Triple-negative Breast Cancer Group Research Articles

Expression of PD-L1 and BRCA1 in Triple-Negative Breast Cancer Patients and Relationship with Clinicopathological Characteristics.

Objective To study the expression of programmed cell death ligand-1 (PD-L1) and breast cancer susceptibility gene 1 (BRCA1) in triple-negative breast cancer (TNBC) patients and analyze their relationship with clinicopathological characteristics. Methods 76 TNBC tissues were collected as the research object, while 60 adjacent tissues were used as controls. All patients underwent surgical treatment, and the expression of PD-L1 and BRCA1 in cancer tissues and adjacent tissues was detected by immunohistochemistry. At the same time, the relationship between PD-L1, BRCA1, and clinicopathological characteristics of patients with TNBC (including patient age, menopausal status, tumor size, lymph node metastasis, histological grade, Ki-67 expression, and p53 expression) were analyzed by univariate and logistic multivariate analysis. Results The positive expression rate of PD-L1 in the TNBC group was 64.47%, which was higher than the control group by 41.67%. The positive expression rate of BRCA1 was 27.63%, which was lower than the control group by 48.33%. PD-L1 expression has no significant relationship with age, menopausal status, and p53 expression in TNBC patients. TNBC patients with tumors ≥2 cm, histological grade III, lymph node metastasis, and Ki-67 expression ≥20% had higher PD-L1 positive expression rates. The tumor size, Ki-67 expression, and PD-L1 expression of TNBC patients have independent effects. The expression of BRCA1 has no significant relationship with menopausal status, tumor size, Ki-67 expression, etc. TNBC patients with age <45 years, histological grade I or II, no lymph node metastasis, and high p53 expression positive rate had higher BRCA1 positive expression rate. The age of TNBC patients, p53 expression, and BRCA1 expression have independent effects. Conclusion In TNBC cancer tissues, there is a high expression of PD-L1 and low expression of BRAC1. The tumor size, Ki-67 expression, and PD-L1 expression of TNBC patients have independent effects. The age of TNBC patients, p53 expression, and BRCA1 expression have independent effects.

Automated breast volume scanning combined with shear wave elastography for diagnosis of triple-negative breast cancer and human epidermal growth factor receptor 2-positive breast cancer.

To explore the values of automated breast volume scanning (ABVS) combined with shear wave elastography (SWE) in the differential diagnosis of triple-negative breast cancer (TNBC) and human epidermal growth factor receptor 2-positive breast cancers (HER2+BC). In this study, 28 patients with TNBC and 32 patients with HER2+BC were enrolled. The characteristics of ABVS and virtual touch quantification (VTQ) in SWE of all patients were reviewed. The multivariate logistic regression analysis was carried out and the receiver operating characteristic curves of ABVS and ABVS+VTQ were drawn. In ABVS imaging, the microcalcification, posterior echo, internal echo, shape, and edge had significant difference between TNBC and HER2+BC groups (p<0.05). The regular shape was the independent factor for TNBC (p=0.04, odds ratio [OR]=4.479), and the microcalcification in mass was the independent factor for HER2+BC (p=0.01, OR=2.997). In VTQ imaging, the shear wave velocity (SWV)max, SWVmin, and SWVmean in TNBC group were significantly lower than those in HER2+BC group (p<0.001). The sensitivity, specificity, and accuracy of ABVS+VTQ in diagnosing TNBC were higher than those of ABVS alone. ABVS combined with SWE has certain advantages in differentiating TNBC from HER2+BC, which is helpful for the treatment planning and prognosis judgment.

The importance of androgen receptors in breast cancer.

Breast cancer (BC) is the most common malignancy among women worldwide, and one of the leading causes of cancer-related deaths in females. For the breast malignant tumors there are numerous targeted therapies, depending on the receptors expressed. Regulating the process of epithelial-mesenchyme transcription, the steroid nuclear receptors are important in invasion and progression of BC cells. Till now, it is known that androgen receptor (AR) is present in about 60-80% of BC cells but, unfortunately, there is no targeted therapy available yet. We revised the recent literature that included the AR mechanism of action in patients diagnosed with breast cancer, the preclinical, retrospective and clinical studies and the aspects related to the prognosis of these patients, depending on the molecular subtype. A total of 12 articles were eligible for this review. AR positivity was assessed using immunohistochemistry. Herein, neither 1 nor 10% cut-points were robustly prognostic. AR was an independent prognostic marker of BC outcome, especially in triple negative BC group. AR is a potential targeted pathway which can improve the prognostic of AR positive patients with BC. Further preclinical and clinical studies are necessary to clarify the mechanism of action and to establish the drugs which can be used, either alone or in combination.

Application value of conventional ultrasound combined with shear wave elastography in diagnosing triple negative breast cancer

At present, most ultrasound (US) studies on triple negative breast cancer (TNBC) are limited to conventional US features, so it is necessary to develop new joint diagnostic methods. The study aims to explore the values of conventional US and shear wave elastography (SWE) in differential diagnoses of TNBC and non-TNBC. A total of 120 breast cancer (BC) patients involving 120 lesions that were pathologically verified were retrospectively analyzed in this study. All participants had received both conventional US and SWE before surgery. Meanwhile, the participants were divided into a TNBC group or a non-TNBC group according to their immunohistochemical (IHC) results. The differences between the conventional US features (including lesion shape, growth location, margin, boundary, internal echo, micro-calcification, posterior echo, and internal blood supply) and the SWE image features [including mean lesion hardness (Emean), maximum (Emax), minimum (Emin), standard deviation (SD), and ratio to normal gland (Eratio)] of 2 groups were compared. The receiver operating characteristic (ROC) curve of the diagnosed lesion was calculated by the area under the curve (AUC). According to the findings of conventional US, the TNBC group mostly manifested as a micro-lobulated margin, with a clear boundary and no internal micro-calcification; the non-TNBC group mainly manifested as marginal angulation or burr, and hyper-echo halo in the boundary, accompanied with internal micro-calcification, and the difference was statistically significant (P<0.05); the internal thrombolysis in myocardial infarction (TIMI) and resistance index between the TNBC group and non-TNBC group were similar, and the differences were not statistically significant (P>0.05). The findings of SWE were as follows: differences in Emax, Emean, and Eratio values between 2 groups were statistically significant (P<0.05); and the areas under the ROC curve (AUC) of these three in diagnosing the lesions were 0.811, 0.781 and 0.770, respectively. Conventional US combined with SWE can comprehensively analyze the morphological, blood supply, and hardness features of breast lesions, and provide more reliable information for the differential diagnosis between TNBC and non-TNBC.

Strong impact of pathological node-negative on long-term overall survival of patients with triple-negative breast cancer receiving neoadjuvant chemotherapy.

Triple-negative breast cancer (TNBC) has a high pathological complete response (pCR) rate; however patients without a high pCR are reported to have a poor prognosis. The current study investigated the long-term overall survival of patients with TNBC who received neoadjuvant chemotherapy (NAC) and analyzed various prognostic factors including basal marker and claudin expressions. Between November 2005 and March 2012, the current study retrospectively reviewed the records of 323 patients with breast cancer who received anthracycline followed by taxane as NAC at the Jikei University Hospital Basal marker and claudin expression was determined via immunohistochemistry. The median age of the patients was 53.0 years. Of the 323 patients, 26 (8%) achieved a pCR, including 13 patients (19.7%) with TNBC and 13 (5.1%) with non-TNBC (P<0.001). Of the 66 patients with TNBC, 13 (19.7%) demonstrated recurrence and 8 (12.1%) died after a median follow-up time of 111.5 months [10-year disease-free survival (DFS), 80.3%; 95% confidence interval (CI), 0.68-0.88; 10-year overall survival (OS), 84.8%; 95% CI, 0.72-0.92]. Of the 257 patients with non-TNBC, 45 (17.5%) patients demonstrated recurrence and 26 (10.1%) died (10-year DFS, 82.1%; 95% CI, 0.76-0.87; 10-year OS, 88.6%; 95% CI, 0.83-0.92). There was no statistical difference between the patients with and without TNBC. In the TNBC group, patients with pathological node-negative status survived without distant recurrence. Additionally, negative lymphovascular infiltration was another favorable prognostic factor. Patients with TNBC who received NAC demonstrated comparably high prognoses to non-TNBC patients. Overall, pathological node status after NAC had a strong impact on the prognosis of patients with TNBC.

THE CORRELATION OF THE BRCA-IRIS GENE EXPRESSION AND BIOMARKER GENES IN BREAST CANCER PATIENTS: CORRELATION WITH TUMOR PROGRESSION AND RESPONSE TO TREATMENT

Background: Triple-negative breast cancer (TNBC) is an aggressive phenotype with bad prognosis and poor survival. The prognostic and predictive values of BRCA1-Iris, octamer-binding transcription factor 4 (oct 4), cyclin D1andsurvivin were assessed in TNBC patients compared with N-TNBC patients. Method: RNA expression levels of BRCA1-IRIS, Oct4, Cyclin D1 and Survivin gene were tested in for 100 breast cancer patients by qRT-PCR. Result: overexpression ofBRCA1-IRIS RNA was found in 49 % of breast cancer patients [(35(71 %) of TNBC cases vs 14(28.6% of N-TNBC] (p < 0.001). There was significantly correlated between the BRCA1-IRIS and tumor stage, ER and PR (p=0.036, p < 0.001 and p=0.006; respectively) in breast cancer patients. In TNBC or N-TNBC, no statistically significance between BRCA1-IRIS and any clinicopathological features of patients. There was significant association between BRCA1-IRIS positive and the expression of oct4, cyclin D1 and survivin gene in TNBC patients (p < 0.001, p=0.001 and p=0.002; respectively). There was a strong significant correlation between the expression of oct4 and the BRCA1-IRIS negative (p < 0.001) in TNBC group. No significant association between BRCA1-IRISpositive in N-TNBC and any of the gene expression. BRCA1-IRISnegative was correlated significantly with oct4 and cyclin D1expression (p=0.001each).No significant correlation between the expression RNA level of oct4, cyclin D1 and survivin gene and the expression RNA level of BRCA1-IRIS except lymphnode with survivin gene expression (p=0.05). No significant relation between the expression level of oct4, cyclin D1 and survivin gene and the relevant clinicopathological features with or without BRCA1-IRIS expression. Expression level of survivin and cyclin D1 was significantly correlation with the response to treatment (p=0.001 and p < 0.001). No significant different between overall survival and BRCA1-IRIS expression (p=0.291 long rang). However, disease-free survival was increased in BRCA1-IRIS positive cases (p = 0.052 long rang)

THE RNA EXPRESSION LEVEL OF SURVIVIN, OCT 4 AND CYCLIN D1 GENES IN TRIPLE NEGATIVE AND NON-TRIPLE NEGATIVE BREAST CANCER

Background: the purpose of the present study to investigate the RNA expression level of Oct4, Survivin and Cyclin D1 genes in Triple-negative breast cancer (TNBC) and compared to non-triple negative breast cancer (non-TNBC) group. Also, correlate the results of gene expression with clinicopathological features of patients. Method, RNA expression levels of Oct4, Survivin and Cyclin D1 was tested for 100 breast cancer (BC) patients [formalin fixed paraffin embedded tissue (FFPE)] diagnosed as invasive duct carcinoma in Pathology Department, National Cancer Institute (NCI), Cairo University. The qRT-PCR technique is use and the results correlated to clinico-pathological characteristics of patients and survival rates. Result, in (TNBC) patients, oct4, survivin and cyclin D1 showed positive expression in 30 (60%), 32(64%) and 32 (64%) (p < 0.001, p= 0.001 and p= 0.003;respectively). In non-TNBC patients, the positive expression of oct4, survivin and cyclin D1 gene was 11(22%), 15(30%) and 17(34%) (p < 0.001, p= 0.001 and p= 0.003;respectively). All patients in TNBC have negative ER, PR and HER-2 receptor (p=0.001). There was highly statistically significant difference between oct4 gene expression and all the clinico-pathological features except the family history. A significant difference between cyclin D1gene expression and all the clinico-pathological features except the tumor size, tumor grade and lymphnode status. Statistically difference was found between survivin gene expression and all the clinico-pathological features except the menopause state, tumor stage and tumor grade. Triple negative breast cancer patients showed significantly decreased DFS (p < 0.001, log rank) and OS (p= 0.002, log rank) when compared to those with non-TNBC patients. In TNBC group, patients with positive expression of survivin was significantly associated with decreased OS (p= 0.03, log rank). TNBC tumors with positive oct4 and cyclin D1 had reduced OS compared to those negative to oct4 and cyclin D1 but without significant difference (p= 0.8 and p= 0.09, respectively, log rank). In non-TNBC group, with positive oct4, cyclin D1 and survivin did not significantly differ in terms of DFS and OS when compared to those with negative expression. Conclusion: Oct4 and survivin expression gene are better marker used for diagnosis in TNBC and for molecular targeting therapy of TNBC treatment. Cyclin D1 expression used as a marker for aggressive TNBC. The TNBC tumor possibly respond to treatment that downregulates cyclin D1 amplifica tion. More studied and large sample size is needed.

Abstract PS4-37: A pro-tumorigenic mechanism of M2 tumor-associated macrophages (TAM) in triple-negative breast cancer

Abstract Introduction:Triple-Negative Breast Cancer (TNBC) comprises approximately 30% of all breast cancers in Indian women. Given their aggressive nature, TNBCs have high rates of systemic metastasis and mortality with only chemotherapy available for treatment. The success of immunotherapy in solid tumors has raised the hope for their utility in TNBCs as well. But only a subset of patients have a clinical response to check-point inhibitors. The cellular and molecular mechanisms that mediate the immunological response or tolerance are just beginning to be understood. Compared to ER/PR+ breast cancer, TNBC features a unique tumor microenvironment (TME) characterized by a large number of tumor-infiltrating lymphocytes (TILs) and tumor-associated macrophages (TAMs). The density of TILs in and of themselves do not accurately predict response to neoadjuvant chemotherapy or survival. M2 tumor-associated macrophages (M2-TAMs) have been reported to associate with solid tumors to facilitate epithelial to mesenchymal transition (EMT), tumor invasiveness, metastasis, and resistance to therapy. In this study, we have characterized presence of M2-TAM in the TNBC immune environment by examining expression of several biomarkers. Methods:Surgically excised tumor specimens from 88 Indian women with TNBC were accessed from the longitudinal observational series of SJNAHS tissue bank under IERB approved protocols and assayed on nCounter® PanCancer Immune Profiling Panel which comprised of 740 genes and characterises 14 different immune cell types. Transcriptome analysis using Non-negative Matrix factorization (NMF) based unsupervised clustering was done to arrive at stable subtypes characterized by different tumour microenvironments within TNBC. CIEBERSORT tool was used to analyse for distribution of cell types. Identification of macrophages was done by Immunohistochemistry (IHC) for CD68 and CD163 markers in TNBCs and a control group of ER+HER2-.Results:NMF analysis with an enriched immune gene signature of 111 genes, yielded 3 subtypes (ST) (37%, 27% and 36% with relative proportions of ST1, ST2 and ST3) within the TNBC. The three subtypes had distinct survival patterns with ST1 having the best prognosis with enriched TH1 gene signature and ST3 had poorest (log rank p=0.5). On application of this gene signature to TNBC groups in METABRIC and TCGA data, similar pattern emerged with a significant survival pattern of ST 3 having the poorest outcome (p=0.005). A closer examination of ST 3 revealed a signature enriched for M2 macrophages also known as TAM. Immunohistochemistry for Pan macrophage marker CD68 and an M2 specific marker CD163 between TNBC and ER/PR+ revealed difference in the two groups was significantly different (Mann Whitney p=0.03), with TNBC exhibiting 2.4 fold higher expression of CD163. CD 68, was enriched in ST2 and ST3. CD163 which is an M2 specific marker was highest in ST3 as compared to ST 1&amp; 2 (p=0.04). This group also correlated with signal molecules secreted by macrophages containing growth factors, cytokines and chemokines, such as TGF-β, VEGF, IL-10 and CXCL and interleukins like IL4 and IL6 suggestive of TAM recruitment and polarization. It is likely that the TAM enriched subgroup is likely to be unresponsive to PD1/PDL1 inhibitors but could be targeted by novel therapeutic strategies to directly target M2-TAM. In vitro and In vivo analysis to target M2-TAM to evaluate the response to these therapeutic compounds in cell lines as well as a mouse syngeneic model is underway.Conclusions and future directions: As a cell type within the tumor microenvironment, that promotes invasion, M2-TAMs makes an ideal therapeutic target in TNBC. In addition, this sub-type lends itself to easy identification by a simple IHC assay. Citation Format: Aruna Korlimarla, Jyothi Prabhu, Chantirika Ragulan, Gnanapriya Shivakumar, Krisha Desai, Maggie Cheang, Srinath BS, Ravi Diwakar, Sandhya Apachu, Rekha Kumar, TS Sridhar, Savitha RajaRajan, Rohini Kaluve, Annie Alexander, Anguraj Sadanandam. A pro-tumorigenic mechanism of M2 tumor-associated macrophages (TAM) in triple-negative breast cancer [abstract]. In: Proceedings of the 2020 San Antonio Breast Cancer Virtual Symposium; 2020 Dec 8-11; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2021;81(4 Suppl):Abstract nr PS4-37.

Hsa_circ_0000199 facilitates chemo-tolerance of triple-negative breast cancer by interfering with miR-206/613-led PI3K/Akt/mTOR signaling.

Increasing attentions have been paid to the role of circRNAs in the etiology of triple-negative breast cancer (TNBC), and we strived to figure out the association of circRNA AKT3/miRNA axis with TNBC chemo-resistance. Altogether 207 BC patients were divided into TNBC group (n=83) and non-TNBC group (n=124), and MCF-10A, MDA-MB-231, MDA-MB-468, SK-BR-3 and MCF-7 cell lines were prepared in advance. Expressions of AKT3-derived circRNAs and relevant miRNAs in the TNBC tissues and cell lines were determined by employing real-time polymerase chain reaction (PCR). It was indicated that hsa_circ_0000199 expression was higher in TNBC tissues than in non-TNBC tissues, and high hsa_circ_0000199 expression was predictive of large tumor size, advanced TNM grade, high Ki-67 level and poor 3-year survival of TNBC patients (all P<0.05). Furthermore, miR-613 and miR-206 were sponged and negatively regulated by hsa_circ_0000199 (P<0.001), and PI3K/Akt/mTOR signaling was depressed by si-hsa_circ_0000199 in TNBC cell lines (P<0.01). Ultimately, miR-206/miR-613 inhibitor reversed impacts of si-hsa_circ_0000199 on PI3K/Akt/mTOR signaling, proliferation, migration, invasion, chemo-sensitivity and autophagy of TNBC cells (all P<0.01). Conclusively, silencing of hsa_circ_0000199 enhanced TNBC chemo-sensitivity by promoting miR-206/miR-613 expression and deactivating PI3K/Akt/mTOR signaling, which was conducive to improving chemotherapeutic efficacy of TNBC patients.

Clinical Significance of Receptor-Interacting Protein 3 and Parkin, Essential Molecules for Necroptosis, in Breast Cancer.

PurposeReceptor-interacting protein 3 (RIP3) is the main initiator of necroptosis. Parkin prevents the formation of the RIP1–RIP3 complex by promoting polyubiquitination of RIP3. However, the mechanism by which necroptosis affects the clinical features of breast cancer and prognosis is not known. Here, we aimed to study the effect of necroptosis on the clinical features and prognosis of breast cancer by assessing the expression of RIP3 and Parkin.MethodsTissue microarrays (TMAs) were constructed from 257 cases of breast cancer. Immunohistochemistry was performed on 4-μm tissue sections from each TMA block. The χ2 test, Kaplan-Meier survival analysis with log-rank test, and Cox regression proportional hazard model were used for statistical analysis.ResultsLow RIP3 expression resulted in a large tumor size and high nuclear grade. Low RIP3 expression was correlated with human epidermal growth factor receptor 2 positivity, short overall survival (OS), and short disease-free survival (DFS). The triple negative breast cancer group with low RIP3 expression and lymph node (LN) positive group with low RIP3 expression had the shortest OS. High Parkin expression was associated with high histological grade, estrogen and/or progesterone receptor negativity, and lymphatic emboli, but was not correlated with OS and DFS. OS was correlated with LN metastasis and RIP3 loss and DFS with large tumor size, LN metastasis, and RIP3 loss.ConclusionLow RIP3 and high Parkin expression are associated with aggressive clinical features in breast cancer. RIP3, a molecular marker of necroptosis, is an independent factor associated with survival in breast cancer. Further in-depth studies are needed to investigate the role of necroptosis in breast cancer development, metastasis, and treatment in the future.

Predictive Biomarkers for Immune Checkpoint Inhibitors in Metastatic Breast Cancer.

We examined a large dataset of female metastatic breast cancers (MBCs) profiled with comprehensive genomic profiling (CGP) to identify the prevalence and distribution of immunotherapy responsiveness‐associated biomarkers. DNA was extracted from 3831 consecutive MBCs: 1237 (ERpos/HER2 neg), 1953 ERneg/HER2 amp, and 641 triple‐negative breast cancer (TNBC). CGP was performed using the FoundationOne® or FoundationOne®CDx NGS assay. Tumor mutational burden (TMB) and microsatellite instability (MSI) were determined in a subset of cases. PD‐L1 expression in immunocytes in a subset of cases was determined by immunohistochemistry using the companion diagnostic VENTANA PD‐L1 SP142 Assay. The median age of the cohort was 54 years (range 20–89). Genomic alterations (GAs)/tumor were similar (range: 5.9–7.3). Markers of potential immune checkpoint inhibitor (ICPI) benefit included: CD274 (PD‐L1) amplification (1%–3%), BRAF GA (1%–4%), TMB of ≥10 mutations/Mb (8%–12%), MSI‐high (0.1%–0.4%), PBRM1 GA (1%), and positive PD‐L1 staining of immunocytes ranging from 13% in ERpos/HER2 neg and 33% in ERneg/HER2 amp to 47% in the TNBC group. Potential markers of ICPI resistance included inactivating STK11 GA (1%–2%) and MDM2 amplification (3%–6%). MTOR pathway targets were common with lowest frequency in TNBC. ERBB2 short variant mutations were most frequent ERpos/HER2 neg and absent in TNBC. BRCA1/2 GA were least frequent in ERneg/HER2 amp. The demonstrations of clinical benefit of immunotherapy in MBC support the need for development and utilization of biomarkers to guide the use of ICPIs for these patients. In addition to guiding therapy selection, CGP shows potential to identify GA linked to response and resistance to ICPI in MBC.

Profile and Outcome of Triple Negative Breast Cancer at a Tertiary Care University Hospital in Nepal

Background: TNBC (Triple negative breast cancer) subtype (ER-/PR-/HER2-) of breast cancers are known for aggressive tumor biology and poor survival prospects, with high early relapse rate. However, little is known about the prevalence and characteristics of TNBC breast cancer in Nepal. Objectives: To clarify the geographical distribution, clinical profile and outcome of TNBC patients when compared with non-TNBC patients managed in tertiary care university hospital in Nepal. Materials and Methods: This is a study on prospective observational analyses of TNBC and non-TNBC patients managed at Tribhuvan University Teaching Hospital, Kathmandu from October 2015-March 2018. We collected and analyzed data on clinical profile, pathological tumor features and outcome of the two patient groups. Results: A total of 108 breast cancer patients were included in our study, 38 (35.2%) of which were TNBCs. Mean age at diagnosis was 49±12 years. Majority of TNBCs (29%) were concentrated in Terai districts compared to non-TNBCs (18.6%). Among TNBCs, 15.2% had tumor size 5 cm or more while only 11.9% had such finding in non-TNBCs. Majority of TNBCs and non-TNBCs were of invasive ductal carcinoma of NST histology (76.3 vs 90%). TNBC tumors were significantly of grade 3 (P value=0.003). Perineural invasion was seen more (15.8%) in TNBCs compared to 8.96% in non-TNBCs. On average, 23.6% of total nodes retrieved from axilla were positive for tumor in TNBCs compared to 21% in non-TNBCs. Three patients developed metastases in TNBCs of which two were to brain while 5 had metastases in non-TNBC with none to brain. Higher percentage of patients died in TNBC group (13.2 vs 7.1%). Conclusions: TNBCs are quite common, higher grade tumors with brain metastasis without particular geographic distribution.

Machine Learning for Precision Breast Cancer Diagnosis and Prediction of the Nanoparticle Cellular Internalization.

In the field of theranostics, diagnostic nanoparticles are designed to collect highly patient-selective disease profiles, which is then leveraged by a set of nanotherapeutics to improve the therapeutic results. Despite their early promise, high interpatient and intratumoral heterogeneities make any rational design and analysis of these theranostics platforms extremely problematic. Recent advances in deep-learning-based tools may help bridge this gap, using pattern recognition algorithms for better diagnostic precision and therapeutic outcome. Triple-negative breast cancer (TNBC) is a conundrum because of the complex molecular diversity, making its diagnosis and therapy challenging. To address these challenges, we propose a method to predict the cellular internalization of nanoparticles (NPs) against different cancer stages using artificial intelligence. Here, we demonstrate for the first time that a combination of machine-learning (ML) algorithm and characteristic cellular uptake responses for individual cancer cell types can be successfully used to classify various cancer cell types. Utilizing this approach, we can optimize the nanomaterials to get an optimum structure-internalization response for a given particle. This methodology predicted the structure-internalization response of the evaluated nanoparticles with remarkable accuracy (Q2 = 0.9). We anticipate that it can reduce the effort by minimizing the number of nanoparticles that need to be tested and could be utilized as a screening tool for designing nanotherapeutics. Following this, we have proposed a diagnostic nanomaterial-based platform used to assemble a patient-specific cancer profile with the assistance of machine learning (ML). The platform is composed of eight carbon nanoparticles (CNPs) with multifarious surface chemistries that can differentiate healthy breast cells from cancerous cells and then subclassify TNBC cells vs non-TNBC cells, within the TNBC group. The artificial neural network (ANN) algorithm has been successfully used in identifying the type of cancer cells from 36 unknown cancer samples with an overall accuracy of >98%, providing potential applications in cancer diagnostics.

ATM kinase regulates tumor immunoreactions in lymphocyte-predominant breast cancer through modulation of NKG2D ligand and TNF cytokines on tumor cells.

To explore impact of Ataxia telangiectasia mutated (ATM) kinase on immunoreactions in lymphocyte-predominant breast cancer (LPBC), particularly its role in triple negative breast cancer (TNBC), 194 cases of LPBC were identified with pertinent clinical information retrieved. The expressions of ATM, activated ATM (P-ATM), Fas ligand (FASL), tumor necrosis factor-related apoptosis-induced ligand (TRAIL), major histocompatibility complex class I chain-related protein A (MICA), CD8, and Forkhead box P3 (FOXP3) were assessed by immunohistochemically. We found that ATM expressed on tumor cells was correlated with upregulated expression of P-ATM and MICA (P < 0.05), down-regulated expression of FASL and TRAIL (P < 0.01), and decreased Ki-67 tumor labeling (P < 0.05). However, within the TNBC group, only a negative correlation with FASL expression was found (P = 0.001). ATM and MICA expressions were significantly down -regulated in TNBC (P < 0.01) compared to non-TNBC, while TRAIL was significantly upregulated (P < 0.01). Tregs were increased in TNBC (P < 0.05), with CD8 + TILs decreased (P < 0.01). Ki-67 index was higher in TNBC than in non-TNBC (P < 0.01). ATM may play an important role in immunoreaction of LPBC, probably through upregulation of MICA and down-regulation of FASL and TRAIL. The down-regulated ATM expression in TNBC might be responsible for impaired tumor immunoactivity, rapid tumor growth, and aggressive clinical course.

Clinicopathological, Treatment and Event-Free Survival Characteristics in a Moroccan Population of Triple-Negative Breast Cancer

Introduction:Triple-negative breast cancer (TNBC) is a group of breast carcinoma characterized by the lack of expression of estrogen and progesterone hormone receptors (ER, PgR) and HER2. This form is also characterized by its aggressiveness, a low survival rate, and the absence of targeted therapies. This study was planned to evaluate the clinical features, treatment, and prognosis characteristics of TNBC in a population of Moroccan patients.Methods:In this retrospective study, a total of 905 patients diagnosed with breast cancer at the National Institute of Oncology in Rabat, Morocco, have been included. Based on molecular subtype, patients were divided into 2 categories: TNBC and non-TNBC patients. Data were recorded from patients’ medical files and analyzed using SPSS 13.0 software (IBM).Results:Overall, 17% of the patients had TNBC. At diagnosis, the median age of TNBC cases was 47 years, with extreme ages of 40 and 55 years. The median follow-up time was 30 months (10-53 months) and the 3-year survival rate was 76%. No significant difference was observed among the patients in terms of age at diagnosis, age at menarche, age at the time of first birth, nulliparity, oral contraception, and family history of breast cancer. Menopausal status and the number of pregnancy were significantly higher in the non-TNBC group. The percentage of grade 3 (G3) tumors was higher in the TNBC group (P < .001). Using neoadjuvant, adjuvant chemotherapy and radiotherapy, a net benefit in the event-free survival was registered for the 2 groups.Conclusions:This retrospective study was very informative and showed that women with TNBC had a less favorable prognosis than non-TNBC cases. Clinical data demonstrated that risk factors including age, premenopausal status, parity, hormonal contraceptive use, advanced disease, and a high histologic grade were independently associated with TNBC. However, large tumors and high Scarff-Bloom and Richardson grade prevail in TNBC cases with a higher incidence of lymph node metastases.

CircUSP42 Is Downregulated in Triple-Negative Breast Cancer and Associated With Poor Prognosis.

We previously showed that microRNA-182 (miR-182) might promote cell proliferation and migration in triple-negative breast cancer (TNBC). This study aimed to investigate circular RNAs (circRNAs) that interact with miR-182 and play important roles in TNBC. Thirty patients with TNBC were enrolled. One pair of tumor and adjacent tissue samples (control) were submitted for circRNA sequencing to establish the expression profile of circRNAs. Concomitantly, circRNAs aberrantly expressed between TNBC and control groups were identified, and these differentially expressed circRNAs (DEcircRNAs) were subjected to Gene Ontology and KEGG pathway enrichment analyses, as well as prediction of interactions with miRNAs. The expression levels of 5 circRNAs interacting with miR-182 were validated using qRT-PCR. Associations between the expression of circUSP42 and clinicopathological features and prognosis were evaluated. A total of 825 upregulated and 1127 downregulated DEcircRNAs were identified between tumor and control groups. Upregulated DEcircRNAs were significantly involved in proteoglycans in cancer, and endocytosis. Downregulated DEcircRNAs were involved in the pathway of resistance to EGFR tyrosine kinase inhibitors. Prediction of circRNA-miRNA interactions showed that hsa_circ_0002032, chr6:131973682-132047340+, hsa_circ_0005982, hsa_circ_0007823 (circUSP42), and hsa_circ_0001777 might act as miRNA sponges for miR-182. qRT-PCR showed consistent results with circRNA sequencing data (P < 0.05). Downregulation of circUSP42 was significantly associated with lymph node metastasis (P = 0.005) and advanced clinical stage (P = 0.032). Furthermore, Kaplan-Meier plots showed that low expression of circUSP42 was closely associated with poor outcome (log-rank test, P < 0.001). Our data suggested that dysregulation of circUSP42 might contribute to the development and progression of TNBC.

T1‑2N1M0 triple‑negative breast cancer patients from the SEER database showed potential benefit from post‑mastectomy radiotherapy

The effects of post-mastectomy radiotherapy (PMRT) on different subtypes of T1-2N1M0 breast cancer remain controversial. Patients with T1-2N1M0 breast cancer treated by mastectomy or mastectomy and PMRT were identified from the 2010–2013 dataset from the Surveillance, Epidemiology and End Results (SEER) registry. A total of 7,466 patients with the 7th American Joint Committee on Cancer stage (Tumor-Node-Metastasis stages 1–2, 1 and 0, respectively) including 2,760 cases (36.97%) treated by mastectomy and PMRT and 4,706 cases (63.03%) treated by mastectomy alone were analyzed in this study. The follow-up time for patients in the dataset used from the SEER registry was 0–59 months. The breast cancer-specific survival (BCSS) of the patients was derived from the SEER dataset and stratified by treatment approach. A propensity score matching (PSM) analysis (experimental group: Control group ratio, 1:1) was conducted. Using univariate and multivariate analyses Cox proportional hazards analyses, PMRT was identified as an independent prognostic factor for triple-negative breast cancer (TNBC). Before PSM analysis, the BCSS favored PMRT in the hormone receptor (HR)+/human epidermal growth factor receptor 2 (HER2)+ (P=0.025) and HR−/HER2− groups (P=0.010) but not in the HR+/HER2− (P=0.346) and HR−/HER2+ (P=0.288) groups. Following PSM analysis, BCSS favored PMRT alone in the TNBC (HR−/HER2−) group (P=0.025). Patients with T1-2N1M0 TNBC may benefit from radiotherapy post-mastectomy.

Human Epidermal Growth Factor-2 Receptor [HER2] Status in Patients Aged 70 years or more with Operable Early Breast Cancer — Multicentre Based Study with Review of 150 Cases

Background: Breast cancer is the most common invasive cancer in females but often has more favourable tumour biology in the older age group. More than 30% of those diagnosed with breast cancer worldwide are aged ≥ 65. Some report this percentage to be even higher, reaching up to more than 50% in developed countries that have a longer overall life expectancy. Within this group, hormone receptor-positive and human epidermal growth factor-2 (HER-2) negative are the most favourable tumour biological patterns. In contrast, the triple-negative breast cancer group has the worst prognosis. Invasive breast cancers in younger age groups have a more poorly differentiated histological grade, more hormone receptor-negative status, a remarkable extent of lympho-vascular invasion and a greater expression/amplification of HER-2 than older age groups. HER-2 receptor is amplified and over-expressed in about 20–30% of invasive breast cancers within all age groups. To analyse the status of HER-2 receptors early invasive breast cancer in relation to the histological subtype in the older age group. Methods: The study group includes 150 patients with early breast cancer . The patho-morphological data and immune-histochemistry results of invasive cancer have been analysed, this included histology subtype, grade, oestrogen/progesterone receptor status as well as HER-2 expression. Results: HER-2 expression was only seen in 17% of cases, it was seen mainly in Grade 3 cancers (9%), followed by Grade 2 (6%), with Grade 1 cancer accounting for only 1% of the study group. Conclusions: Patients with breast cancer aged ≥ 70 are seen to have less aggressive biology and expression of HER-2 receptors.

Abstract P2-09-01: Subgroups analysis of a multicenter, prospective, randomized, blinded phase 2b trial of trastuzumab + nelipeptimut-S (NeuVax) vs trastuzumab for prevention of recurrence in breast cancer patients

Abstract Background:HER2 low-expressing (LE) (IHC 1-2+, FISH non-amplified) breast cancer (BC) patients (pts) have not benefited from HER2-directed therapy despite HER2 antigen availability. Triple negative BC (TNBC), in particular, is immunogenic and in need of additional therapeutic options. We have previously shown the HER2-derived nelipeptimut-S (E75) + GM-CSF (NeuVax) to be synergistic with trastuzumab (Tz) in pre-clinical and pilot clinical studies. In a planned interim analysis of a multi-center, prospective, randomized, single-blinded, placebo-controlled phase 2b trial of Tz + NeuVax vs Tz to reduce recurrence in HER2 LE, node-positive (NP) and/or triple negative BC (TNBC) pts, we previously reported that the NeuVax + Tz was safe without added cardiac toxicity and demonstrated a significant reduction of recurrences in TNBC pts. This analysis examines additional subsets in this trial. Methods:HER2 LE, NP and/or TNBC pts who were clinically disease-free after standard therapy were randomized to receive Tz+NeuVax (vaccine group; VG) or Tz+GM-CSF (control group; CG). All pts received 1 yr of Tz per label. NeuVax or GM-CSF was given every 3 weeks x 6 starting with the 3rdTz dose, and then boosted every 6 months x 4. This pre-specified interim analysis was triggered 6 months after last enrollment. The primary endpoint is intention-to-treat 24 month disease-free survival (DFS) evaluated by log rank. Results: Of 275 pts randomized in the study (VG n=136, CG n=139), 98 had TNBC (VG=53, CG=45). In the interim analysis, estimated disease-free survival (DFS) was assessed with a median follow up of 18.8 months. No significant clinicopathologic differences were seen between treatment groups. In the TNBC group, estimated DFS was higher overall in VG vs CG (91.9% v 69.9%, p=0.023; hazard ratio [HR] 0.29, 95% confidence interval [CI] 0.09-0.90). On TNBC subgroup analysis, estimated DFS was higher in VG vs CG among pts who received neoadjuvant chemotherapy (VG n=35, CG n=31; HR 0.26, CI 0.07-0.93; p=0.03), HER2 IHC 1+ BC (VG n=34, CG n=28; HR 0.20, CI 0.04-0.96; p=0.03), pts who were AJCC 7thedition stage I/II (VG n=37, CG n=27; HR incalculable, no recurrences in the VG, p=0.008), and pts 351yr of age (VG n=32 &amp; CG n = 26; HR 0.26 CI 0.07,0.94; p=0.009). HRs did not appreciably vary based on the histologic grade or presence of lymphovascular invasion. Conclusion:Examining the subgroups from the pre-specified interim analysis demonstrates a highly significant clinical benefit in TNBC pts overall. Within the TNBC cohort, specific benefit was seen in pts who received chemotherapy neoadjuvantly, expressed lower HER2, were earlier stage, and were older in age. These factors may help enrich the TNBC population targeted in a definitive Phase 3 study in TNBC patients with residual disease after neoadjuvant chemotherapy. Citation Format: Clifton GT, Kemp Bohan PM, Hale DF, Myers JW, Brown TA, Holmes JP, Vreeland TJ, Litton JK, Murthy RK, Mittendorf EA, Peoples GE. Subgroups analysis of a multicenter, prospective, randomized, blinded phase 2b trial of trastuzumab + nelipeptimut-S (NeuVax) vs trastuzumab for prevention of recurrence in breast cancer patients [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 P2-09-01.

Abstract P1-16-05: Prognosis and survival in metastatic breast cancer – Ten years in review, a population-based analysis

Abstract Background: The rapidly evolving landscape of systemic treatment for metastatic breast cancer (MBC) during the 1990s led to meaningful improvements in the overall survival (OS) of MBC patients[1]. Despite ongoing and expanded access to new treatments, it remains unclear if this has translated into further advances in survival. Moreover, the prognosis of MBC patients based on subtype, over time, are also important to differentiate. Methods: The BC Cancer Breast Cancer Outcomes Unit (BCOU) database was utilized to identify patients referred to BC Cancer who were diagnosed with MBC during 3 time cohorts (cohort 1:2003-2005; cohort 2:2007-2009; cohort 3:2011-2013), to reflect changes in MBC treatment over these separate time periods. Baseline clinical and pathological criteria were compiled, in addition to adjuvant treatments received, as well as number of lines of treatment in the metastatic setting. OS was compared across time cohorts for all patients and then between subtypes using Kaplan-Meier survival curves. Results: A total of 3,953 patients met the inclusion criteria, consisting of 2,440 (61.7%) estrogen-receptor positive (ER+), 778 (19.7%) HER2 positive and 542 (13.7%) triple-negative breast cancer (TNBC) patients. One hundred and ninety-three patients (4.9%) were unable to be subtyped and were therefore excluded from the analysis . A total of 2,205 (90.4%) ER+ patients received at least 1 line of systemic therapy, with 80.0% receiving at least 1 line of hormonal therapy. The median time on hormonal treatment was 8.9 months (range 0.03 - 156.7) for first-line and 6.1 months (range 0.1 – 173.3) for second-line. In the HER2+ group, 665 (85.5%) patients received at least 1 line of treatment, with a median of 2 lines of treatment (range 1-16). Median duration of anti-HER2 treatment was 6.7 months (range 0.03 - 163.8) with a median of 1 line of anti-HER2 directed treatment (range 1-5). For TNBC patients, 357 (65.9%) received at least 1 line of treatment, with a median of 2 (range 1-10). No significant differences in OS were observed between the 3 time cohorts, with a median overall survival (mOS) of 1.63 years, 1.37 years and 1.57 years in cohorts 1-3, respectively (p=0.12).When comparing across subtypes, the ER+ group faired best with a mOS of 1.96 years (95% CI 1.8-2.1), consistent across time cohorts (p=0.72). This was followed by the HER2+ group with a mOS of 1.53 years (95% CI 1.3-1.7), also consistent across time cohorts (p=0.31). The TNBC group faired worst, with a mOS of 0.67 years (95% CI 0.6-0.8) over time (p=0.87). Conclusions: Despite advances in systemic therapy since the early 2000s, no meaningful improvements in overall survival were observed over time, regardless of subtype. It remains to be seen if developments since 2013 will lead to gains in overall survival for MBC patients, at a real life, population-based level. [1]Chia SK, Speers CH, D'yachkova Y, et al. Cancer 2007;110:973-979. Citation Format: LeVasseur N, Fiorino L, Speers CH, Aparicio M, Lohrisch C, Chia SK. Prognosis and survival in metastatic breast cancer – Ten years in review, a population-based analysis [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 P1-16-05.