Plasma miR-21 Research Articles

Is microRNA-451 expression a useful hemolytic marker in children with β-thalassemia? A pilot study

Background Beta-thalassemia (BT) is the most prevalent inherited hemoglobin disorder in Egypt. MicroRNA-451 (miR-451), an erythroid cell-specific miRNA, is upregulated during erythroid maturation and suggested to be a hemolytic marker. Aim The aim was to investigate plasma miR-451 value as a hemolytic marker, the authors conducted a pilot study on patients with BT attending a hematology clinic in Egypt. Patients and methods The patients with BT were categorized as transfusion dependent (TDT) and nontransfusion-dependent (NTDT). A total of 29 patients were included: 13 TDT (4.5–14.8 years), 16 NTDT (3.2–17.6 years), and 10 controls aged 3–14 years. The miR-451 expression in the plasma of patients was compared with the controls using real-time quantitative polymerase chain reaction. Results Plasma miR-451 expression was higher among patients with BT compared with controls (P=0.272). Similarly, there was a higher expression among the TDT subgroup than the NTDT subgroup (P=0.288). Patients with a severe disease phenotype showed a higher level of miR-451 expression when compared with mild and moderate cases (P=0.301) and among splenectomized patients (P=0.10). Plasma miR-451 did not significantly differentiate between patients with BT and normal controls (area under curve=0.621; P=0.17). In controls, the hemoglobin, red blood cells, and platelet count were significantly correlated with the plasma miR-451 (P=0.03, 0.008, and 0.008, respectively). However, in patients with BT, the alkaline phosphatase significantly correlated with miR-451 (P=0.007). Conclusion The expression of miR-451 was more in patients with TDT than NTDT and controls, but this was not statistically significant. Plasma miR-451 failed to differentiate between controls and patients with BT. More studies are needed to determine its significance in the clinical practice.

Prognostic Implications of microRNA-155, -133a, -21 and -205 in Breast Cancer Patients' Plasma.

Breast cancer, being a heterogenous disease at the intra-tumoral and intertumoral levels, presents challenges in following the progress of the disease. Tumour-secreted aberrantly expressed miRNAs obtained from peripheral blood represent a non-invasive alternative resource for detecting and monitoring the development of the disease. This study evaluates the expression of miR-155, miR-133a, miR-21 and miR-205 as non-invasive, prognostic and follow-up markers for breast cancer. Plasma expression levels of miR-155, miR-133a, miR-21 and miR-205 were measured using real-time PCR in breast cancer patients (n=63) at presentation, healthy controls (n=25), and in post-treatment samples of 31 patients. A meta-analysis was performed using 43 studies identified from PubMed, Google Scholar and Scopus databases. Hedge's g values were used to calculate the overall effect size. Plasma miR-21 levels were higher in breast cancer patients at presentation compared to controls, while no difference was observed for miR-155, miR-133a and miR-205. These results were further supported by the meta-analysis. The altered levels of miR-155 during tamoxifen treatment indicated a potential role for miR-155 in monitoring treatment response. Further, high expressions of at least three miRNAs correlated with poor overall survival in the breast cancer patients. Plasma levels of miR-155, miR-133a, miR-21 and miR-205 may be useful as prognostic and follow-up markers for breast cancer with further validation in a large cohort of patients.

Plasma expression of microRNA-425-5p and microRNA-451a as biomarkers of cardiovascular disease in rheumatoid arthritis patients

To validate in a cohort of 214 rheumatoid arthritis patients a panel of 10 plasmatic microRNAs, which we previously identified and that can facilitate earlier diagnosis of cardiovascular disease in rheumatoid arthritis patients. We identified 10 plasma miRs that were downregulated in male rheumatoid arthritis patients and in patients with acute myocardial infarction compared to controls suggesting that these microRNAs could be epigenetic biomarkers for cardiovascular disease in rheumatoid arthritis patients. Six of those microRNAs were validated in independent plasma samples from 214 rheumatoid arthritis patients and levels of expression were associated with surrogate markers of cardiovascular disease (carotid intima-media thickness, plaque formation, pulse wave velocity and distensibility) and with prior cardiovascular disease. Multivariate analyses adjusted for traditional confounders and treatments showed that decreased expression of microRNA-425-5p in men and decreased expression of microRNA-451 in women were significantly associated with increased (β = 0.072; p = 0.017) and decreased carotid intima-media thickness (β = −0.05; p = 0.013), respectively. MicroRNA-425-5p and microRNA-451 also increased the accuracy to discriminate patients with pathological carotid intima-media thickness by 1.8% (p = 0.036) in men and 3.5% (p = 0.027) in women, respectively. In addition, microRNA-425-5p increased the accuracy to discriminate male patients with prior cardiovascular disease by 3% (p = 0.008). Additionally, decreased expression of microRNA-451 was significantly associated with decreased pulse wave velocity (β = −0.72; p = 0.035) in overall rheumatoid arthritis population. Distensibility showed no significant association with expression levels of the microRNAs studied. We provide evidence of a possible role of microRNA-425-5p and microRNA-451 as useful epigenetic biomarkers to assess cardiovascular disease risk in patients with rheumatoid arthritis.

Mastiha has efficacy in immune-mediated inflammatory diseases through a microRNA-155 Th17 dependent action

Mastiha is a natural nutritional supplement with known anti-inflammatory properties. Non-alcoholic fatty liver disease (NAFLD) and Inflammatory bowel disease (IBD) are immune mediated inflammatory diseases that share common pathophysiological features. Mastiha has shown beneficial effects in both diseases. MicroRNAs have emerged as key regulators of inflammation and their modulation by phytochemicals have been extensively studied over the last years. Therefore, the aim of this study was to investigate whether a common route exists in the anti-inflammatory activity of Mastiha, specifically through the regulation of miRNA levels. Plasma miR-16, miR-21 and miR-155 were measured by Real-Time PCR before and after two double blinded and placebo-controlled randomized clinical trials with Mastiha. In IBD and particularly in ulcerative colitis patients in relapse, miR-155 increased in the placebo group (p = 0.054) whereas this increase was prevented by Mastiha. The mean changes were different in the two groups even after adjusting for age, sex and BMI (p = 0.024 for IBD and p = 0.042). Although the results were not so prominent in NAFLD, miR-155 displayed a downward trend in the placebo group (p = 0.054) whereas the levels did not changed significantly in the Mastiha group in patients with less advanced fibrosis. Our results propose a regulatory role for Mastiha in circulating levels of miR-155, a critical player in T helper-17 (Th17) differentiation and function.

The relation between plasma miR-126 levels and cerebral collateral circulation in patients with intracranial arterial stenosis.

This study aimed to investigate the correlation between the circulating miR-126 regulation pathway and the cerebral collateral circulation (CCC), and to test whether miR-126 could serve as a potential biomarker for CCC formation in patients with intracranial arterial stenosis or occlusion. This single-centre cross-sectional study enrolled patients who underwent cerebral angiography with severe stenosis (≥70%) or occlusion in at least one major intracranial artery. Collateral degree was graded according to the ASITN/SIR classification. The patients were divided into a good CCC group (grade 3-4) or a poor CCC group (grade 0-2). We investigated the plasma levels of miR-126, VEGF, Spred-1 and PIK3R2 by using qRT-PCR, ELISA and Western blot methods, respectively. In addition, we assessed the correlations of plasma miR-126 with VEGF, Spred-1, PIK3R2 and ASITN/SIR grade using the Spearman correlation test and investigated its predictive power for CCC status by using the receiver operating characteristic curve. A total of 68 patients were enrolled (44 with good CCC and 24 with poor CCC). Data showed that plasma miR-126 and VEGF were significantly higher in the good CCC group than in the poor CCC group. Plasma Spred-1 and PIK3R2 level were lower in the good CCC group than in the poor CCC group. In addition, miR-126 and VEGF were positively correlated with ASITN/SIR (miR-126: R = 0.595, P < 0.01; VEGF: R = 0.595, P < 0.01), whereas Spred-1 and PIK3R2 were negatively correlated with ASITN/SIR (Spred-1: R = -0.817, P < 0.01; PIK3R2: R = -0.513, P=0.01). However, the area under the curve of miR-126 level for CCC status was only 0.328 (95% CI: 0.158-0.498; p = 0.067). Plasma miR-126 level may be related to better CCC formation, one of the mechanisms that may be explained by upregulation of VEGF and reduction of Spred-1 and PIK3R2 protein expression. However, miR-126 might not be an independent predictor for CCC, given its low predictive value.

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.

Insulin micro-secretion in Type 1 diabetes and related microRNA profiles

The aim of this cross-sectional study was to compare plasma C-peptide presence and levels in people without diabetes (CON) and with Type 1 diabetes and relate C-peptide status to clinical factors. In a subset we evaluated 50 microRNAs (miRs) previously implicated in beta-cell death and associations with clinical status and C-peptide levels. Diabetes age of onset was stratified as adult (≥ 18 y.o) or childhood (< 18 y.o.), and diabetes duration was stratified as ≤ 10 years, 10–20 years and > 20 years. Plasma C-peptide was measured by ultrasensitive ELISA. Plasma miRs were quantified using TaqMan probe-primer mix on an OpenArray platform. C-peptide was detectable in 55.3% of (n = 349) people with diabetes, including 64.1% of adults and 34.0% of youth with diabetes, p < 0.0001 and in all (n = 253) participants without diabetes (CON). C-peptide levels, when detectable, were lower in the individuals with diabetes than in the CON group [median lower quartile (LQ)–upper quartile (UQ)] 5.0 (2.6–28.7) versus 650.9 (401.2–732.4) pmol/L respectively, p < 0.0001 and lower in childhood versus adult-onset diabetes [median (LQ–UQ) 4.2 (2.6–12.2) pmol/L vs. 8.0 (2.3–80.5) pmol/L, p = 0.02, respectively]. In the childhood-onset group more people with longer diabetes duration (> 20 years) had detectable C-peptide (60%) than in those with shorter diabetes duration (39%, p for trend < 0.05). Nine miRs significantly correlated with detectable C-peptide levels in people with diabetes and 16 miRs correlated with C-peptide levels in CON. Our cross-sectional study results are supportive of (a) greater beta-cell function loss in younger onset Type 1 diabetes; (b) persistent insulin secretion in adult-onset diabetes and possibly regenerative secretion in childhood-onset long diabetes duration; and (c) relationships of C-peptide levels with circulating miRs. Confirmatory clinical studies and related basic science studies are merited.

Relationship of Circulating Mirnas With Insulin Resistance and Metabolic Dysfunction in Adolescents

Abstract Objectives To evaluate the relationship of plasma expression of miR-122, -375, -139, 15-a and -let7c with insulin resistance and factors related to metabolic syndrome (MetS) in adolescents. Methods Data from 188 adolescents aged 12–19 years participating in a cross-sectional population-based study were analyzed, all non-smokers, without acute inflammatory diseases and cancer, and without the use of medication that would interfere in the outcome. The quantification of circulating miRNA was performed by RT-PCR, using the Fluidigm chip technique. The miRNA expression was compared among adolescents grouped by the absence (n = 94) or presence of insulin resistance (n = 94)—categorized according to with the HOMA-IR index -, and the absence (n = 92) or presence of 1 (n = 59), 2 (n = 23), 3 (n = 10) or 4 (n = 4) factors related to MetS. The comparison of means was performed using the Mann-Whitney test for two samples or Kruskal-Wallis followed by Dunn's post-test to compare three or more samples. Results Insulin resistance was identified in 50% of the individuals (HOMA-IR &amp;gt; 2.7). In adolescents with insulin resistance, the plasma expression of miR-122 was higher in comparison to normoglycemic individuals and was positively associated with fasting insulinemia (r = 0.157; P = 0.031), HOMA-IR index (r = 0.160; P = 0.028) and plasma hepcidin (r = 0.209; P = 0.005) levels. When evaluating the expression according to the number of factors related to MetS, the expression of miR-let7c, -139 and -15a increased according to the increase of the factors related to MetS. The opposite effect was observed in relation to miR-375. Individuals with 3 or more factors differed from groups whith 1 or 2 factores for plasma miR-375, -139, 15-a and -let7c expression. MiR-375 showed a negative association with body weight (r = −0.169; P = 0.019), while miR-139 was associated with systolic (r = 0.153; P = 0.055) and diastolic (r = 0.193; P = 0.016) blood pressure. Plasma miR-15a expression had a positive correlation with diastolic blood pressure (r = 0.198; P = 0.0064). MiR-let7c showed a positive correlation plasma TG (r = 0.156; P = 0.029). Conclusions Plasma miR 122 expression was related to insulin resistance, while plasma miR 375, 139, 15 a and let7c expressions were associated with MetS in adolescents. Funding Sources From Sao Paulo Research Foundation and National Council for Scientific and Technological Development

MiR-223 or miR-126 predicts resistance to dual antiplatelet therapy in patients with ST-elevation myocardial infarction.

ObjectiveTo explore the role of miR-223 and miR-126 in predicting treatment responses to dual antiplatelet therapy (DAPT) in patients with ST-elevation myocardial infarction (STEMI).MethodsPlasma miR-223 and miR-126 levels were measured before treatment. Treatment responses and 2-year survival were determined. In vitro experiments were performed to explore the mechanism of action.ResultsPatients with resistance to DAPT had a lower level of miR-223 and miR-126. Cardiac-event-free survival was shorter in patients with lower miR-223 or miR-126 levels. MiR-223 and miR-126 independently predicted DAPT resistance. Modulating miR-223 or miR-126 in platelets in vitro significantly changed the response to clopidogrel by regulating platelet aggregation.ConclusionMiR-223 and miR-126 play a role in DAPT resistance and may provide potential biomarkers in patients with STEMI.

MiR-126 Is an Independent Predictor of Long-Term All-Cause Mortality in Patients with Type 2 Diabetes Mellitus.

MicroRNAs are endogenous non-coding RNAs that are involved in numerous biological processes through regulation of gene expression. The aim of our study was to determine the ability of several miRNAs to predict mortality and response to antiplatelet treatment among T2DM patients. Two hundred fifty-two patients with diabetes were enrolled in the study. Among the patients included, 26 (10.3%) patients died within a median observation time of 5.9 years. The patients were receiving either acetylsalicylic acid (ASA) 75 mg (65%), ASA 150 mg (15%) or clopidogrel (19%). Plasma miR-126, miR-223, miR-125a-3p and Let-7e expressions were assessed by quantitative real time PCR and compared between the patients who survived and those who died. Adjusted Cox-regression analysis was used for prediction of mortality. Differential miRNA expression due to different antiplatelet treatment was analyzed. After including all miRNAs into one multivariate Cox regression model, only miR-126 was predictive of future occurrence of long-term all-cause death (HR = 5.82, 95% CI: 1.3–24.9; p = 0.024). Furthermore, miR-126, Let-7e and miR-223 expressions in the clopidogrel group were significantly higher than in the ASA group (p = 0.014; p = 0.013; p = 0.028, respectively). To conclude, miR-126 expression is a strong and independent predictor of long-term all-cause mortality among patients with T2DM. Moreover, miR-223, miR-126 and Let-7e present significant interactions with antiplatelet treatment regimens and clinical outcomes.

MicroRNAs as Biomarkers of Charcot-Marie-Tooth Disease Type 1A.

To determine whether microRNAs (miRs) are elevated in the plasma of individuals with the inherited peripheral neuropathy Charcot-Marie-Tooth disease type 1A (CMT1A), miR profiling was employed to compare control and CMT1A plasma. We performed a screen of CMT1A and control plasma samples to identify miRs that are elevated in CMT1A using next-generation sequencing, followed by validation of selected miRs by quantitative PCR, and correlation with protein biomarkers and clinical data: Rasch-modified CMT Examination and Neuropathy Scores, ulnar compound muscle action potentials, and motor nerve conduction velocities. After an initial pilot screen, a broader screen confirmed elevated levels of several muscle-associated miRNAs (miR1, -133a, -133b, and -206, known as myomiRs) along with a set of miRs that are highly expressed in Schwann cells of peripheral nerve. Comparison to other candidate biomarkers for CMT1A (e.g., neurofilament light) measured on the same sample set shows a comparable elevation of several miRs (e.g., miR133a, -206, -223) and ability to discriminate cases from controls. Neurofilament light levels were most highly correlated with miR133a. In addition, the putative Schwann cell miRs (e.g., miR223, -199a, -328, -409, -431) correlate with the recently described transmembrane protease serine 5 (TMPRSS5) protein biomarker that is most highly expressed in Schwann cells and also elevated in CMT1A plasma. These studies identify a set of miRs that are candidate biomarkers for clinical trials in CMT1A. Some of the miRs may reflect Schwann cell processes that underlie the pathogenesis of the disease. This study provides Class III evidence that a set of plasma miRs are elevated in patients with CMT1A.

Altered endothelial dysfunction-related miRs in plasma from ME/CFS patients

Myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) is a complex disease characterized by unexplained debilitating fatigue. Although the etiology is unknown, evidence supports immunological abnormalities, such as persistent inflammation and immune-cell activation, in a subset of patients. Since the interplay between inflammation and vascular alterations is well-established in other diseases, endothelial dysfunction has emerged as another player in ME/CFS pathogenesis. Endothelial nitric oxide synthase (eNOS) generates nitric oxide (NO) that maintains endothelial homeostasis. eNOS is activated by silent information regulator 1 (Sirt1), an anti-inflammatory protein. Despite its relevance, no study has addressed the Sirt1/eNOS axis in ME/CFS. The interest in circulating microRNAs (miRs) as potential biomarkers in ME/CFS has increased in recent years. Accordingly, we analyze a set of miRs reported to modulate the Sirt1/eNOS axis using plasma from ME/CFS patients. Our results show that miR-21, miR-34a, miR-92a, miR-126, and miR-200c are jointly increased in ME/CFS patients compared to healthy controls. A similar finding was obtained when analyzing public miR data on peripheral blood mononuclear cells. Bioinformatics analysis shows that endothelial function-related signaling pathways are associated with these miRs, including oxidative stress and oxygen regulation. Interestingly, histone deacetylase 1, a protein responsible for epigenetic regulations, represented the most relevant node within the network. In conclusion, our study provides a basis to find endothelial dysfunction-related biomarkers and explore novel targets in ME/CFS.

Correlation between the Expression Levels of miR-21 and miR-192 and Clinicopatological Features in Plasma of Patients with Gastric Cancer in Iran

Background: Gastric cancer (GC) is the most prevalent malignancy worldwide and a common cause of death in Iran. Studies have proved that a variety of dysregulated microRNAs is involved in the development and progression of gastric cancer. The present study aimed to evaluate the expression levels of plasma circulating oncogenic miR-21 and miR-192 and their association with clinical phenotypes of patients with gastric cancer in the north of Iran. Material and Methods: Clinico-pathological analysis was conducted using a standard protocol and pathological tests. The expression levels of miR-21 and miR-192 were measured using quantitative reverse transcription-polymerase chain reaction in the plasma of twenty pre/post-operative gastric cancer patients and twenty healthy subjects. The receiver operating characteristic (ROC) curve of these microRNAs was analyzed to investigate their diagnosis properties. Results: The study results indicated that plasma miR-21 expression was significantly associated with tumor stage and helicobacter pylori infection status (P=0.024, P=0.0004, respectively). However, no association was observed between clinic-pathological characteristics and miR-192 expression. The results showed that the plasma levels of miR-21 (P=0.0001) and miR-192 (P=0.0007) were significantly higher in GC patients compared to those in healthy individuals. Furthermore, the ROC analyses yielded the area under the curve (AUC) values of 0.9525±0.03 (P&lt;0.0001) and 0.5925±0.09 (P=0.316) for miR-21and miR-192, respectively. Pearson regression analysis showed that there was no significant correlation between the expression of miR-21 and miR-192 (P=0.1507). Conclusion: Based on the obtained results, the expression of the plasma level of miR-21 was significantly higher in gastric cancer patients compared to that in the healthy group. Furthermore, the higher levels of AUC in miR-21 indicated the potential role of miR-21 as a noninvasive biomarker for the prognosis of gastric cancer in the population of the north of Iran.

Upregulation of miR-21 and miR-106b in plasma and tissues as a possible prognostic marker in aggressive breast cancer

Background: The miRNAs are referred to small non-coding RNAs (consisting of 18 to 25 nucleotides). Functional studies have shown their functions to be oncogenes or tumor suppressor genes in different types of cancers. The miR-106b and miR-21 have been identified to participate in the biological behaviors of cells. This study aimed to evaluate the tissue and plasma levels of miR-21 and miR-106b in patients with breast cancer who were diagnosed with ductal carcinoma. Methods: In total, 40 cases of breast cancer patients 180 samples were examined in this project. Samples included ductal carcinoma breast tumors (n=40), normal breast tissues of the margin of the tumor (n=40) and 20 samples from unaffected mammary tissue of females undergoing reduction mammoplasty (control group), plasma samples of patients with breast cancer (n=40), and plasma of non-affected individuals (n=40). The expression levels of miR-106b and miR-21 were determined using SYBR Green real-time RT-PCR assay in breast tissues and plasma of cancerous patients in comparison to the controls. Results: MiR-106b and miR-21 revealed much higher expression in tissues and plasma of patients with breast cancer in comparison to that in the group of control (P&lt;0.001). High levels of mir-106b and miR-21 expression in plasma and tumor tissues were highly correlated with tumors in higher stages and lymph node involvement (P&lt;0.0001). Conclusions: Based on the obtained results, upregulation of miR-106b and miR-21 in the plasma of patients with breast cancer can act as a possible non-invasive biomarker for breast cancer prognosis. Further follow-up studies are required to confirm this.

Associations between Circulating MicroRNAs and Measures of Insulin/Glucose Dynamics in People Living with HIV with Dysglycemia

People living with HIV (PLWH) have near normal life expectancies with advances in antiretroviral therapy, but with the increasing average age, comorbid conditions create a new health burden for this population. Insulin resistance and type II diabetes are significantly higher in PLWH. microRNAs (miRs) are small non-coding RNAs that alter protein expression, and dysregulated expression of multiple miRs are implicated in altered insulin/glucose dynamics. There is increasing interest in using miRs as non-invasive biomarkers of pathophysiological states since most miRs are found in circulation and can be traced back to the cell type where they are either actively secreted or released during cell death. In this study, we aimed to identify circulating miRs that are associated with measures of insulin/glucose dynamics in PLWH with dysglycemia. PLWH (N=98, 68 male) with fasting plasma glucose between 95-124 mg/dL were enrolled in the parent ALIVE-Ex study provided their informed consent. Anthropometric measurements were taken along with adiponectin and an oral glucose tolerance test (OGTT) where glucose, insulin, and C-peptide were measured at 0, 1, and 2 hours. Average participant age was 52 ± 10 years, and average BMI was 27.7 ± 6.8 kg/m2. Total RNA was isolated from plasma (0h) and absolute miR expression was determined. miRs were selected based on their role in skeletal muscle (miR133a, miR-206), pancreatic β-cells (miR-375), adipose tissue (miR-let-7b), and liver (miR-20a). miR expression was correlated to measures of insulin/glucose dynamics using Spearman correlations. Multiple regression analyses were completed to control for sex, age, and BMI. Additional regression analyses combining miRs that significantly added to models (p<0.05) were used to improve predictions of insulin/glucose measures. miR-20a was positively correlated with glucose-2h (p<0.01) and this association remained significant when controlling for sex, age, and BMI. miR-375 was positively correlated with adiponectin (p<0.05); however, the association was not significant when controlling for sex, age, and BMI which were all significantly associated (p<0.01) with adiponectin. miR-206, 133a, and let-7b were not significantly correlated with any insulin/glucose measures. Combining miR-20a and miR-375 improved the prediction (R=0.42, p<0.0001) of increased glucose levels (2h) compared with miR-20a alone (R=0.30, p<0.01). In this cohort of PLWH with dysglycemia, our results indicate that only miR-375 and miR-20a significantly correlated with glucose/insulin measures. miR-375 is important for maintaining β-cell mass and was positively correlated with adiponectin, but sex, age, and BMI are stronger predictors of adiponectin. miR-20a is implicated in liver function and regulates hepatic glycogen synthesis, and the increased plasma miR-20a seen with higher plasma glucose levels could indicate hepatic dysregulation. Combining miR-20a and miR-375 led to improved predictions indicating that combining expression of multiple circulating miRs into a composite score could be utilized as an indicator of altered insulin/glucose dynamics in PLWH.

The value of circulating microRNA-1 in the early diagnosis of coronary atherosclerotic plaque rupture in patients with stable coronary heart disease

To evaluate the diagnostic value of circulating microRNA-1 (miR-1) in early coronary artery plaque rupture in patients with stable coronary artery disease (SCAD). A prospective cohort study was conducted. Sixty-seven patients with SCAD admitted to the department of cardiology of the Third Affiliated Hospital of Soochow University from January to June in 2019 were enrolled. All patients had completed coronary angiography (CAG), percutaneous coronary intervention (PCI) single stent implantation or only CAG was performed according to the CAG results. Blood samples were collected before (0 hour) and 3 hours after the procedure. The expression of plasma miR-1 was detected by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR), and electrocardiogram was used to detect cardiac troponin I (cTnI) levels. The difference of miR-1 and cTnI levels in PCI or CAG patients before and after procedure were compared, and the value for early diagnosis of coronary artery plaque rupture in SCAD patients was evaluated. The diagnostic efficacy was evaluated by the receiver operating characteristic curve (ROC curve). There were 38 CAG patients and 29 PCI patients. There were no significant differences in gender, age, previous history (without hypertension history) and baseline data of cardiac function between the two groups. The expression of miR-1 after PCI was significantly higher than that before PCI [2-ΔΔCt: 2.11 (1.56, 2.73) vs. 1.26 (1.07, 1.92), P < 0.01], and there was no significant difference in cTnI level before and after PCI [μg/L: 0.00 (0.00, 0.02) vs. 0.00 (0.00, 0.02), P > 0.05]. There were no significant differences in miR-1 and cTnI levels before and after procedure in the CAG group [miR-1 (2-ΔΔCt): 1.09 (1.00, 1.40) vs. 1.21 (1.00, 1.71), cTnI (μg/L): 0.00 (0.00, 0.02) vs. 0.00 (0.00, 0.02), both P > 0.05]. ROC curve analysis showed that the area under ROC curve (AUC) and 95% confidence interval (95%CI) of miR-1 in the diagnosis of coronary plaque rupture were 0.794 (0.687-0.900), P < 0.01, the sensitivity was 82.8%, the specificity was 68.4%, and the optimal cut-off value was 1.51. The AUC and 95%CI of the difference of miR-1 before and after operation (ΔmiR-1) were 0.704 (0.567-0.842), P = 0.004, the sensitivity was 62.1%, the specificity was 84.2%, and the optimal cut-off value was 0.39. The efficancy of miR-1 and ΔmiR-1 after procedure to diagnose coronary plaque rupture in patients with SCAD was similar (Z = 1.287, P = 0.198). However, baseline miR-1 might not predict whether patients with SCAD need PCI or not (AUC = 0.630, P > 0.05). Multivariate binary Logistic regression analysis showed that increased postoperative miR-1 expression was an independent risk factor for coronary plaque rupture in SCAD patients [odds ratios (OR) = 2.887, 95%CI was 1.044-7.978, P = 0.041]. Circulating miR-1 might have the value for early diagnosis of coronary artery plaque rupture in SCAD patients.

A High-Accuracy Model Based on Plasma miRNAs Diagnoses Intrahepatic Cholangiocarcinoma: A Single Center with 1001 Samples

Objectives: Intrahepatic cholangiocarcinoma (iCCA) is a highly malignant cancer. More than 70% of patients are diagnosed at an advanced stage. The aim of this study was to evaluate the diagnostic value of plasma miR-21, miR-122, and CA19-9, hoping to establish a novel model to improve the accuracy for diagnosing iCCA. Materials and methods: Plasma miR-21 and miR-122 were detected in 359 iCCA patients and 642 controls (healthy, benign liver lesions, other malignant liver tumors). All 1001 samples were allocated to training cohort (n = 668) and validation cohort (n = 333) in a chronological order. A logistic regression model was applied to combine these markers. Area under the receiver operating characteristic curve (AUC) was used as an accuracy index to evaluate the diagnostic performance. Results: Plasma miR-21 and miR-122 were significantly higher in iCCA patients than those in controls. Higher plasma miR-21 level was significantly correlated with larger tumor size (p = 0.030). A three-marker model was constructed by using miR-21, miR-122 and CA19-9, which showed an AUC of 0.853 (95% CI: 0.824–0.879; sensitivity: 73.0%, specificity: 87.4%) to differentiate iCCA from controls. These results were subsequently confirmed in the validation cohort with an AUC of 0.866 (0.825–0.901). The results were similar for diagnosing early (stages 0–I) iCCA patients (AUC: 0.848) and CA19-9negative iCCA patients (AUC: 0.795). Conclusions: We established a novel three-marker model with a high accuracy based on a large number of participants to differentiate iCCA from controls. This model showed a great clinical value especially for the diagnosis of early iCCA and CA19-9negative iCCA.

Circulating miR-145 as a marker of therapeutic response to anti-TNF therapy in patients with ankylosing spondylitis

Circulating miRNAs appear promising therapeutic and prognostic biomarkers. We aimed to investigate the predictive value of circulating miRNAs on the disease outcome following anti-TNF therapy in patients with ankylosing spondylitis (AS). Our study included 19 AS patients assessed at baseline (M0), after three (M3) and twelve months (M12) of therapy. Total RNA was isolated from plasma. A comprehensive analysis of 380 miRNAs using TaqMan Low Density Array (TLDA) was followed by a single assay validation of selected miRNAs. All AS patients had high baseline disease activity and an excellent response to anti-TNF therapy at M3 and M12. TLDA analysis revealed the dysregulation of 17 circulating miRNAs, including miR-145. Single assay validation confirmed that miR-145 is significantly downregulated at M3 compared to baseline. The decrease in the levels of miR-145 from M0 to M3 negatively correlated with the change in BASDAI from M0 to M3; and positively correlated with disease activity improvement from M3 to M12 as per BASDAI and ASDAS. The predictive value of the early change in miR-145 and levels of miR-145 at M3 were further validated by Receiver operating curves analysis. We show thatthe early change in circulating miR-145 may be a predictor for the future outcome ofAS patients treated with TNF inhibitors. Patients with a more significant decrease in miR-145 levels may show further significant improvement of disease activity after 12 months. Monitoring the expression of miR-145 in plasma in AS patients may, therefore, influence our therapeutic decision-making.

Role of serum miR-21 and miR-92a in colorectal cancer diagnosis as novel molecular biomarkers

Background: Our study aimed to evaluate the clinical utility of detecting plasma microRNAs (miR-21 & miR-92a) for diagnosis of colorectal cancer patients and its relation to tumor staging. Patients and Methods: Quantitative real-time RT-PCR was applied to determine the relative expression level of miR-21 and miR-92a in serum. The sensitivity and specificity of these markers were evaluated by receiver operating characteristic (ROC) curve analysis. Final staging of colorectal cancer cases was assigned according to results of histopathologic examination of surgically resected specimens. Results: This study included 52 cases of colorectal cancer (CRC), 20 cases of precancerous colorectal lesions, and 20 healthy controls. Both Plasma miR-21 and Plasma miR-92a were significantly higher in CRC group compared to both the control group and precancerous group. Also, they were significantly higher in advanced CRC stages than early CRC stages. The sensitivity and specificity of miR-21 for discriminating CRC from controls were found to be 90.38% and 100.0%, respectively. However, for miR-92a, sensitivity and specificity were found to be 94.23% and 100.0%, respectively. For discriminating CRC cases from precancerous lesions, the sensitivity and specificity of miR-21 were found to be 75.08% and 95.0%, respectively. However, for miR-92a, sensitivity and specificity were found to be 80.77% and 100.0%, respectively. Conclusions: both plasma miR-21 and miR-92a have significant value for early detection of CRC as non-invasive screening molecular biomarkers with high sensitivity and specificity. They also help for differentiation between patients with benign and malignant colorectal lesions and those with early and advanced CRC.

A preliminary study of micro-RNAs as minimally invasive biomarkers for the diagnosis of prostate cancer patients

BackgroundA prostate cancer diagnosis is based on biopsy sampling that is an invasive, expensive procedure, and doesn’t accurately represent multifocal disease.MethodsTo establish a model using plasma miRs to distinguish Prostate cancer patients from non-cancer controls, we enrolled 600 patients histologically diagnosed as having or not prostate cancer at biopsy. Two hundred ninety patients were eligible for the analysis. Samples were randomly divided into discovery and validation cohorts.ResultsNGS-miR-expression profiling revealed a miRs signature able to distinguish prostate cancer from non-cancer plasma samples. Of 51 miRs selected in the discovery cohort, we successfully validated 5 miRs (4732-3p, 98-5p, let-7a-5p, 26b-5p, and 21-5p) deregulated in prostate cancer samples compared to controls (p ≤ 0.05). Multivariate and ROC analyses show miR-26b-5p as a strong predictor of PCa, with an AUC of 0.89 (CI = 0.83–0.95;p < 0.001). Combining miRs 26b-5p and 98-5p, we developed a model that has the best predictive power in discriminating prostate cancer from non-cancer (AUC = 0.94; CI: 0,835-0,954).To distinguish between low and high-grade prostate cancer, we found that miR-4732-3p levels were significantly higher; instead, miR-26b-5p and miR-98-5p levels were lower in low-grade compared to the high-grade group (p ≤ 0.05).Combining miR-26b-5p and miR-4732-3p we have the highest diagnostic accuracy for high-grade prostate cancer patients, (AUC = 0.80; CI 0,69-0,873).ConclusionsNoninvasive diagnostic tests may reduce the number of unnecessary prostate biopsies.The 2-miRs-diagnostic model (miR-26b-5p and miR-98-5p) and the 2-miRs-grade model (miR-26b-5p and miR-4732-3p) are promising minimally invasive tools in prostate cancer clinical management.