusionsOur findings suggest that the loss of HLA-I expression is associated with poor prognosis in breast cancer patients, highlighting the role of HLA-I alterations in immune evasion mechanisms of breast cancer. HLA-I could be a promising marker that enables the application of more effective and precise immunotherapies for patients with advanced breast cancer.

Comorbidity Assessment in the National Cancer Database for Patients With Surgically Resected Breast, Colorectal, or Lung Cancer (AFT-01, -02, -03).

Accurate comorbidity measurement is critical for cancer research. We evaluated comorbidity assessment in the National Cancer Database (NCDB), which uses a code-based Charlson-Deyo Comorbidity Index (CCI), and compared its prognostic performance with a chart-based CCI and individual comorbidities in a national sample of patients with breast, colorectal, or lung cancer. Through an NCDB Special Study, cancer registrars re-abstracted perioperative comorbidities for 11,243 patients with stage II to III breast cancer, 10,880 with stage I to III colorectal cancer, and 9,640 with stage I to III lung cancer treated with definitive surgical resection in 2006-2007. For each cancer type, we compared the prognostic performance of the NCDB code-based CCI (categorical: 0 or missing data, 1, 2+), Special Study chart-based CCI (continuous), and 18 individual comorbidities in three separate Cox proportional hazards models for postoperative 5-year overall survival. Comorbidity was highest among patients with lung cancer (13.2% NCDB CCI 2+) and lowest among patients with breast cancer (2.8% NCDB CCI 2+). Agreement between the NCDB and Special Study CCI was highest for breast cancer (rank correlation, 0.50) and lowest for lung cancer (rank correlation, 0.40). The NCDB CCI underestimated comorbidity for 19.1%, 29.3%, and 36.2% of patients with breast, colorectal, and lung cancer, respectively. Within each cancer type, the prognostic performance of the NCDB CCI, Special Study CCI, and individual comorbidities to predict postoperative 5-year overall survival was similar. The NCDB underestimated comorbidity in patients with surgically resected breast, colorectal, or lung cancer, partly because the NCDB codes missing data as CCI 0. However, despite underestimation of comorbidity, the NCDB CCI was similar to the more complete measures of comorbidity in the Special Study in predicting overall survival.