Expression In Breast Carcinoma Research Articles

KDM7A-DT induces genotoxic stress, tumorigenesis, and progression of p53 missense mutation-associated invasive breast cancer.

Stress-induced promoter-associated and antisense lncRNAs (si-paancRNAs) originate from a reservoir of oxidative stress (OS)-specific promoters via RNAPII pausing-mediated divergent antisense transcription. Several studies have shown that the KDM7A divergent transcript gene (KDM7A-DT), which encodes a si-paancRNA, is overexpressed in some cancer types. However, the mechanisms of this overexpression and its corresponding roles in oncogenesis and cancer progression are poorly understood. We found that KDM7A-DT expression is correlated with highly aggressive cancer types and specific inherently determined subtypes (such as ductal invasive breast carcinoma (BRCA) basal subtype). Its regulation is determined by missense TP53 mutations in a subtype-specific context. KDM7A-DT transcribes several intermediate-sized ncRNAs and a full-length transcript, exhibiting distinct expression and localization patterns. Overexpression of KDM7A-DT upregulates TP53 protein expression and H2AX phosphorylation in nonmalignant fibroblasts, while in semi-transformed fibroblasts, OS superinduces KDM7A-DT expression in a TP53-dependent manner. KDM7A-DT knockdown and gene expression profiling in TP53-missense mutated luminal A BRCA variant, where it is abundantly expressed, indicate its significant role in cancer pathways. Endogenous over-expression of KDM7A-DT inhibits DNA damage response/repair (DDR/R) via the TP53BP1-mediated pathway, reducing apoptosis and promoting G2/M checkpoint arrest. Higher KDM7A-DT expression in BRCA is associated with KDM7A-DT locus gain/amplification, higher histologic grade, aneuploidy, hypoxia, immune modulation scores, and activation of the c-myc pathway. Higher KDM7A-DT expression is associated with relatively poor survival outcomes in patients with luminal A or Basal subtypes. In contrast, it is associated with favorable outcomes in patients with HER2+ER- or luminal B subtypes. KDM7A-DT levels are coregulated with critical transcripts and proteins aberrantly expressed in BRCA, including those involved in DNA repair via non-homologous end joining and epithelial-to-mesenchymal transition pathway. In summary, KDM7A-DT and its si-lncRNA exhibit several intrinsic biological and clinical characteristics that suggest important roles in invasive BRCA and its subtypes. KDM7A-DT-defined mRNA and protein subnetworks offer resources for identifying clinically relevant RNA-based signatures and prospective targets for therapeutic intervention.

An Observational Study of Stromal CD10 Expression in Breast Carcinoma and Its Association with Clinicopathological Parameters at SMS Medical College and Hospital

Introduction: Female breast cancer is now more prevalent than lung cancer worldwide, with an estimated 2.3 million new cases worldwide, or 11.7% of all cancer cases. Breast cancer is an epithelial malignancy, but tumor microenvironment i.e. stroma plays remarkable role in invasion and metastasis. The cross talk between tumor cells and stromal cells is critical step in tumor progression as a result there is increased interest in developing novel therapies targeting the microenvironment. Aim and Objectives: To study the frequency and intensity of stromal CD10 expression in invasive breast cancer and to correlate it with various clinicopathological parameters.Materials and Methods: The study was conducted in Department of Pathology at S.M.S. Medical College, Jaipur. CD 10 expression in 70 patients diagnosed with carcinoma breast during the period of 2021-2022, assessed by immunohistochemistry and scored as negative (<10% stromal positivity), weak (10-30% stromal positivity) and strong (>30% stromal positivity) and statistically analyzed with other parameters. Results and Discussion: Out of the 70 cases, 51 (72.86%) showed CD10 positivity in the stroma, with 28 (40%) showing strongly positive and 23 (32.86%) with weakly positive results. With respect to histopathological grade (P value = 0.0234), molecular subtype (P value = 0.0006), and PR negativity (P value = 0.0480), CD10 expression in the stroma showed a positive correlation. Conclusion: The correlation between stromal expression of CD10 and histopathological grade, molecular subtype, and PR negativity was statistically significant. To confirm CD10 expression in invasive breast cancer as a predictor of overall survival rates and disease-free survival, more research is required..

PRAME expression and its prognostic significance in invasive breast carcinoma

ObjectivesPRAME (PReferentially expressed Antigen in MElanoma) is a carcinoma testis antigen expressed in numerous tumour types. The aim of this study was to assess PRAME expression in different surrogate subtypes of breast carcinoma and its correlation with other prognostic factors. Material and methodsA total of 220 cases of invasive breast carcinoma were selected and categorized according to ER, PgR, HER2 status, and Ki67 proliferation index in luminal A like, luminal B HER2+ like, luminal B HER2- negative like, HER2 positive like and triple-negative or basal-like. All cases were examined for PRAME expression by immunohistochemistry (IHC). ResultsA PRAME-high profile was detected in 53 (24,1 %) of all examined breast carcinoma samples. A significantly higher expression of PRAME was detected in HER2-positive carcinomas (50 %) and TN breast carcinomas (40,54 %) compared to ER-positive (luminal-like) subtype of breast carcinomas (3,38 % luminal A and 15,38 % luminal B). Percentage of PRAME positive tumour cells showed positive correlation with tumor size, Ki67 proliferation index, HER2 status, nuclear grade, TILs and presence of metastasis, and negative correlation with ER status and disease-free survival (DFS). ConclusionOur study showed that HER2 positive and TN breast carcinomas more commonly express PRAME than ER positive carcinomas and that PRAME expression shows positive correlation with certain prognostic factors, however PRAME wasn't revealed as an independent prognostic factor in our study. The importance of PRAME expression in breast carcinoma lies in its potential use as an immunotherapeutic target, particularly in patients with limited therapeutic options.

Immunohistochemical study of CD74 biomarker in normal and malignant breast tissues.

Aim: To detect the role of CD74 expression in breast carcinoma as a predictive marker for identifying the biological behavior of malignancy in Iraqi women.. Materials and Methods: The study used technique of immunohistochemistry for detection CD74 protein role in breast cancer, and its expression in breast cancer tissue samples. Samples were collected in Al-Najaf city in Iraq, from Al-Forat Al-Awsat Oncology Center. The study was achieved at the Laboratories of the Faculty of Science in the University of Kufa. Fifty samples of breast cancer tissue, and twenty controls benign tissue were included in the study. The study has investigated relationship between expression of biomarker with grade, age of patient and tumor size. Results: The study showed that the cytoplasmic expression of CD74 with more clear and intensive staining in the cytoplasm, and reported that CD74 positivity rate was 52%. A significant association between CD74 expression and grade and size of tumor, so CD74 can be considered as a biomarker for prediction of breast cancer in women. No association was found between CD74 expression and each of patients' age and node metastasis. Conclusions: The study represents an important step in our region because there are a few studies about this topic; more efforts are required to approve the function of this biomarker.

The Study on BCL2 Expression in Breast Carcinoma: Correlation with Predictive and Prognostic Factors

The Study on BCL2 Expression in Breast Carcinoma: Correlation with Predictive and Prognostic Factors

Vascular Endothelial Growth Factor Expression in Breast Carcinoma and its Association with Expression of Estrogen Receptors, Progesterone Receptors and HER-2/neu and Molecular Subtypes

Background: The management of breast carcinoma depends on several molecular markers and tumor stages. In the last decades, estrogen receptors (ER), progesterone receptors (PR), and HER-2/neu have shown good therapeutic responses. Among other molecular markers, vascular endothelial growth factor (VEGF) is becoming more widely used as a prognostic indicator in patients with breast carcinoma. Anti-VEGF therapy already has been proven as an effective chemotherapeutic agent in some other carcinomas. The study aimed to find out the immunohistochemical expression of Vascular Endothelial Growth Factor (VEGF) in breast carcinoma and its possible correlation with the expression of ER, PR, and HER-2/neu and molecular subtypes to evaluate its prognostic value. Material & Methods: This study was conducted in the Department of Pathology, BIRDEM General Hospital, Dhaka, from March 2018 to January 2020. In this study, 45 diagnosed cases of breast carcinoma were enrolled. Slides of all cases were stained with ER, PR, HER-2/neu, and VEGF antibodies following the avidin-biotin-peroxidase staining method. Results: Among 45 cases, 60% showed positive immunohistochemical expression of VEGF. Most of these cases (71.1%) were ER/PR positive. VEGF did not show a significant association with other molecular markers or molecular subtypes. Conclusion: Although, the potential prognostic value of VEGF has not been confirmed. Based on the findings of the current study, it can be assumed that VEGF plays an important role in the pathogenesis of breast cancer. So, it may serve as a useful biomarker for immuno-targeting therapy in patients with breast cancer.

Variable Landscape of PD-L1 Expression in Breast Carcinoma as Detected by the DAKO 22C3 Immunohistochemistry Assay.

In 2020, pembrolizumab was approved as a therapy for triple-negative breast cancer (TNBC) with the companion diagnostic DAKO 22C3 programmed death ligand-1 (PD-L1) immunohistochemistry assay. The study aimed to determine the landscape of PD-L1 expression as detected by the DAKO 22C3 PD-L1 assay in breast cancer subtypes and compare the clinicopathologic and genomic characteristics of PD-L1 positive and negative TNBC. PD-L1 expression using the DAKO 22C3 antibody was scored using a combined positive score (CPS) and positive status was defined as CPS ≥10. Comprehensive genomic profiling was performed using the FoundationOne CDx assay. Of the 396 BC patients stained with DAKO 22C3, the majority were HR+/HER2- and TNBC (42% and 36%, respectively). Median PD-L1 expression and frequency of CPS ≥10 was highest in TNBC cases (median: 7.5, 50% CPS ≥10) and lowest in the HR+/HER2- group (median: 1.0, 15.5% CPS ≥10) (P < .0001). A comparison of PD-L1 positive and PD-L1 negative TNBC demonstrated no significant differences in clinicopathologic or genomic characteristics. TNBC tissue samples from the breast did have an observed enrichment for PD-L1 positivity compared to TNBC tissue samples from a metastatic site (57% vs. 44%), but this was not statistically significant (P = .1766). In the HR+/HER2- group, genomic alterations in TP53, CREBBP, and CCNE1 were more prevalent and genomic loss of heterozygosity was higher in the PD-L1(+) group compared to the PD-L1(-) group. The subtypes of breast cancer have distinct patterns of PD-L1 expression, supporting that further research of immunotherapies may include specific evaluation of optimum cutoffs for non-TNBC patients. In TNBC, PD-L1 positivity is not associated with other clinicopathologic or genomic features and should be integrated into future studies of immunotherapy efficacy.

Immunohistochemical study of proline, glutamic acid and leucine-rich protein 1 (PELP1) in correlation with guanine adenine thymine adenine family member 3 (GATA-3) receptors expression in breast carcinomas

Abstract Background Breast cancer is the most common cancer affecting Egyptian women. Guanine adenine thymine adenine family member 3 (GATA-3) and Proline, glutamic acid and leucine-rich protein 1 (PELP1) are both expressed in breast carcinomas. Aim To evaluate GATA-3 and PELP1 proteins expression in breast carcinoma, assess association of both immunohistochemical markers with variable prognostic factors, correlate between GATA-3 and PELP1 expression and assess the diagnostic utility of both markers in breast carcinomas. Subjects and methods The current study was a cross-sectional descriptive study, performed in the Pathology laboratory of Suez Canal University Hospital on 86 samples of formalin fixed paraffin embedded blocks of breast carcinoma during the period from January 2016 to May 2019. Results GATA-3 and PELP1 showed positive nuclear expression in breast carcinoma cases (74.4% and 82.6%, respectively). PELP1 expression showed statistically significant association with older age cases. GATA-3 expression showed statistically significant association with positive ER, PR and HER-2 expression and molecular subtypes. A statistically significant correlation was found between GATA-3 and PELP1 expression in breast carcinomas (n=86, P value &lt;0.05). PELP1 showed the highest sensitivity in stage III breast cancer samples (84.9%) compared to GATA-3 (75.8%), ER (66.7%), PR (54.6%) and HER2 (39.4%). PELP1 is more sensitive (80%) than GATA-3 (60%) in grade III breast carcinomas. GATA-3 is more sensitive (89.7%) than PELP1 (86.2%) in luminal breast cancer subtype. the combined parallel use of GATA-3 and PELP1 raised their sensitivity to 92% in Grade III and 98.6% in luminal breast cancers. Conclusions GATA-3 and PELP1 may be a promising diagnostic markers of breast carcinoma. Further studies are needed to fully evaluate the prognostic significance of GATA-3 and PELP1 expression in invasive breast carcinomas.

Stromal CD10 Expression in Breast Carcinoma and its Association with ER, PR, and HER2/neu Using Immunohistochemistry: A Cross-sectional Study

Introduction: Breast cancer is a common and deadly malignancy affecting women worldwide. Various immune markers, such as Estrogen Receptor (ER), Progesterone Receptor (PR), and Human Epidermal growth factor Receptor 2 (HER2/neu), are commonly used to assess prognosis. Currently, ongoing research aims to evaluate molecular pathways that contribute to invasion and metastasis. One important immunomarker under investigation is CD10, a zinc-dependent Matrix Metalloproteinase (MMP) that degrades bioactive peptides. CD10 expression in the tumour stroma has been associated with the biological aggressiveness of several epithelial malignancies, including breast carcinoma. Aim: To analyse the association between stromal CD10 expression and different prognostic factors, such as age, histological grade, and status of ER, PR, and HER2/neu markers, in patients with breast cancer. Materials and Methods: This institutional-based, crosssectional study was conducted at the Department of Pathology, Nil Ratan Sircar Medical College and Hospital (NRSMC) in Kolkata, West Bengal, India over a period of one and a half years (February 2021 to July 2022). It included 120 cases of breast carcinoma diagnosed through histopathological examination of formalin fixed paraffin embedded sections, which were prepared from trucut biopsies and resection specimens referred from the Department of General Surgery, NRSMC. CD10 expression was assessed by Immunohistochemistry (IHC) in all cases and scored as negative, weak, or strong. The study examined the association between CD10 expression and the following parameters: age, histopathological grade, and the status of ER, PR, and HER2/neu markers. Data were entered into Microsoft Excel (MS) for statistical analysis. The significance of the study was determined using the Chi-square test, and data were analysed using Statistical Package for the Social Sciences (SPSS) version 23.0 (IBM, Illinois, US). A p-value of &lt;0.05 was considered statistically significant. Results: The study included a total of 120 cases, with 119 cases of female breast cancer (99.16%) and one case of male breast cancer (0.84%). The mean±SD age of the patients was 54±5.038 years (range 44 to 67 years). The majority of cases 102 (85.00%) were diagnosed as Invasive Ductal Carcinoma (IDC), Not Otherwise Specified (NOS), followed by IDC-special types 18 (15.00%). CD10 expression was evaluated in all cases, and stromal CD10 positivity was observed in 79 out of 120 cases (65.80%), with 39 individuals (49.40%) showing weak positivity and 40 cases (50.60%) showing strong positivity. The remaining 41 cases (34.16%) were CD10 negative. Grade 3 cancers were predominant in this study 62 (51.66%). It was noted that CD10 stromal positivity increased with higher grade. Most of the cancers in this study were negative for ER, PR, and HER2/ Neu (78, 65.00%; 84, 70.00%; and 67, 55.83%, respectively). Stromal CD10 expression showed a significant association with ER (p-value=0.00001), HER2/neu (p-value=0.000089), tumour grade (p-value=0.0012), and an insignificant association with age (p-value0.264) and PR (p-value=0.256). Conclusion: Therefore, CD10 expression is strongly associated with well-established prognostic markers, namely higher tumour grade, HER2/neu negativity, and ER negativity. This indicates that CD10 can not only be used as an independent marker of poor prognosis but also as a target for the development of novel therapies.

Expressions and Prognostic Values of Notch3 and DLL4 in Human Breast Cancer.

Background: Notch signaling played a critical role in promoting breast tumorigenesis and progression. However, the role and prognostic value of Notch3 combined with DLL4 expression in breast carcinoma had not been explored. Methods: The retrospective study enrolled 90 breast cancer tissues and 60 noncancerous tissues from (conceal). The expression and prognostic value of Notch3 and DLL4 in patients with breast carcinoma were investigated using Oncomine and UALCAN database. Notch3 and DLL4 expression levels were detected by quantitative real-time polymerase chain reaction, western blotting, and immunohistochemistry. We analyzed the correlation between both proteins expression and clinicopathological parameters and survival data, respectively. Results: The expressions of Notch3 and DLL4 were increased, and Notch3 expression was significantly positively associated with DLL4 in breast carcinoma. The 2 proteins dramatically correlated with advanced stage, high grade and negative Her2 status. The overexpressing of single or both Notch3 and DLL4 resulted in shortened survival of breast cancer patients. And Notch3 overexpression was one of independent risk predictors to poor prognosis. Conclusion: The interaction of Notch3 receptor and DLL4 ligand accelerates oncogenesis, progression, and poor prognosis of breast cancer patients. Notch3 protein may serve as one of biomarker to independently predict prognosis of patients.

Study of Interleukin-10 Gene Expression in Breast carcinoma in Telangana Population

Background: Carcinoma breast is the second most common malignancy next to carcinoma cervix in India andabroad. Hence, the expression of IL-10 m-RNA gene in tumour can be a novel prognostic indicator.Method: 30 mastectomy specimens of carcinoma Breast patients aged between 30 to 70 years were studied. 2to 5 gram of tumour tissue and equivalent sample from normal peri-tumoral breast were taken from resectedbreast immediately after surgery and preserved in RNA-later solution. IL-10 m-RNA expression was detectedand quantified by RT-PCR analysis through TaqMan chemistry, on 30 samples and 30 controls. After fixation withformalin 2 micro sections of 4-5micron thickness were taken from tumor area and corresponding paraffin blockswere prepared, mounted, on albumin coated slide for H &amp; E staining and results were observed and recorded.Results: Histologically, 08 specimens were diagnosed as grade-I (26.5%), 16 were grade-II (53.5%) and 06 weregrade-III (20%). 10 specimens (33.5%) had reactive lymph node, and the remaining 20 (66.5%) had metastaticlymph node. Mean RQ values for IL-10 were calculated by the formula for comparative ct (cycle threshold)method. 26 neoplastic tissue specimens (86.5%) showed strong expression of IL-10 mRNA, while only 2 (6.5%)normal peritumoral surrounding tissue showed IL-10 expression. 4 (13.5%) neoplastic tissue and 28 (93.5%) peritumoraltissue showed weak expression of IL-10. All grade 3 tumours showed strong expression of IL-10.Conclusion: Present pragmatic study had evidence that, IL-10 m-RNA gene expression was significant in neoplasticbreast tissue and may have potential prognostic significance.

Leptin and leptin receptor expression in breast carcinomas and their relationship with clinicopathological features

ContextBreast cancer is the most common cancer and the most common cause of death from cancer among women. Leptin is thought to play role in cell proliferation. ObjectiveIn this study, we evaluated the expression of leptin and leptin receptor in breast cancer and normal breast tissue, analyzed the relationship between expression of leptin, leptin receptor and clinicopathological features in breast cancers. Materials and MethodsThe expression of leptin and leptin receptor were investigated by immunohistochemistry in 205 breast cancer and 30 normal breast tissue. ResultsBreast cancers showed higher expression of leptin (p = 0,003) and leptin receptor (p < 0,001) than normal breast tissues. The expression of leptin showed significant correlation with leptin receptor expression (r = 0,644; p < 0,001). Higher rates of ER and PR positivity are found in leptin positive tumors(p < 0,001). Expression levels of leptin were positively corelated with ER (r = 0,336; p < 0,001) and PR (r = 0,359; p < 0,001) expression. HER2 positivity rates were lower in leptin positive tumors (p = 0,020). Negative correlation between leptin expression levels and histological grade was found (r = -0,168; p = 0,016). Higher rates of leptin receptor positivity were detected in invasive ductal carcinoma NSTs than invasive lobular carcinomas. ConclusionElevated expressions of leptin and leptin receptor levels in breast cancer tissues suggest that leptin may have a promoting effect on the carcinogenesis of breast cancer, possibly in an autocrine manner. Inhibition of leptin may be effective for the treatment of breast cancer.

Alpha-methylacyl-CoA racemase (AMACR) protein is upregulated in early proliferative lesions of the breast irrespective of apocrine differentiation

Alpha-methylacyl-CoA racemase (AMACR/P504S) is a mitochondrial and peroxisomal enzyme involved in the branched-chain fatty acid and bile acid metabolism. AMACR is a useful diagnostic biomarker for prostate carcinomas and several other malignancies. Its expression in apocrine breast lesions had been previously reported, but its role in breast cancer progression has not been fully investigated. One hundred fifty breast samples (80 with invasive carcinomas) were studied. The expression of AMACR protein was analyzed using the immunohistochemical method (IHC). Lesions were considered positive if AMACR was detected in ≥10% of the cells at any intensity comprising a histologically defined normal epithelial structure or a pathologic lesion. In addition, AMACR mRNA relative expression was calculated from the whole-transcript RNA-Seq performed on >20,000 diverse tumor samples using a 20,000+ hybrid-capture NGS assay with the transcript capture panel based on the Agilent SureSelect Human All ExonV7. Expression of AMACR protein was restricted to epithelia. It was uncommon in the normal breast (7/81 samples, 9%). Increasing AMACR expression was observed with proliferative epithelial lesions (18% of usual ductal hyperplasias/adenosis, 70% of atypical lesions and 72% of DCIS/LCIS). Invasive ductal carcinomas NST and invasive lobular carcinomas expressed AMACR in 64% and 46%, respectively. The highest AMACR expression was observed in luminal B and HER2-positive breast carcinomas (86-100%). Triple-negative breast carcinomas exhibited AMACR in 50% of the cases. Apocrine lesions showed strong, nearly uniform overexpression of AMACR (100% of metaplasias, hyperplasias and in situ carcinomas and 88% of invasive apocrine carcinomas were positive). RNA-Seq analysis also confirmed AMACR expression in breast carcinomas, although its median value was substantially lower with a lower standard deviation than in prostate carcinomas. Over-expression of AMACR characterizes various proliferative, preinvasive and invasive breast lesions and is not specific to the apocrine morphology. It points to altered lipid metabolism (branched fatty acids) as one of the general characteristics of breast carcinogenesis, like several other malignancies. Its early detection may represent a potential target for cancer progression intervention.

Study of GATA-3 Expression in Breast Carcinoma in a Tertiary Care Hospital in Eastern India

Background: Transcription factor GATA3 is a relatively new and specific biomarker for breast carcinoma that regulates luminal cell differentiation. However, further studies that elucidate its role and diagnostic utility in breast cancer is desired. Therefore, this study was conducted to assess GATA3 expression in breast carcinoma and determine its association with various known prognostic and predictive factors of breast carcinoma. Methods: This is a cross-sectional, observational study where 110 breast carcinoma cases were examined histologically on H&amp;E stained slides for routine parameters such as tumor morphological type and histologic grade.ER/PR/Her-2-neu/ki67 and GATA3 expression was determined using immunohistochemistry. All cases were assessed to determine the association of GATA3 with different established clinicopathological parameters in breast cancer such as patient age, axillary lymph node status, tumor grade, lymphovascular invasion, molecular subtype etc. The results were statistically analyzed and P value &lt; 0.05 was considered to be significant. Results: 97 cases (88.1%) expressed GATA3 and it was significantly associated with most of the established clinico-pathological parameters like family history of breast cancer, tumour size and histology grade, lymph node and margin status, presence of lymphovascular invasion and carcinoma in-situ component .GATA3 expression was also significantly associated with ER/PR/Her2neu/ki67 expression. Conclusion: GATA3 expression is significantly associated with the different established clinicopathological parameters of breast cancer. Therefore, it may be considered as a relevant marker more routinely. Further studies of GATA3 in breast cancer may refine prognostic models, predict clinical outcomes and modify treatment guidelines in future

Abstract 991: OVO-like genes are master regulators of the epithelial fate in breast cancer

Abstract The OVO-like genes are evolutionary conserved C2H2 zinc finger transcription factors (TFs) with SNAG repressor domains involved in the maintenance of epithelial homeostasis. From the three Ovo-like family genes identified in mammals, Ovol-like1 (Ovol1) and Ovol-like2 (Ovol2) are the most studied members with multiple reports pinpointing relevant roles in development and disease. In cancer, one of the hallmarks of invasive carcinoma involves the loss of E-cadherin expression by the repression of SNAG transcription factors. Interestingly, contrary to the other SNAG domain family members the OVO-like genes regulate epithelial cell fate. We performed data analysis in breast cancer cell lines and patient data obtained from GEO, CCLE, TCGA and METABRIC to assess the potential of the OVO-like genes as relevant regulators of gene expression in breast carcinoma. In addition, we stablished breast cancer models with inducible expression or repression of the OVO-like genes. We also analyzed the cellular systems we generated using naïve high-throughput methods such as RNA-Seq. Likewise, we studied the effect of OVO-like genes on tumor progression and metastasis using inducible expression systems in both xenograft and syngeneic breast cancer models. We observed that Ovol1 and Ovol2 are constitutively expressed in human breast cancer cell lines, and they are upregulated due to chromosomal gains in breast cancer patients. We observed how the introduction of Ovol1 and Ovol2 into mesenchymal-like breast cancer cell lines modified their transcriptome in a way that promoted expression of epithelial marker genes (e.g., Amphiregulin) and downregulated mesenchymal markers (e.g., Vimentin). In addition, introducing Ovo-like genes in mesenchymal-like cells greatly affected their viability and function. Surprisingly, we observed a similar effect by downregulating the Ovo-like genes in epithelial-like cells. We propose a model where a common epigenetic machinery is shared between structurally related transcription factors with opposing effects, such as OVO-like proteins and other SNAG-containing transcription factors, such as Snail and Slug, such that the abundance of the transcription factors and their co-repressors defines the state of the system as either epithelial or mesenchymal. Citation Format: Diana Drago Garcia, Yosef Yarden. OVO-like genes are master regulators of the epithelial fate in breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 991.

Expression of novel neuroendocrine markers in breast carcinomas: a study of INSM1, ASCL1, and POU2F3

INSM1, ASCL1, and POU2F3 are novel transcription factors involved in neuroendocrine (NE) differentiation of neoplasms in several organs, but data on their expression in breast carcinomas (BCs) are limited. We retrospectively evaluated the expression of these markers in a series of 97 BCs (58 with NE morphology and 39 with otherwise uncommon morphology) tested prospectively using immunohistochemistry (IHC). Nuclear staining in >50% of the cells was used as the positive cut-off. Thirty-two of the 97 BCs (33%) were INSM1-positive. INSM1-positivity correlated significantly with histologic type and presence of stromal mucin. INSM1 also correlated with synaptophysin and chromogranin, established markers of NE differentiation (P<.0001 and P=.0023, respectively). In BC with NE morphology, the expression of INSM1 supported NE differentiation, and INSM1 was more specific than synaptophysin and more sensitive and specific than chromogranin. INSM1 was the most expressed NE marker in 17BCs. INSM1-positive BCs included 56% of solid papillary BCs, 88% of BCs with solid papillary features, and 75% of high-grade NE carcinomas. Of 35 BCs tested for POU2F3 and ASCL1, only 1 and 4 cases were positive, respectively. Our results show that INSM1 is a sensitive marker of NE differentiation in BC and should be included with synaptophysin and chromogranin in the IHC panel used to evaluate NE differentiation in BC with NE morphology. ASCL1 and POU2F3 are uncommon in BC and their routine assessment is not warranted.

MTHFD2 Expression Profile and Its Prognostic Importance in Invasive Breast Carcinoma

Background: Breast cancer is the most common cancer among women. Therefore, it is very important to determine prognostic parameters and biomarkers related to these parameters. Recent studies have shown that folate metabolism is effective on cell division and invasiveness. One of the most important regulators of folate metabolism is the methylene tetrahydrofolate dehydrogenase enzyme (MTHFD2). Method: This study, immunohistochemical staining with MTHFD2 was performed in 194 cases diagnosed with invasive breast carcinoma. The relationship between the immunohistochemical expression of MTHFD2 in normal ductal areas and invasive tumoral areas of these cases and some clinicopathological parameters was investigated. Results: While no expression was observed in non-tumor areas with MTHFD2, varying degrees of expression were observed in all tumor areas (p&lt;0.0001). Significant correlations were also observed between important clinicopathological and prognostic parameters such as MTHFD2 nuclear grade, hormonal receptor, Ki67 and etc. (all p&lt;0.05). Conclusion: High MTHFD2 expression in breast carcinomas with invasive and poor prognostic values ​​was determined to be very significant. Therefore, it can be predicted that detecting and lowering MTHFD2 levels in these cases may prevent tumor development, invasion and metastasis. We think that more studies on the subject are needed, including the development of chemotherapeutics targeting MTHFD2.

The Valuable Role of ARMC1 in Invasive Breast Cancer as a Novel Biomarker.

Background Invasive breast carcinoma (BRCA) is a common type of breast cancer with a high clinical incidence. Thus, it is significant to find effective biomarkers for BRCA diagnosis and treatment. Although some members of armadillo (ARM) repeat family of proteins are confirmed to be biomarkers in cancers, the role of armadillo repeat-containing 1 (ARMC1) in BRCA remains unknown. Methods We firstly analyzed the ARMC1 expression in normal breast tissues and BRCA samples and its association with overall survival by the public database. Next, the χ2 test was used to evaluate the prognostic significance of ARMC1 expression in TCGA-BRCA patient samples. The ARMC1 mutations in BRCA were explored in the cBioportal database. Besides, the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were used to explore the biological functions of ARMC1 in BRCA. Finally, immunohistochemistry and immunofluorescence staining were performed to validate the ARMC1 expression in BRCA. Results ARMC1 expression in tumor samples was significantly higher than that in normal tissues, and higher expression of ARMC1 was related to lower survival. Moreover, the tumor stage and histology of BRCA patients were associated with ARMC1 expression. ARMC1 genetic mutations occurred in 32% of BRCA patients, and the amplification and high expression of ARMC1 accounted for most of them. Furthermore, functional enrichment analysis suggested that ARMC1 might be involved in the cell cycle in BRCA. Ultimately, increased ARMC1 expression was found in clinical breast carcinoma tissues by our confirmatory experiments. Conclusions ARMC1 may play a significant role in BRCA and act as a biomarker, which provides valuable clues for the treatment and diagnosis of BRCA.

Prognostic value of programmed death ligand-1 expression in breast carcinomas

ObjectivesTo assess the prognostic value of programmed death ligand-1 (PDL-1) expression in patients with nonmetastatic breast carcinoma patients both in tumor cells and tumor-infiltrating lymphocytes (TILs).BackgroundPDL-1 has emerged as important immune biomarkers in breast cancer. It can be targeted with immunotherapy, which represents a significant advancement in the management of this disease.Patients and methodsParaffin blocks of patients with breast cancer will be tested for the expression of PDL-1 both on TILs and malignant epithelial cells and correlated with patients and disease features.ResultsThis retrospective study included 60 patients. PDL-1 expression was positive in 33 (55%) patients on TILs and was positive in tumor cells in 27 (45%) patients of the studied patients. There was no significant relation between PDL-1 expression on TILs and patients and disease features. There was significant relation between positive PDL-1 expression on epithelial cells and triple-negative breast cancer, estrogen receptor negative, progesterone receptor negative status (P = 0.006, 0.006, and 0.006). PDL-1 intensity in intratumoral lymphocytes is significantly related to tumor stage (P = 0.006). PDL-1 H score in malignant epithelial cells was significantly related to disease stage (P = 0.021). Tumor size, HER-2neu status, and PDL-1 H score on malignant epithelial cells were independent risk factors for disease progression.ConclusionPDL-1 expression is common in both malignant tumor cells and TILs. A higher PDL-1 H score on malignant epithelial cells was associated with longer time to progression.

TRPS1 Expression in Breast Carcinomas: Focusing on Metaplastic Breast Carcinomas.

TRPS1 has been recently demonstrated as a highly sensitive and specific marker for breast carcinomas. To further explore TRPS1's utility in breast carcinoma, we systematically evaluated TRPS1 expression on tissue microarrays from 160 estrogen receptor (ER)-positive/human epidermal growth factor receptor 2 (HER2)-positive, 94 ER-/HER2+, 117 triple-negative breast carcinomas, and 618 other primary carcinomas (cholangiocarcinoma, endometrial, colorectal, and hepatocellular carcinomas), and whole tissue sections from 64 HER2+, 76 triple-negative, and 67 metaplastic breast carcinomas. The results showed TRPS1 was highly expressed in breast carcinomas (100% of HER2+ and 97.4% of triple negative on whole tissue sections), but almost completely negative in other tested tumor types. TRPS1 was also highly expressed in metaplastic carcinoma (91%), significantly higher than GATA3 (55.2%). The different expression between TRPS1 and GATA3 was most prominent in chondroid/mesenchymal subtypes (100% vs. 36.4%), followed by spindle cell carcinoma (66.7% vs. 44.4%). In addition, TRPS1 was expressed in normal breast ductal epithelial cells with less staining than in carcinoma cells, and TRPS1 showed aberrant membranous staining in HER2+ breast carcinomas that suggests a potential cross-reactivity with HER2 protein.