Triple Negative Group Research Articles

Role of P-53 Gene in Preventing Breast Cancer

Breast cancer affects more than one million patients annually in the world and is a leading cause of mortality. Histological type, grade, tumor size, lymph node involvement, and estrogen receptor and HER-2 receptor status, all influence prognosis and the probability of response to systemic therapies. P53 gene is mutated in about 30% of breast cancers/.The possible links between alterations of p53 and clinical or pathological features of breast tumors have been widely investigated. The first study to examine gene-expression patterns of breast cancer suggested that at least four major molecular classes of breast cancer exist: luminal-like, basal-like, normal like, and HER-2 positive. Basal-like breast cancer account for 15% of breast cancers and are often described as triple negative breast cancers (TNBCs). In fact, TNBCs, defined by lack of expression of estrogen receptor, progesterone receptor, and HER2, probably include both basal-like breast cancers and some poorly differentiated luminal breast cancers. They are also associated with a younger age and a poor prognosis. TNBCs also have an increased frequency of TP53 mutations. Recently, it was shown that mutant p53 status was a strongly unfavorable prognostic factor for relapse-free survival and overall survival only in a triple negative group in patients treated with adjuvant anthracycline-containing chemotherapy. Inflammatory breast cancer (IBC) is a clinical diagnosis known as the T4d category in the TNM classification. It is a distinct clinical subtype of locally advanced breast cancer (LABC), with a particularly aggressive behavior and poor prognosis. TP53 mutations are more frequent in inflammatory breast cancer (50%) than in non-inflammatory breast cancer (20–30%). The results from these studies served as the justification for attempts to vaccinate patients using p53-derived peptides, and a number of clinical trials are in progress. The most advanced work used a long synthetic peptide mixture derived from p53 (p53-SLP; Netherlands).The vaccine is delivered in the adjuvant setting and induces T helper type I response in the majority of patients. However, the response may not be potent enough to result in clinical benefit as a mono-therapy because most patients had T-helper cells that failed to produce key cytokines, indicating that this treatment should also be associated with another type of conventional or immunotherapy therapy.

Abstract 4341: Large mutational spectrum of cutaneous melanoma in a Brazilian population: The experience of A.C. Camargo Cancer Center

Abstract Introduction: Cutaneous melanoma (CM) is an aggressive type of malignancy, with elevated mortality rates. The Cancer Genome Atlas Network (TCGA) recently reported the molecular classification of CM and divided these lesions according to their somatic mutation profile in 4 categories: BRAF, NRAS or NF1 mutated, and also the triple-negative group. The aim of this study is to describe the spectrum of molecular alterations from a Brazilian population of CM patients. Methods: DNA was extracted from formalin-fixed, paraffin-embedded (FFPE) tissue from primary and metastatic melanoma samples from A.C. Camargo Cancer Center molecular pathology section. Next-generation sequencing (Ion Torrent®) targeting a 14-gene panel was performed, targeting the following genes: BRAF (códon 600), NRAS (códons 12, 13, 61, 117 and 146) e KIT (éxon 11, 13, 17), KRAS, CTNNB1, IDH1, EGFR, HRAS, PDGFR-a, PIK3CA, RET, JAK2, MET, and CDH1. Results: 131 cutaneous melanoma samples from different patients were sequenced. The majority of the samples (57%) showed mutation in at least one gene. BRAF was the most frequent detected mutation, found in 40% of the cases, and p.V600E, p.V600K and p.V600R represented 90%, 8% and 2% of BRAF mutations, respectively. NRAS mutation was detected in 10% of the cases, and among those 88% involved codon 61. KIT mutation was identified in 10% of the cases. Mutations in KRAS, CTNNB1, PDGFRa, PIK3CA, EGFR and HRAS were identified in 5%, 4%, 4%, 3%, 2% and 1% of the cases, respectively. No mutations in IDH1, JAK2, MET or RET was identified. Most of the mutated cases showed mutation in only one tested gene (71%), while 14%, 10% and 5% showed mutations in 2, 3 or 4 genes respectively. None of the cases showed simultaneous mutation involving combinations of BRAF, NRAS or KIT mutations. Discussion and Conclusion: The BRAF, NRAS and KIT mutation frequency in this population with CM diagnosis is similar to the one described in the world literature for this disease, as well as the distribution of the most frequent mutated regions and specific types of mutations. These results add the mutational spectrum of CM of a large Brazilian cohort to the world literature. Treatment of CM with targeted therapies aiming the MAP-Kinase pathway, and especially blocking BRAF, are a reality in the clinical oncology routine for treating CM patients, but there are still many obstacles in order to fully understand the resistance mechanism and the possibility of drug combinations. Additionally, new targeted agents are under development, such as the ones for NRAS mutated cases, and in this way the understanding of the large mutational spectrum of melanoma is of extreme importance in the clinical scenario. Citation Format: Mariana Petaccia de Macedo, Felipe Fidaldo, Clovis Lopes Pinto, João Pedreira Duprat, Maria Dirlei S.f Begnami, Dirce Maria Carraro, Isabela Werneck Cunha. Large mutational spectrum of cutaneous melanoma in a Brazilian population: The experience of A.C. Camargo Cancer Center [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4341.

Quantitative analysis of peri-tumor fat in different molecular subtypes of breast cancer

Background and purposesThe aim of this study was to develop morphological analytic methods to analyze the tumor-fat interface and in different peritumoral shells away from the tumor, and to compare the results among three molecular subtypes of breast cancer. Materials and methodsA total of 102 women (mean age 48.5 y/o) with solitary well-defined breast cancers were analyzed, including 46 human epidermal growth factor receptor 2 (HER2) (+), 46 HER2(−) hormonal receptor (HR) (+), and 10 triple negative (TN) breast cancers. The tumor lesion, the breast, the fibroglandular and fatty tissue were segmented using well-established methods. The whole breast fat percentage and the peri-tumor interface fat percentage were measured. Three shells (SH1, SH2, SH3) surrounding the convex hall of the three dimensional (3D) tumor were defined and in each shell the volumetric percentage of fat was calculated. The peri-tumor interface fat percentage and the volumetric percentage of fat in the three peri-tumoral shells were compared among different subtypes. ResultsIn the TN group, the fat percentage on the tumor boundary was 43 ± 20% and 78 ± 12% for two dimensional (2D) and 3D measurement, respectively, which were the highest among the three subtypes but not significantly different. The fat percentage in SH2 and SH3 in the TN group was 82 ± 7% and 85 ± 7%, which was significantly higher compared to the two other two subtypes. The results remained after controlling for the whole breast fat percentage. ConclusionsThis study provided a feasible method for quantitative analysis of peri-tumoral tissue characteristics. Because of small patient number, the finding that TN tumors had the highest peri-tumor fat content among the three subtypes needs to be further verified with a large cohort study.

The Relationship of Reproductive Risk Factors and Histologic Patterns with Molecular Subtypes of Breast Cancer

Background: The close link between molecular subtypes and different histological types of breast cancer has recently been taken into consideration. Objectives: The present study aimed at evaluating the reproductive risk factors in relation to molecular subtypes and histological features of breast cancer in a large group of Iranian patients. Methods: This historical cohort study was conducted on 1988 women diagnosed as different subtypes of breast cancers recruited in 2011 to 2016 from cancer research center in Shahid Beheshti University of Medical Sciences, Iran. Data on molecular markers were obtained from hospital files obtaining originally from immunohistochemical staining technique. Based on the pathological reports in hospital recorded files, the histological patterns of the cancer was also determined. The patients were followed for 5 years to assess the 5-year survival and compared the survival across the different molecular subtypes. Results: The highest mean age was found for the group with HER2-overexpression and the lowest for those with luminal A (P = 0.045). The most and the least tumor size was revealed in triple negative group and luminal A group, respectively (P = 0.035). The mean number of lymph nodes involved in breast cancer was significantly higher in luminal B subtypes compared to luminal A and triple negative subtypes (P = 0.004). The tumor stages III-IV were found in 31.6% of patients with luminal A subtype, 42.2% in patients with luminal B, 34.3% in patients with HER2 overexpression, and 26.0% in those with triple negative subtype (P = 0.006). The histological patterns of the tumor were powerfully different in terms of the molecular subtypes of tumor so that luminal A subtype was found more in ILC pattern, luminal B subtype was found more in DCIS pattern, HER2-overexpression subtype was revealed more in DCIS pattern, and triple negative subtype was found more in IDC pattern. Based on the long-term survival analysis, 5-year survival was found to be 98.3% in luminal A group, 98.3% in luminal B group, 100% in HER2 overexpression, and 98.1% in triple negative with no difference between different molecular subtypes. The lowest 5-year survival was found in the patients aged higher than 30 years at first pregnancy and live birth with triple negative subtype (survival rate of 75.0%). The long-term survival was adversely associated with the tumor stage but independent to tumor molecular subtypes. Conclusions: Age at first live birth, tumor size, lymph node involvement, tumor stage, and histological pattern of breast cancer are linked to its molecular subtype. The lower long-term survival of breast cancer can be predicted by advanced age (especially in triple negative subtype) and by higher tumor stage independent to molecular subtype.

P-53 Gene As Marker in Breast Cancer

Purpose: Aim of this current review is to emphasis a possible links between alterations of P-53 gene together with its protein in pathological features of breast cancer. Design: The study examines the gene-expression patterns of breast cancer suggested that at least four major molecular classes of breast cancer exist: luminal-like, basal-like, normal like, and HER-2 positive. Basal-like breast cancer account for 15% of breast cancers and are often described as triple negative breast cancers (TNBCs). In fact, TNBCs, defined by lack of expression of estrogen receptor, progesterone receptor, and HER2, probably include both basal-like breast cancers and some poorly differentiated luminal breast cancers. They are also associated with a younger age and a poor prognosis. TNBCs also have an increased frequency of TP53 mutations. Inflammatory breast cancer (IBC) is a clinical diagnosis known as the T4d category in the TNM classification. It is a distinct clinical subtype of locally advanced breast cancer (LABC), with a particularly aggressive behavior and poor prognosis. Results: TP53 mutations are more frequent in inflammatory breast cancer (50%) than in non-inflammatory breast cancer (20-30%). Histological type, grade, tumor size, lymph node involvement, and estrogen receptor and HER-2 receptor status, all influence prognosis and the probability of response to systemic therapies. P53 gene is mutated in about 30% of breast cancers. Conclusion: Recently studies shown that mutant P-53 gene status was a strongly unfavorable prognostic factor for relapse-free survival and overall survival only in a triple negative group in patients treated with adjuvant anthracycline-containing chemotherapy. The results from these studies served as the justification for attempts to vaccinate patients using p53-derived peptides, and a number of clinical trials are in progress.

Wide Expression and Significance of Alternative Immune Checkpoint Molecules, B7x and HHLA2, in PD-L1-Negative Human Lung Cancers.

Purpose: Immunotherapy targeting the PD-1/PD-L1 pathway has changed the treatment landscape of non-small cell lung carcinoma (NSCLC). We demonstrated that HHLA2, a newly identified immune inhibitory molecule, was widely expressed in NSCLC. We now compared the expression and function of PD-L1 with alternative immune checkpoints, B7x and HHLA2.Experimental Design: Expression was examined in tissue microarrays consisting of 392 resected NSCLC tumors. Effects of PD-L1, B7x, and HHLA2 on human T-cell proliferation and cytokine production were investigated.Results: PD-L1 expression was identified in 25% and 31% of tumors in the discovery and validation cohorts and was associated with higher stage and lymph node involvement. The multivariate analysis showed that stage, TIL status, and lymph node involvement were independently associated with PD-L1 expression. B7x was expressed in 69% and 68%, whereas HHLA2 was positive in 61% and 64% of tumors in the two sets. The coexpression of PD-L1 with B7x or HHLA2 was infrequent, 6% and 3%. The majority (78%) of PD-L1-negative cases expressed B7x, HHLA2, or both. The triple-positive group had more TIL infiltration than the triple-negative group. B7x-Ig and HHLA2-Ig inhibited TCR-mediated proliferation of CD4 and CD8 T cells more robustly than PD-L1-Ig. All three significantly suppressed cytokine productions by T cells.Conclusions: The majority of PD-L1-negative lung cancers express alternative immune checkpoints. The roles of the B7x and HHLA2 pathway in mediating immune evasion in PD-L1-negative tumors deserve to be explored to provide the rationale for an effective immunotherapy strategy in these tumors. Clin Cancer Res; 24(8); 1954-64. ©2018 AACR.

Abstract P5-07-12: miR-34a as a prognostic biomarker for overall survival triple negative breast cancer within Tunisian patients

Abstract Introduction: miR-34a has been recently introduced to clinical trials especially in breast cancer depending on its crucial role as microRNA suppressor tumors since it regulates several oncogenes. We explore here the prognostic effect of miR-34a on triple negative breast cancer (TNBC) overall survival compared to no triple negative (NTNBC) group and its effect on PARP-1 and BRCA1 protein expression. Methods: 76 patients with TNBC and NTNBC were recruited for this study. miR-34a was quantified using qPCR Syber Green technique. PARP-1 and BRCA1 were analyzed by IHC using Tissue Micro-Array method. Results: PARP-1 is more expressed in TNBC compared to NTNBC; P = 0.04; RR= 1.8 (1.1-3.1), however, BRCA1 is less expressed in TNBC; P = 0.03; RR= 2 (1.2-12.6). Furthermore, we evaluated the relationship between miR-34a and clinicopathological features with survival rate of both studied groups, we found that miR-34a is more expressed among patients with distance metastases in TNBC group P = 0.001; RR = 6.5 (2.1-19.6). Survival analysis showed that miR-34a low expression in TNBC was remarkably related to overall survival; P=0.012. Furthermore, multivariate survival test suggested that the absence of miR-34a was linked to the poor survival within TNBC according to the presence of distance metastasis; P=0.048; HR= 2.69. Our results showed that miR-34a is decreased in triple negative breast cancer patients with a very poor prognosis. Conclusion:Our results give mechanistic insights in miR-34a as a prognostic biomarker for overall survival Triple Negative Breast Cancer within Tunisian patients through its effect on PARP-1 and BRCA1 protein expression which may open potential new strategies towards prevention and therapeutic inhibition of PARP-1 positive within mammary tumors. Citation Format: Medimegh I, Ayari H, Baraudi O, Majoul H, Omrane I, Yousef E, Benayed F, Benromdhan K, Gaboury L, Mader S, Benammarelgaaied A. miR-34a as a prognostic biomarker for overall survival triple negative breast cancer within Tunisian patients [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 P5-07-12.

Outcome evaluation in pre-trastuzumab era between different breast cancer phenotypes: a population-based study on Italian women.

Based on estrogen receptor (ER), progesterone receptor (PgR) and Her2/neu (HER2) expression, four breast cancer subtypes have been distinguished: luminal A (ER and/or PgR/HER2-, Ki67 <14%), luminal B (ER and/or PgR/HER2-, Ki67 ≥14% or ER and/or PgR/HER2), triple-negative (ER-/PgR-/HER2-), and HER2 (ER-/PgR-/HER2). Our aim was to evaluate the prognosis of these phenotypes in the pre-trastuzumab era in a large cohort of Italian women. We studied 2347 breast cancer patients, in stage I-II, registered by the Modena Cancer Registry from 1999 to 2006 in the Modena province, Italy. Overall survival, disease-free survival and second non-mammary tumors were evaluated. A total of 1868 luminal A (79.6%), 195 luminal B (8.3%), 205 triple-negative (8.7%) and 79 HER2 (3.4%) patients were identified. A better prognosis was observed for luminal A than for luminal B, HER2 and triple-negative subtypes (5-year overall survival, 91% vs 89% vs 87% vs 86%, respectively, P <0.001). Disease-free survival for pT1a and pT1b tumors was worse in HER2 (82%) than in triple-negative (90%), luminal B (95%) and luminal A (97%) (P = 0.013). Finally, luminal B patients had a significantly higher rate of second non-mammary tumors than the other groups. In the pre-trastuzumab era, luminal A patients showed a better 5-year overall survival than luminal B, HER2 and triple-negative patients, but in terms of disease-free survival, HER2 subtype represented an unfavorable group over time, whereas the triple-negative group had an increased risk of relapse in the first 42 months and then decreased. Among each prognostic factor, ER <10%, Ki67 >14% and HER2 overexpression are considered as risk factors, but only HER2 positivity seems to preserve the role over time.

Breast cancer stem cell markers CD44 and ALDH1A1 in serum: distribution and prognostic value in patients with primary breast cancer.

Background: CD44 and ALDH1 have been recognized as the most widely used markers to identify breast cancer stem cells (BCSCs). However, limited to tissue sample and rare population, BCSCs have always been not easily detected. We aimed to measure CD44 and ALDH1A1 (major contributor to ALDH1 activity) levels in serum and explore the prognostic value in primary breast cancer patients.Methods: This study included 140 primary breast cancer patients with stage I-III. Serum samples were collected before surgery and stored at -80 degrees. CD44 and ALDH1A1 were measured by chemiluminescent assay.Results: High serum CD44 levels (≥ 417.4 ng/mL) were correlated with postmenopausal status (P = 0.006), estrogen receptor negativity (P = 0.025), progesterone receptor negativity (P = 0.002) and adjuvant chemotherapy (P = 0.003). The mean serum CD44 levels of luminal group (406.4 ± 68.3 ng/mL) were significantly lower than triple negative group (506.8 ± 175.5 ng/mL) (P < 0.001). There was no correlation between serum ALDH1A1 levels and molecular subtypes. Multivariate analysis revealed that high serum CD44 level (≥ 417.4 ng/mL), was an independent factor for PFS (P = 0.019) and OS (P = 0.008). However, serum ALDH1A1 has no impact on either PFS (P = 0.613) or OS (P = 0.441).Conclusion: Serum CD44 was an independent prognostic indicator in primary breast cancer. However, serum ALDH1A1 has no impact on survivals and might not be an appropriate candidate to predict prognosis for breast cancer.

Study of prognostic and predictive factors in locally advanced breast cancer in Egypt

Objective We aimed to evaluate the prognosis in locally advanced breast cancer (LABC), and the efficacy of neoadjuvant chemotherapy in inducing pathological response in LABC. Background Despite efforts at early detection, LABC remains a clinical challenge, as distant metastasis develops in most patients, and they will experience disease relapse and eventual death. Patients and methods To study the hematoxylin and eosin-stained slides of pretreatment and post-treatment specimens and correlate them with prognostic and predictive factors. Results LABC in Egypt showed poor prognostic factors such as a high proportion of the cases have a large tumor size (46%) and axillary lymph node (LN) metastasis (87%) and younger age at time of presentation (median = 53 year). These results indicate a more aggressive biology of LABC in Egypt compared with the same group in developed countries. Negative preoperative LN predicts complete response in postoperative LN. Prolonged survival was observed within the triple-negative (P = 0.004) and progesterone receptor negative groups (P = 0.02). The disease-free survival was shortened in positive lymphovascular invasion (P = 0.04) and high histologic grade (0.03). Conclusion LABC in Egypt showed poor prognostic outcome. Triple-negative and progesterone-negative cases that received neoadjuvant therapy showed significant prolonged survival.

Respuesta al Tratamiento Neoadyuvante en pacientes con Cáncer de Mama en SOLCA-Guayaquil.

Introducción: La quimioterapia neoadyuvante es ampliamente aceptada como tratamiento de elección en cáncer de mama localmente avanzado. El objetivo de la presente comunicación corta es conocer la tasa de respuesta completa patológica (pRC) luego de neoadyuvancia, la frecuencia de cirugía conservadora, así como también el porcentaje de la sobrevida. Métodos: Ingresaron al estudio pacientes con cáncer de mama que recibieron tratamiento de quimioterapia neoadyuvante y luego sometidas a cirugía, fueron analizadas retrospectivamente usando historias clínicas desde enero 2009 hasta diciembre 2011 en el Instituto Oncológico Nacional Dr. ¨Juan Tanca Marengo¨ Solca-Guayaquil, se excluyeron pacientes sin suficiente información clínica y aquellas tratadas en otros centros. El procesamiento de datos se realizó mediante un sistema estadístico SPSS v20. Resultados: 1367 pacientes con cáncer de mama fueron diagnosticadas desde el año 2009-2011, se excluyeron aquellas que no reunieron los criterios de inclusión. 200 pacientes evaluables recibieron quimioterapia neoadyuvante y fueron operadas. La edad promedio al diagnóstico fue de 51 años (rango 26-79 años), el tipo histológico ductal Infiltrante fue el más frecuente 185 (92.5 %), Lobular 10 (5 %), Medular 4 (2 %) y metaplásico 1 (0.5 %). El Grado histológico II 143 (71.5 %), Grado histológico III 30 (15 %). En Estadio I: 1 (0.5 %), IIA: 25 (12.5 %), IIB: 42 (21 %), IIIA: 58 (28 %), IIIB: 69 (34.5 %), IIIC: 5 (2.5 %). Conclusión: El cáncer de mama es un grupo heterogéneo, la mayoría de pacientes acuden con enfermedad avanzada (65 %), respuesta patológica completa (16.5 %) luego de neoadyuvancia es muy similar a la obtenida en otros estudios, como es conocido el grupo triple negativo obtuvo los mejores resultados (25 %) y una mejoría en el porcentaje de la sobrevida global en este subgrupo, es importante completar el protocolo de neoadyyuvancia previo a la cirugía para aumentar la tasa de pRC y así como también la cirugía conservadora (10 %) que es el objetivo primario en neoadyuvancia.

Biological subtypes and survival outcomes in breast cancer patients with brain metastases in the targeted therapy era.

There is recognition that breast cancer is a collection of heterogeneous diseases divided in subtypes based on combined molecular features such as hormonal receptors (HR) and human epidermal growth factor receptor 2 (HER2) status. We aimed to study clinical differences among biological subtypes in brain metastasis from breast cancer after targeted therapy introduction. This was a retrospective study with 406 consecutive patients with brain metastasis from breast cancer treated at MD Anderson Cancer Center from 1998 to 2013. Overall, 315 of these patients met the study criteria and were analyzed. Subtypes were classified as HER2-/HR+ (96 patients), HER2+/HR+ (57 patients), HER2+/HR- (63 patients), and triple negative (HER2-/HR-) (99 patients). End points were time to development of brain metastasis (TDBM), brain metastasis-free survival (BMFS), and overall survival from start of treatment of brain metastasis (OSBM). Univariate and multivariate Cox proportional hazard regression models were used to analyze the data. TDBM was 41 months for HER2-/HR+; 58 months for HER2+/HR+; 30 months for HER2+/HR-; and 27 months for triple negative (P < .001). BMFS was 9 months for HER2-/HR+; 24 months for HER2+/HR+; 9 months for HER2+/HR-; and 7 months for triple negative (P = .06). OSBM was 20 months for HER2-/HR+; 22 months for HER2+/HR+; 24 months for HER2+/HR-; and 9 months for triple negative (P < .001). On multivariate analyses, triple negative showed lower OSBM compared with other subtypes, with a hazard ratio of 1.9 (P < .001). Comparing all breast cancer subgroups we noticed that HR and HER2 are the most significant biomarkers in brain metastasis behavior. Patients who received targeted therapy had better outcomes, but not in the triple negative group. Prospective studies with different treatment modalities for each subgroup are recommended.

Treg depletion potentiates checkpoint inhibition in claudin-low breast cancer.

Claudin-low breast cancer is an aggressive subtype that confers poor prognosis and is found largely within the clinical triple-negative group of breast cancer patients. Here, we have shown that intrinsic and immune cell gene signatures distinguish the claudin-low subtype clinically as well as in mouse models of other breast cancer subtypes. Despite adaptive immune cell infiltration in claudin-low tumors, treatment with immune checkpoint inhibitory antibodies against cytotoxic T lymphocyte-associated protein 4 (CTLA-4) and programmed death receptor 1 (PD-1) were ineffective in controlling tumor growth. CD4+FoxP3+ Tregs represented a large proportion of the tumor-infiltrating lymphocytes (TILs) in claudin-low tumors, and Tregs isolated from tumor-bearing mice were able to suppress effector T cell responses. Tregs in the tumor microenvironment highly expressed PD-1 and were recruited partly through tumor generation of the chemokine CXCL12. Antitumor efficacy required stringent Treg depletion combined with checkpoint inhibition; delays in tumor growth were not observed using therapies that modestly diminished the number of Tregs in the tumor microenvironment. This study provides evidence that the recruitment of Tregs to the tumor microenvironment inhibits an effective antitumor immune response and highlights early Treg recruitment as a possible mechanism for the lack of response to immune checkpoint blockade antibodies in specific subtypes of cancer that are heavily infiltrated with adaptive immune cells.

Abstract 3286: Correlation between vitamin D levels and DNA repair capacity in breast cancer patients stratified by molecular subtypes

Abstract Vitamin D exists as vitamin D2 and D3, which are metabolized to 25-hydroxyvitamin D [25(OH)D], the major circulating vitamin D metabolite. Besides its physiological functions, vitamin D levels have also been studied as a risk factor for several hormonal cancers including breast cancer (BC). Worldwide, BC accounts for nearly a quarter of all cancers in women. Nutritional studies report that vitamin D intake is associated with a lower BC risk. Several discrepancies exist regarding the role of serum vitamin D in BC risk. While some studies report BC risk reduction by vitamin D only in premenopausal, others propose that it only occurs in postmenopausal women. BC tumors may (+) or may not (-) have three hormonal receptors: estrogen (ER), progesterone (PR), and HER2. Based on their status, four principal molecular BC subtypes have been identified: luminal A (ER+/−, PR+/−, HER2-), luminal B (ER+/−, PR+/−, HER2+), HER2+ (ER−, PR−, HER2+), and triple negative (TN) (ER−, PR−, HER2−). If BC is analyzed in terms of molecular subtypes, low vitamin D levels have been associated with aggressive phenotypes and worse prognosis. Vitamin D also influences estrogen synthesis. Since we have previously shown that a low DNA repair capacity (DRC), measured through the nucleotide excision repair pathway, is a risk factor for BC and vitamin D has also been found to affect DNA repair, the focus of this study is to examine the role of plasma vitamin D levels and DRC in BC. The main aim is to elucidate whether there is an association between vitamin D and DRC levels among the four molecular BC subtypes. We hypothesize that a negative correlation between 25(OH)D and DRC levels will be observed among these subtypes. As an initial effort, 47 BC cases and 20 controls without BC were selected from our large BC cohort. DRC was measured in lymphocytes of untreated women using the host cell reactivation assay. Pathology reports were examined to divide BC cases according to their molecular BC subtype: luminal A (n=13), luminal B (n=11), HER2+ (n=10), and TN (n=13). Plasma 25(OH)D levels were measured using the UniCel DxC System at a CLIA-certified lab. Our results show a negative correlation between 25(OH)D and DRC levels (p=0.04). Statistically significant differences were found for vitamin D levels among the different groups (p=0.0019, ANOVA). Moreover, higher 25(OH)D levels (47.97±2.4 ng/mL) were found in ER- BC cases (p=0.03, t-test). When comparing vitamin D levels in BC subtypes, a significant difference was found in HER2+ and TN groups when compared with the control group (p&amp;lt;0.05, Kruskal-Wallis). Based on these preliminary results we conclude that plasma vitamin D levels may be correlated with DRC levels in women with BC. Ongoing studies with a larger sample size are aimed at elucidating more precisely the potential association between vitamin D and DRC in these molecular BC subtypes. Citation Format: Carmen Ortiz, Jarline Encarnación, Ralphdy Vergne, Wanda Vargas, Jaime Matta. Correlation between vitamin D levels and DNA repair capacity in breast cancer patients stratified by molecular subtypes [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 3286. doi:10.1158/1538-7445.AM2017-3286

Abstract 3822: Expression of androgen receptor and CD24 correlates with improved 5-year survival in Pakistani patients with invasive breast cancer

Abstract Introduction: Androgen receptor (AR) expression has been shown to correlate with improved survival in luminal A, B and triple negative (TN) breast cancer (BCa). Cancer stem cells (CSC) with CD44+/CD24- and ALDH1+ phenotype is associated with pro-invasive gene signature in BCa. Aim of our study was to evaluate prognostic significance of AR expression in relation to CD44, CD24 and ALDH1 in invasive BCa. Methodology: Immunohistochemical expression of AR, CD44, CD24 and ALDH1 was evaluated in 180 FFPE blocks of BCa cases from one institution. Clinico-pathological details and follow-up data were collected from medical charts. Data was analyzed on SPSS 19. Five-year survival was estimated by Kaplan Meier. Results: Median age of patients was 55 years (range: 28-87). Fifteen percent of patients were diagnosed with stage I, 52.2% with stage II, 25% with stage III and 7.8% with stage IV disease. Mean duration of follow-up was 4.5 years (SD+ 2.7) and 29% mortalities were attributed to BCa. AR was expressed in 63.9%, ER in 61%, PR in 51% &amp; HER-2/neu in 30% of tumors. CD44, CD24 and ALDH1 were expressed in 60%, 43.3% &amp; 28.3% of tumors respectively. AR expression was associated (p-value&amp;lt;0.001) with grade II tumors expressing ER, PR and CD24 but lacking ALDH1. Multivariable analysis confirmed AR (adjusted HR 0.43, 95% CI 0.19-0.95) and PR (adjusted HR: 0.20, 95% CI: 0.07-0.60) to be independently associated with better outcome. Five year survival of patients with AR+ versus AR- expressing tumors, showed that patients with AR+ tumors had significantly better outcome (p-value=0.03) with survival advantage of 39.6 months. Survival advantage decreased by 9.6 months in tumors expressing CD44+/CD24-/AR- (p-value=0.01) phenotype and by 12 months in patients whose tumors were phenotypically ALDHI+ /AR- (p-value=0.12). We stratified TN tumors into 2 groups: A. TN/CD24+/AR+; B. TN/CD24-/AR- and found that group A tumors had a trend towards better 5 survival (p-value=0.09). Conclusions: Three individual markers (CD44, CD24, AR) may afford prognostic information. Combinations of CD44, CD24 and AR permitted patient stratification into 3 risk categories: poor risk CD24-/AR-; intermediate risk CD44+/CD24-/AR- and good risk CD24+/AR+. Stratification of TN group showed that loss of AR and CD24 was associated with poorest outcome. These results underscore the relevance and importance of AR and CD24 as prognostic markers in invasive BCa. Citation Format: Nazia Riaz, Romana Idress, Sadia Habib, Azhar Hussain, El-Nasir Lalani. Expression of androgen receptor and CD24 correlates with improved 5-year survival in Pakistani patients with invasive breast cancer [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 3822. doi:10.1158/1538-7445.AM2017-3822

The Diagnostic Value of miR-155 Expression in the Serum of Patients with Breast Cancer According to Molecular Subtypes of Breast Cancer

Background: microRNAs (miRNAs) have been highlighted as potential circulating biomarkers and therapeutic targets for breast cancer (BC). miRNAs have emerged as an extremely promising new class of biomarkers. They have been demonstrated to be remarkably stable in human blood. miR-155 has more than 4 hundred predicted gene targets. The miR-155 plays an essential role in the pathogenesis of breast cancer. The aim of this study was to assess the diagnostic value of miR-155 expression in the serum of patients with breast cancer according to the molecular subtypes of BC. Methods: 90 samples were classified into different groups with respect to their immunohistochemical (IHC) characteristics: estrogen receptor (ER) positive and/or progesterone receptor (PR) positive group (Luminal A), human epidermal growth factor receptor2 (HER2) positive group (as Luminal B), and Triple negative group (TNBC). The complementary DNA (cDNA) was synthesized in line with the guidelines of the Kit Company. A real-time PCR method was performed as the expression assay. Results: A substantial difference was observed between the phrase standard of miR-155 in patients with the molecular subtypes of breast cancer. The subtypes of Luminal A, Luminal B, and TNBC showed respectively 6 fold, 7 fold, and 14 fold increased expression (P < 0.001). Receiver operating characteristic (ROC) analysis revealed that miR-155 had a considerable diagnostic accuracy for Luminal A with area under the curve (AUC) of 0.875 (sensitivity 100%, specificity 68.63%), for Luminal B with AUC of 0.886 (sensitivity 100%, specificity 62.75%), and for TNBC with AUC of 0.982 (sensitivity 100%, specificity 92.16%) Conclusions: The study found that increased mir-155 expression was significantly associated with BC. The study results showed the mir-155 expression has extremely high sensitivity and specificity for the diagnosis of subtypes of breast tumor. Therefore, checking the serum levels of miR-155 expression in patients with breast cancer may be helpful. Yet, more studies are required to investigate this relationship.

Outcomes of patients with inflammatory breast cancer by hormone receptor- and HER2-defined molecular subtypes: A population-based study from the SEER program.

BackgroundThe aim of this study was to evaluate the outcomes of patients with inflammatory breast cancer (IBC), with emphasis on the role of molecular subtypes and radiotherapy.MethodsA retrospective cohort study to investigate overall survival (OS) and breast cancer-specific mortality (BCSM) in patients with IBC was conducted using data obtained by the Surveillance, Epidemiology, and End Results (SEER) program from 2010–2013. Cox multivariate regression was used to calculate the adjusted Hazard Ratios (aHR).Results403 patients were eligible for this study. Patients in the group with hormone receptors (HR)+/HER2- subtype had an OS of 79.6% compared with 89.0 % in the group with (HR)+/HER2+ subtype and 76.8% in the HR-/HER2+ group and 62.9% in the triple-negative (TN) group. BCSM was 16.3% for the HR+/HER2- group, 9.8% for the HR+/HER2+ group, 21.7% for the HR-/HER2+ group, and 30.5% for the TN group. For distant metastases, the results showed that there was a high probability of bone metastasis in HR-positive groups, brain and liver metastasis in HER2-positive groups, and lung metastasis in the TN group. Multivariate analysis demonstrated that estrogen receptor and HER2 positivity were associated with better survival and that the TN subtype had a poorer OS and BCSM compared with other subtypes (P<0.05). Furthermore, patients who received radiotherapy were more likely to have improved survival (P< 0.05).ConclusionInflammatory breast cancer appears to alter the prognosis in association with the receptor status and molecular subtypes. Radiotherapy was still considered to be a crucial treatment for patients with IBC.

Failure patterns according to molecular subtype in patients with invasive breast cancer following postoperative adjuvant radiotherapy: long-term outcomes in contemporary clinical practice

Although gene expression profiling provides critical information, knowledge remains limited regarding the differential effects of molecular subtype on clinical course. This study evaluated the impact of molecular status on long-term patterns of failure in patients with non-metastatic breast cancer. We analyzed data from 1181 individuals with invasive breast cancer undergoing surgery plus PORT from 2003 to 2011. Molecular subtypes were defined as luminal A (LA), luminal B (LB)-HER2(-), LB-HER2(+), HER2, and triple-negative (TN) based on the 2013 St. Gallen Consensus criteria. Competing risks analysis and baseline hazard rate function plots were used to explore subtype-specific recurrence patterns. The 10-year overall survival rates of LA, LB-HER2(-), LB-HER2(+), HER2, and TN groups were 96, 93, 94, 84, and 85%, respectively (P<0.001). Distant metastatic events differed significantly according to molecular subtype (P<0.001). In competing risks regression analysis, initial development of distant metastasis was the highest with TN tumors, followed by HER2, LB-HER2(-), and LB-HER2(+) subtypes (P=0.005). Regarding preferential sites of distant metastasis, the risk of initial brain metastasis was significantly higher with HER2 tumors, followed by TN tumors (P=0.001). A low-level but sustained metastatic risk increment was observed in luminal tumors, whereas TN and HER2 subtypes showed a short-term risk surge within 5years. From the significant impact of molecular profile on distant metastasis, subtype-specific individualization of systemic treatment and close surveillance are suggested. The preferential and long-term risk of brain metastasis in the HER2 subtype underlines the importance of alternative anti-HER2 therapies.

Abstract P6-09-12: A consecutive series of early breast cancers with a low estrogen receptor expression

Abstract P6-09-12: A consecutive series of early breast cancers with a low estrogen receptor expression

Abstract P2-05-12: The ASCO-recommended prognostic factors uPA/PAI-1 in a multicenter cohort study (PiA)

Abstract Introduction The PiA-study (Prognostic assessment in routine Application, NCT 01592825) was designed as a representative cohort of breast cancer patients to estimate the proportions of traditional and modern prognostic factors. The ASCO-recommended biomarkers uPA (urokinase-type plasminogen activator) and its inhibitor PAI-1 were used for biological risk assessment particularly for intermediate risk breast cancer patients and disease-free survival of the patients after 5 yrs of follow-up (F/U) was calculated. Material &amp; Methods Between 2009 and 2011, 1,074 non-metastasized, primarily operated breast cancer patients from six centers in Germany were included. From 815 patients, fresh frozen tissue was obtained and processed for central testing uPA/PAI-1 by ELISA (FEMTELLE®, Sekisui Diagnostics GmbH). Low uPA/PAI-1 status is defined by uPA and PAI-1 concentrations below the published cut-offs, high status means one or both were higher than the corresponding cut-offs. Tumor characteristics were based on local pathology. The centers had to follow the national guidelines. In low-risk patients, adjuvant chemotherapy was spared. The median F/U is 56 months (range 0-78). Results In the total cohort of 1,074 patients, 166 had G1- and 237 had G3-tumors. Of the 671 patients with a G2-tumor, the following were allocated to the high-risk group: node-positive (n=371), younger than 35 yrs (n=17), and triple-negative (TN) or HER2-positive (n=118). For 253 tumors of the remaining 355 patients with an intermediate risk of recurrence (pN0, G2, HR positive, HER2-negative, ≥35 yrs), uPA/PAI-1 status was available. 126 (49.8%) were allocated to the low-risk group, one patient had a recurrence. At 5 yrs, in the total cohort 90.6% (95% CI, 89.5-91.7) of the patients were free of invasive disease. Of 114 HER2-positive tumors, 94 (82.4 %) had a high uPA/PAI-1 status, only one of the 38 HR negative/HER2 positive tumors had a low uPA/PAI-1 status. In the TN group, the majority of tumors had a high uPA/PAI-1 status (66 of 81; 81.5 %). In 30 patients lymph nodes were involved, 18.5% (n=15) had a low uPA/PAI-1 status, one event was detected. In N pos. patients with an high uPA/PAI-1 6 events were observed. Conclusion Testing for uPA/PAI-1 in the daily routine is feasible, fresh frozen tissue has been prepared from 76% of the tumors of the recruited patients, 37% of them had a low risk status. Using uPA/PAI-1, about half of the node-negative patients with an intermediate risk of recurrence were allocated to a group with an extremely low risk of recurrence and thus chemotherapy could be spared. Also in node-positive disease, uPA/PAI-1 has a prognostic impact. Tab 1: Proportion of the subgroups according to IHC, grading and uPA/PAI-1-statusTumor typetotallow uPA/PAI-1 statushigh uPA/PAI-1 statusn=815 (100%)n=304 (37%)n=511 (63%)Luminal A-like tumors:HRpos., HER2neg., G1, G2515 (63.2%)240 (78.9%)275 (53.9%)Luminal B/HER2-negative-like tumors:HRpos., HER2neg., G3104 (12.8%)29 (9.5%)75 (14.7%)Luminal B/HER2-positive-like tumors:HR pos., HER2 pos., all grades77 (9.4%)19 (6.2%)58 (11.4%)HER2-positive (nonluminal) - like tumors:HRneg., HER2pos., all grades38 (4.7%)1 (0.3%)37 (7.3%)TN tumors:HRneg., HER2neg., all grades81 (9.9%)15 (4.9%)66 (12.9%) Citation Format: Vetter M, Hartung C, Hanf V, Lantzsch T, Uleer C, Peschel S, John J, Buchmann J, Bürrig K-F, Weigert E, Thomssen C, Kantelhardt EJ. The ASCO-recommended prognostic factors uPA/PAI-1 in a multicenter cohort study (PiA) [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 P2-05-12.