Various Subtypes Of Breast Cancer Research Articles

The therapeutic effect of traditional Chinese medicine on breast cancer through modulation of the Wnt/β-catenin signaling pathway.

Breast cancer, the most prevalent malignant tumor among women globally, is significantly influenced by the Wnt/β-catenin signaling pathway, which plays a crucial role in its initiation and progression. While conventional chemotherapy, the standard clinical treatment, suffers from significant drawbacks like severe side effects, high toxicity, and limited prognostic efficacy, Traditional Chinese Medicine (TCM) provides a promising alternative. TCM employs a multi-targeted therapeutic approach, which results in fewer side effects and offers a high potential for effective treatment. This paper presents a detailed analysis of the therapeutic impacts of TCM on various subtypes of breast cancer, focusing on its interaction with the Wnt/β-catenin signaling pathway. Additionally, it explores the effectiveness of both monomeric and compound forms of TCM in the management of breast cancer. We also discuss the potential of establishing biomarkers for breast cancer treatment based on key proteins within the Wnt/β-catenin signaling pathway. Our aim is to offer new insights into the prevention and treatment of breast cancer and to contribute to the standardization of TCM.

Downregulation of long noncoding RNA B4GALT1-AS1 is associated with breast cancer development

The misregulation of long non-coding RNAs (lncRNAs) is related to the progressive evolution of various human cancers, such as Breast cancer (BC). The role of lncRNA B4GALT1-AS1 has been investigated in some human cancers. Therefore, studying B4GALT1-AS1 expression was aimed for the first time in the tumor and marginal tissues of BC in this study. The cancer genome atlas (TCGA) database was utilized to evaluate the relative expression of B4GALT1-AS1 in BC and other cancers. RNA was extracted from twenty-eight paired BC and marginal tissues, and cDNA was synthesized. The quantitative expression level of B4GALT1-AS1 was evaluated using real-time PCR. The bioinformatics analyses were performed to identify co-expression genes and related pathways. B4GALT1-AS1 was significantly downregulated in BC specimens compared to tumor marginal samples. The TCGA data analysis confirmed the downregulation of B4GALT1-AS1 in BC. The bioinformatics analysis discovered the correlation between 700 genes and B4GALT1-AS1 and identified GNAI1 as the high degree gene which was positively correlated with B4GALT1-AS1 expression. It seems B4GALT1-AS1 provides its function, at least partly, in association with one of the hippo pathway components, YAP, in other cancers. This protein has the opposite role in BC and its loss of function can result in poor survival in BC. Further research is needed to investigate the interaction between B4GALT1‐AS1 and YAP in various subtypes of BC.

Targeting P53 Gene in Breast Cancer

Infectious diseases and breast cancer rely heavily on p53, a tumor suppressor that protects genetic integrity. It is present on chromosome 17p13 and controls angiogenesis, programmed cell death, and cell proliferation. The expression of p53 and its isoforms is more predictive of treatment outcomes and responses to chemotherapy for breast cancer patients than the presence or absence of TP53 mutations. Subtype-dependent mutations of p53's negative regulator MDM2 can both be signs of p53 deactivation in breast cancer. Targeting both wild-type and p53 mutant across various subtypes of breast cancer is one possible treatment strategy. Notably, through paracrine signaling, mutant p53 in stromal fibroblasts speeds up the formation of breast tumors. In the progression and trajectory of breast cancer, the pivotal involvement of the p53 gene, alongside the bcl-2 proto-oncogene, is noteworthy. Addressing the role of p53 is crucial in the context of breast cancer development and its course.

Intra-tumoral microbial community profiling and associated metabolites alterations of TNBC.

Triple-negative breast cancer (TNBC) presents significant challenges to female health owing to the lack of therapeutic targets and its poor prognosis. In recent years, in the field of molecular pathology, there has been a growing focus on the role of intra-tumoral microbial communities and metabolic alterations in tumor cells. However, the precise mechanism through which microbiota and their metabolites influence TNBC remains unclear and warrants further investigation. In this study, we analyzed the microbial community composition in various subtypes of breast cancer through 16S rRNA MiSeq sequencing of formalin-fixed, paraffin-embedded (FFPE) tissue samples. Notably, Turicibacter, a microbe associated with cancer response, exhibited a significantly higher abundance in TNBC. Similarly, mass spectrometry-based metabolomic analysis revealed substantial differences in specific metabolites, such as nutriacholic, pregnanetriol, and cortol. Furthermore, we observed significant correlations between the intra-tumoral microbiome, clinicopathological characteristics, and human epidermal growth factor receptor-2 expression(HER2). Three microbial taxa (Cytophagaceae, Conexibacteraceae, and Flavobacteriaceae) were associated with tumor-infiltrating lymphocytes(TILs), which are indicative of antitumor immunity. This study creatively utilized FFPE tissue samples to assess intra-tumoral microbial communities and their related metabolic correlations, presenting avenues for the identification of novel diagnostic biomarkers, the development of therapeutic strategies, and the early clinical diagnosis of TNBC.

Repurposing of approved drugs for targeting CDK4/6 and aromatase protein using molecular docking and molecular dynamics studies.

Breast cancer is a leading cause of cancer-related morbidity and mortality worldwide, with the highest incidence among women. Among the various subtypes of breast cancer, estrogen-receptor positive (ER+) is the most diagnosed. Estrogen upregulates cyclin D1, which in turn promotes the activity of CDK4/6 and facilitates cell cycle progression. To address this, the first-line treatment for ER+ breast cancer focuses on inhibiting estrogen production by targeting aromatase, the enzyme responsible for the rate-limiting step in estrogen synthesis. Thus, combining CDK4/6 inhibitors with aromatase inhibitors has emerged as a crucial treatment strategy for this type of breast cancer. This approach effectively suppresses estrogen biosynthesis and controls uncontrolled cell proliferation, significantly improving overall survival rates and delayed disease progression. This study aimed to identify compounds that are likely to inhibit CDK4/6 and aromatase simultaneously by using a structure-based drug design strategy. 12,432 approved and investigational drugs were prepared and docked into the active site of CDK6 using HTVS and XP docking modes of Glide resulting in 277 compounds with docking scores ≤ -7 kcal/mol. These compounds were docked into aromatase enzyme using XP mode to give seven drugs with docking scores≤ -6.001 kcal/mol. Furthermore, the shortlisted drugs were docked against CDK4 showing docking scores ranging from -3.254 to -8.254 kcal/mol. Moreover, MM-GBSA for the top seven drugs was calculated. Four drugs, namely ellagic acid, carazolol, dantron, and apomorphine, demonstrated good binding affinity to all three protein targets CDK4/6 and aromatase. Specifically, they exhibited favourable binding free energy with CDK6, with values of -51.92, -53.90, -50.22, and -60.97 kcal/mol, respectively. Among these drugs, apomorphine displayed the most favourable binding free energy with all three protein targets. To further evaluate the stability of the interaction, apomorphine was subjected to a 100 ns molecular dynamics simulation with CDK6. The results indicated the formation of a stable ligand-protein complex. While the results obtained from the MM-GBSA calculation of the binding free energies of the MD conformations of apomorphine showed less favourable binding free energy compared to that obtained post-docking. All these computational findings will provide better structural insight for the development of CDK4/6 and aromatase multi-target inhibitors.

Relation between morphological features of initial breast MRI and breast cancer molecular subtypes

BackgroundBecause different gene receptors might cause each case of breast cancer, the disease is classified as a heterogeneous form because it can be subdivided into molecular subtypes. These molecular subtypes are different in disease manifestation, therapeutic response, and prognosis. Magnetic resonance imaging (MRI) has many applications in breast cancer's initial diagnosis and assessment of treatment response. The purpose of this research was to determine whether or not there is a correlation between specific morphological aspects of breast MRI in breast cancer and specific breast cancer subtypes and their impact on treatment decisions.ResultsThere was an insignificant difference between different mass shapes and different molecular subtypes (P > 0.05). One hundred percent of triple-negative breast cancers (TNBC) were rounded. Molecular subtypes and the spiculated mass border diverged significantly statistically (P = 0.023). The percentage of the hormonal receptor (HR)-positive breast cancers with a spiculated border (80%) was greater than TNBC (0%) and distinctively different (P = 0.044). Surgical alternatives were observed to correlate significantly with the MRI mass border type (P = 0.030). There was an insignificant difference between molecular subtypes and tumor size (P = 0.602), lymph nodes (P = 0.283), multicentricity (P = 0.386), and curve type (P = 0.107).ConclusionsMRI breast imaging has an important role in diagnostic and prognostic settings. The morphological results of MRI can be a helpful tool in distinguishing between the various subtypes of breast cancer. There was a statistically significant difference between different molecular subtypes and the spiculated mass border (P = 0.023). There was a statistically significant difference between the type of breast cancer surgery and the mass border (P = 0.030).

Neuroendocrine Carcinoma of the Breast: Report of A Case

Objective: Breast cancer is the most prevalent malignant disease among women and ranks among the top three most common cancers globally, alongside lung and colon cancer. Various subtypes of breast cancer have been identified. Primary neuroendocrine breast cancer, a rare and distinct type of breast carcinoma, lacks specific radiological findings. The definitive diagnosis is achieved through the expression of Synaptophysin and Chromogranin A on tumor biopsy, and it necessitates the absence of tumor detection in other regions. Staging and treatment are recommended to follow a similar approach as that of conventional breast cancer. It is believed that the prognosis of neuroendocrine breast tumors is generally poorer compared to invasive cancers.

Abstract 5563: Androgen receptor (AR) is frequently found in human cancers and inversely linked to patient outcome in breast and renal cell carcinomas

Abstract Background: Androgen receptor (AR) is a nuclear transcription regulator which mediates the growths stimulating effect of androgens. Inhibition of AR signaling is the most important first line therapy in prostate cancer. Clinical trials are ongoing for molecularly defined subsets of breast cancers expressing AR. In surgical pathology, AR immunohistochemistry is used as a marker for prostate cancer. Design: To comprehensively determine AR expression in normal and neoplastic tissues, a tissue microarray containing 18,234 samples from 141 different tumor types and subtypes as well as 608 samples of 76 different normal tissue types was analyzed by immunohistochemistry. Results: AR positivity was found in 116 of 141 tumor categories, and 66 of these tumor categories contained at least one case with strong AR staining. AR positivity was most seen in different categories of prostate cancer (87.8-100%), various subtypes of breast cancer (34.3-94.9%) and ovarian cancers (48.7-97.1%), in endometrial cancers (53-61.1%), salivary duct carcinomas (70%), teratomas (60%), renal cell carcinomas (41.8-62.7%), urinary bladder cancers (43.2-48.7%), granular cell tumors (40.7%), leiomyosarcomas (33.3%), gastrointestinal stroma tumors (31.1%) and urothelial carcinomas of the kidney pelvis (30.4%). Seventy-seven additional tumor types showed AR expression in up to 30% of the analyzed samples, but AR expression was typically only weak in these tumors. Among 1,430 evaluable invasive breast carcinomas of no special type (NST), absent or low AR immunostaining was significantly linked to high BRE grade (p<0.0001), advanced pT stage (p<0.0001), presence of nodal and distant metastasis (p<0.0001 each), HER2 overexpression (p=0.005), reduced estrogen and progesterone receptor expression (p<0.0001 each), triple negative status (p<0.0001) and shortened patient survival (p=0.0024). Among 1,149 clear cell renal cell cancers (RCC), low AR immunostaining was linked to high ISUP, Fuhrman and Thoenes grades (p<0.0001 each), high UICC and pT stage (p<0.0001 each), presence of distant metastases (p=0.0075), and reduced time to recurrence (p=0.0007), overall survival (p=0.0097) and tumor-specific survival (p=0.0129). In 297 papillary RCC, low AR staining was associated with high ISUP grade (p=0.0179), advanced pT stage (p=0.0055) and nodal metastasis (p=0.0044). Low AR expression was linked to invasive growth (p<0.0001) and nodal metastasis in urothelial carcinoma (p=0.0052). Conclusion: The results of our study demonstrate AR expression in a wide range of cancers. AR expression occurs most frequent in cancers of the prostate, breast, and ovary, but can be found in many types of non-prostate and non-gynecological neoplasms. The poor prognosis of breast and kidney cancers with reduced AR expression argues against a tumor promoting role of AR in these tumor types. Citation Format: Florian Viehweger, David Dum, Anne Menz, Ria Uhlig, Andrea Hinsch, Doris Höflmayer, Tim Mandelkow, Christoph Fraune, Christian Bernreuther, Patrick Lebok, Sören Weidemann, Guido Sauter, Maximilian Lennartz, Frank Jacobsen, Till S. Clauditz, Till Krech, Andreas H. Marx, Ronald Simon, Stefan Steurer, Eike Burandt, Sarah Minner. Androgen receptor (AR) is frequently found in human cancers and inversely linked to patient outcome in breast and renal cell carcinomas. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5563.

Abstract 2164: Patterns of estrogen receptor expression across 18,500 tumor samples obtained from 149 cancer types

Abstract Estrogen receptor alpha (ER) is a nuclear transcription factor with a pivotal role in the function of the reproductive organs and of other organ systems. It constitutes a well-established therapeutic target for various small molecules. ER immunohistochemistry is used in surgical pathology for the distinction of tumors of the breast and gynecological organs. To comprehensively determine ER expression in normal and neoplastic tissues, a tissue microarray containing 18,560 samples from 149 different tumor types and subtypes as well as 608 samples of 76 different normal tissue types was analyzed by immunohistochemistry. ER positivity was found in 55 of 149 tumor categories, and 27 of these tumor categories contained at least one case with strong ER staining. ER positivity was most seen in various subtypes of breast cancer (78.9-95.7%) and in several different tumor categories of the ovary and the endometrium (8.5-75.9%). Other commonly ER positive tumor entities included leiomyoma (65.9%), neuroendocrine tumor of the appendix (43.8%), leiomyosarcoma (22.9%), Sertoli Leydig cell tumor of the ovary (1 of 3 positive), rhabdomyosarcoma (16.7%), angiomyolipoma (10%), colorectal neuroendocrine carcinoma (9.1%), myoepithelial carcinoma of the salivary gland (8.3%), granulosa cell tumor of the ovary (8.3%), epithelial-myoepithelial carcinoma of the salivary gland (7.7%), squamous cell carcinomas of various sites of origin (0.8-6.7%), testicular teratoma (5.3%), Brenner tumor of the ovary (5.1%), Ewing sarcoma (5%), adenocarcinoma NOS of the salivary gland (4.8%), papillary thyroid carcinoma (3.3%), sarcoma, not otherwise specified (NOS) (2.8%), paraganglioma (2.6%), non-invasive papillary urothelial carcinoma, pTa G3 (2.4%), pulmonary adenocarcinoma (2.1%), neuroendocrine tumor of the ileum (2.1%), and adenocarcinoma of the prostate, Gleason 5+5 (1.2%). Among 1,411 evaluable invasive breast carcinomas of no special type (NST), absent or low ER immunostaining was significantly linked to high BRE grade (p<0.0001), advanced pT stage (p<0.0001), distant metastasis (p=0.0012), HER2 overexpression and reduced progesterone receptor expression (p<0.0001 each) but not to nodal metastasis and patient survival. Among 378 high grade serous ovarian cancer, low ER immunostaining was linked to nodal metastasis (p=0.0112). Our data provide a comprehensive overview on the pattern of ER expression in cancer. The data show that ER expression predominated in gynecological and breast tumors. However, occasional (strong) ER expression can also occur in a broad variety of non-breast and non-gynecological neoplasms. Citation Format: Florian Viehweger, David Dum, Anne Menz, Ria Uhlig, Andrea Hinsch, Doris Hoeflmayer, Christoph Fraune, Christian Bernreuther, Patrick Lebok, Soeren Weidemann, Guido Sauter, Maximilian Lennartz, Frank Jacobsen, Till Sebastian Clauditz, Till Krech, Andreas H Marx, Ronald Simon, Stefan Steurer, Sarah Minner, Eike Burandt, Natalia Gorbokon. Patterns of estrogen receptor expression across 18,500 tumor samples obtained from 149 cancer types [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2164.

Abstract P6-13-01: EPITHELIAL-MESENCHYMAL TRANSITION AND TUMOR STEM CELLS NETWORK IDENTIFICATION IN BREAST CARCINOMAS: IN SILICO STUDY

Abstract INTRODUCTION: Breast cancer (BC) is the most prevalent malignant tumor in the female population, except for skin tumors. Despite therapeutic advances, over 20% of patients with localized disease will relapse. The research of epithelial-mesenchymal transition (EMT), tumor stem cells (TSC), and the biological mechanisms related to refractoriness and metastasis development is, therefore, an opportunity for new therapeutic strategies and better results. OBJECTIVES: To identify breast cancer cellular markers related to EMT and TSC according to their histological subtypes, stage and to analyze their relationship with clinical outcomes. METHODS: After selecting a public breast cancer patients database with interactome, we identified differentially expressed genes, their associated processes, coexpression networks and interactions with pathways related to the stem phenotype and epithelial-mesenchymal transition. The characterization of key genes and the correlation with histological subtypes and clinical outcome allowed us to determine a group of genes as potential breast cancer EMT/TSC prognostic markers. RESULTS: In the 989 patients studied, 1033 differentially expressed genes (DEGs) were found, categorized according to histological subtype (hormone receptor positive, HER2 positive, triple negative) and stage IV. Seven communities of gene coexpression were found, with the gray community showing greater interaction with hormone positive tumors, the green community with HER2 positive, and the turquoise community with the triple negative one. The hormone positive disease was related to extracellular matrix processes and neuronal communication, the HER2 positive to the extracellular matrix interaction and the triple negative tumors to mitotic processes. Investigation networks with EMT and TSC related genes demonstrated a strong correlation with HER2-positive and triple-negative tumors; being eight genes in HER2 positive subtypes correlated with survival (SYNDIG1, ​​COL10A1, SLC24A2, LINC00922, KLKP1, MMP11 and ECM2); and one of the hormone positive subtype (ITIH5). ECM2 was highlighted in terms of EMT/TSC connectivity and survival. CONCLUSION:The EMT/TSC processes are significant in the various subtypes of breast cancer and impact on survival, especially in the HER2 positive subtype. Citation Format: Vanessa Dybal, Bruno R. Cavalcante, Gisele V. Rocha, Clarissa Gurgel. EPITHELIAL-MESENCHYMAL TRANSITION AND TUMOR STEM CELLS NETWORK IDENTIFICATION IN BREAST CARCINOMAS: IN SILICO STUDY. [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P6-13-01.

Dysregulation of miR-577, miR-505-3p, miR-3682-3p, and miR-4661 in Breast Cancer Patients Based on Estrogen Receptor Status

Background: Breast cancer is one of the most common malignancies and the second leading cause of cancer-related death in women. Approximately 75% of all breast cancers are estrogen receptor-positive (ER+ ) and highly responsive to endocrine therapy. MicroRNAs (miRNAs) are short non-coding RNA with a pivotal role in mammal cells by regulating gene expression. Hence, this study aimed to evaluate the miRNAs expression in various breast cancer subtypes. Materials and Methods: In this study, after total RNA extraction and cDNA synthesis, expressions of miR-577, miR-505-3p, miR-3682-3p, and miR-4661-5p were investigated in 36 breast cancer samples of ER+ and ER- types and compared with 18 normal adjacent tissues by real-time polymerase chain reaction. Also, diagnostic values of miRNAs were determined based on receiver operating characteristic (ROC) by calculating the area under the curve (AUC). Results: Downregulation of miR-577 and miR-505-3p were detected in breast cancer samples, significantly in the ER+ subtype compared to ER- subtype (P<0.001). Also, we showed upregulation of miR-3682-3p and miR-4661-5p in breast cancer tissues compared to normal tissues. Compared to the ER+ subtype, the miR-3682-3p expression significantly decreased in the ER- subtype (P<0.001). However, there was no significant difference between ER+ and ER- subtypes in the term of miR-4661-5p (P˃0.05). The ROC analysis demonstrated that miR-577 and miR-505-3p have acceptable diagnostic values, and miR-3682-3p has a relatively proper diagnostic value in diagnosing breast cancer. Conclusion: Our results revealed that miR-577 and miR-505- 3p could be used as biomarkers for the diagnosis of breast cancer, especially in ER+ subtype.

Impact of Body Mass Index on Pathological Complete Response Following Neoadjuvant Chemotherapy in Locally Advanced Breast Cancer – A South Indian Single-institute Experience

Context: Obesity is a risk factor for the development of breast cancer. Neoadjuvant chemotherapy (NACT) is now increasingly being used in its management. Pathological complete response to NACT has proven to be a surrogate marker for improved outcomes in various molecular subtypes of breast cancer. Aims: The primary objective of this prospective observational study was to analyze the impact of body mass index (BMI) on pathological complete response (pCR) rates for locally advanced breast cancer (LABC) after NACT. The secondary endpoint was to assess the histopathological features of the surgical specimen in response to NACT and to investigate the relationship with prechemotherapy BMI taking into account the various molecular subtypes of breast cancer. Subjects and Methods: Biopsy-proven LABC patients who received NACT and underwent surgery were included. Patients were categorized based on BMI classification for Asian population prechemotherapy. Molecular subtyping of breast cancer was done using immunohistochemistry and fluorescence in situ hybridization as necessary. An analysis of the association between BMI and pCR in various subtypes of breast cancer, based on hormone receptors and HER2 status, was performed. Results: The study included 205 patients. The pCR detection rate in this study was 15.6% (n = 32) which was highest in under/normal weight patients (30.2%) in comparison to overweight (12.5%) and obese (7.8%) patients (P = 0.002). For the patients who achieved pCR, the breast-conservative surgery rates were seen as higher than modified radical mastectomy rates (P = 0.025). The molecular subtype triple-negative breast cancer patients were more likely to achieve pCR (32.5%) than other molecular subtype patients (P < 0.001). Stage II patients were more likely to achieve pCR than Stage III and IV patients (P = 0.005). Conclusions: This prospective study established that overweight and obese patients suffering from breast carcinoma had a reduced chance of achieving pCR following NACT in comparison with those who had an under-/normal BMI.

Tools for Contralateral Prophylactic Mastectomy Decision Making.

Women with unilateral breast cancer are increasingly opting for the removal of not only the involved breast, but also for the removal of the opposite uninvolved breast (contralateral prophylactic mastectomy [CPM]), although the risk of contralateral breast cancer (CBC) has decreased in recent years. Models to predict the absolute risk of CBC can help a woman decide whether to undergo CPM. Our objective is to illustrate that a better decision can be made if the patient and doctor also have estimates of the absolute risks of regional and distant recurrences and mortality from non-breast cancer causes. We based our analyses on two published models for CBC and published information on the hazards of regional and distant recurrences and non-breast cancer mortality. Assuming that CPM eliminates CBC but has no effect on other events, we calculated how much CPM reduces a woman's CBC risk and total risk from all these events for 10 hypothetical women with various subtypes of breast cancer and risk factors. The risk of CBC and total risk vary greatly, depending on the breast cancer subtype. In some cases, a decision for or against CPM can be based on CBC risk alone, but in others, additional consideration of total risk may cause a woman to decline CPM. There is a potential to develop more informative tools for deciding on CPM. Realizing this potential will require more and better data to validate existing models of absolute CBC risk and to characterize the hazards of regional and distant recurrences and deaths from non-breast cancer causes for women with various subtypes of breast cancers and risk factors.

Low CRIM1 Levels Predict Poor Prognosis in Breast Cancer Patients.

BackgroundCRIM1 is involved in the development and preservation of the nervous system, capillary development, and vascular maintenance. Although CRIM1 was reported to involve in multiple cancers, its role in breast cancer is unclear.MethodsWe investigated CRIM1 expression levels using Oncomine, HPA, and immunohistochemistry analyses. BC-GenExMiner was employed to evaluate the relationship of CRIM1 expression with the clinicopathological characteristics of breast cancer. Its association with breast cancer prognosis was assessed by Kaplan-Meier analysis and PrognoScan. The correlation of the expression of CRIM1 with tumor immune infiltration was explored via TIMER. Gene set enrichment analysis (GSEA) was utilized to determine the cascades that are linked to CRIM1 in breast cancer. Finally, we explored CRIM1 and its co-expressed genes using R (3.6.3).ResultsHere, we find that CRIM1 expression was downregulated in various subtypes of breast cancer, and it was lowest in triple-negative breast cancers. ER and PR status were positively correlated with CRIM1 expression, while HER-2 expression was negatively correlated with CRIM1 expression. But in our immunohistochemical results in breast cancer specimens collected from our laboratory, HER-2 expression was positively correlated with CRIM1 expression. The expression of CRIM1 was correlated with menopause status, T stage, pathologic stage, histological type, and P53 status but not with age, N-stage, M-stage, Radiation therapy, and BRCA1/2 status. Survival analysis found that low CRIM1 expression was correlated with poorer DMFS, RFS and OS. Notably, CRIM1 expression was positively linked to the level of infiltration by CD8+ T-cells, endothelial cells, and neutrophils, and negatively linked to NK, B-cells, CD4+ T-cells, tumor purity, macrophage M1, and Tregs. Besides, DIXDC1 and PFDN6 were correlated to CRIM1 possibly.ConclusionOur findings demonstrated that low CRIM1 expression predict poor prognosis of breast cancer and CRIM1 might be used as a possible treatment target or prognostic marker in breast cancer. More researches are needed to better understand the prognostic value of CRIM1 in breast cancer.

O-Acetyl-GD2 as a Therapeutic Target for Breast Cancer Stem Cells.

SynopsisA sugar-lipid molecule called OAcGD2 is a novel marker for breast cancer stem cells. Treatment with anti-OAcGD2 mAb8B6 may have superior anticancer efficacy by targeting cancer stem cells, thereby reducing metastasis and recurrence of cancer.BackgroundCancer stem cells (CSCs) that drive tumor progression and disease recurrence are rare subsets of tumor cells. CSCs are relatively resistant to conventional chemotherapy and radiotherapy. Eradication of CSCs is thus essential to achieve durable responses. GD2 was reported to be a CSC marker in human triple-negative breast cancer, and anti-GD2 immunotherapy showed reduced tumor growth in cell lines. Using a specific anti-OAcGD2 antibody, mAb8D6, we set out to determine whether OAcGD2+ cells exhibit stem cell properties and mAb8D6 can inhibit tumor growth by targeting OAcGD2+CSCs.MethodOAcGD2 expression in patient-derived xenografts (PDXs) of breast cancer was determined by flow cytometric analyses using mAb8D6. The stemness of OAcGD2+ cells isolated by sorting and the effects of mAb8B6 were assessed by CSC growth and mammosphere formation in vitro and tumor growth in vivo using PDX models.ResultWe found that the OAcGD2 expression levels in six PDXs of various molecular subtypes of breast cancer highly correlated with their previously defined CSC markers in these PDXs. The sorted OAcGD2+ cells displayed a greater capacity for mammosphere formation in vitro and tumor initiation in vivo than OAcGD2− cells. In addition, the majority of OAcGD2+ cells were aldehyde dehydrogenase (ALDH+) or CD44hiCD24lo, the known CSC markers in breast cancer. Treatment of PDXs-bearing mice with mAb8B6, but not doxorubicin, suppressed the tumor growth, along with reduced CSCs as assessed by CSC markers and in vivo tumorigenicity. In vitro, mAb8B6 suppressed proliferation and mammosphere formation and induced apoptosis of OAcGD2+ breast cancer cells harvested from PDXs, in a dose-dependent manner. Finally, administration of mAb8B6 in vivo dramatically suppressed tumor growth of OAcGD2+ breast CSCs (BCSCs) with complete tumor abrogation in 3/6 mice.ConclusionOAcGD2 is a novel marker for CSC in various subtypes of breast cancer. Anti-OAcGD2 mAb8B6 directly eradicated OAcGD2+ cells and reduced tumor growth in PDX model. Our data demonstrate the potential of mAb8B6 as a promising immunotherapeutic agent to target BCSCs.

Иммуногистохимическая характеристика сторожевых лимфатических узлов при разных молекулярно-биологических подтипах рака молочной железы

Introduction. Breast cancer in women is a very common malignant tumor. The prognosis of the development and management of the disease depend on the clinical stage and biological subtype of the tumor. The aim of the research was to study immunohistochemical characteristics of sentinel lymph nodes in various molecular and biological subtypes of breast cancer. Materials and methods. We studied 44 lymph nodes of females with a diagnosed breast cancer using histological and immunohistochemical methods. Regional axillary lymph nodes without signs of metastatic lesions were taken into the investigation. Hematoxylin and eosin staining was performed; monoclonal antibodies to S100 protein and cluster of lymphocyte differentiation 4 and 8 were used. We used light microscopy to assess the results. Results. We determined molecular and biological subtypes of breast cancer. In 45.5% of cases (n=20) women were diagnosed with luminal cancer (Lum); in 25% of cases (n=11), with Her2+ variant; and in 29.5% of cases (n=13), with Tr– cancer (triple-negative). A more pronounced expression of S100-positive cells was observed in the paracortical zone of lymph nodes in triple-negative compared with the luminal one. We revealed uneven distribution of СD8+ lymphocytes in various subtypes of breast cancer, with an increase in their area in the following sequence: Lum (18.6%), Her2+ (19.8%), and Tr– (20.1%). The lowest number of СD4+ lymphocytes was found in the luminal breast cancer. The largest number of CD4+ cells was observed in the Her2+ subtype. Conclusion. The research demonstrated no reliable differences in the reaction of various sub-populations of T-lymphocytes in early-stage breast cancer. At the same time, we revealed a reliable increase in the number of intrafollicular S100+ cells that indicates dendrite cells activation in Tr– cancer. Keywords: breast cancer, immunohistochemistry, regional lymph node, cellular immunity, lymphocytes

Prognostic Value of a Glycolytic Signature and Its Regulation by Y-Box-Binding Protein 1 in Triple-Negative Breast Cancer.

Triple-negative breast cancer (TNBC) is the most malignant subtype of breast cancer as it shows a high capacity for metastasis and poor prognoses. Metabolic reprogramming is one of the hallmarks of cancer, and aberrant glycolysis was reported to be upregulated in TNBC. Thus, identifying metabolic biomarkers for diagnoses and investigating cross-talk between glycolysis and invasiveness could potentially enable the development of therapeutics for patients with TNBC. In order to determine novel and reliable metabolic biomarkers for predicting clinical outcomes of TNBC, we analyzed transcriptome levels of glycolysis-related genes in various subtypes of breast cancer from public databases and identified a distinct glycolysis gene signature, which included ENO1, SLC2A6, LDHA, PFKP, PGAM1, and GPI, that was elevated and associated with poorer prognoses of TNBC patients. Notably, we found a transcription factor named Y-box-binding protein 1 (YBX1) to be strongly associated with this glycolysis gene signature, and it was overexpressed in TNBC. A mechanistic study further validated that YBX1 was upregulated in TNBC cell lines, and knockdown of YBX1 suppressed expression of those glycolytic genes. Moreover, YBX1 expression was positively associated with epithelial-to-mesenchymal transition (EMT) genes in breast cancer patients, and suppression of YBX1 downregulated expressions of EMT-related genes and tumor migration and invasion in MDA-MB-231 and BT549 TNBC cells. Our data revealed an YBX1-glycolysis-EMT network as an attractive diagnostic marker and metabolic target in TNBC patients.

Identification of differentially expressed genes-related prognostic risk model for survival prediction in breast carcinoma patients.

Since the imbalance of gene expression has been demonstrated to tightly related to breast cancer (BRCA) genesis and growth, common genes expressed of BRCA were screened to explore the essence in-between. In current work, most common differentially expressed genes (DEGs) in various subtypes of BRCA were identified. Functional enrichment analysis illustrated the driving factor of deactivation of the cell cycle and the oocyte meiosis, which critically triggers the development of BRCA. Herein, we constructed a 12-gene prognostic risk model relative to differential gene expression. Subsequently, the K-M curves, analysis on time-ROC curve and Cox regression were performed to assess this risk model by determining the respective prognostic value, and the prediction performance were ascertained for both training and validation cohorts. In addition, multivariate Cox regression was analysed to reveal the independence between risk score and prognostic stage, and the accuracy and sensitivity of prognosis are particularly improved after clinical indicators are included into the analysis. In summary, this study offers novel insights into the imbalance of gene expression within BRCA, and highlights 12 selected genes associated with patient prognosis. The risk model can help individualize treatment for patients at different risks, and propose precise strategies and treatments for BRCA therapy.

Geographical disparities of incidence and prevalence of triple-negative breast cancers: Multistate study data analysis.

e12599 Background: Higher prevalence of triple negative breast cancer (TNBC) in black women with associated poor outcomes due to various disparities is well documented within a single state. We examine multiple states to better understand the state effect on such differences in incidence and prevalence of TNBC in black women. Methods: Female patients of ages 19 years old and above with breast cancer from the Surveillance, Epidemiology and End Results (SEER) Program across 13 states (608 counties) from 2015 (n = 66,444) and 2016 (n = 66,122) were examined. The relationships between the proportion of black and white women and the rate of patients with different tumor subtypes (luminal A, luminal B, HR-HER2+, and triple negative) were examined at the county level using ordinary least-square regression models. In parallel, due to consideration of various state-specific healthcare policies, socio-cultural norms, and socio-economic disparities, multi-level regression models were applied to examine the nested, random effect of each state on TNBC prevalence in each county. Bonferroni correction was applied to reduce the Type I error caused by repeated use of the same variables in multiple tests. Results: The baseline breast cancer rates between black and white women were similar in the population (0.171% for black and 0.168% for white). Consistent to previous studies, we demonstrate a significant positive correlation (p < 0.001) in TNBC in black females in both years. Surprisingly, when accounted for the random effects on states, 38.2% (2015) and 34.3% (2016) increase in incidence of TNBC in black females were seen, suggestive of state-specific disparity affecting race-specific health. In 2015, other subtypes of breast cancer in both black and white females did not result in significant relationship. Interestingly, in 2016, there was a significant relationship seen between the TNBC rate in white females and the white female population rate only after adjusting for the state effect (p = 0.026). This indicates the impact of non-biological factors such as state-wide health policies. Additionally, HR-HER2+ black females had a significant relationship against respective population rate only after adjusting for the state effect as well (p = 0.0394). For luminal A white females, a 15% decrease in incidence was seen after adjusting for state effect (p = 0.0424). Conclusions: This is the first known across-state examination of breast cancer subtypes by race with random effects on state. This study shows the role of state-specific factors affecting incidence in black and white females and potentially indicates the importance of state-level management for breast cancer on health disparities in addition to race-driven effects. Further studies are needed to elucidate comparable differences between states affecting the rates of various subtypes of breast cancer and thus health outcomes.

Abstract PO-001: HDAC6 inhibitors sensitize non-mesenchymal triple-negative breast cancer cells to cysteine deprivation

Abstract Triple-negative breast cancer (TNBC) accounts for 15% ~ 20% of overall breast cancer cases and exhibits earlier age of onset, high metastasis, and aggressiveness with poor clinical outcomes shown by higher relapse and lower survival rates than other types of breast cancer. TNBC contributes to the major mortality of breast cancer patients. Treatments of TNBC patients are still limited to surgery, chemotherapy, or radiation since the absence of cell receptors makes targeted hormonal therapies impossible. Targeting cysteine-dependence is an emergent strategy to treat the mesenchymal TNBC. However, many TNBC cells are non-mesenchymal and irresponsive to cysteine deprivation. To overcome such resistances, three selective HDAC6 inhibitors (Tubacin, CAY10603, and Tubastatin A), identified by epigenetic compound library screening, can synergize with cysteine deprivation to induce cell death in the non-mesenchymal TNBC. Despite the efficacy of the HDAC6 inhibitor, knockout of HDAC6 did not mimic the synthetic lethality induced by its inhibitors, particularly tubacin, indicating that HDAC6 is not the actual target of HDAC6 inhibitor in this context. Instead, transcriptomic profiling showed that tubacin triggers an extensive gene transcriptional program in combination with erastin, a cysteine transport blocker. Notably, the zinc-related gene response and an increase of labile zinc were induced in cells by the combination treatment. We found that the disturbance of cellular zinc homeostasis was driven by PKCγ activation, which revealed that the PKCγ signaling pathway is required for HDAC6 inhibitor-mediated synthetic lethality. Overall, our study identifies a novel function of HDAC6 inhibitors that function as potent sensitizers of cysteine deprivation. HDAC6 inhibitors are capable to overcome cysteine independence in non-mesenchymal tumor cells, which allows the application of targeted cysteine-dependence therapy in various subtypes of breast cancer. Citation Format: Tahiyat Alothaim, Morgan Charbonneau, Xiaohu (Mark) Tang. HDAC6 inhibitors sensitize non-mesenchymal triple-negative breast cancer cells to cysteine deprivation [abstract]. In: Proceedings of the AACR Virtual Special Conference on Radiation Science and Medicine; 2021 Mar 2-3. Philadelphia (PA): AACR; Clin Cancer Res 2021;27(8_Suppl):Abstract nr PO-001.