The Elevated Serum Interleukin-40 Levels and Gene Expression as Novel Biomarkers in Iraqi Women with Breast Cancer

Amplification of a 280-Kilobase Core Region at the ERBB2 Locus Leads to Activation of Two Hypothetical Proteins in Breast Cancer

BACKGROUND: MicroRNAs have emerged as new diagnostic and therapeutic biomarkers for breast cancer. Herein, we analyzed miR-99a-5p expression levels in primary tumours and plasma of breast cancer patients to evaluate its usefulness as a minimally invasive diagnostic biomarker. METHODS: MiR-99a-5p expression levels were determined by quantitative real-time PCR in three indepent cohorts of patients: I) Discovery cohort: breast cancer tissues (n=103) and healthy breast tissues (n=26). II) Testing cohort: plasma samples from 105 patients and 98 healthy donors. III) Validation cohort: plasma samples from 89 patients and 85 healthy donors. Receiver operating characteristic (ROC) curve analyses were applied to evaluate the diagnostic potential of miR-99a-5p expression levels in tissue and plasma samples. RESULTS: MiR-99a-5p was significantly downregulated in breast cancer tissues compared to healthy breast tissues (p < 0.0001), being able to discriminate BC from healthy breast tissues with an AUC of 0.85, 87.38% sensitivity, 76.92% specificity, and 85.27% accuracy. Conversely, miR-99a-5p levels were significantly higher in breast cancer patients than in healthy controls in plasma samples from both testing and validation cohorts (p < 0.0001). ROC curve analysis revealed that miR-99a-5p has good diagnostic potential, with an AUC of 0.76, 63.81% sensitivity, 79.59% specificity, and 71.43% accuracy. The value of circulating miR-99a-5p levels as a breast cancer biomarker was further validated in an independent cohort, where it was able to identify breast cancer with 57.30% sensitivity, 67.06% specificity, and 62.07% accuracy. Besides, we also confirmed that circulating miR-99a-5p levels were able to discriminate early breast cancer from healthy controls with a 66.67% accuracy, 68.80% sensitivity, 65.28% specificity, and an AUC of 0.69 (p < 0.0001). CONCLUSION: MiR-99a-5p’s deregulated expression distinguished healthy individuals from breast cancer patients in two different types of samples (tissues and plasma). Interestingly, plasma expression levels were significantly lower in healthy controls than in early-stage breast cancer patients. Our findings suggest that circulating miR-99a-5p as a novel promising non-invasive biomarker for breast cancer detection. Citation Format: Iris Garrido-Cano, Vera Constâncio, Anna Adam-Artigues, Ana Lameirinhas, Soraya Simón, Belen Ortega, María Teresa Martínez, Cristina Hernando, Begoña Bermejo, Ana Lluch, Paula Lopes, Rui Henrique, Carmen Jerónimo, Juan Miguel Cejalvo, Pilar Eroles. Circulating miR-99a-5p expression in plasma: A potential biomarker for early diagnosis of breast cancer [abstract]. In: Proceedings of the 2020 San Antonio Breast Cancer Virtual Symposium; 2020 Dec 8-11; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2021;81(4 Suppl):Abstract nr PS2-31.

Targeting Androgen Receptor in Estrogen Receptor-Negative Breast Cancer

Purpose: Currently, there are no molecular biomarkers for the early detection of breast cancer (BC). This study focused on identifying surface proteins found on circulating extracellular vesicles (EVs) for detecting early-stage BC. Experimental Design: Circulating EVs, isolated from the plasma of 10 patients with BC (stages I and II), and 5 healthy controls, were analyzed using LC-MS/MS. Developmental endothelial locus-1 protein (Del-1) was selected as a candidate biomarker. Two different enzyme-linked immunosorbent assays (ELISAs) were used to measure Del-1 in plasma samples from healthy controls (n = 81), patients with BC (n = 269), BC patients after surgical resection (n = 50), patients with benign breast tumors (n = 64), and patients with non-cancerous diseases (n = 98), in two cohorts. Results: Plasma Del-1 levels were significantly higher (P < 0.0001) in patients with BC than in all controls and returned to almost normal after tumor removal. The diagnostic accuracy of Del-1 was area under the curve (AUC), 0.961 [95% CI, 0.924-0.983], sensitivity of 94.70% and specificity of 86.36% in test cohort, and 0.968 [0.933-0.988], 92.31% and 86.62% in validation cohort for early-stage BC by one type of ELISA. Furthermore, Del-1 maintained diagnostic accuracy for patients with early-stage BC using the other type of ELISA (0.946 [0.905-0.972], 90.90%, and 77.14% in the test cohort; 0.943 [0.900-0.971], 89.23%, and 80.99% in the validation cohort). Conclusions: Del-1 on circulating EVs is a promising marker to improve identification of patients with early-stage BC and distinguish BC from benign breast tumors and non-cancerous diseases. Citation Format: Pyong-Gon P. Moon, Jeong-Eun Lee, Young-Eun Cho, Soo Jung Lee, Jin Hyang Jung, Yee Soo Chae, Han-Ik Bae, Young-Bum Kim, In-San Kim, Hoyong Park, Moon-Chang Baek. Identification of developmental endothelial locus-1 on circulating extracellular vesicles as a novel biomarker for early breast cancer detection. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3147.

Background: Breast cancer is the most common type of cancer in Iraq, and the incidence of it has increased among Iraqi women in the past two decades, as it represents one of the most important threats to women's health. Aim: this study aimed to investigate the miR-148a gene expression as a biomarker in different age groups of Iraqi women with breast cancer, as well as its association with patient characteristics such as body mass index, disease stage, and tumor location. Method: Fifty women diagnosed with breast cancer were enrolled in this study in addition to 25 healthy individuals as a control group. The fold expression (FE) of miRNA-148a was detected by quantitative polymerase chain reaction (qPCR) techniques. Results: The results showed the highest significant difference in the gene expression of miR-148a in breast cancer patients in the age group (50) compared to the control group, and the difference was significant in the age group (<40) compared to the control group, highly significant differences was found in the age group (40-49) compared with the control group. The results also showed no significant differences in the fold level of gene expression for miR-148a in the grades (I, II, and III) of breast cancer patients, and that the highest level was recorded in the second grade, and the highest level was recorded in the grade. Keywords: Breast Cancer, miR-148a, BMI

Prognostic Gene Expression Signatures Can Be Measured in Tissues Collected in RNAlater Preservative

BMI is an independent prognostic factor for late outcome in patients diagnosed with early breast cancer: A landmark survival analysis

Content: Identification of panel of SEREX-defined antigens for breast cancer autoantibodies profile detection.Objective: To create panel of antigens that can differentiate breast cancer patients and healthy individuals.Methods: SEREX (serological analysis of cDNA expression libraries) method, ELISA (enzyme-linked immunosorbent assay), qPCR (quantitative polymerase chain reaction).Results: In large-scale screening of 16 SEREX-antigens by sera of breast cancer patients and healthy donors, a combination of six antigens (RAD50, PARD3, SPP1, SAP30BP, NY-BR-62 and NY-CO-58) was identified, which can differentiate breast cancer patients and healthy donors with 70% sensitivity and 91% specificity. Elevated mRNA expression of SPP1 gene was revealed in breast tumors (2–7-fold) that correlated with SPP1 antigen immunoreactivity in autologous patients’ sera.Conclusions: The new panel of six SEREX-antigens was proposed, which enables creation of serological assay for breast cancer diagnostics and/or prognosis.

Gene Expression Analysis of Immune-Mediated Arrest of Tumorigenesis in a Transgenic Mouse Model of HER-2/neu-Positive Basal-Like Mammary Carcinoma

The aim of this study is to investigate clinical implications of human leukocyte antigen-G (HLA-G) expression in breast cancer. HLA-G expression in 235 primary breast cancer tissues was investigated using immunohistochemistry, and plasma soluble HLA-G (sHLA-G) was measured in 44 breast cancer patients using a specific HLA-G enzyme-linked immunosorbent assay (ELISA). Effects of estradiol/progesterone and their antagonists tamoxifen/RU486 on HLA-G expression in cultured breast cancer MCF-7 cells were determined by real-time polymerase chain reaction (PCR) and the ELISA. Alterations of HLA-G expression by the hormone treatments on subsequent allocytotoxic lymphocyte (allo-CTL) response were also examined. In the study, approximately 66% of neoplasm lesions were identified to have positive HLA-G expression. This expression was significantly correlated with tumor size, nodal status, and clinical disease stage (P = 0.0001, 0.012, and 0.0001, respectively). Patients with positive HLA-G expression had a lower survival rate than those with negative expression (P < 0.028). Plasma sHLA-G levels were significantly higher in breast cancer patients than in healthy controls (P < 0.001), with the area under the receiver-operating characteristic (ROC) curve being 0.95. HLA-G expression in breast cancer MCF-7 cells was enhanced by estradiol/progesterone but reduced by their antagonists. Cytotoxicity studies showed that allo-CTL response in MCF-7 cells was inhibited by prior treatment with estradiol/progesterone, but was amplified by their antagonists. The effects could be restored or further strengthened by the addition of anti-HLA-G antibodies. Our findings suggest that HLA-G may have potential clinical implications in diagnosis, prognosis, and immunotherapy of patients with breast cancer.

A cross-sectional study was carried out in Kirkuk city from 15th of June 2018 to 15th of December 2018 to detect of Toxoplasma gondii DNA in sera of women with breast cancer by real-time PCR and comparing the results with IgG and IgM toward T. gondii. The number of breast cancer women understudy were 35. The ages of the patients were between 20-75 years old. These patients admitted to Kirkuk oncology center. The control group were included 50 healthy individuals. blood was collected from each patients and control for molecular tests of T. gondii using real-time PCR and serological testing for detection specific Toxoplasma gondii IgM and IgG by using ELISA technique. The study showed that the highest rate of anti T. gondii IgM+ IgG- antibodies 8.57% was recorded among women with breast cancer comparing with 8% in healthy control while 14.29% of women with breast cancer were IgM+IgG+ comparing 6% of the healthy control group. The highest rate of total T. gondii antibodies by ELISA 68.57% was noted among women with breast cancer and the lowest rate was among healthy control 20%. The study revealed that 54.17% of women with breast cancer with positive ELISA was positive by PCR comparing with 9.09% of patients with negative ELISA results. The result was highly significant with sensitivity and specificity of 54.17% and 90.09% respectively. The highest rate of anti T. gondii IgM+ IgG- antibodies 66.67% women with breast cancer was positive by PCR followed by 40% of patients with IgM+ IgG+ antibodies. It was concluded that Toxoplasma gondii more frequently associated with breast cancer and PCR was more accurate in detection of the parasite.

Background: Breast cancer (BC) is a heterogeneous disease that can be classified into many subtypes according to histopathological and molecular characteristics. Forkhead box protein A1 (FOXA1) is a transcriptional pioneer factor that opens chromatin allowing estrogen receptor (α-ER) access to its genomic targets. FOXA1 expression is related to luminal BC with a good prognosis. Objectives: The present study is sought to determine the FOXA1 expression in Iraqi women with ER+ BC. Methods: Forty-eight fresh malignant breast tissues were analyzed by immunohistochemistry assay to choose ER+ samples, and then by RT-qPCR to evaluate FOXA1 gene expression. Results: The ER-positive samples were (72.91%) of the total samples, and the molecular subtype of luminal A was the most common with a percentage of 56.25%. It was also noted that the high expression of the FOXA1 gene is highly significant (p&lt;0.05) in Iraqi women with BC when compared with healthy controls. Conclusions: Highly significant FOXA1 expression was found in Iraqi women with BC makes it eligible to be a good predictor or a biomarker for BC.

Background: Breast cancer risk has been linked to obesity, especially in postmenopausal women. Adipocyte fatty acid binding protein (A-FABP) is found in adipose tissue, and preliminary evidence suggests that its expression in serum is associated with obesity and breast cancer risk. Surgery for morbid obesity appears effective in decreasing morbidity and/or mortality from a variety diseases of adulthood, including cancer. We evaluated the association of A-FABP expression with 1) body mass index (BMI), 2) breast cancer and 3) change in BMI after obesity surgery. Hypothesis: Increased A-FABP expression is associated with 1) BMI level, 2) breast cancer, and 3) change in BMI after obesity surgery. Methods: Serum was collected under an institutional review board approved protocol from two cohorts of obese women: 1) those with or without breast cancer, and 2) those undergoing surgery for morbid obesity. In the first cohort, samples were collected from 101 women prior to surgery to diagnose and treat a concerning breast lesion. In the second, samples were collected from 82 healthy obese women at baseline, and whenever possible, at 3, 6, and 12 months after obesity surgery. A-FABP levels in serum were measured using human A-FABP4 ELISA kits. The student's t-test was used to compare A-FABP levels in the two groups. Linear mixed effect models were used to examine the relationship between A-FABP and BMI or change of BMI over time, controlling for age, menopause status and history of breast cancer. Results: A-FABP levels were significantly higher in women with cancer than healthy controls among obese women (p=0.039), but not among non-obese women (p &amp;gt; 0.05). A-FABP levels were higher in obese than non-obese women without (p=0.038) and with breast cancer (p&amp;lt;0.001). Among healthy obese controls, A-FABP was associated with BMI at baseline (p=0.016) and its change over time after surgery (p=0.005). The association of A-FABP with BMI was significant for women of normal breast cancer risk regardless of their menopause status (p=0.012 for pre-menopause and p&amp;lt;0.001 for post-menopause), but not for those with a family history of breast cancer (p&amp;gt;0.05). A-FABP was also more predictive of change in BMI in postmenopausal normal risk women (p&amp;lt;0.001) than in other groups (p&amp;gt;0.05). Impact: Circulating levels of A-FABP are associated with cancer in obese women, with baseline BMI in healthy women and with change in BMI in normal risk women undergoing surgery for morbid obesity. These findings all point to a clinically important mechanistic role for A-FABP in obesity influenced breast cancer. They also suggest that downregulation of A-FABP may be involved in the decreased risk of cancer, including breast cancer, that has been observed in follow-up after obesity surgery. Citation Format: Sauter E, Hao J, Yan X, Kong M, Li B. Adipocyte fatty acid binding protein levels predict risk of obesity associated breast cancer [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P1-02-04.

Background: Breast cancer is the most common malignant tumor and the second most significant cause of death for women in Iraq, behind cardiovascular diseases. Obesity has been linked to a substantial increase in the risk of breast cancer. Adipose tissue functions as an endocrine gland, controlling the body's metabolism by secreting adipokines, which play a significant role in metabolism and inflammatory reactions. Methods: Overall, 90 postmenopausal women participated in this research. Of these, 60 patients with breast cancer were recruited at Baghdad's Oncology Teaching Hospital between October 2021 and February 2022: 30 were obese with a body mass index (BMI) of &gt; 30 kg/m2 (group 1), and 30 were not obese (group 2). The third group consisted of 30 participants without breast cancer or obesity (group 3). Each person donated five milliliters of venous blood. The blood levels of adiponectin and leptin are determined using enzyme-linked immunosorbent assay (ELISA) kits. Results: Control individuals who were not obese (group 3) had greater blood adiponectin levels than patients with cancer who were both obese and non-obese (groups 1 and 2), with no significant difference in serum adiponectin levels seen between groups 1 and 2. The findings also showed that group 1 (patients with breast cancer and obesity) had greater serum leptin levels than both group 2 (patients with breast cancer without obesity) and the control group (group 3), with no significant difference in serum leptin levels between groups 2 and 3. Conclusions: Adiponectin levels in the blood of women with breast cancer and obesity were low which may be due to high BMI, which reduces adiponectin's protective effects. Conversely, Leptin levels were more significant in the blood of women with breast cancer and obesity than in the control group, which may be due to its pro-inflammatory effects in obesity, among other variables.

The state of HER-2 status