Recurrence In Early Breast Cancer Research Articles

Abstract P3-08-14: Prediction of distant recurrence in low risk early breast cancer by RT-qPCR based subtyping using MammaTyper®

Abstract Background: Estrogen receptor (ER/ESR1), progesterone receptor (PR/PGR) and epidermal growth factor receptor 2 (HER2/ERBB2) are routinely assessed by immunohistochemistry (IHC) during workup of breast cancer samples. The routine use of Ki67 (MKI67) IHC assessment in the context of breast cancer subtyping, however, remains controversial, due to poor reproducibility and lack of standardization. The MammaTyper® test is an in-vitro diagnostic (IVD) test which measures the expression status of the four breast cancer biomarkers ERBB2, ESR1, PGR and MKI67 on the mRNA level via reverse transcription-quantitative PCR (RT-qPCR) and has demonstrated a high degree of reproducibility in the assessment of the four markers. In this retrospective analysis we assessed the prognostic power of molecular subtyping by MammaTyper® in archived samples from low risk early breast cancers treated with adjuvant endocrine therapy only. Methods: ER+/HER2- (according to initial diagnosis) FFPE breast cancer samples from patients treated with adjuvant endocrine therapy only were obtained from 6 different centers. Tumor cellularity was assessed by H&E staining and RNA was extracted from samples with a tumor cell content of ≥20% using a bead-based RNA purification kit (RNXtract®). Total RNA was then used as input for MammaTyper® RT-qPCR. Expression values were classified as positive or negative for each marker based on predefined cutoff values. Tumor subtypes were assigned to each sample based on the combination of binary (pos/neg) single marker expression status according the St Gallen surrogate subtype definition. Distant disease free survival of Luminal A-like samples vs. samples with other subtypes was assessed by Kaplan Meier analysis and Cox regression using SAS version 9.4. Results: The final analysis included 319 samples with sufficient tumor cellularity and RNA content for reliable analysis. The rate of distant recurrence in the analyzed set was 8.5%. Median follow up was 7.8 years. The MammaTyper® test called 60% (192) of samples as Luminal-A-like (4.7% (9) distant events), 37% (119) as Luminal B-like (HER2 negative) (13.4% (16) distant events), 1.3% (4) as Triple negative (ductal) (25% (1) distant events), 0.6% (2) as “not defined according to St Gallen” (ESR1-/PGR+) (50% (1) distant events) and 0.6% (2) as Luminal B-like (HER2 positive) (no distant events). When comparing Luminal A-like samples with the samples of the other subtypes in survival analysis, Luminal A-like samples had a significantly better distant disease free survival when assessing samples from patients with pN0 status (278) (p=0.0177, HR=0.344 (95% CI 0.137-0.866), pN1 status (0-3 affected nodes) (314) (p=0.0153, HR=0.374 (95% CI 0.163-0.855) as well as for all samples (p=0.0032, HR=0.319 (95% CI 0.143-0.711). Conclusion: Determination of HER2, ER, PR and Ki67 mRNA levels allows molecular subtyping according to the St Gallen surrogate subtype definition. Low risk of distant recurrence could be confirmed for the MammaTyper® Luminal A-like samples suggesting that for this patient group endocrine treatment alone may be sufficient. The high degree of standardization of mRNA measurement may drive the use of the Ki67/MKI67 biomarker in routine breast cancer pathology. Citation Format: Laible M, Hartmann K, Gürtler C, Anzeneder T, Weber S, Keller T, Sahin U. Prediction of distant recurrence in low risk early breast cancer by RT-qPCR based subtyping using MammaTyper® [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 P3-08-14.

Abstract P1-07-19: Protective effect of 27-hydoxycholesterol on disease recurrence in HER2-negative luminal type of early breast cancer

Abstract Background: 27-hydoxycholesterol (27-HC) is a metabolite of cholesterol (Chol) and known as endogeneous selective estrogen receptor modulator (SERM). Laboratory researches showed high 27-HC could stimulate cancer cell proliferation and promote metastasis processes in hormone receptor positive breast cancer. However, there is no clinical data about the effect of 27-HC on disease recurrence in early breast cancer. Methods: A total of 539 early breast cancer patients who underwent surgery at the National Cancer Center Hospital between 2001 and 2010 with the deposition of blood samples at Tumor Bank were accrued. The blood samples were taken with overnight fasting at surgery day. Serum 27-HC was analyzed by liquid chromatography–mass spectrometry (LC-MS). 27-HC and Chol were categorized into two groups (low vs high) by median, respectively. Results : Hormone receptor was positive in 441 patients and negative in 98 while HER2 was positive in 81, negative in 325 and unknown in 133 patients. The median follow up time was 92 months (range 24 -153.7). The 27-HC (median, 115.73 ng/ml with the range of 29.36- 234.0) and Chol (median, 170.0 mg/dl with the range of 91- 290) showed weak correlation (ρ=0.24, P<0.001). While 27-HC was not significant in analysis for RFS in all cases, it showed marginal significance for RFS in the subset analysis with HER2-negative luminal breast cancer patients (HR, 0.463; 95% CI, 0.208-1.031; P = 0.059). When the patients were grouped according to 27-HC and Chol level (group27HC/Chol) in luminal subgroup, it showed a trend for increasing RFS in the order of grouplow/high , grouplow/low, grouphigh/low and grouphigh/high(Ptrend= 0.009). Conclusion : Serum 27-HC was not associated with poor RFS. High serum 27-HC could be related to good prognosis in HER2-negative luminal type of breast cancer, which may be related to the protective effect of 27-HC from disease recurrence. Further study is warranted. Citation Format: Sung Hoon S, In Hae P, So-Youn J, Seeyoun L, Sun-Young K, Han-Sung K, Eun Sook L, Keun Seok L. Protective effect of 27-hydoxycholesterol on disease recurrence in HER2-negative luminal type of early 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 P1-07-19.

Aromatase inhibitor use during ovarian stimulation suppresses growth of uterine endometrial cancer in xenograft mouse model.

Could aromatase inhibitors (AI) be used to reduce risks of uterine endometrial cancer growth or recurrence during ovarian stimulation? In a xenograft mouse model of endometrial cancer, concomitant AI administration suppressed the growth of endometrial cancer during ovarian stimulation. Recurrence and mortality rates of estrogen receptor-positive early breast cancer are reduced by long-term AI administration. Concomitant AI use for ovarian stimulation in patients with breast cancer is recommended for reducing estrogen-related potential risks. However, the efficacy of concomitant AI use for estrogen receptor-positive endometrial cancer have not been demonstrated conclusively by clinical or experimental animal studies. Forty nude mice xenografted with uterine endometrial cancer cells were allocated to four groups. Group 1: no ovarian stimulation (control). Group 2: ovarian stimulation. Group 3: AI administration + ovarian stimulation. Group 4: ovariectomy and ovarian stimulation. Tumor growth was evaluated during the 6-week treatment period. Ishikawa 3-H-12 uterine endometrial cancer cells (estrogen and progesterone receptors-positive) were transplanted into 6-week-old BALB/cSlc-nu/nu nude mice, followed by interventions 2 weeks later. Compared to ovarian stimulation alone (Group 2), significant suppressions of tumor growth were observed in other three groups (Groups 1, 3 and 4, all at P < 0.05) and correlated with estrogen levels. AI administration had no apparent impact on embryo development. In this study, we examined the growth of endometrial cancer tumors using one endometrial cancer cell line. Clinical endometrial cancer or hyperplasia cells can have diverse origins and AI may not be effective against other cancer cell types. Concomitant AI use may provide a chance for safer childbirth by for patients with endometrial cancer or hyperplasia. This study was supported by the Graduate Student Aid from the St. Marianna University School of Medicine. The authors declare no competing interests.

Caveolin-1 in tumor microenvironment: A review

Caveolins are a member of membrane-bound scaffolding proteins which could negatively regulate and compartmentalize signaling in caveloae. Loss of caveolin-1 (Cav-1) expression occurs in the pathogenesis of tumors, and loss of Cav-1 expression in cancer associated fibroblasts contributes to the activation of tumor microenvironment, thus advancing early recurrence and metastasis in prostate and breast cancer. Recent studies have indicated loss of stromal Cav-1 expression is a predictor of clinical outcome in cancers. Finally, we discuss the mechanism of loss of stromal Cav-1 facilitating the cancer progression, including extracellular matrix remodeling, fibrosis and origin of cancer associated fibroblasts. Key words: Caveolin-1; Tumor microenvironment; Cancer associated fibroblast

66P - Prognostic and clinical values for prediction of breast cancer recurrence in HER2 positive early breast cancer after surgery in King Chulalongkorn Memorial Hospital 2005-2016

Background: Human epidermal growth factor receptor 2 (HER-2) overexpressed breast cancer occurs in 15-20% and associated with diminished disease-free and overall survival. After surgery, the standard treatment in HER-2 positive early breast cancer is systemic chemotherapy combined with trastuzumab. However, one fifth of these patients had disease recurrence in five years after treatment. So, the study was aimed to identify prognostic and clinical values that associated with survival outcomes especially in term of recurrence free survival. Methods: One hundred and sixty-nine patients with early stage HER-2 positive breast cancer who were diagnosed and treated at King Chulalongkorn Memorial Hospital from January 2005 to December 2016 were reviewed retrospectively. Clinico-pathologic features and survival outcomes were analyzed. Results: In initial analysis, median age was 50 years (26-83), 49.1% had premenopausal status, 93.4% had invasive ductal carcinoma, 52.6% had T2 tumor, 58.6% had node negative, 46.1% had lymphovascular invasion, 79.2% had free margin and 46.7% had positive estrogen and progesterone receptor. For the treatment, 69.8% underwent modified radical mastectomy, 96.4% received neoadjuvant/adjuvant chemotherapy, 55.6% received adjuvant trastuzumab, 48.5% received adjuvant hormonal therapy and 68% received post-operative radiotherapy. 35 out of 169 (20.7%) patients had recurrence disease. Distant metastasis was identified in 62.8%. From univariate and multivariate analysis, trastuzumab used (HR 0.44 [95%CI 0.21-0.9]; p = 0.007 from multivariate) and surgical margin (HR 1.02 [95%CI 1.0-1.03]; p = 0.012 from multivariate) were associated with increase risk of recurrences. After median follow up 4.9 years, the 5-years recurrence free survival was 81.2%, 89.2% and 70.7% in overall populations, trastuzumab used subgroup and non-trastuzumab used subgroup, respectively. Overall survival will be further analyzed. Conclusions: Trastuzumab used and surgical margin status were statistically significant prognostic values for predicting disease recurrence in HER-2 positive early breast cancer. Legal entity responsible for the study: Division of Medical Oncology, Department of Medicine, Faculty of Medicine, Chulalongkorn University and the King Chulalongkorn Memorial Hospital. Funding: None Disclosure: All authors have declared no conflicts of interest.

Oncotype Score as a Predictor of Local-Regional Recurrence in Early Stage Breast Cancer (BC)

Local-regional therapy in BC is largely guided by clinico-pathologic features, whereas molecular biology and intrinsic gene signatures of the cancer are used to prescribe risk-tailored systemic therapy. The objective of this study was to evaluate the incidence of local-regional relapse in early stage BC. Further, to evaluate the correlation of Oncotype Dx score, in addition to the clinico-pathologic features with risk of relapse. This was an institutional review board approved retrospective study from our institutional database. We identified 1355 patients with ER+ Stage I and II BC who had undergone Oncotype Dx testing. For this study, we included only those patients with unilateral BC and minimum follow-up of 2 years (n = 792). Patients were treated with mastectomy, or lumpectomy and radiation (RT) + systemic therapy at the discretion of treating physician. We scored patients based on the first site of failure: local-regional relapse (LRR) only, distant metastases (DM) only, and both (LRR+DM). The association between clinical covariates (Age: ≥50, <50; T-size: ≤2 cm, >2 cm; Grade), Oncotype Dx score (<18, ≥18), local treatment (mastectomy vs. lumpectomy/RT), and failure events was evaluated by univariable and multivariable Cox proportional hazards models. Survival was estimated using the Kaplan-Meier and log-rank test method. Among 792 patients that met the study criteria, 73.5% (n = 582) underwent lumpectomy/RT and 26.5% (n = 210) were treated with mastectomy. Systemic therapy was administered in 50% of patients. Majority (81%) had T-size ≤2 cm. Median age was 56 years (range, 25-84 years), and median follow-up was 55 months (range 24-137 months). Five-year OS was 99.1% (95% CI = 97.9-99.6%) and 5-year DFS was 93.8% (95% CI = 91.3-95.6%). The incidence of LRR was 2.2%, DM was 2.6%, and LRR+DM was 0.25% of patients. On univariable analysis, the risk of LRR was significantly associated T-size >2 cm, and Oncotype Dx score ≥18. The factors significantly associated with risk for DM were T-size > 2 cm, grade 2/3, and Oncotype Dx score ≥18. On multivariable analysis both T-size> 2 cm, and Oncotype Dx score >18 remained significant risk factors for LRR and DM (Table 1). Both factors also remained significant upon subset analysis of patients treated with lumpectomy/RT + systemic therapy.Tabled 1Abstract 116; Table 1 Factors Influencing LRR and DM on multivariate analysisRisk FactorsLocal-regional relapseHazard RatioDistant MetastasisHazard RatioT-size >2 cm2.77 (95% CI = 1.07-7.15; P = 0.034)*6.7 (95% CI 2.8-16.3; P<0.0001)*Oncotype Score ≥182.8 (95% CI = 1.08-7.36; P = 0.03)*5.2 (95% CI 1.7-15.5; P = 0.003)* Open table in a new tab Our study suggests that in addition to clinicopathologic variables Oncotype Dx score may provide useful information regarding risk of LRR. Further research is needed to establish a role for molecular and gene profiles in providing risk-tailored delivery of radiation.

Hyperinsulineamia as Risk Factor for Early Breast Cancer Recurrence Linked to Hormone Receptors and Human Epidermal Growth Factor Receptor 2

Background: Hyperinsuliunemia is a condition with excess levels of circulating insulin in the blood among non-diabetics. Hyperinsuliunemia may increase breast cancer developing probability.Purpose: to investigate the role of hyperinsuliunemia in early recurrence in breast cancer by estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER2).Methods: 50 patients with invasive breast cancer in stage II and III in age range 17 - 65 years, involved in this study and classified into five subtypes of ER, PR and HER2, and followed prospectively for 2 years from beginning treatment based on insulin serum levels. Results: In newly diagnosed patients the mean concentrations and standard deviation were (17.11 ± 10.17), while the insulin levels in the patients after chemotherapy treatment were (26.06 ± 15.69), when the tow means were compared by t. test the blood concentration of insulin was high significantly in patients as control (p > 0.001). Regarding hormone receptors and HER2, the insulin levels in ER−PR−HER2− and ER− PR−HER2+ increased significantly after chemotherapy (p = 0.005 and 0.047) respectively. 10 of triple negative breast cancer patients suffered recurrence.Conclusion: Hyperinsuliunemia and weight change after chemotherapy may consider as risk factors for breast cancer recurrence among patients with triple-negative breast cancer.

2435

OBJECTIVES/SPECIFIC AIMS: The goal of the study is to evaluate the prognostic importance and accuracies of a biomarker, human epidermal receptor 2 (Her2), for breast cancer recurrence in a cohort study, namely Lifetime after Cancer Epidemiology (LACE). We specifically interested in the role that Her2 plays in prognosis of breast tumor recurrence for women after a previously diagnosed and treated breast cancer. METHODS/STUDY POPULATION: The study cohort includes 2267 women enrolled in LACE who had previously diagnosed breast cancer. Patients were enrolled from each of the 2 LACE registries in California and Utah. The main endpoint of the study is the right-censored time to breast cancer recurrence. Patients’ enrollments were, on average, 2 years after diagnosis of the first breast cancer. The patients’ characteristics at baseline were obtained through self-administered questionnaires. Cox proportional hazard model with time-varying covariates was used to relate the Her2 status (Her2+ and Her2−) to the primary end point (time to breast cancer recurrence). Hazard ratios (HRs) and their 95% confidence interval comparing Her2+ and Her2− arms were estimated. Time-dependent sensitivity and specificity were used to investigate the performance of using Her2 for classifying patients into high and low risk (Her2 + is classified as hi risk and Her− as low risk) of future breast cancer recurrence at time points after baseline. The time-dependent sensitivity was calculated as the proportion of patients being classified as high risk of recurrence who had breast cancer recurrence before a series of pre-specified time points after baseline, and the time-dependent specificity was calculated as the proportion of subjects being classified as low risk of recurrence who did not have breast cancer recurrence at the same time points. RESULTS/ANTICIPATED RESULTS: The average patient follow-up time was 9.8 years, and 18% of the women got positive Her2 test results at baseline. Among 2267 patients in the study cohort, 2031 had records on their Her2 status, among whom 326 (16.1%) patients were Her2+ and 1705 (83.9%) were Her2−. The mean tumor size among the 2031 patient was 2.10±1.22 cm. A majority of the patients (78.9%) were White. Over one-half of these patients were neither current nor past smokers. Only 3% of the patient had a baseline stage IIIA or higher. About 49% of the patients underwent a mastectomy. Radiation therapy was used by 63.5% of the patients, and Tamoxifen users accounted for 78% of the study cohort. We found a statistically significant association between Her2 and breast cancer recurrence (HR=1.33, log-ran p-value=0.006). However, the HRs of breast cancer recurrence comparing Her2+ and Her2− patients decreased over time. We also investigate the effect of combined Her2, estrogen (ER), and progesterone (PR) on breast cancer recurrence and found that patients with Her2+/ER+/PR− had the highest risk of breast cancer recurrence. The hazard of recurrence for this group of patients was 85% higher than patients with Her2−/ER+/PR+. We also investigate the prognostic accuracies of Her2 in terms of time-dependent sensitivity and specificity. Using Her2 as the prognostic biomarker resulted in a specificity consistently over 80% from baseline up until 15 years post-baseline. The time-dependent sensitivity of Her2 was above 90% between baseline and 1.5 years. Then, the sensitivity dropped gradually to 40% from 1.5 years to 3 years post-baseline. For prognosis of breast cancer over 3 years from baseline, the sensitivity was between 30% and 40%. DISCUSSION/SIGNIFICANCE OF IMPACT: As a single biomarker and risk factor, Her2 was statistically significantly associated with the recurrence of breast cancer among patients in the LACE cohort. A composite biomarker by combining Her2, ER, and PR status was also significantly associated with the breast cancer recurrence. However, the HRs of breast cancer recurrence comparing Her2+ and Her2− patients decreased over time, implying that the Her2 status had a high impact on early recurrent breast tumors. Single biomarkers, usually, have very limited ability for prognosis of future events. However, we found that using HER2 as a single biomarker can give a relatively larger specificity consistently over 15 years of the study period. The sensitivity of Her2 is high for detecting early breast cancer recurrence. However, after 2.5 years from baseline, using Her2 for breast cancer recurrence detection is not reliable. Due to the relatively high accuracies of using Her2 status for prognosis of breast cancer recurrence, we conclude that Her2 should be considered in clinical studies related to prognosis of breast cancer recurrence. Future studies will investigate if prognostic accuracies can be improved by combining Her2 with baseline clinical risk factors such as age, tumor size and lymph nodes. In conclusion, our study has the clinical impact on prognosis (or early detection) of breast cancer recurrence among women with previously diagnosed and treated breast cancers.

Abstract 90: STING colocalizes with gamma-H2AX upon treatment of breast cancer cells with genotoxics: A new role in DNA repair

Abstract One of the current tumor immunotherapy strategies involves the use of STING agonist, a well-known inducer of interferon (IFN) signaling in the immune system, to promote tumor-rejection. However, recent evidence also indicates that constitutive activation of IFN signaling in the tumor may lead to a bad outcome. For instance, increased expression of IFN- stimulated genes (ISGs) at time of surgery was associated with early breast cancer recurrence, and an IFN-related DNA damage resistance signature (IRDS) was identified as a predictive marker of recurrence after radiotherapy. Thus, IFNs secreted into the tumor microenvironment may have complex opposite effects on tumor behavior and response to treatment. Using patient-derived xenograft (PDX) models, we previously showed that the IFN/STAT1 pathway was activated within breast cancer cells in response to chemotherapy and that this pathway may be involved in treatment resistance and recurrence. The aim of this study was to elucidate the mechanisms by which IFN-signaling is triggered in breast cancer cells following chemotherapy and how its activation leads to tumor survival and recurrence. To this aim, the breast cancer cell line MCF7 was treated in vitro with mafosfamide and the activity of different IFN pathway effectors was monitored using western blot, immunofluorescence and cell fractionation techniques. We found that in breast cancer cells, similarly to what is observed in immune cells, type I IFN expression is triggered in a STING-dependent manner. STING silencing abrogated chemotherapy-induced type I IFN production and signaling, while potentiating genotoxic treatment efficacy by promoting cell death and delaying cell colony regrowth. Surprisingly, while STING is described as an endoplasmic reticulum resident protein that relocalizes to perinuclear vesicles upon activation in immune cells, we showed that STING is constitutively present in the nucleus of breast cancer cells and, under activation, forms nuclear clusters that co-localize with gH2AX at DNA breaks. This study provides the first demonstration of STING activation in breast cancer cells and describes a potential new role for the nuclear form of STING in DNA damage response. Our data suggest that genotoxic stress-induced STING activation is a cell-intrinsic mechanism of breast cancer cell survival and regrowth. Whether STING pro-survival role is IFN-dependent (through the activation of given ISGs), IFN-independent (through its involvement in the DNA damage response) or both is currently under investigation. Citation Format: Laura Cheredame, Julie Gaston, Vanessa Yvonnet, Olivier Deas, Marie-Françe Poupon, Jean-Gabriel Judde, Vincent Goffin, Stefano Cairo. STING colocalizes with gamma-H2AX upon treatment of breast cancer cells with genotoxics: A new role in DNA repair [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 90. doi:10.1158/1538-7445.AM2017-90

Abstract 5710: PGRMC1 is a biomarker for breast cancer cell proliferation and early relapse

Abstract Background: Breast cancers may recur following a long latency period, implying a dormant or quiescent phase, however little is known about the biology of cellular quiescence and few markers exist for this clinically important state. The purpose of this study was to investigate whether progesterone receptor membrane component family 1 (PGRMC1), which is expressed in both proliferating and quiescent tumor cells, is a prognostic biomarker in human breast cancer and to define its expression in breast cancer subtypes. Methods: Seventeen publicly available data sets were combined to analyze PGRMC1 expression in 4,463 invasive breast cancers as a function of established molecular and phenotypic markers, estimates of cellular proliferation, and recurrence-free survival data. A gene expression signature-based assay was utilized to estimate cellular proliferation. Association between gene expression and relapse-free survival was assessed using Cox proportional hazards regression while controlling for the effect of proliferation. Results: PGRMC1 expression was analyzed stratified by immunohistochemical and molecular subtype, tumor grade and size and compared with a known triple negative breast cancer biomarker, BCL11a. PGRMC1 and BCL11a each exhibited a robust positive correlation of comparable magnitude with proliferation across all breast cancer subtypes (r=0.27, p=6.5x10-17 and r=0.29, 1.7x10-41, respectively), and PGRMC1 was strongly associated with proliferation within the basal subtype (r=0.42, p=2.4x10-37 PGRMC1 versus 0.31, p=1.1x10-8 BCL11a). PGRMC1 expression was associated with a higher risk of early breast cancer recurrence (HR=1.25, 95% CI [1.12,1.39], p=6.4x10-5) and this association was dependent upon its association with proliferation. Conclusions: PGRMC1 is a breast cancer proliferation biomarker. Further study of the function of this protein is warranted. Citation Format: Elizabeth S. McDonald, Dhruv K. Pant, Tien-chi Pan, David A. Mankoff, Robert H. Mach, Lewis A. Chodosh. PGRMC1 is a biomarker for breast cancer cell proliferation and early relapse [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5710. doi:10.1158/1538-7445.AM2017-5710

Revealing determinant factors for early breast cancer recurrence by decision tree

Early breast cancer recurrence is indicative of poor response to adjuvant therapy and poses threats to patients’ lives. Most existing prediction models for breast cancer recurrence are regression-based models and difficult to interpret. We apply a Decision Tree algorithm to the clinical information of a cohort of non-metastatic invasive breast cancer patients, to establish a classifier that categorizes patients based on whether they develop early recurrence and on similarities of their clinical and pathological diagnoses. The classifier predicts for whether a patient developed early disease recurrence; and is estimated to be about 70% accurate. For an independent validation cohort of 65 patients, the classifier predicts correctly for 55 patients. The classifier also groups patients based on intrinsic properties of their diseases; and for each subgroup lists the disease characteristics in a hierarchal order, according to their relevance to early relapse. Overall, it identifies pathological nodal stage, percentage of intra-tumor stroma and components of TGFβ-Smad signaling pathway as highly relevant factors for early breast cancer recurrence. Since most of the disease characteristics used by this classifier are results of standardized tests, routinely collected during breast cancer diagnosis, the classifier can easily be adopted in various research and clinical settings.

Erratum to: Post-relapse survival in patients with the early and late distant recurrence in estrogen receptor-positive HER2-negative breast cancer.

Erratum to: Post-relapse survival in patients with the early and late distant recurrence in estrogen receptor-positive HER2-negative breast cancer.

Regional Nodal Irradiation After Breast Conserving Surgery for Early HER2-Positive Breast Cancer: Results of a Subanalysis From the ALTTO Trial

Two randomized trials recently demonstrated that regional nodal irradiation (RNI) could reduce the risk of recurrence in early breast cancer; however, these trials were conducted in the pretrastuzumab era. Whether these results are applicable to human epidermal growth factor receptor 2 (HER2)-positive breast cancer patients treated with anti-HER2-targeted therapy is unknown. This retrospective analysis was performed on patients with node-positive breast cancer who were enrolled in the Adjuvant Lapatinib and/or Trastuzumab Treatment Optimization phase III adjuvant trial and subjected to BCS. The primary objective of the present study was to examine the effect of RNI on disease-free survival (DFS). A multivariable cox regression analysis adjusted for number of positive lymph nodes, tumor size, grade, age, hormone receptors status, presence of macrometastatis, treatment arm, and chemotherapy timing was carried out to investigate the relationship between RNI and DFS. One thousand six hundred sixty-four HER2-positive breast cancer patients were included, of whom 878 (52.8%) had received RNI to the axillary, supraclavicular, and/or internal mammary lymph nodes. Patients in the RNI group had higher nodal burden and more frequently had tumors larger than 2 cm. At a median follow-up of 4.5 years, DFS was 84.3% in the RNI group and 88.3% in the non-RNI group. No differences in regional recurrence (0.9 % vs 0.6 %) or in overall survival (93.6% vs 95.3%) were observed between the two groups. After adjustment in multivariable analysis, there was no statistically significant association between RNI and DFS (hazard ratio = 0.96, 95% confidence interval = 0.71 to 1.29). Our analysis did not demonstrate a DFS benefit of RNI in HER2-positive, node-positive patients treated with adjuvant HER2-targeted therapy. The benefit of RNI in HER2-positive breast cancer needs further testing within randomized clinical trials.

Differences in Ki67 expressions between pre- and post-neoadjuvant chemotherapy specimens might predict early recurrence of breast cancer

The prognosis of breast cancer patients not obtaining a pathological complete response (pCR) with neoadjuvant chemotherapy (NAC) is poorer than that of pCR patients. Identifying new prognostic factors for non-pCR patients is important because fractions of this population might benefit from novel adjuvant treatments currently under development. High Ki67 expression in remnant disease after NAC has been described as a poor prognostic factor. Studies have shown that a reduction in Ki67 expression is more often observed in good responders to chemotherapy. We hypothesized that the change in Ki67 expression might be useful for predicting patient outcomes and thus retrospectively examined pairs of biopsy and surgical specimens of breast tissue from individual patients. One hundred sixteen patients with remnant invasive disease in the breast, who received NAC and underwent surgery at our institution, were retrospectively examined. Differences in Ki67 expression between pre- and post-NAC specimens were analyzed in relation to patient outcomes. The mean Ki67 expression value after NAC was higher in patients who developed metastasis than in those without metastasis (P<.01). Tumors showing higher Ki67 expression in the surgical than in the biopsy specimen were more frequent in patients with metastasis (P<.01). This trend was more obvious in patients who developed metastasis within 1 year after surgery. Our results indicate that a difference in Ki67 expressions after versus before NAC might be an important predictor of early metastasis. Evaluating not only absolute Ki67 values, but also any changes in response to NAC, may improve the prediction of patient outcomes.

Abstract P4-12-07: CCND1 amplification in early breast cancer patients: Correlation with subtypes and prognosis

Abstract Background: Mechanisms of progress and recurrence of early breast cancer (BC) have gained importance since several targeted therapeutic options in metastatic breast cancer have been introduced over the last years, especially in hormone receptor positive BC. This study investigates the amplification of cell cycle regulator cyclin D1 gene (CCND1) amplification as one possible progression mechanism. Patients &amp; Methods: Patients from an unselected cohort of early BC patients were included into this study. A tissue microarray (TMA) was available for n=832 patients with early breast cancer. CCND1 amplification was assessed after FISH (Abbott Vysis LSI CCND1 SpectumOrange/CEP11 SpectrumGreen Probes Kit). A CCND1/CEP11-ratio ≥ 2.0 was considered as amplification. Staining was successful in 545 tumor cores. Amplification results were correlated with clinical patient and tumor characteristics and survival analyses were performed with regard to distant disease free survival and overall survival. Results: CCND1 amplification was found in 13.6% of patients. Triple negative, luminal A like, luminal B like and HER2 positive tumors were amplified in 7.5%, 7.8%, 18.6% and 15.7% respectively (p = 0.010). CCDN1 amplification was significantly associated with a higher grading and a higher body mass index. Furthermore an amplification was seen more frequently in lobular BC and ductal BC than other histological subtypes. Survival analysis showed a reduced DDFS for patients with CCDN1-amplification. 5 year DDFS rates were 90.6% for non-amplified tumors and 86.0% for amplified tumors (p, log-rank =0.066 ). 5 year OS rates were 93.0% for non-amplified tumors and 90.1% for amplified tumors (p, log-rank =0.119). Adjusted for age, tumor size, nodal status and molecular subtype, cox regression showed HR of 1.3 (95% CI: 0.76-2.5, p=0.46) for DDFS and a HR of 1.33 (95% CI: 0.7-2.53, p=0.38) for OS. Conclusion: With a 13.6% prevalence in all breast cancer patients, mainly present in luminal B like cancers, CCND1-amplification is a genetic aberration associated with an unfavorable prognosis. Within hormone receptor positive women CCDN1 amplification might play a role in treatment resistance mechanisms in early breast cancer patients. Citation Format: Hanf DC, Fasching PA, Villalobos IE, Gasparyan A, Wachter D, Santiago A, Guzman R, Weihbrecht S, Hanf V, Hartmann A, Beckmann MW, Press MF. CCND1 amplification in early breast cancer patients: Correlation with subtypes and prognosis [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 P4-12-07.

Oncotype Dx recurrence score among BRCA1/2 germline mutation carriers with hormone receptors positive breast cancer.

Gene expression assays are widely used to predict risk of recurrence in early breast cancer (BC). We report the 21-gene expression assay (Oncotype Dx) recurrence score (RS) distribution of 27 BRCA carriers with estrogen receptor (ER) positive BCs, identified at Hadassah Medical Center, combined with 2 previous studies. Treatment decision and outcomes of the 27 BRCA carriers were compared with an Israeli cohort of 1594 patients published recently. We found Oncotype Dx RS low (<18), intermediate (18-30) and high (>30) among 12 (21.4%), 23 (41.1%) and 21 (37.5%) of 56 BRCA1 carriers compared with 15 (17.2%), 49 (56.3%) and 23 (26.4%) of 87 BRCA2 carriers (p = 0.2). The corresponding distribution in a population of 82,434 women published by Genomic Health was 53.4%, 36.3% and 10.3% for low, intermediate and high RS (p < 0.001 for BRCA1 and BRCA2). Treatment decision regarding chemotherapy according to RS was similar in BRCA1, BRCA2 and the control group. Two of 27 carriers had distant recurrence: a BRCA1 carrier with RS of 18 and a BRCA2 carrier with RS of 22; both have an excellent response to chemotherapy. We found an approximately ∼3 fold increased rate of high RS among BRCA1 and 2 carriers with ER positive BC compared with the general BC population. These data might indicate that hormone positive BC in BRCA carriers are molecularly unique. The surprisingly good response to chemotherapy in the metastatic setting in 2 patients may suggest that the predictive value of low-intermediate RS in carriers merits further studies.

Post-relapse survival in patients with the early and late distant recurrence in estrogen receptor-positive HER2-negative breast cancer

Few studies have been performed on post-relapse survival in patients with the early and late distant recurrence in estrogen receptor (ER)-positive, HER2-negative breast cancer. A total of 205 patients with the early distant recurrence and 134 patients with the late distant recurrence of ER-positive, HER2-negative breast cancer who had undergone breast surgery or neoadjuvant chemotherapy between January 2000 and December 2004 were registered from nine institutions. Prognostic factors for post-relapse survival in patients with the early and late recurrence were analyzed. Post-relapse survival was significantly longer in patients with the late recurrence than in patients with the early recurrence. Predictive factors for post-relapse survival in patients with the early recurrence were lack of adjuvant chemotherapy, a long disease-free interval, and long durations of endocrine therapies and chemotherapies after relapse. In patients with the late recurrence, post-relapse survival was significantly improved for those individuals with one metastatic organ at relapse and individuals who were treated with the first-line and subsequent endocrine therapies for prolonged periods. Moreover, ER expression in primary breast tumors of late recurrence patients was significantly higher with a duration of the first-line endocrine therapy >6months than in those with a duration ≤6months. Predictors for prognosis after relapse differed between patients with the early and late distant recurrence. Endocrine responsiveness after relapse is a key factor for improved post-relapse survival, and it is thus important to establish whether metastatic tumors are endocrine-resistant in ER-positive, HER2-negative recurrent breast cancer.

IHC4 score plus clinical treatment score predicts locoregional recurrence in early breast cancer

PurposeImmunohistochemical 4 (IHC4) score plus Clinical Treatment Score (CTS) is an inexpensive tool predicting risk of distant recurrence in women with early breast cancer (EBC). IHC4 score is based on ER, PR, HER2 and Ki67 index. This study explores the role of the combined score (IHC4 + CTS) in predicting risk of locoregional recurrence (LRR) in women with EBC who had breast conservation surgery (BCS) without adjuvant radiotherapy (study group). The secondary objective was to evaluate the clinicopathological differences between our study group and women who had adjuvant radiation following BCS (control group). Methods and materialsPatients were selected from the local database over a 13-year period. IHC testing was done where results were missing. Combined scores were calculated using the appropriate formulae. ResultsPatients in the study group (81 patients) had favorable clinicopathological features compared to the control group (1406 patients).The Cox regression indicated a statistically significant association between the combined score and the risk of LRR (p = 0.03). The incidence of LRR was zero, 20% and 33.3% in the low, intermediate and high risk groups respectively (p = 0.007).Margin status was the only variable not included in the combined score. The Cox regression analysis demonstrated that the combined score (p = 0.02) and the ordinal measure of margins (p = 0.03) were significant independent predictors of LRR. ConclusionThis is the first study of its kind. The IHC4 score + CTS can be used to identify low risk women who can potentially avoid adjuvant radiotherapy.

Regional nodal irradiation after breast conserving surgery for HER2-positive breast cancer: Results of a sub analysis from the ALTTO trial.

604Background: The optimal treatment of regional nodal areas following breast-conserving surgery (BCS) is a source of debate. Two randomized trials recently demonstrated that regional nodal irradiation (RNI) can reduce the risk of recurrence in early breast cancer; however,these trials were conducted in the pre-trastuzumab era.Whether these results are applicable to HER2-positive breast cancer patients treated with anti-HER2 targeted therapy, is unknown. Methods: This retrospective analysis was performed on patients with node-positive breast cancer, who were enrolled in the ALTTO phase III adjuvant trial and subjected to BCS. The primary objective of the present study was to examine the effect of RNI on disease-free survival (DFS). Secondary objectives included regional recurrences and overall survival (OS). A multivariate cox regression analysis adjusted for number of positive lymph nodes, tumor size, grade, age, hormone receptors status, presence of macrometastatis, treatment arm, and chemotherapy timin...

A miRNA-based signature predicts development of disease recurrence in HER2 positive early breast cancer after adjuvant trastuzumab-based treatment.

e12070Background: Adjuvant trastuzumab is the standard treatment for early HER2 positive breast cancer. However, 15%-22% of patients still suffered disease recurrence after adjuvant anti-HER2 treatment. A molecular prognostic model that can reliably stratify patients by risk of developing disease recurrence is urgently needed. Methods: All the patients included were diagnosed as HER2-postive breast cancer and received radical surgery followed by adjuvant trastuzuamb treatment. Using miRNA microarrays, we analyzed the FFPE tumor tissue from14 patients (7 recurrence vs.7 non-recurrence) and identified nine miRNAs that were differentially expressed. Then, we validated the expression of these miRNAs using qRT-PCR in training set (n = 101), and built the miRNA-based classifier using the Cox regression model. We further confirmed the prognostic accuracy of this signature in an internal testing set (n = 57), and an external independent testing set (n = 53) Results: We identified a two-microRNA signature that cla...