Plasma miRNA Expression Levels Research Articles

Preliminary Evidence Supports that Long-Term Consumption of Higher-Protein Breakfast Promotes Higher Expression of Select miRNA Associated with Cardiometabolic Health in Adolescents

BackgroundIncreased dietary protein at breakfast promotes cardiometabolic health; however, whether these improvements occur at the molecular level is unknown. ObjectivesThe objective was to examine whether long-term consumption of breakfast, varying in protein quantity, alters the expression of circulating microRNAs (miRNAs) associated with cardiometabolic health in “breakfast-skipping” adolescents. MethodsThirty adolescents (age: 19 ± 1 y; body mass index: 25.4 ± 3 kg/m2) completed a 6-mo tightly controlled breakfast trial in which participants consumed 350 kcal normal-protein (NP, 10 g protein) or higher-protein (HP, 30 g protein) breakfasts or continued to BS for 6 mo. Fasting blood samples were collected at baseline (PRE) and 6 mo (POST) for assessment of 12 a priori circulating plasma miRNA expression levels (real-time quantitative polymerase chain reaction), glucose, insulin, IL-6, and C-reactive protein. ResultsNo main effects of group were observed for any miRNAs; however, a time-by-group interaction was detected for the expression of miR-126-3p (P = 0.05). HP breakfast tended to increase miR-126-3p expression throughout the study (POST-PRE, P = 0.09) leading to greater expression at POST compared with BS (P = 0.03), whereas NP breakfast did not. Additionally, several miRNAs predicted fasting concentrations of IL-6: miR-320a-3p, -146a-5p, -150-5p, -423-5p, -122-5p, glucose: miR-24-3p, -126-3p; insulin: miR-24-3p, -126-3p, -15b-5p; insulin sensitivity: miR-24-3p, -126-3p, -199a-5p, -15b-5p; and β-cell function: miR-15b-5p (R2 between 0.2 and 0.39; P < 0.05) from PRE and POST samples across groups. ConclusionsThese data support the daily consumption of a HP breakfast to promote cardiometabolic health, potentially through changes in miRNA expression, in a sensitive life-stage where early intervention strategies are critical to reduce the risk of adult-onset chronic disease. Trial registration numberNCT03146442.

Expression Analysis of Circulating microRNAs in Saliva and Plasma for the Identification of Clinically Relevant Biomarkers for Oral Squamous Cell Carcinoma and Oral Potentially Malignant Disorders.

This study aims to evaluate the expression of salivary and plasmatic miRNAs as diagnostic biomarkers in patients with oral squamous cell carcinoma (OSCC) and oral potentially malignant disorders (OPMDs). A total of 25 patients were divided into three groups, according to their diagnosis: OSCC patients (n = 14); OPMDs patients (n = 6); and healthy controls (n = 5). At the time at diagnosis/enrolment, patients underwent salivary and plasmatic collection. The expression of miRNA -21, -31, -138, -145, -184, and -424 were evaluated by real-time PCR. An F-test and ANOVA test were performed to evaluate the miRNA levels (significance at p < 0.05). By comparing miRNA expression levels from saliva, a statistically significant difference emerged in the expression of miR-138 and miR-424 between the three groups (p < 0.05). In particular, these two miRNAs showed decreased expression levels in saliva samples from OSCC and OPMD patients compared to those from healthy controls. On the other hand, miRNA expression levels in plasma were low in all the groups, and no statistically significant differences were found. Overall, our results showed that liquid biopsy from saliva may be a useful tool for the identification of diagnostic molecular biomarkers in OSCC and OPMDs.

MicroRNA in refined diagnosis of choroidal melanoma

Epigenetic studies of the level of microRNAs in human oncogenesis indicate their signifi cant role in the development and growth of malignant tumors of various origins. The fi rst works on the role of microRNAs in patients with uveal melanoma appeared in 2008.The aim: to analyze the expression level of miRNA-126 and miRNA-223 in the plasma blood of patients and to determine their signifi cance in the refi ned diagnosis of choroidal melanoma. Materials and methods. We examined 84 patients with choroidal melanoma (CM), mean age – 63.4 ± 1.2 (35–86 y.o.). Localization – a single CM node with a thickness of 0.77–17.19 mm. The control group consisted of 28 volunteers, age – 62.9 ± 1.42 (45–78 y.o.). Plasma miRNA expression levels were determined by real-time PCR.Results. An increase in the level of expression of miRNA-223 and miRNA-126 in blood plasma was confi rmed in all 84 patients with choroidal melanoma N0M0 compared with the control group. An increase in the expression of miRNA-223 and miRNA-126 was proved with an increase in tumor prominence.Conclusion. The obtained results of an increase in the expression of miRNA-223 indicate an increase in cell proliferation, and an increase in the expression of miRNA-126 on the activation of angiogenesis in a growing tumor, which makes it possible to recommend a study of the level of miRNA-223 and miRNA-126 for a more accurate diagnosis of small CM in cases of difficulty of differential diagnosis with other tumor-like diseases of the choroid.

MicroRNAs Regulating Tumor Immune Response in the Prediction of the Outcome in Patients With Breast Cancer.

MicroRNAs (miRNAs) are key regulators in immune surveillance and immune escape as well as modulators in the metastatic process of breast cancer cells. We evaluated the differential expression of plasma miR-10b, miR-19a, miR-20a, miR-126 and miR-155, which regulate immune response in breast cancer progression and we investigated their clinical relevance in the outcomes of breast cancer patients. Plasma samples were obtained from early (eBC; n = 140) and metastatic (mBC; n = 64) breast cancer patients before adjuvant or first-line chemotherapy, respectively. Plasma miRNA expression levels were assessed by qRT-PCR. We revealed a 4-miRNA panel consisted of miR-19a, miR-20a, miR-126, and miR-155 able to discriminate eBC from mBC patients with an AUC of 0.802 (p < 0.001). Survival analysis in eBC patients revealed that low miR-10b and miR-155 expression was associated with shorter disease free survival (disease free survival; p = 0.012 and p = 0.04, respectively) compared to high expression. Furthermore, miR-126 expression was associated with shorter overall survival (overall survival; p = 0.045). In multivariate analysis the number of infiltrated axillary lymph nodes and low miR-10b expression independently predicted for shorter DFS (HR: 2.538; p = 0.002 and HR: 1.943; p = 0.033, respectively) and axillary lymph nodes and low miR-126 for shorter OS (HR: 3.537; p = 0.001 and HR: 2.558; p = 0.018). In the subgroup of triple negative breast cancer (TNBC) patients, low miR-155 expression independently predicted for shorter DFS (HR: 5.056; p = 0.037). Accordingly in mBC, patients with low miR-10b expression had shorter progression free survival and OS compared to patients with high expression (p = 0.0017 and p = 0.042, respectively). In multivariate analysis, recurrent disease and low miR-10b expression independently predicted for shorter PFS (HR: 2.657; p = 0.001 and HR: 1.920; p = 0.017, respectively), whereas performance status two independently predicted for shorter OS (HR: 2.031; p = 0.03). In summary, deregulated expression of circulating miRNAs involved in tumor and immune cell interactions evaluated before adjuvant and 1st-line chemotherapy can distinguish disease status and emerge as independent predictors for outcomes of breast cancer patients.

Relationship Between Metabolic Syndrome and Circulating microRNA Expression in Adults

Abstract Objectives To verify the relationship between circulating miRNA (16, 363, 375, 486, and let7c) expression and risk factors for metabolic syndrome (MetS) in adults. Methods A cross-sectional population-based study including 192 adults without acute inflammatory diseases, cancer, or using medication that would affect the main outcomes. The BioMarkTM Fluidigm Dynamic Array for real-time qPCR was used to quantify miRNA expression levels in plasma. The miRNA expression levels categorized by the MetS status were analyzed by the Mann-Whitney test. The Kruskal-Wallis with post hoc Dunn's tests were used to compare miRNA expression levels categorized by the number of MetS risk factors (0, 1–2, and ≥3). Spearman's correlation between miRNA expression and MetS risk factors was performed. Results The sample included individuals with median age of 41 (20–59) years, 54.7% were women, and 58.3% overweight/obesity. MetS was observed in 33.9% of individuals (15.6% men, and 18.2% women), being increased waist circumference (WC) (67.7%), and reduced plasma HDL-c levels (52.1%) the most prevalent risk factors. Individuals with MetS showed reduced plasma expression of miR-16, −363, −375, and −486, but increased miR-let7c expression compared to the group without MetS (P &amp;lt; 0.05). Analysis of miRNA expression levels categorized according to the number of MetS risk factors demonstrated that individuals without risk factors did not differ from those with 1–2 risk factors. Individuals with 3 or more risk factors differed from all groups, except for the miR-16, which presented difference only in comparison to the group without risk factors. The miR-16 expression correlated negatively with body mass index (BMI) (r = −0.169; P = 0.019), WC (r = −0.153; P = 0.034), diastolic blood pressure (r = −0.144; P = 0.046), and glucose levels (r = −0.171; P = 0.017). The miR-let7c correlated positively with triacylglycerol (r = 0.306; P &amp;lt; 0.001), and negatively with HDL-c (r = −0.173; P = 0.016) levels. The miR-375 correlated negatively with BMI (r = −0.203; P &amp;lt; 0.001), WC (r = −0.187; P &amp;lt; 0.001), and positively with HDL-c levels (r = 0.261; P &amp;lt; 0.001). The miR-486 and −363 correlated negatively with BMI (r = −0.155; P = 0.031 | r = −0.142; P = 0.049, respectively). Conclusions The circulating miRNA evaluated were associated with the risk factors for MetS in adults. Funding Sources São Paulo Research Foundation - FAPESP

MicroRNAs Regulating Tumor and Immune Cell Interactions in the Prediction of Relapse in Early Stage Breast Cancer.

MicroRNAs (miRNAs) are involved in the regulation of immune response and hold an important role in tumor immune escape. We investigated the differential expression of the immunomodulatory miR-10b, miR-19a, miR-20a, miR-126, and miR-155 in the plasma of healthy women and patients with early stage breast cancer and interrogated their role in the prediction of patients’ relapse. Blood samples were obtained from healthy women (n = 20) and patients with early stage breast cancer (n = 140) before adjuvant chemotherapy. Plasma miRNA expression levels were assessed by RT-qPCR. Relapse predicting models were developed using binary logistic regression and receiver operating curves (ROC) were constructed to determine miRNA sensitivity and specificity. Only miR-155 expression was lower in patients compared with healthy women (p = 0.023), whereas miR-155 and miR-10b were lower in patients who relapsed compared with healthy women (p = 0.039 and p = 0.002, respectively). MiR-155 expression combined with axillary lymph node infiltration and tumor grade demonstrated increased capability in distinguishing relapsed from non-relapsed patients [(area under the curve, (AUC = 0.861; p < 0.001)]. Combined miR-19a and miR-20a expression had the highest performance in discriminating patients with early relapse (AUC = 0.816; p < 0.001). Finally, miR-10b in combination with lymph node status and grade had the highest accuracy to discriminate patients with late relapse (AUC = 0.971; p < 0.001). The robustness of the relapse predicting models was further confirmed in a 10-fold cross validation. Deregulation of circulating miRNAs involved in tumor-immune interactions may predict relapse in early stage breast cancer. Their successful clinical integration could potentially address the significance challenge of treatment escalation or de-escalation according to the risk of recurrence.

MicroRNA-125b-5p is a promising novel plasma biomarker for alveolar echinococcosis in patients from the southern province of Qinghai

BackgroundAlveolar echinococcosis (AE) is caused by parasitic infection by Echinococcus multilocularis. Its diagnosis is usually based on clinical symptoms, ultrasound, and other imaging methods. MicroRNAs (miRNAs) play important roles in disease processes and can exist in a highly stable cell-free form in body fluids. It is important to identify specific, sensitive diagnostic markers for early diagnosis and evaluation of AE. In this study, we examined hsa-miR-125b-5p as a potential plasma biomarker of E. multilocularis infection.MethodsPlasma samples from patients with AE and healthy individuals were screened for the presence of five miRNAs using miRNA chips. We used quantitative polymerase chain reaction to measure miRNA expression levels in plasma and liver tissue samples from patients with AE.Resultshsa-miR-125b-5p was stably upregulated in the plasma and liver tissue samples from patients with AE.ConclusionsThe results suggest that hsa-miR-125b-5p may be a promising biomarker for early, non-invasive diagnosis of AE.

MicroRNAs involved in immune response as prognostic markers in early and metastatic breast cancer.

e15528 Background: MicroRNAs (miRNAs) are involved in the regulation of immune response and have an important role in immune escape. We analyzed the expression levels of plasma miR-10b, miR-126, miR-19α, miR-20α and miR-155, which regulate tumor-immune interactions and investigated their prognostic implications in patients with early (eBC) and metastatic (mBC) breast cancer. Methods: Blood samples were obtained before treatment from 140 and 64 patients with eBC and mBC, respectively. Plasma miRNA expression levels were assessed by qRT-PCR and expression was classified as high or low according to the median values. Results: A panel of four miRNAs (miR-19a, miR-20a, miR-126 and miR-155) could discriminate eBC from mBC (AUC 0.802, p &lt; 0.001). In early disease, miR-10b (p = 0.022) and miR-155 (p = 0.005) expression was lower in relapsed (n = 46) compared to non-relapsed (n = 94) patients; miR-155 expression along with lymph node infiltration and tumor grade had increased ability to predict relapse (AUC = 0.775; p = 0.003). In addition, miR-10b (p = 0.015), miR-19a (p = 0.003), miR-20a (p = 0.012), miR-126 (p = 0.001) and miR-155 (p &lt; 0.001) expression levels were lower in patients with early relapse (relapse at ≤2 years). MiR-155 and miR-19a had the highest performance in discriminating early relapse (AUC 0.855; p &lt; 0.001 and AUC = 0.729; p = 0.003, respectively), whereas, combined mir-155 and miR-19a expression further increased the accuracy of prediction (AUC = 0.867; p &lt; 0.001). In eBC, the number of infiltrated lymph nodes and low miR-10b independently predicted for shorter DFS (p = 0.001 and p = 0.03, respectively) and axillary lymph nodes for shorter OS (p = 0.003). In the triple negative subgroup, low miR-155 strongly predicted for shorter DFS (p = 0.037). In mBC, recurrent disease and low miR-10b expression independently predicted for shorter PFS (p = 0.001 and p = 0.017, respectively), whereas performance status of 2 independently predicted for shorter OS (p = 0.03). Conclusions: Deregulated expression of circulating miRNAs involved in tumor-immune interactions can discriminate disease status in BC and independently predicts for patients’ outcome in early and mBC. Our results further support the notion that circulating miRNAs represent a useful prognostic tool in patients with BC.

MON-LB82 Plasma Expression Level of MiRNA -21 Is Related With the Presence of Papillary Thyroid Cancer

Introduction.: There is no effective and reliable biomarker to distinguish benign thyroid nodules (BTN) from papillary thyroid carcinomas (PTC). In this study we analyzed a set of four miRNA molecules in plasma of patients with papillary thyroid cancer, benign nodules and healthy controls to identify miRNA molecules that may be markers of PTC.Aim.: We aimed to investigate the dysregulation of plasma miRNAs in PTC and evaluate the diagnostic value for differentiation of PTC from BTN. Methods.: The expression levels of 4 miRNAs (miR-221, miR-222, miR-146b, miR-21) were measured in 48 PTC patients before thyroidectomy and again after thyroidectomy in a subgroup of 36 patients. Preoperative and postoperative plasma miRNA expression levels were compared with baseline levels established in plasma from the heathy controls group (N=57) and patients with BTN (N=22). MicroRNA-222 and miR-146b, miR-221, miR-21 were included in a panel because they all reportedly were overexpressed in PTC compared to benign nodules or normal thyroid tissue.Results.: Compared with baseline levels in the healthy controls group, miR-221, miR-222, miR-146b, miR-21 levels were significantly higher in the preoperative PTC group (P <0.0001, P=0.002, P=0.028, P =0.021, respectively). A significant reduction in miR-21 expression was observed in postoperative PTC patients. MiR-21 decreased by 5.98-fold (P=0.046) in post- operative samples compared with preoperative samples in the PTC patients.In comparison MiRNRs expression levels in BTN group with healthy controls, miR-221, miR-21 expression levels were significantly higher in the BTN group (P=0.003, P=0.048, respectively). No significant difference was observed between the preoperative PTC group and the preoperative BTN group with regard to the expression of these four miRNA’s. Conclusions: The expression levels of miR-222, miR-146b in plasma were significantly higher in patients who had PTC than in healthy volunteers, whereas levels of miR-221, miR-21 in plasma were significantly higher in patients who had either PTC or BTN before thyroidectomy than in healthy volunteers. Furthermore, miR-21 showed a significant reduction of expression levels after thyroidectomy in PTC patients. However, value of these four miRNAs is still limited in differential diagnosis of PTC and benign nodules.

Plasma and Tissue Specific miRNA Expression Pattern and Functional Analysis Associated to Colorectal Cancer Patients.

An increasing number of studies suggest the implication of microRNAs (miRNAs) in colorectal (CRC) carcinogenesis and disease progression. Nevertheless, the basic mechanism is not yet clear. We determined plasma miRNA expression levels using Agilent microarray technology followed by overlapping with The Cancer Genome Atlas (TCGA) tissue data and a qRT-PCR validation step and analysis of the altered miRNA signatures to emphasize new mechanistic insights. For TGCA dataset, we identified 156 altered miRNAs (79 downregulated and 77 upregulated) in colorectal tissue samples versus normal tissue. The microarray experiment is based on 16 control samples, 38 CRC plasma samples from colorectal cancer patients who have not undergone chemotherapy, and 17 chemo-treated samples. In the case of the analysis of CRC cancer versus healthy control we identified 359 altered miRNAs (214 downregulated and 60 upregulated), considering as the cutoff value a fold-change of ±1.5 and p < 0.01. An additional microarray analysis was performed on plasma from untreated colorectal cancer (n = 38) and chemotherapy-treated colorectal cancer patients (n = 17), which revealed 15 downregulated miRNAs and 53 upregulated miRNAs, demonstrating that the plasma miRNA pattern is affected by chemotherapy and emphasizing important regulators of drug resistance mechanisms. For the validation of the microarray data, we selected a panel of 4 miRNAs from the common miRNA signatures for colon and rectal cancer (miR-642b-3p, miR-195-5p and miR-4741). At the tissue level, the expression levels were in agreement with those observed in colorectal plasma. miR-1228-3p, the top upregulated miRNA in CRC, was chosen to be validated on tissue and plasma samples, as it was demonstrated to be downregulated at tissue level in our patient cohort. This was confirmed by TCGA data and was one example of ta ranscript that has a different expression level between tumor tissue and plasma. Developing more efficient investigation methods will help explain the mechanisms responsible for miRNAs released in biofluids, which is the most upregulated transcript in colorectal plasma samples and which can function as a prediction tool within the oncological field.

Abstract P1-05-07: microRNAs regulating tumor and immune cell interactions in the prediction of early relapse in breast cancer

Abstract Background: Metastasis remains the major lethal consequence in the progression of early breast cancer (BC). Effective immunosurveillance can inhibit metastasis, while a dysfunctional immune system can create a favorable environment for tumor spread. MicroRNAs (miRNAs) are involved in the regulation of immune response and there is evidence that they play an important role in immune escape. We investigated the effect of plasma miR-10b, miR-126, miR-19α, miR-20α and miR-155, which regulate the interaction between cancer and immune cells during immunosurveillance and immune evasion in patients with early BC. Methods: Blood samples were obtained from relapsed (n=52) and non-relapsed (n=100) patients with early BC before adjuvant chemotherapy. Plasma miRNA expression levels were assessed by qRT-PCR and expression was classified as high or low according to the median values. Receiver operating curves (ROC) were constructed to determine miRNA sensitivity and specificity. Results: miRNA expression was not correlated with patients’ characteristics or outcome. No differences in miRNA expression were observed between relapsed and non-relapsed patients. In multivariate analysis the number of infiltrated axillary lymph nodes and stage III disease independently predicted for shorter DFS (HR: 2.591; p=0.002) and OS (HR: 2.344; p=0.035), respectively. miR-126, miR-20a, miR-19a and miR-155 expression levels were lower in patients with early relapse (defined as relapse at ≤ 2 yrs; n=19) compared to those who either relapse later than 2 years (n=33) or those who remained disease-free during follow up (n=100). ROC curve analysis showed that miR-155 had the highest performance to discriminate patients with early relapse [AUC 0.825; sensitivity 90%, specificity 67% (p&amp;lt;0.001; 95% CI: 0.742-0.909)]. Multivariate analysis revealed that ER negative status and miR-155 low expression independently predicted for shorter DFS (HR: 2.616; p=0.039 and HR: 9.104; p=0.003, respectively). For the triple negative subgroup (n=31), low miR-155 expression was associated with both shorter DFS and OS (p=0.02 and p=0.032, respectively) and low miR-155 expression was an independent predictor of shorter DFS (HR: 5.056; p=0.037). Conclusions: Deregulated expression of circulating miRNAs involved in tumor and immune cell interactions evaluated before adjuvant chemotherapy is associated with early relapse in patients with primary BC. Low miR-155 expression has high accuracy in the prediction of early relapse in the whole group of patients and independently predicts for shorter DFS in the triple negative subgroup. The role of miR-155 in the pathogenesis and management of early relapse in BC merits further investigation. Citation Format: Konstantina Thomopoulou, Chara Papadaki, Alexia Monastirioti, George Koronakis, Lambros Vamvakas, Maria A Papadaki, Sofia Agelaki, Dimitrios Mavroudis. microRNAs regulating tumor and immune cell interactions in the prediction of early relapse in breast cancer [abstract]. In: Proceedings of the 2019 San Antonio Breast Cancer Symposium; 2019 Dec 10-14; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2020;80(4 Suppl):Abstract nr P1-05-07.

Plasma expression of miRNA-21, \u2212\u2009214, \u221234a, and -200a in patients with persistent HPV infection and cervical lesions

BackgroundTo examine differences in the plasma levels of miRNA-21, − 214, −34a, and -200a in patients with persistent high-risk human papillomavirus (hr-HPV) infection or with cervical lesions of different grades.MethodsVenous blood was collected from 232 individuals to measure the plasma expression levels of miRNA-21, − 214, −34a, and -200a. The subjects included normal controls and patients with persistent hr-HPV infection, CIN1, CIN2, CIN3, or cervical cancer (n = 42, 31, 19, 54, 71, and 15 patients, respectively). Cervical conization specimens were collected from all the women. To ensure the accuracy of histopathology, three consecutive tissue sections with an identical diagnosis were selected, and dissection samples were taken from them for miRNA detection. Eligible cases met the inclusion criteria based on sample observation using the middle slice of sandwich tissue sections from the pathological tissue in accordance with the diagnosis of CIN1, CIN2 and CIN3 in 8, 29, and 26 cases, respectively. The miRNA-21, − 214, −34a, and -200a expression levels in the paraffin-embedded tissue samples were determined. The percentage of patients with a CIN2+ diagnosis at 30–49 years old was significantly different from that of those diagnosed with CIN1. The incidence of CIN2+ patients exposed to passive smoking was significantly different from that of CIN1- patients. The percentage of CIN2+ patients with three pregnancies was significantly different from that of those with CIN1, and the percentage of CIN2+ subjects with ≥4 pregnancies was significantly different from that of CIN1- patients. The number of CIN2+ patients with two or more induced abortions was significantly different from that of patients with CIN1. The percentage of CIN2+ patients who underwent a caesarean section was significantly different from that of patients with CIN. The percentage of CIN2+ patients with first-degree relatives with cancer was significantly different from that of those with CIN1. Among CIN2+ patients, the percentage with a first sexual encounter at ≤20 years old was significantly different from that of those with CIN1. The percentage of CIN2+ patients with ≥2 sexual partners was significantly different from that of CIN1- patients.ResultsThe plasma miRNA-214, −34a, and -200a expression levels were decreased in patients with more severe cervical lesions. Plasma miRNA levels in CIN1- patients were significantly different from those in CIN2+ patients. The kappa values for miRNA-21, − 214, −34a and -200a in tissue versus plasma were 0.7122, 0.9998, 0.8986 and 0.7458, respectively. The sensitivity of each biomarker for detecting CIN2 was calculated, and ROC curves of the four miRNA biomarkers were drawn. The AUC of the four plasma miRNAs was greater than 0.5, with the AUC of miRNA-21 being the largest at 0.703. The plasma miRNA expression levels exhibited at least one tie between CIN1 and CIN2. The AUCs for miRNA-21, −34a, −200a and − 214 were 0.613, 0.508, 0.615 and 0.505, respectively.ConclusionsChanges in plasma miRNA-21, − 214, −34a and -200a levels were associated with cervical lesion severity. The plasma miRNA levels in CIN1- subjects were significantly different from those in CIN2+ subjects. This analysis may help in detection of high-grade cervical lesions.

Circulating miRNAs as a marker of metastatic disease and prognostic factor in metastatic breast cancer.

BackgroundCirculating miRNAs (miRs) are increasingly recognized as potential biomarkers in cancer. We aimed to evaluate the differential expression of miR-23b and miR-190 which are involved in tumor dormancy, miR-21 involved in metastasis and miR-200b and miR-200c involved in epithelial-mesenchymal transition (EMT) and metastasis, in the plasma of patients with early and metastatic breast cancer (MBC). We also aimed to identify associations of the expression levels with patient and disease characteristics and outcomes in metastatic patients treated with first-line chemotherapy.ResultsmiR-21 (p < 0.001), miR-23b (p = 0.033), miR-200b (p < 0.001) and miR-200c (p < 0.001) expression was higher in metastatic compared to early breast cancer. ROC curve analysis showed that miR-21 (AUC = 0.722; p < 0.001) and miR-200b (AUC = 0.720; p < 0.001) distinguished with high accuracy among the two disease states, whereas the combination of miR-21, miR-190, miR-200b and miR-200c, further improved accuracy (AUC = 0.797; p < 0.001). High miR-200b expression independently predicted for shorter OS (p = 0.026) in MBC. High expression of both miR23b and miR-190 emerged as a strong independent factor associated with shorter PFS (p = 0.001) in de novo metastatic patients and high miR-200b independently predicted for decreased OS in the HER2-negative subgroup (p = 0.007).Materials and MethodsBlood samples were obtained from patients with early (n = 133) and MBC (n = 110) before adjuvant or first-line chemotherapy, respectively. Plasma miRNA expression levels were assessed by RT-qPCR and were classified as high or low according to the median values.ConclusionsOur results are in support of the concept that circulating miRNAs represent a tool with significant diagnostic and prognostic implications in breast cancer.

Robust expression of tumor suppressor miRNA\u2019s let-7 and miR-195 detected in plasma of Saudi female breast cancer patients

BackgroundFemale breast cancer is frequently diagnosed at a later stage and the leading cause of cancer deaths world-wide. Levels of cell-free circulating microRNAs (miRNAs) can potentially be used as biomarkers to measure disease progression in breast cancer patients in a non-invasive way and are therefore of high clinical value.MethodsUsing quantitative RT-PCR, circulating miRNAs were measured in blood samples collected from disease-free individuals (n = 34), triple-negative breast tumours (TNBC) (n = 36) and luminal tumours (n = 57). In addition to intergroup comparisons, plasma miRNA expression levels of all groups were analyzed against RNASeq data from cancerous breast tissue via The Cancer Genome Atlas (TCGA).ResultsA differential set of 18 miRNAs were identified in the plasma of breast cancer patients and 10 miRNAs were uniquely identified based on ROC analysis. The most striking findings revealed elevated tumor suppressor let-7 miRNA in luminal breast cancer patients, irrespective of subtype, and elevated miR-195 in plasma of TNBC breast cancer patients. In contrast, hsa-miR-195 and let-7 miRNAs were absent from cancerous TCGA tissue and strongly expressed in surrounding non-tumor tissue indicating that cancerous cells may selectively export tumor suppressor hsa-miR-195 and let-7 miRNAs in order to maintain oncogenesis.ConclusionsWhile studies have indicated that the restoration of let-7 and miR-195 may be a potential therapy for cancer, these results suggested that tumor cells may selectively export hsa-miR-195 and let-7 miRNAs thereby neutralizing their potential therapeutic effect. However, in order to facilitate earlier detection of breast cancer, blood based screening of hsa-miR-195 and let-7 may be beneficial in a female patient cohort.

Differential expression and correlation analysis of miRNAs in thyroid nodule and plasma

Objective The variation of miRNAs profiling was identified in thyroid nodule tissue and patient plasma samples, the correlation between the expression of miRNA was analyzed in thyroid nodules and plasma. Furthermore, the importance of miRNA in plasma as a marker in thyroid nodule diagnosis was evaluated. Methods Case-control method was used. 5 female patients with thyroid lobectomy in Beijing Dongzhimen Hospital were enrolled during May and July in 2015. 16 female patients with thyroid nodules by ultrasound diagnosis, also15 healthy female without thyroid nodules were recruited during the same time. miRNA array was used to identify a subset of differentially expressed miRNAs between thyroid nodule and adjacent tissue (n=5). These miRNAs were further validated by real-time RT-PCR in a cohort of 16 thyroid nodule patients and 15 healthy controls. The miRNA expression difference between patients with thyroid nodules and healthy controls was analyzed by t test analysis. The correlation of miRNA expression between thyroid tissue and plasma was analyzed by Pearson correlation analysis. Results The miRNA array identified 57 miRNAs dysregulation, including 34 upregulation ones and 23 downregulation ones(t=3.637, 3.821, 4.907, 3.383,-5.516, 4.332,-4.993, 3.980, 5.208,-2.981, 2.840, 4.945, 4.945,-2.927,-3.561,-4.665, 3.161, 3.363, 4.846, 3.834, 2.826,-3.868, 3.816, 2.802, 5.511, 3.772,-4.424, 3.420, 3.199, 4.320, 4.053, 4.997,-3.292, 5.304,-3.281,-3.262,-4.128,-2.963, 4.228, 2.949, 4.436, 2.796, 3.199,-4.636, 4.091, 3.633, 3.481,-5.344, 4.753,-3.766,-5.340, 4.888,-4.631, 5.511, 2.921,-3.042,-4.935, P 1.5) and miR-199a-5p, miR-31 are downregulated (fold change <0.6) in thyroid nodule tissue comparing with normal tissue. Their dysregulation was further validated by real time RT-PCR in plasma samples. The significant miRNAs were confirmed with miR-181b, miR-494 and miR-106a* upregulation in thyroid nodule patients plasma. And the plasma miRNA expression levels were correlated with miRNA array results according to Pearson correlation analysis (r=0.713, 0.878 and 0.751 respectively, P<0.05). Conclusion The study highlighted that miR-181b, miR-494 and miR-106a* were differentially expressed in thyroid nodule patients not only in tissue but also in plasma, which could provide potential value as a marker for early diagnosis of thyroid nodule. (Chin J Lab Med, 2016, 39: 837-842) Key words: Thyroid nodule; microRNAs; Gene expression profiling; Biological markers

Plasma microRNA profiling of children with idiopathic dilated cardiomyopathy

Context: Dilated cardiomyopathy (DCM) is the most common cardiomyopathy in children. MicroRNAs (miRNA) are small RNAs which have regulatory functions in many biological processes.Objective: We aimed to determine miRNA expression levels in plasma of children with DCM.Materials and methods: Plasma expression levels of 379 miRNAs were compared between 23 DCM and 26 healthy children.Results: The expression levels of miR-618, miR-875-3p, miR-205, miR-194, miR-302a, miR-147, and miR-544 were found decreased. The expression levels of miR-518f and miR-454 were found increased in DCM patients.Discussion: miRNA level differences may provide the chance of using these miRNAs as new biomarkers.

MicroRNA-22-3p is down-regulated in the plasma of Han Chinese patients with premature ovarian failure

To determine whether plasma microRNAs (miRNAs) are differentially expressed between women with and without premature ovarian failure (POF), and to uncover the association of miRNAs with risk of POF. Microarray with real-time polymerase chain reaction validation. University hospital. A total of 140 individuals with premature ovarian failure (POF) and 140 age- and body mass index-matched control subjects of Han Chinese ancestry. None. Relative miRNA expression levels in plasma of POF and control group. Fifty-one differentially expressed miRNAs were identified by chip-based discovery stage between ten patients with POF and ten control subjects, among which nine miRNAs (let-7b-5p, let-7c, miR-15b-5p, miR-22-3p, miR-23a-3p, miR-23b-3p, miR-24-3p, miR-151a-5p, and miR-151b) were selected and validated. The relative expression level of miR-22-3p was significantly down-regulated in POF compared with control subjects. MiR-22-3p yielded a receiver operating characteristic (ROC) curve area of 0.668 (95% confidence interval 0.602-0.733) in discriminating POF from controls. In addition, logistic binary regression analysis and linear regression analysis showed the miR-22-3p to be a protective factor for POF (odds ratio 0.766, 95% CI 0.643-0.912) and negatively associated with serum FSH. Furthermore, bioinformatics analysis indicated that the target function of miR-22-3p was involved in apoptosis, endocytosis, and tumorigenesis. Mir-22-3p showed a lower expression level in POF and was modestly effective in distinguishing POF from control subjects. The decreased expression of miR-22-3p in plasma of POF may reflect the diminished ovarian reserve and be a consequence of the pathologic process of POF.

Cell-Free miR-27a, a Potential Diagnostic and Prognostic Biomarker for Gastric Cancer.

MicroRNAs (miRNAs) have been demonstrated to play an important role in carcinogenesis. Previous studies revealed that miRNAs are present in human plasma in a remarkably stable form that is protected from endogenous RNase activity. In this study, we measured the plasma expression levels of three miRNAs (miR-21, miR-27a, and miR-155) to investigate the usefulness of miRNAs for gastric cancer detection. We initially examined plasma miRNA expression levels in a screening cohort consisting of 15 patients with gastric cancer and 15 healthy controls from Korean population, using TaqMan quantitative real-time polymerase chain reaction. We observed that the expression level of miR-27a was significantly higher in patients with gastric cancer than in healthy controls, whereas the miR-21 and miR-155a expression levels were not significantly higher in the patients with gastric cancer. Therefore, we further validated the miR-27a expression level in 73 paired gastric cancer tissues and in a validation plasma cohort from 35 patients with gastric cancer and 35 healthy controls. In both the gastric cancer tissues and the validation plasma cohort, the miR-27a expression levels were significantly higher in patients with gastric cancer. Receiver-operator characteristic (ROC) analysis of the validation cohort, revealed an area under the ROC curve value of 0.70 with 75% sensitivity and 56% specificity in discriminating gastric cancer. Thus, the miR-27a expression level in plasma could be a useful biomarker for the diagnosis and/or prognosis of gastric cancer.

Circulating microRNA Profile throughout the Menstrual Cycle

Normal physiological variables, such as age and gender, contribute to alterations in circulating microRNA (miRNA) expression levels. The changes in the female body during the menstrual cycle can also be reflected in plasma miRNA expression levels. Therefore, this study aimed to determine the plasma miRNA profile of healthy women during the menstrual cycle and to assess which circulating miRNAs are derived from blood cells. The plasma miRNA expression profiles in nine healthy women were determined by quantitative real time PCR using Exiqon Human Panel I assays from four time-points of the menstrual cycle. This platform was also used for studying miRNAs from pooled whole blood RNA samples at the same four time-points. Our results indicated that circulating miRNA expression levels in healthy women were not significantly altered by the processes occurring during the menstrual cycle. No significant differences in plasma miRNA expression levels were observed between the menstrual cycle time-points, but the number of detected miRNAs showed considerable variation among the studied individuals. miRNA analysis from whole blood samples revealed that majority of miRNAs in plasma are derived from blood cells. The most abundant miRNA in plasma and blood was hsa-miR-451a, but a number of miRNAs were only detected in one or the other sample type. In conclusion, our data suggest that the changes in the female body during the menstrual cycle do not affect the expression of circulating miRNAs at measurable levels.

Abstract 161: Plasma-based biomarkers for breast cancer diagnosis

Abstract Introduction It has been demonstrated that there are abundant, stable microRNAs (miRNAs) in plasma/serum, which can be detected and is potentially disease-specific. In this study, we aimed to assess whether serum/plasma miRNAs may be used as potential biomarkers for breast cancer diagnosis. Methods We used systematically Solexa sequencing as well as TaqMan Low Density Array followed by individual quantitative reverse transcription polymerase chain reaction assays to screen plasma miRNAs in 48 breast cancer patients and 48 age-matched controls of Chinese origin for the discovery stage and validated in an internal validation set (50 patients and 50 controls in Chinese) and in an external validation set (24 patients and 24 controls in Caucasians). Results Four miRNAs (miR-16, miR-25, miR-222 and miR-324-3p) were differentially expressed between breast cancer cases and controls and subsequently validated in the internal and external validation stage samples. Taken together, Chinese patients with high plasma miRNA expression levels had a significantly increased risk of breast cancer for all the four miRNAs. The Area Under the Receiver Operating Characteristic Curve was 0.917 for the four-miRNA signature in the discovery stage (sensitivity=0.917 and specificity=0.917), 0.930 in the internal validation stage (sensitivity=0.920 and specificity=0.940). Conclusions The four-miRNA signature from plasma may serve as a noninvasive diagnostic biomarker for breast cancer. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 161. doi:10.1158/1538-7445.AM2011-161