Prognosis Of Triple-negative Breast Cancer Research Articles

Prognosis of triple-negative breast cancer associated with pregnancy: A propensity score-matched analysis from the French CALG (Cancer Associé à la Grossesse) network

IntroductionTriple-negative (TN) breast cancer represents one third of pregnancy-associated breast cancers (PABC). The aims of the current study were to describe oncological and obstetrical outcomes of patients with TN-PABC and to compare their prognosis with TN-non-PABC patients using a propensity score. Materials and methodsBetween January 2005 and December 2020, data of patients with histologically proven TN-PABC were collected and compared with data of TN-non-PABC patients under the age of 46 years diagnosed during the same period using a propensity score (PS). ResultsAfter PS matching (tumor size and lymph node involvement),there were 59 patients in each group. The median follow-up was 14 months (IQR 4.8–40.1) for the TN-PABC group and 60 months (IQR 30.7–101.4) for the TN-non-PABC group. Eight recurrences occurred in the TN-PABC group and 10 in the TN-non-PABC group (adjusted OR (AOR) = 0.60 (0.21–1.60), HR (Cox adjusted model- AHR) = 1.25 (0.53–2.94)). Two patients died in the TN-PABC group, and six in the TN-non-PABC group with an AOR = 0.23 (0.03–1.01) and an AHR = 0.58 (0.12–2.69). All the patients diagnosed during the second (n = 17) and third trimesters (n = 28) continued their pregnancies, with a median term at delivery of 38 WG (IQR 36–39). All patients gave birth to healthy newborns. ConclusionAlthough the TN subtype is associated with poor prognosis in pregnant patients due to advanced stage at diagnosis and high lymph node involvement, our PS-matched case-control study showed that pregnancy per se does not worsen the prognosis in terms of recurrence-free and overall survival.

Abstract PO-150: Comparison of RUNX1, RUNX2, RUNX3 and CBFβ gene expression in breast tumors Indicate ethnic differences and similarities by receptor status

Abstract The RUNX family of transcription factors RUNX1, RUNX2 and RUNX3 encode nuclear proteins with a Runt DNA-binding domain that functions in a complex with their obligate partner core-binding factor CBFβ to regulate gene expression. RUNX genes are known for paradoxical roles, with oncogenic as well as tumor suppressor functions, dependent on context. Interestingly, RUNX2 has been shown to play a potentially important functional role in Triple-Negative Breast Cancer (TNBC), and RUNX1 expression associates with poor prognosis in TNBC in a cohort of women of European descent. Women of African descent experience more aggressive TNBC with higher mortality rates than their women of European ancestry counterparts. TNBC is the most difficult breast cancer subtype to treat globally and has a poor prognosis and low survival which remains an important clinical challenge. There is a paucity of genomics studies interrogating TNBC of diverse populations. Hence, RNAseq datasets of 782 breast cancer patients were leveraged from a web-based platform -CbioPortal and published literature. 685 patients were queried for the RUNX1, RUNX2, RUNX3, and CBFβ genes, and an additional 97 patient data were harnessed from research publications. The cohort includes 595 (70%) patients of European ancestry (EA), 90 (11%) of African descent (AD), both from The Cancer Genome Atlas (TCGA), and 97 (13%) patients indigenous African from Nigeria (IAN). The queried genes were compared for race/ethnic disparity by estrogen receptor status (er). Mutual exclusivity was analyzed and P-values were determined by Fisher's extract test with the null hypothesis that a pair of genes alteration frequency of occurrence is proportional to uncorrelated occurrence in each gene. The queried genes enrichment in TP53, PIK3CA, CDH1, MAP3K1, and GATA3 between patients of European ancestry and African descent were analyzed and the Kaplan-Meier plots with P-value from the Long rank test were determined. The queried genes were altered in 69 patients (12%) of EA and 9 patients of AD due to unavailability of data on RUNX2 and RUNX 3 in IAN only RUNX1 and CBfβ was altered in 4 (4%) patients of 97 patients. Ethnic disparities and similarities were indicated in the queried altered genes in RUNX1 and RUNX 2 and showed similar expression in ER-negative correlating with the TNBC poor prognosis however RUNX3 genes were silenced in that of AD and was expressed in EA. They were differences in mutual exclusivity and co-occurrence but were not statistically significant, this may be due to the small data size. Interestingly, there was statistically significant equal gene enrichment of RUNX1 and CBfβ in CDH1mutations of both EA and AD. Larger cohorts in particular the AD cohort is needed, to further elucidate some of these borderline findings. In conclusion, our findings are insightful towards deciphering the characteristic aggressive nature of TNBC in ethnic disparity. Citation Format: Uzoamaka A. Okoli. Comparison of RUNX1, RUNX2, RUNX3 and CBFβ gene expression in breast tumors Indicate ethnic differences and similarities by receptor status [abstract]. In: Proceedings of the AACR Virtual Conference: 14th AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2021 Oct 6-8. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2022;31(1 Suppl):Abstract nr PO-150.

Analysis of single-cell RNA-sequencing data identifies a hypoxic tumor subpopulation associated with poor prognosis in triple-negative breast cancer.

Triple-negative breast cancer (TNBC) is an aggressive subtype of mammary carcinoma characterized by low expression levels of estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER2). Along with the rapid development of the single-cell RNA-sequencing (scRNA-seq) technology, the heterogeneity within the tumor microenvironment (TME) could be studied at a higher resolution level, facilitating an exploration of the mechanisms leading to poor prognosis during tumor progression. In previous studies, hypoxia was considered as an intrinsic characteristic of TME in solid tumors, which would activate downstream signaling pathways associated with angiogenesis and metastasis. Moreover, hypoxia-related genes (HRGs) based risk score models demonstrated nice performance in predicting the prognosis of TNBC patients. However, it is essential to further investigate the heterogeneity within hypoxic TME, such as intercellular communications. In the present study, utilizing single-sample Gene Set Enrichment Analysis (ssGSEA) and cell-cell communication analysis on the scRNA-seq data retrieved from Gene Expression Omnibus (GEO) database with accession number GSM4476488, we identified four tumor subpopulations with diverse functions, particularly a hypoxia-related one. Furthermore, results of cell-cell communication analysis revealed the dominant role of the hypoxic tumor subpopulation in angiogenesis- and metastasis-related signaling pathways as a signal sender. Consequently, regard the TNBC cohorts acquired from The Cancer Genome Atlas (TCGA) and GEO as train set and test set respectively, we constructed a risk score model with reliable capacity for the prediction of overall survival (OS), where ARTN and L1CAM were identified as risk factors promoting angiogenesis and metastasis of tumors. The expression of ARTN and L1CAM were further analyzed through tumor immune estimation resource (TIMER) platform. In conclusion, these two marker genes of the hypoxic tumor subpopulation played vital roles in tumor development, indicating poor prognosis in TNBC patients.

The Characteristics of Tumor Microenvironment in Triple Negative Breast Cancer.

Triple-negative breast cancer (TNBC) is a special subtype of breast cancer, accounting for 10–20% of breast cancers with high intrinsic heterogeneity. Its unique immune microenvironment, including high expression of vascular endothelial growth factors, tumor infiltrating lymphocytes (TILs), tumor-associated macrophages (TAMs), and other molecules that promote the growth and migration of tumor cells, has been shown to play a dual role in the occurrence, growth, and metastasis of TNBC. Understanding the TNBC microenvironment is of great significance for the prognosis and treatment of TNBC. In this article, we describe the composition and function of immune cells in the TNBC microenvironment and summarize the major cytokine growth factors and chemokines in the TNBC microenvironment. Finally, we discuss the progress of TNBC, cytokine-induced killer cell therapy, and immune checkpoint therapy.

Current situation of programmed cell death protein 1/programmed cell death ligand 1 inhibitors in advanced triple-negative breast cancer.

Triple-negative breast cancer (TNBC) has the worst prognosis among all molecular types of breast cancer. Because of the strong immunogenicity of TNBC cells, programmed death 1/programmed death ligand 1 (PD-1/PD-L1) inhibitors, two kinds of immune checkpoint blockade agents, might help improve the prognosis of TNBC. However, how to better use PD-1/PD-L1 inhibitors and select patients who may benefit from treatment options remains controversial. This article summarizes published clinical studies in which PD-1/PD-L1 inhibitors were used in patients with advanced TNBC to explore how to maximize effectiveness of these medications.

Single-Cell Transcriptome Analysis Reveals the M2 Macrophages and Exhausted T Cells and Intratumoral Heterogeneity in Triple-Negative Breast Cancer.

Triple-Negative Breast Cancer (TNBC) is a highly heterogeneous and invasive malignancy that is characterized by high recurrence and mortality rates as well as extremely poor prognosis. The objective of this study is to analyze T cells and Macrophages in the tumor microenvironment with the aim of identifying targets with therapeutic potential. Single-cell sequencing data of TNBC patients from the GSE118389 dataset were analyzed to examine the immune environment and intratumoral heterogeneity of TNBC patients. Polarized alternatively activated macrophages (M2) and exhausted CD8+ T cells were identified in TNBC patients. Immunosuppressive checkpoint analysis revealed that levels of lymphocyte-activation gene 3 (LAG3) and T cell immunoglobulin and mucin domain-containing protein 3 (TIM-3) of exhausted T cells were significantly higher than levels of programmed cell death protein 1 (PD-1) and cytotoxic T-lymphocyteassociated protein 4 (CTLA-4). This indicates that these markers are potential immunotherapy targets. Furthermore, analysis of significantly altered immune cell markers showed that several markers were associated with the prognosis of TNBC. Overall, these findings demonstrate inter-tissue heterogeneity of TNBC, and provides novel therapeutic targets for the treatment of TNBC.

Fibrinogen/Albumin Ratio (FAR) in Patients with Triple Negative Breast Cancer and Its Relationship with Epidermal Growth Factor Receptor Expression.

ObjectiveTo investigate the potential prognostic significance of fibrinogen/albumin ratio (FAR) in triple negative breast cancer (TNBC) patients and its relationship with epidermal growth factor receptor (EGFR) expression.MethodsThere were 164 patients with TNBC enrolled in this study in our hospital from January 2010 to December 2015. The optimal cutoff value of FAR was evaluated by the receiver operating characteristic curve (ROC). The associations between TNBC and clinicopathological variables by FAR were performed by Chi-square test. Kaplan–Meier method and Log rank test were used for survival analysis. The independent prognostic factors were determined by univariate and multivariate Cox’s proportional hazards regression model. The EGFR expression was analyzed by the immunohistochemistry assay.ResultsOne hundred and sixty-four TNBC patients were divided into: low FAR group (FAR < 0.08) and high FAR group (FAR ≥ 0.08) by ROC. The preoperative FAR was associated to BMI, menopause, red blood cell, albumin, fibrinogen (P < 0.05). FAR was an independent prognostic factor for TNBC. In low FAR group, the mean disease-free survival (DFS) and overall survival (OS) were 33.62 months and 52.99 months; in high FAR group, the mean DFS and OS were 30.18 months and 48.27 months, respectively. The DFS and OS survival curve were performed by Log rank assay and were statistically significant (P < 0.05). The mean DFS and OS after operation in patients with EGFR negative expression were longer than that in patients with EGFR positive expression. In EGFR positive group, the mean DFS and OS of low FAR group were higher than that of high FAR group, and the difference was statistically significant (P < 0.05).ConclusionPretreatment FAR is the independent prognostic factor in TNBC, and with low cost, strong repeatability, and high safety. It can be acted as an effective indicator to predict the prognosis of TNBC.

Stress sensing within the breast tumor microenvironment: how glucocorticoid receptors live in the moment.

The classification and treatment of breast cancer is largely defined by the expression of steroid hormone receptors (HRs), namely estrogen receptor (ER) and progesterone receptor (PR), and gene amplification/overexpression of human epidermal growth factor receptor 2 (HER2). More recently, studies of androgen receptor (AR), glucocorticoid receptor (GR), and mineralocorticoid receptor (MR) have revealed that targeting these related HRs may be a promising strategy for a more personalized approach to the treatment of specific subtypes of HR+ breast cancer. For example, GR expression is associated with a good prognosis in ER+ breast cancer, but predicts poor prognosis in triple-negative breast cancer (TNBC). GR, like ER, PRs, and AR, is a ligand-activated transcription factor, but also has significant ligand-independent signaling activities. GR transcriptional activity is classically regulated by circulating glucocorticoids (GCs; ligand-dependent). Recent studies demonstrate that GR transcriptional activity is also regulated by a variety of cellular stress stimuli that input to GR Ser134 phosphorylation via rapid activation of the p38 mitogen activated protein kinase (MAPK) signaling pathway (ligand-independent). Furthermore, ligand-independent GR activation promotes feedforward signaling loops that mediate sustained activation of stress signaling pathways to drive advanced cancer biology (i.e. migration, invasion, chemoresistance, survival, and cellular growth). In this review, we will focus on the role of GR as a key sensor and mediator of physiologic and tumor microenvironment (TME)-derived cellular stress signaling in TNBC and discuss how targeting GR and/or associated signaling pathways may provide a strategy to inhibit deadly TNBC progression.

PROGNOSTIC FACTORS AND SURVIVAL IN TRIPLE NEGATIVE AND NON-TRIPLE NEGATIVE BREAST CANCER: EXPERIENCE OF A TERTIARY CARE CANCER CENTER IN ODISHA.

&#x0D; Background: Because of its high-risk biological features and lack of effective treatment options, triple-negative breast cancer (TNBC) has received greater clinical and experimental interest.&#x0D; &#x0D; &#x0D; Aim and objectives: The aim of this study was to compare and analyze the clinicopathological features, recurrence, metastasis, and prognosis of patients with TNBC and non-triple negative breast cancer (non-TNBC).&#x0D; &#x0D; &#x0D; Material and methods: This single hospital-based retrospective study was conducted on patients who were histopathologically diagnosed with breast cancer and subsequently treated from 2017 to 2018 at the Acharya Harihar Postgraduate Institute of Cancer. The clinical features and prognosis of TNBC and non-TNBC were compared.&#x0D; &#x0D; &#x0D; Results: This study comprised a total of 111 patients, with 36 (32.43%) being TNBC and 75 (67.56%) being non-TNBC. TNBC has 22 patients under the age of 40 (61.1%). Grade III tumors were seen in 47% of TNBC patients and 21% of non-TNBC patients (p-value = 0.05). The disease free survival (DFS) was determined to be 58 % for TNBC and 82% for non-TNBC groups, respectively (p-value = 0.05). These two groups had an overall survival rate (OS) of 72% and 92%, respectively (p-value = 0.05).&#x0D; &#x0D; &#x0D; Conclusion: When compared to non-TNBC, TNBC was related to high-grade malignancies, worse disease-free survival , and overall survival (OS) rates. Understanding the molecular features of TNBC, clarifying its mechanism at the molecular level, interpreting the gene expression profiles of TNBC, and studying and creating new therapeutic targets should be the focus of future research. To enhance the prognosis of TNBC patients, try to find a focused and effective therapy.&#x0D; &#x0D; Keywords:&#x0D; Breast cancer; survival; triple-negative breast cancer.

Role of PTEN, PI3K, and mTOR in Triple-Negative Breast Cancer.

Breast cancer is the most commonly occurring malignancy and the leading cause of cancer-related death in women. Triple-negative breast cancer (TNBC) is the most aggressive subtype and is associated with high recurrence rates, high incidence of distant metastases, and poor overall survival. The aim of this study was to investigate the PI3K/PTEN/Akt/mTOR pathway as one of the most frequently deregulated pathways in cancer. We aimed to explore the impact of PI3K and mTOR oncogenes as well as the PTEN tumor suppressor on TNBC clinical behavior, prognosis, and multidrug resistance (MDR), using immunohistochemistry and copy number analysis by quantitative real-time PCR. Our results revealed that loss of PTEN and high expression of PI3K and mTOR proteins are associated with poor outcome of TNBC patients. PTEN deletions appeared as a major cause of reduced or absent PTEN expression in TNBC. Importantly, homozygous deletions of PTEN (and not hemizygous deletions) are a potential molecular marker of metastasis formation and good predictors of TNBC outcome. In conclusion, we believe that concurrent examination of PTEN/PI3K/mTOR protein expression may be more useful in predicting TNBC clinical course than the analysis of single protein expression. Specifically, our results showed that PTEN-reduced/PI3K-high/mTOR-high expression constitutes a ‘high risk’ profile of TNBC.

High miR-3609 expression is associated with better prognosis in TNBC based on mining using systematic integrated public sequencing data.

MicroRNAs (miRNAs/miRs) are small endogenous RNAs that regulate gene expression post-transcriptionally. Abnormal miR-3609 expression is associated with the occurrence of pancreatic cancer, glioma and other diseases, such as polycystic ovary syndrome. However, the prognostic potential of miR-3609 has been reported in breast cancer. Thus, the present study aimed to investigate the differential expression and prognostic value of miR-3609 in patients with breast cancer from the UALCAN, cBioportal and Kaplan-Meier Plotter databases, respectively. Furthermore, the co-expression genes of miR-3609 in breast cancer were investigated using data from the LinkedOmics database, and functional enrichment analysis was performed using the LinkInterpreter module in LinkedOmics. The co-expression gene network was constructed using the Search Tool for the Retrieval of Interacting Genes/Proteins database, and the cytoHubba plug-in was used to identify the hub genes, which were visualized using Cytoscape software. The prognoses of the hub genes were performed using the Kaplan-Meier Plotter database. The Cell Counting Kit-8 and cell cycle assays were performed to confirm the functions of miR-3609 mimics transfection in MDA-MB-231 cells. Survival analysis using the Kaplan-Meier Plotter database demonstrated that high miR-3609 expression in triple-negative breast cancer (TNBC) was associated with a better prognosis. Furthermore, the experimental results indicated that high miR-3609 expression inhibited the proliferation of TNBC cells and induced cell cycle arrest of TNBC cells in the G0/G1 phase. Taken together, the results of the present study suggest that miR-3609 plays a vital role in mediating cell cycle arrest and inhibiting the proliferation of TNBC cells.

Focal pyroptosis-related genes AIM2 and ZBP1 are prognostic markers for triple-negative breast cancer with brain metastases.

BackgroundExtensive research has shown the role of pyroptosis in the occurrence, progression, and prognosis of breast cancer. The study sought to screen important pyroptosis-related genes and their role in the prognosis of triple-negative breast cancer (TNBC) patients with brain metastasis (BM).MethodsThe Gene Expression Omnibus database was used to obtain transcriptome data from primary TNBC and from TNBC BM patients. Differentially expressed genes (DEGs) between the primary tumors and BMs were analyzed, and the expression, prognostic significance, immune infiltration, function, and drug sensitivity of the pyroptosis genes in the DEGs were analyzed.ResultsIn both data sets, 456 genes differed between primary TNBC and TNBC BM. Absent in melanoma 2 (AIM2) and Z-deoxyribonucleic acid-binding protein 1 (ZBP1) were found to be important pyroptosis genes in DEGs, and significant differences in their expression in primary lesions and BMs were observed. Patients with a high expression of AIM2 had a worse prognosis than low expression, while patients with a high expression of ZBP1 had a better prognosis than low expression. AIM2 and ZBP1 were positively correlated with the infiltration of most immune cells; however, AIM2 was negatively correlated with the infiltration of neural cell adhesion molecule 1 (CD56) bright natural killer cells and central memory cluster of differentiation 8 (CD8) T cells. Increased expression of ZBP1 is negatively correlated with high infiltration levels of central memory CD8 T cells and memory B cells.ConclusionsOur findings suggest that AIM2 and ZBP1 increase immune cell infiltration and may be potential targets for predicting and treating TNBC BM.

Identification of Immune-Related Risk Characteristics and Prognostic Value of Immunophenotyping in TNBC.

Background: Triple-negative breast cancer (TNBC) is not sensitive to targeted therapy with HER-2 monoclonal antibody and endocrine therapy due to lack of ER, PR, and HER-2 receptors. TNBC is a breast cancer subtype with the worst prognosis and the highest mortality rate compared with other subtypes. Materials and Methods: Breast cancer-related data were retrieved from The Cancer Genome Atlas (TCGA) database, and 116 cases of triple-negative breast cancer were identified from the data. GSE31519 dataset was retrieved from Gene Expression Omnibus (GEO) database, comprising a total of 68 cases with TNBC. Survival analysis was performed based on immune score, infiltration score and mutation score to explore differences in prognosis of different immune types. Analysis of differentially expressed genes was conducted and GSEA analysis based on these genes was conducted to explore the potential mechanism. Results: The findings showed that comprehensive immune typing is highly effective and accurate in assessing prognosis of TNBC patients. Analysis showed that MMP9, CXCL9, CXCL10, CXCL11 and CD7 are key genes that may affect immune typing of TNBC patients and play an important role in prediction of prognosis in TNBC patients. Conclusion: The current study presents an evaluation system based on immunophenotyping, which provides a more accurate prognostic evaluation tool for TNBC patients. Differentially expressed genes can be targeted to improve treatment of TNBC.

Identification of Key Genes Affecting the Tumor Microenvironment and Prognosis of Triple-Negative Breast Cancer.

Although the tumor microenvironment (TME) plays an important role in the development of many cancers, its roles in breast cancer, especially triple-negative breast cancer (TNBC), are not well studied. This study aimed to identify genes related to the TME and prognosis of TNBC. Firstly, we identified differentially expressed genes (DEG) in the TME of TNBC, using Expression data (ESTIMATE) datasets obtained from the Cancer Genome Atlas (TCGA) and Estimation of Stromal and Immune cells in Malignant Tumor tissues. Next, survival analysis was performed to analyze the relationship between TME and prognosis of TNBC, as well as determine DEGs. Genes showing significant differences were scored as alternative genes. A protein-protein interaction (PPI) network was constructed and functional enrichment analysis conducted using the DEG. Proteins with a degree greater than 5 and 10 in the PPI network correspond with hub genes and key genes, respectively. Finally, CCR2 and CCR5 were identified as key genes in TME and prognosis of TNBC. Finally, these results were verified using Gene Expression Omnibus (GEO) datasets and immunohistochemistry of TNBC patients. In conclusion, CCR2 and CCR5 are key genes in the TME and prognosis of TNBC with the potential of prognostic biomarkers in TNBC.

Obesity Modulates the Gut Microbiome in Triple-Negative Breast Cancer.

Triple-negative breast cancer (TNBC) is an aggressive, molecularly heterogeneous subtype of breast cancer. Obesity is associated with increased incidence and worse prognosis in TNBC through various potential mechanisms. Recent evidence suggests that the gut microbiome plays a central role in the progression of cancer, and that imbalances or dysbiosis in the population of commensal microbiota can lead to inflammation and contribute to tumor progression. Obesity is characterized by low-grade inflammation, and gut dysbiosis is associated with obesity, chronic inflammation, and failure of cancer immunotherapy. However, the debate on what constitutes a “healthy” gut microbiome is ongoing, and the connection among the gut microbiome, obesity, and TNBC has not yet been addressed. This study aims to characterize the role of obesity in modulating the gut microbiome in a syngeneic mouse model of TNBC. 16S rRNA sequencing and metagenomic analyses were performed to analyze and annotate genus and taxonomic profiles. Our results suggest that obesity decreases alpha diversity in the gut microbiome. Metagenomic analysis revealed that obesity was the only significant factor explaining the similarity of the bacterial communities according to their taxonomic profiles. In contrast to the analysis of taxonomic profiles, the analysis of variation of functional profiles suggested that obesity status, tumor presence, and the obesity–tumor interaction were significant in explaining the variation of profiles, with obesity having the strongest correlation. The presence of tumor modified the profiles to a greater extent in obese than in lean animals. Further research is warranted to understand the impact of the gut microbiome on TNBC progression and immunotherapy.

Periostin Exon-21 Antibody Neutralization of Triple-Negative Breast Cancer Cell-Derived Periostin Regulates Tumor-Associated Macrophage Polarization and Angiogenesis.

Simple SummaryDespite remarkable advances in breast cancer treatment, few strategies other than standard cytotoxic chemotherapy are available for patients with triple-negative breast cancer (TNBC) due to the lack of therapeutic target molecules. TNBC is still the most aggressive subtype, with a high risk of recurrence and metastasis within 2 years after initial treatment. Thus, there is an unmet medical need to develop new treatments for metastatic and recurrent TNBC patients. In this study we tested a new antibody, targeting extracellular periostin protein alternative splicing variants, which are induced by conventional chemotherapy or during the process of endothelial mesenchymal transition. This antibody reduced periostin-secreting TNBC in a mouse xenograft model, accompanied by a decrease in the number of M2 tumor-associated macrophages and tumor vessels. Periostin alternative splicing variants might be a specific and safe therapeutic target in patients with TNBC.Periostin (Pn) is involved in multiple processes of cancer progression. Previously, we reported that Pn expression is correlated with mesenchymal tumor markers and poor prognosis in triple-negative breast cancer (TNBC). In the TNBC xenograft model, chemotherapy increased expression of a Pn alternative splicing variant (ASV) with exon 21, and administration of the neutralizing antibody against Pn with exon 21 (Pn-21 Ab) overcame chemoresistance with a reduction in the mesenchymal cancer cell fraction. In the present study, the role of Pn ASV with exon 21 in TNBC progression has been addressed. We first established a stable cell line carrying a fluorescence-based splicing reporter. Pn-positive TNBC has higher expression of genes related to tumor-associated macrophage (TAM) recruitment and ECM-receptor interaction than Pn-negative cells. In a xenograft model, only Pn-positive cells initiated tumor formation, and the Pn-21 Ab suppressed tumor cell growth, accompanied by decreased M2 TAM polarization and the number of tumor vessels. These data suggest that cancer cell-derived Pn ASV educates TAMs and regulates angiogenesis, which in turn establishes a microenvironmental niche that is supportive of TNBC.

Characterization of Exosome-Related Gene Risk Model to Evaluate the Tumor Immune Microenvironment and Predict Prognosis in Triple-Negative Breast Cancer.

BackgroundAs a kind of small membrane vesicles, exosomes are secreted by most cell types from multivesicular endosomes, including tumor cells. The relationship between exosomes and immune response plays a vital role in the occurrence and development of tumors. Nevertheless, the interaction between exosomes and the microenvironment of tumors remains unclear. Therefore, we set out to study the influence of exosomes on the triple-negative breast cancer (TNBC) microenvironment.MethodOne hundred twenty-one exosome-related genes were downloaded from ExoBCD database, and IVL, CXCL13, and AP2S1 were final selected because of the association with TNBC prognosis. Based on the sum of the expression levels of these three genes, provided by The Cancer Genome Atlas (TCGA), and the regression coefficients, an exosome risk score model was established. With the median risk score value, the patients in the two databases were divided into high- and low-risk groups. R clusterProfiler package was employed to compare the different enrichment ways between the two groups. The ESTIMATE and CIBERSORT methods were employed to analyze ESTIMATE Score and immune cell infiltration. Finally, the correlation between the immune checkpoint-related gene expression levels and exosome-related risk was analyzed. The relationship between selected gene expression and drug sensitivity was also detected.ResultsDifferent risk groups exhibited distinct result of TNBC prognosis, with a higher survival rate in the low-risk group than in the high-risk group. The two groups were enriched by immune response and biological process pathways. A better overall survival (OS) was demonstrated in patients with high scores of immune and ESTIMATE rather than ones with low scores. Subsequently, we found that CD4+-activated memory T cells and M1 macrophages were both upregulated in the low-risk group, whereas M2 macrophages and activated mast cell were downregulated in the low-risk group in patients from the TCGA and GEO databases, respectively. Eventually, four genes previously proposed to be targets of immune checkpoint inhibitors were evaluated, resulting in the expression levels of CD274, CTLA4, LAG3, and TIM3 being higher in the low-risk group than high-risk group.ConclusionThe results of our study suggest that exosome-related risk model was related to the prognosis and ratio of immune cell infiltration in patients with TNBC. This discovery may make contributions to improve immunotherapy for TNBC.

Cytochrome 4Z1 Expression is Associated with Unfavorable Survival in Triple-Negative Breast Cancers.

PurposeTriple-negative breast cancer (TNBC) is characterized by high mortality rate, and its clinical management is difficult and complex. Therefore, there is a need for extensive efforts aimed at accelerating the discovery of novel therapies for TNBC. CYP4Z1 has been implicated in the development of breast cancer. The current study aimed at characterizing the expression of CYP4Z1 on TNBC.Materials and MethodsUsing immunohistochemistry, CYP4Z1 expression was evaluated on 122 TNBC samples, four samples of breast cancers expressing ER, PR, and HER-2, and four samples of normal breast tissues. The association between the enzyme and various histopathological features and survival of patients were determined.ResultsCYP4Z1 was strongly expressed in 83.3% of various histopathological subtypes of TNBC, when compared to negative expression in normal breast tissues. Interestingly, there were marked variations in CYP4Z1 expression with respect to histopathology subtype, histological grade, histological stage and tumor diameter. There was a high incidence of CYP4Z1 expression in patients with advanced grades, late stages and larger tumor sizes. Importantly, CYP4Z1 expression was correlated with the survival of TNBC patients, but it was an independent determinant of the poor prognosis of TNBC (p< 0.05).ConclusionCYP4ZI may be a potential biomarker or target for evolving new CYP4Z1-targeted treatments.

Synergistic effect of anethole and doxorubicin alleviates cell proliferation, cell cycle arrest, and ER stress and promotes ROS-mediated apoptosis in triple-negative breast cancer cells.

The heterogeneity and poor prognosis of triple-negative breast cancer (TNBC) have limited the treatment options and made clinical management challenging. This has nurtured a major effort to discover druggable molecular targets. Currently, chemotherapy is the primary treatment strategy for this disease. Doxorubicin is the most frequently used chemotherapeutic drug for TNBC and due to the fact that chemotherapeutic drugs have a lot of side effects, we evaluated the synergistic effect of the phytocompound anethole and doxorubicin. The cytotoxic effect of anethole in combination with doxorubicin on MDA-MB-231 cells was evaluated by various parameters, including apoptosis, cell cycle analysis, DNA damage, and cell proliferation. Furthermore, mitochondrial membranepotential (MMP), endoplasmic reticulum (ER) stress, and reactive oxygen species (ROS) levels were also evaluated in the cells treated with/without anethole and doxorubicin. Expression of the apoptotic proteins was evaluated by Western blot analysis. Initial evaluation of cytotoxicity of anethole on MDA-MB-231 cells demonstrated preferentialsuppression of cell proliferation and when treated along with doxorubicin it showed enhanced cytotoxicity with a synergistic effect. Cell cycle analysis revealed arrest at different stages of the cell cycle, such as sub G0-G1, G0-G1, S, and G2M in various treatment groups and apoptotic cell death was subsequently evident with propidium iodide (PI) staining. The synergistic action of anethole and doxorubicin effectively induced mitochondrial membrane potential loss, which, in turn, led to a burst of ROS production, which eventually produced unfolded protein response by damaging the ER. Synergistic anticancer effect was observed on exposure of MDA-MB-231 cells to anethole and doxorubicin in inducing cell death.

Co-expression and prognosis analyses of GLUT1\u20134 and RB1 in breast cancer

BackgroundCurrent treatment methods for patients with triple-negative breast cancer (TNBC) are very limited, and the prognosis of TNBC is relatively poor. It has been reported that glucose transporter 1 (GLUT1) is overexpressed in breast cancer cells; however, its association with the prognosis is mostly unclear. Moreover, retinoblastoma gene 1 (RB1) might be used as a biomarker for the sensitivity of breast cancer cells to GLUT1 inhibitors, which brought us to the hypothesis that there might be a close correlation between the expression of GLUT1–4 and the expression of RB1.MethodsIn this study, we systematically analyzed the co-expression of GLUT1–4 and the influence of GLUT1–4 gene expression on the prognosis of breast cancer using data mining methods. We also explored possible relationships between GLUT1–4 and RB1 expression in breast cancer tissues. We used public databases such as ONCOMINE, GEPIA, LinkedOmics, and COEXPEDIA.ResultsAccording to the results, the mRNA expression of SLC2A1 was significantly higher in breast cancer, while the expression levels of SLC2A2–4 were downregulated. The results also indicate that GLUT1 expression does not have significant influence on the overall survival of patients with breast cancer. The mRNA expression of SLC2A1 and RB1 is significantly correlated, which means that tissues with high RB1 mRNA expression might have relatively higher mRNA expression of SLC2A1; however, further study analyzing their roles in the expression regulation pathways with human samples is needed to verify the hypothesis.ConclusionsThe mRNA expression of SLC2A1 was significantly higher in breast cancer. The overall survival of breast cancer patients wasn’t significantly correlated with GLUT1–4 expression. The mRNA expression of SLC2A1 and RB1 is significantly correlated according to the analysis conducted in LinkedOmics. It provides reference for future possible individualized treatment of TNBC using GLUT1 inhibitors, especially in patients with higher mRNA expression of RB1. Further study analyzing the roles of these two genes in the regulation pathways is needed.