Plasma miR-21 Research Articles

433 Is miR let-7c protective against Acute Chest Syndrome in Sickle Cell Disease?

OBJECTIVES/GOALS: We have shown that small extracellular vesicles (exosomes) isolated from patients with a history of ACS disrupt the endothelium in vitro. Sequencing of miRNA contents of these vesicles suggested that miR let-7c was differentially expressed. The current study was designed to determine the relationship between miR let-7c levels and ACS. METHODS/STUDY POPULATION: We identified 16 subjects from the SCD Lungomics biobank at the University of Chicago Comer and La Rabida Childrens Hospitals who had samples obtained at baseline. Among them, 9 had a history of ACS (ACS(+)) and 7 did not (ACS(-)). For all subjects, we reviewed clinical data relevant to their SCD and laboratory data (including hemoglobin, absolute reticulocyte count, white blood cell count) obtained at the same time as the baseline samples. RNA was isolated from the plasma and miR let7c was quantified using quantitative RT-PCR. RESULTS/ANTICIPATED RESULTS: Subjects were similar clinically, except that those with a history of ACS were more likely to be on hydroxyurea (p<0.05) and to have obstructive sleep apnea (p<0.05). Hematologic laboratory values were similar irrespective of ACS history. The mean miR let-7c level was 2-fold less in subjects with a history of ACS than in those who had never had ACS (p<0.05). A plasma miR let-7c level < 1 (normalized to the control subjects) had a positive predictive value of 0.78 for history of ACS and a sensitivity of 78% and specificity of 71%. Among subjects with a history of ACS, the let7c levels did not correlate with time since the last ACS event. However, among subjects who developed ACS following the baseline samples, higher miR let-7c levels correlated with increased length of time to next ACS event (R=0.8). DISCUSSION/SIGNIFICANCE: Our results in a group of subjects with SCD show that plasma miR let-7c levels are decreased in subjects with a history of ACS. They suggest that miR let-7c may be protective against development of ACS and that measurement of its levels could be a useful biomarker to assess or predict risk for this complication of SCD.

Associations between circulating microRNAs and coronary plaque characteristics: potential impact from physical exercise.

microRNAs (miRs) are involved in different steps in the development of atherosclerosis and are proposed as promising biomarkers of coronary artery disease (CAD). We hypothesized that circulating levels of miRs were associated with coronary plaque components assessed by radiofrequency intravascular ultrasound (RF-IVUS) before and after aerobic exercise intervention. Thirty-one patients with CAD treated with percutaneous coronary intervention (PCI) previously included in a randomized trial with aerobic interval training (AIT) or moderate continuous training (MCT) as post-PCI intervention were included. Coronary plaque characteristics by grayscale and RF-IVUS and predefined circulating candidate miRs in plasma were analyzed at baseline and follow-up. Associations between miRs and coronary plaque composition, and the potential effect from exercise, were analyzed using linear regression. Circulating levels of miR-15a-5p, miR-30e-5p, miR-92a-3p, miR-199a-3p, miR-221-3p, and miR-222-3p were associated with baseline coronary necrotic core volume. Following exercise intervention, decreased levels of miR-15a-5p, miR-93-5p, and miR-451a, and increased levels of miR-146a-5p were associated with an observed regression of coronary plaque burden. A mirPath prediction tool identified that genes regulated by miR-15a-5p, miR-199a-3p, and miR-30e-5p were significantly overrepresented in pathways related to fatty acid biosynthesis and fatty acid metabolism. This exploratory study demonstrated six miRs associated with coronary necrotic core, a marker of plaque vulnerability. In addition, changes in four miRs were associated with a regression of coronary plaque burden following exercise intervention. These novel findings may identify potential future biomarkers of CAD and coronary plaque composition.

LiKidMiRs: A ddPCR-Based Panel of 4 Circulating miRNAs for Detection of Renal Cell Carcinoma.

Simple SummaryEarly detection of renal cell carcinoma (RCC) significantly increases the likelihood of curative treatment, avoiding the need of adjuvant therapies, associated side effects and comorbidities. Thus, we aimed to discover circulating microRNAs that might aid in early, minimally invasive, RCC detection/diagnosis.Background: Decreased renal cell cancer-related mortality is an important societal goal, embodied by efforts to develop effective biomarkers enabling early detection and increasing the likelihood of curative treatment. Herein, we sought to develop a new biomarker for early and minimally invasive detection of renal cell carcinoma (RCC) based on a microRNA panel assessed by ddPCR. Methods: Plasma samples from patients with RCC (n = 124) or oncocytomas (n = 15), and 64 healthy donors, were selected. Hsa-miR-21-5p, hsa-miR-126-3p, hsa-miR-155-5p and hsa-miR-200b-3p levels were evaluated using a ddPCR protocol. Results: RCC patients disclosed significantly higher circulating levels of hsa-miR-155-5p compared to healthy donors, whereas the opposite was observed for hsa-miR-21-5p levels. Furthermore, hsa-miR-21-5p and hsa-miR-155-5p panels detected RCC with high sensitivity (82.66%) and accuracy (71.89%). The hsa-miR-126-3p/hsa-miR-200b-3p panel identified the most common RCC subtype (clear cell, ccRCC) with 74.78% sensitivity. Conclusion: Variable combinations of plasma miR levels assessed by ddPCR enable accurate detection of RCC in general, and of ccRCC. These findings, if confirmed in larger studies, provide evidence for a novel ancillary tool which might aid in early detection of RCC.

Abstract WP137: Mirna 206 Expression Is Associated With Perihematomal Edema Volume And Functional Outcomes After Intracerebral Hemorrhage

Introduction: Circulating micro-RNAs (miR) are reported in the pathophysiology of intracerebral hemorrhage. Previous reports suggested that miR-206, miR-486, miR-150 and miR-146a are associated with ICH outcomes. In this, study we explored the associated of miRNA expression with radiological predictors of outcome hematoma expansion (HE) and perihematomal edema volume (PHEV) and functional outcomes. Methods: Patients (n=96) with acute spontaneous ICH were consented and blood samples were collected within 48 hours of ICH onset. Plasma miR were used to screen 752 well-characterized miR using locked nucleic acid (LNA) probe-based real-time PCR (ver3.3.5). Cycle threshold data were obtained using the regression method and global mean normalization to a universally expressed miR-92a was applied. Significance levels were assessed using the Mann-Whitney’s U-test and the Welch’s t-test (GenEx). Demographic, clinical data and Modified Rankin Score (MRS) were collected with follow-up calls at 30, 90, and 365-days from ICH onset. Hematoma and perihematomal edema volumes were calculated based on the ABC/2 formula at presentation, 24hrs, and 96hrs from ICH onset. Spearman’s rank correlation was used to determine the association of miRNA expression with HE (>30% increase in volume) and PHEV at onset, 24hrs and 96hrs. Simple logistic regression was used to determine the odds of survival at 30-days and favorable outcome at 90- and 365-days, defined as MRS < 3. Results: Overall, cohort mean age 60.2years (IQR 52-73), 57.1% were deep nuclei (thalamic and basal ganglia ICH), mean ICH vol. 38.4 (SD23.2). We observed a positive correlation with miR-206 and PHEV at 48-96hrs (rho 0.451, p=0.003). Expression of miR-206 was associated with a higher odds of survival at 30-days (OR 1.149 95% CI 1.016-1.299 p=0.0266) and a favorable outcome at 365-days (OR 1.135 95% CI 1.007-1.278 p=0.038). There was no association of miR-206 expression with HE. miR-486, miR-150, and miR-146a were not associated with radiological or functional outcomes. Conclusion: In our cohort, miR-206 was associated with PHEV at 96hrs, survival at 30days and favorable outcome at 1 year. Further studies are ongoing to further delineate the role of miR206 in ICH related biological mechanisms.

Decreased Levels of miR-126 and miR-132 in Plasma and Vitreous Humor of Non-Proliferative Diabetic Retinopathy Among Subjects with Type-2 Diabetes Mellitus.

PurposeDiabetic retinopathy (DR), the leading cause of blindness among working adults, is an urgent public health problem as diabetes mellitus (DM) is increasing at an alarming rate. Hyperglycemia-induced endothelial dysfunction is the principal contributing factor leading to the development of microangiopathy. Altered levels of microRNA (miR), the negative regulator of protein-coding genes, have been observed and considered to be markers for DR. Present study aimed to find out whether miR levels in plasma could be effective biomarkers to differentiate between type 2 diabetes mellitus (T2DM) with non-proliferative retinopathy (NPDR) from T2DM with no-DR (DNR).MethodsWe recruited 50 T2DM subjects comprising 31 NPDR and 19 DNR individuals. Surrogate markers of systemic oxidative stress and vascular endothelial growth factor (VEGF) were measured in plasma. Levels of miR-126 and miR-132 were determined in plasma and vitreous fluid using real-time PCR.ResultsWe observed that levels of miR-126 and miR-132 were decreased in NPDR subjects in comparison to DNR. Plasma levels of miRs were inversely correlated with secreted levels of VEGF and oxidative stress marker. The levels of these miRs showed discriminating ability between NPDR and DNR.ConclusionCirculating miRs 126 and 132 in plasma or vitreous may serve as biomarkers for early diabetic retinopathy risk prediction, provided validated in a larger cohort and other forms of retinal vasculopathy or retinopathy in the future.

Ingestion of orange juice prevents hyperglycemia and increases plasma miR-375 expression

The intake of high-fat, high-carbohydrate (HFHC) meals is associated with an increased risk of type 2 diabetes. There is evidence that the association of orange juice to a HFHC meal can modulate the expression of microRNAs (miRNAs) linked to pancreatic β-cell function such as miR-375. We evaluated the effect of a commercial orange juice intake with HFHC meal on plasma miRNAs expression in twelve healthy subjects in a crossover design study. Subjects ingested water, orange juice, or an isocaloric beverage along with a 1037kcal HFHC meal. Blood glucose and miRNAs were evaluated at baseline and 1, 3, and 5h after the intake. The area under the curve (AUC) for glycemia after ingestion of HFHC+orange juice did not differ from ingestion of HPHC+glucose or HFHC+water. However, the AUC was higher in HFHC meal+glucose compared to HFHC meal+water (p=0.034). Glucose and insulin concentrations were significantly higher in HFHC meal+glucose group after 1h, when compared with other groups and times (p<0.001). There was an increase in plasma miR-375 expression after 3h of ingestion of HFHC+orange juice versus water (p=0.026), and a decrease in plasma miR-205-5p expression after HFHC meal+glucose versus water (p=0.023). A single HFHC meal+orange juice modulated plasma miR-375 expression, which is a biomarker of pancreatic β-cell function, and contributed to preventing hyperglycemia.

Alterations in plasma miR-21, miR-590, miR-192 and miR-215 in idiopathic pulmonary fibrosis and their clinical importance.

Many studies have revealed that microRNA (miRNA) molecules may take part in idiopathic pulmonary fibrosis (IPF). But, the role of miRNAs in the development of IPF is not yet clear. We investigated the plasma levels of miR-21, miR-590, miR-192, and miR-215 in IPF (n = 88) and healthy control (n = 20) groups in this study. We compared the expression levels of target miRNAs in patients with IPF and healthy participants. We grouped the patients with IPF according to age, forced vital capacity, carbon monoxide diffusing capacity (DLCO), gender-Age-pulmonary physiology (GAP) score, the presence of honeycombing and compared the expression levels of target miRNAs in these clinical subgroups. 82 (93.18%) of the patients with IPF were male and the mean age was 66.6 ± 8.6years. There was no significant difference between the gender and age distributions of IPF and the control group. The mean plasma miR-21 and miR-590 levels in IPF group were significantly higher than in the control group (p < 0.0001, p < 0.0001, respectively). There was no significant difference between the miR-192 and miR-215 expression levels of the IPF and control group. Both miR-21 and miR-590 correlated positively with age (p = 0.041, p = 0.007, respectively) while miR-192 and miR-215 displayed a negative correlation with age (p = 0.0002, p < 0.0001, respectively). The levels of miR-192 and miR-215 increased as the GAP score decreased. The levels of miR-192 in patients with honeycombing were significantly lower than in those without honeycombing (p = 0.003). Our study showed that both miR-21 and miR-590 were overexpressed in IPF. The miR-21 and miR-590 were associated with DLCO, while miR-192 and miR-215 were associated with the GAP score and honeycombing.

Preliminary evidence that lectins in infant soy formula apparently bind bovine milk exosomes and prevent their absorption in healthy adults

BackgroundMilk exosomes and their microRNA (miR) cargos are bioavailable. The content of exosomes and miRs is negligible in infant formulas compared to human milk, and dietary depletion of exosomes led to changes in bacterial communities and impaired gut health in juvenile mice. Adverse effects of formula feeding may be compounded by using soy formulas due to exosome binding by abundant lectins in that matrix. The purpose of this study was to assess the bioavailability of milk exosomes and their miR cargos added to soy formula in adults, as well as the potential role of soy lectins in exosome bioavailability.MethodsEleven healthy adults (6 men, 5 women) enrolled in this randomized crossover study. Participants consumed 1.0 l of soy formula without (SF) or with (SFE) bovine milk exosomes added. Concentration-time curves of six plasma miRs were analyzed using reverse transcription quantitative PCR. Lectin affinity chromatography was used to assess the binding of exosomes by soy lectins. Data were analyzed by using paired t test. P < 0.05 was considered statistically significant.ResultsConsumption of SF and SFE did not elicit postprandial increases in plasma miRs. Approximately 39% of bovine milk exosome particles were retained by lectin columns.ConclusionsWe conclude that fortification of soy formulas with milk exosomes, in the absence of removing lectins, is not a viable strategy for delivering bioavailable exosomes and their miR cargos. Lectins in soy formulas bind glycoprotein on the surfaces of milk exosomes, thereby preventing exosome absorption.Trial registrationISRCTN registry ID: 16329971. Retrospectively registered on February 7th, 2019.

Dysregulated circulating microRNA-126 in chronic obstructive pulmonary disease: linkage with acute exacerbation risk, severity degree, and inflammatory cytokines.

BackgroundMicroRNA‐126 (miR‐126) is engaged in respiratory diseases via regulating airway tissue injury and pulmonary inflammation, while its relation with chronic obstructive pulmonary disease (COPD) is not reported. The study aimed to evaluate the value of miR‐126 for estimating COPD acute exacerbation risk and its relation to disease severity and inflammatory cytokines in COPD patients.MethodsThis study was a case–control study. Seventy acute exacerbation COPD (AECOPD) patients, 70 stable COPD (SCOPD) patients, and 70 healthy controls (HCs) were consecutively recruited. Plasma miR‐126 expression was detected by reverse transcription quantitative polymerase chain reaction. Plasma tumor necrosis factor α (TNF‐α), interleukin‐1β (IL‐1β), interleukin‐6 (IL‐6), and interleukin‐17 (IL‐17) in COPD patients were further determined by enzyme‐linked immunosorbent assay.ResultsMiR‐126 was higher in AECOPD patients compared to SCOPD patients and HCs (both P adj < 0.001). Receiver operating characteristic curves revealed miR‐126 distinguished AECOPD patients from SCOPD patients (area under curve (AUC): 0.805, 95%CI: 0.733–0.877) and HCs (AUC: 0.884, 95%CI: 0.829–0.939) and also distinguished SCOPD from HCs (AUC = 0.656, 95%CI: 0.566–0.747). MiR‐126 positively related to GOLD stage in both AECOPD patients (p < 0.001) and SCOPD patients (p < 0.001). Furthermore, miR‐126 positively linked with TNF‐α (p < 0.001), IL‐1β (p = 0.002), IL‐6 (p = 0.009), and IL‐17 (p < 0.001) levels in AECOPD patients; but miR‐126 only positively related to TNF‐α and IL‐17 levels (all p < 0.050), instead of IL‐1β or IL‐6 level (all p > 0.050) in SCOPD patients and HCs.ConclusionDysregulated circulating miR‐126 not only relates to COPD susceptibility and its acute exacerbation risk but also links with disease severity and inflammatory cytokines in COPD patients.

Overexpression of miR-126 Protects Hypoxic-Reoxygenation-Exposed HUVEC Cellular Injury through Regulating LRP6 Expression.

The aim of the study was to explore the clinical impact of circulatory miR-126 as a candidate for novel biomarker in patients with coronary artery disease (CAD) and its protective role against hypoxia/reoxygenation- (H/R-) exposed HUVEC cellular injury. A total of 278 subjects, which included 153 subjects with angiographically confirmed CAD, 70 unstable angina subjects, and 55 healthy individuals, along with 18-hour HR-induced HUVECs were recruited in this study. Plasma miR-126 levels were significantly downregulated in stable and unstable CAD patients as well as 18-hour HR-exposed HUVECs as compared with controls. Stable and unstable CAD subjects were significantly differentiated from healthy individuals with a predictive value of AUC 0.903 and 0.923, respectively. Moreover, peripheral circulatory miR-126 expressions in elderly (71-90 years) stable and unstable CAD patients were comparatively lower than younger (30-50 years) subjects. The caspase-3 activity, intracellular ROS concentrations, and cellular viabilities were evidently increased in 18-hour HR-exposed HUVECs than in normal cells (P < 0.001). On the contrary, mimic expressions of miR-126 prominently reduced caspase-3 activity and intracellular ROS levels and markedly enhanced HUVEC cellular viabilities (P < 0.001). LRP6 expressions were significantly elevated in HR-induced HUVECs, whereas overexpression of miR-126 remarkably decreased LRP6 expressions (P < 0.001). Plasma miR-126 could be used as a novel biomarker for early prediction of CAD subjects. Overexpression of miR-126 significantly improved HUVEC cellular viabilities by downregulation of LRP6 protein expression, suggesting a potential therapeutic target for CAD patients.

MicroRNA Expression in Plasma of Esophageal Squamous Cell Carcinoma Patients.

BackgroundPatients with esophageal squamous cell carcinoma (ESCC) have a poor prognosis and there are no effective clinical biomarkers. Recently, stable microRNAs detected in the blood have been suggested as potential biomarkers in various cancers. Therefore, we investigated whether plasma microRNAs could be feasible biomarkers for ESCC.MethodsPeripheral blood samples were obtained from 16 healthy volunteers and 66 ESCC patients before treatment between May 2016 and April 2021. Plasma miR-18b, miR-21, miR-31, and miR-375 expression levels were measured using reverse transcription-quantitative polymerase chain reaction.ResultsCompared with those in healthy controls, the expression levels of plasma miR-21 were significantly higher (P = 0.022) and those of plasma miR-31 and miR-375 were significantly lower in ESCC patients (both P < 0.001). Plasma miR-18b expression levels increased in ESCC patients, but the difference was not significant (P = 0.164). The sensitivities and specificities of miR-21, miR-31, and miR-375 for differentiating ESCC patients from healthy controls were 87.5% and 61.9%, 87.5% and 98.4%, and 87.5% and 100%, respectively. There was no difference in expression levels of plasma miR-21, miR-31, and miR-375 according to clinicopathological characteristics of sex, age, tumor size and location, histologic grade, and tumor-node-metastasis stage.ConclusionOur study demonstrated that plasma miR-21, miR-31, and miR-375 could be potential biomarkers for the diagnosis of ESCC. Particularly, plasma miR-31 and miR-375 showed high sensitivity and specificity for differentiating ESCC patients from healthy controls.

Plasma miR-21 as a potential predictor in prediabetic individuals with a positive family history of type 2 diabetes mellitus.

Type 2 diabetes mellitus (T2DM) is a heritable metabolic perturbation, rapidly growing across the world. Primary recognition of susceptible individuals with a family history of type 2 diabetes (FHD) in the prediabetes stage could delay the onset of T2DM or reduce complications induced by diabetes. This study aims to evaluate the expression levels of miR‐21, miR‐126 as noninvasive predictive biomarkers in individuals with genetic predisposition and investigate the correlation of miRNAs and cardiometabolic risk factors. Our study demonstrated that miR‐21 expression has a notable elevate in both groups of T2DM and pre‐T2DM. miR‐21 expression was distinguished in the pre‐T2DM and T2DM from the nondiabetic individuals by ROC curve analysis with AUC of 0.77 (95% CI 0.65–0.90; p = 0.0004) and AUC of 0.78 (95% CI 0.64–0.92; p = 0.0042), respectively. The relative gene expression of miR‐126 was nearly equal among groups. miR‐21 expression was positively associated with glycosylated hemoglobin (HbA1c), fasting blood sugar (FBS), and triglyceride (TG) and might have diagnostic value for T2DM and pre‐T2DM. This study has revealed that the expression level of miR‐21 can be considered as a non‐invasive and rapid tool for distinguishing pre‐T2DM and T2DM counterparts from healthy individuals.

The Expression and Clinical Value of miR-221 and miR-320 in the Plasma of Women with Gestational Diabetes Mellitus.

The goal was to investigate the expression of plasma miR-221 and miR-320 in gestational diabetes mellitus (GDM) and to further explore the relationship between miRNA and risk factors for GDM. This study included 85 GDM and 85 age-matched normal pregnant women who visited our hospital from January 2019 to January 2020. Real-time polymerase chain reaction (RT-qPCR) was used to determine the expression of miR-221 and miR-320 in the plasma of pregnant women. The correlation analysis was used to detect the relationship between miR-221, miR-320, and risk factors of GDM, including homeostatic model assessment for insulin resistance (HOMA-IR), percentage of glycosylated hemoglobin (HbA1c), and the prepregnancy BMI. The receiver operating characteristic curve (ROC) was used to determine the diagnostic value of miR-320 and miR-221 in GDM. Compared with normal pregnant women, the expression of miR-221 and miR-320 in GDM was significantly higher (p < 0.05). The results also demonstrate that the expression of miR-221 and miR-320 increases grad-ually with the development of pregnancy in GDM at 24 weeks, 28 weeks, and 32 weeks (p < 0.05). Spearman's correlation analysis confirmed that the expression level of miR-221 and miR-320 in the plasma of GDM is positively correlated with the HOMA-IR and HbA1c, but has no significant correlation with pre-pregnancy BMI. The area under the curve (AUC) values of miR-221 and miR-320 were 0.862 and 0.853, respectively. Meanwhile, the area under the combined detection curve is 0.904. Plasma miR-221 and miR-320 are significantly elevated in GDM, and are positively correlated with HbA1c and HOMA-IR. The high expression of miR-221 and miR-320 in the peripheral plasma of pregnant women may directly or indirectly participate in the occurrence and development of GDM or may become a new target for the diagnosis, treatment, and prognosis of GDM.

Decrease in Plasma miR-27a and miR-221 After Concussion in Australian Football Players

Introduction:Sports-related concussion (SRC) is a common form of brain injury that lacks reliable methods to guide clinical decisions. MicroRNAs (miRNAs) can influence biological processes involved in SRC, and measurement of miRNAs in biological fluids may provide objective diagnostic and return to play/recovery biomarkers. Therefore, this prospective study investigated the temporal profile of circulating miRNA levels in concussed male and female athletes.Methods:Pre-season baseline blood samples were collected from amateur Australian rules football players (82 males, 45 females). Of these, 20 males and 8 females sustained an SRC during the subsequent season and underwent blood sampling at 2-, 6- and 13-days post-injury. A miRNA discovery Open Array was conducted on plasma to assess the expression of 754 known/validated miRNAs. miRNA target identified were further investigated with quantitative real-time PCR (qRT-PCR) in a validation study. Data pertaining to SRC symptoms, demographics, sporting history, education history and concussion history were also collected.Results:Discovery analysis identified 18 candidate miRNA. The consequent validation study found that plasma miR-221-3p levels were decreased at 6d and 13d, and that miR-27a-3p levels were decreased at 6d, when compared to baseline. Moreover, miR-27a and miR-221-3p levels were inversely correlated with SRC symptom severity.Conclusion:Circulating levels of miR-27a-3p and miR-221-3p were decreased in the sub-acute stages after SRC, and were inversely correlated with SRC symptom severity. Although further studies are required, these analyses have identified miRNA biomarker candidates of SRC severity and recovery that may one day assist in its clinical management.

Correlation of Plasma MiR-146a, MiR-223 Levels with the Prognosis of Childhood with Acute Lymphoblastic Leukemia

To observe the expression of plasma microRNA (miR)-146a and miR-223 in children with acute lymphoblastic leukemia (ALL), so as to analyze the relationship between the two factors and the prognosis of children with ALL. 100 children with ALL treated in the hospital from January 2015 to December 2017 were selected, according to the standard of Chinese Children's Leukemia Group (CCLG)-ALL-2008 program, the children were performed standardized treatment in our hospital according to different risk degree, the follow-up results were obtained, the follow-up time was ≥36 months, and the follow-up time was till to March 2021, the recurrence and mortality of the children were used as prognostic indicators; the baseline data of the children at admission were inquired and recorded, the plasma miR-146a and miR-223 levels were analyzed at admission, and their correlation with the prognosis of children with ALL was analyzed. During the follow-up period, 4 cases of children died while 18 cases recurred, which means 22(22.00%) children showed the poor prognosis; the plasma miR-146a level of the children in poor prognosis group at admission was higher than those in good prognosis group, while the plasma miR-223 level was lower than those in good prognosis group, the differences showed statistically significantly (P<0.05); the results of regression analysis showed that the over expression of plasma miR-146a and low expression of plasma miR-223 at admission might be associated with poor prognosis in ALL children, and might be a risk factor for poor prognosis in children (P<0.05); the ROC curve showed that the AUC of plasma miR-146a and miR-223 at admission alone or in combination showed the predictive value for the risk of poor prognosis in children with ALL(AUC >0.80); the results of correlation test showed that there was a negative correlation of plasma miR-146a with miR-223 levels at admission (r=-0.239, P=0.016). Plasma miR-146a is overexpressed and miR-223 is low-expressed in children with ALL, the abnormal expression of the two factors is related to the prognosis of children with ALL.

Differences in plasma miRNA levels in inferior petrosal sinus samples of patients with ACTH-dependent Cushing's syndrome

For the last decades microRNAs (miR) have proven themselves as novel biomarkers for various types of diseases. Identification of specific circulating microRNA panel that differ patient with Cushing's disease (CD) and ectopic ACTH syndrome (EAS) could improve the diagnostic procedure. to evaluate the differences in miR levels in plasma samples drained from inferior petrosal sinuses in patients with CD and EAS. single-center, case-control study: we enrolled 24 patients with ACTH-dependent Cushing's syndrome (CS) requiring bilateral inferior petrosal sinus sampling (BIPSS). Among them 12 subjects were confirmed as CD (males=2, females=10; median age 46,5 [IR 33,8;53,5]) and 12 as EAS (males=4, females=8, median age54 [IR 38,75;60,75]). BIPSS was performed through a percutaneous bilateral approach. Once catheters were properly placed, blood samples were withdrawn simultaneously from each petrosal sinus and a peripheral vein. Plasma samples from both sinuses were centrifuged and then stored at -80 C. MiRNA isolation from plasma was carried out by an miRneasy Plasma/Serum Kit (Qiagen, Germany) on the automatic QIAcube station according to the manufacturer protocol. To prevent degradation, we added 1 unit of RiboLock Rnase Inhibitor (Thermo Fisher Scientific, USA) per 1 μL of RNA solution. The concentration of total RNA in the aqueous solution was evaluated on a NanoVue Plus spectrophotometer (GE Healthcare, USA). The libraries were prepared by the QIAseq miRNA Library Kitfollowing the manufacturer standard protocols. MiR expression was then analyzed by sequencing on Illumina NextSeq 500 (Illumina, USA). 108 miRNAs were differently expressed (p&lt;0,05) in inferior petrosal sinus samples of patients with CD vs EAS. We divided these miRNAs into 3 groups based on the significance of the results. The first group consisted of samples with thehighest levels of detected miR in both groups. Four miRNAs were included:miR-1203 was downregulated inCD vs EAS- 36.74 (p=0,013), and three other were upregulated in CD vs EAS: miR-383-3p46.36 (p=0,01), miR-42906.84 (p=0,036),miR-6717-5p 4.49 (p=0,031). This miRs will be validated in larger cohorts using RT-qPCR. Plasma miR levels differ in inferior petrosal samples taken from patients with CD vs EAS. These miRs need to be validated by different methods and in peripheral plasma samples in order to be used as potentially non-invasive biomarkers to differentiate ACTH-dependent CS.

Circulating plasma miR-155 is a potential biomarker for the detection of SARS-CoV-2 infection

Identification of human miRNAs involved in coronavirus-host interplay is important due to the current COVID-19 pandemic. Therefore, this study aimed to measure the circulating plasma miR-155 expression level in COVID-19 patients and healthy controls to investigate its roles in the pathogenesis and severity of COVID-19 disease and to assess its usefulness as a clinical biomarker for the detection of COVID-19 disease and the severity of infection.A total of 150 COVID-19 patients and 50 controls were enrolled into our study. Beside the routine laboratory work and chest computed tomography (CT) scans of COVID-19 patients, plasma miR-155 expression level was measured using reverse transcription quantitative real-time PCR (RT-qPCR) technique.Our results demonstrated increased miR-155 expression level in COVID-19 patients compared to controls, in severe compared to moderate COVID-19 patients, and in non-survival compared to survival COVID-19 patients. miR-155 expression level also had significant correlation with clinicopathological characteristics of COVID-19 patients such as chest CT findings, CRP, ferritin, mortality, D-dimer, WBC count, and lymphocytes and neutrophils percentages. Also, our results showed that the area under the curve (AUC) for miR-155 was 0.986 with 90% sensitivity and 100% specificity when used as a biomarker for the detection of COVID-19 disease; while in detection of severity of COVID-19 disease, AUC for miR-155 was 0.75 with 76% sensitivity and specificity.From these results we can conclude that miR-155 has a crucial role in the pathogenesis and severity of COVID-19; also, it could be a good diagnostic clinical biomarker for the detection of COVID-19 disease and the severity of infection.

Increased Let-7e expression is associated with long-term all-cause mortality and antiplatelet treatment in patients with type 2 diabetes mellitus

Abstract Background In the light of growing prevalence of type 2 diabetes mellitus (T2DM), efforts are made to discover novel biomarkers. MicroRNAs (miRNAs-miR) are non-coding RNAs used in various processes involved in regulating gene expression which play a role in platelet function. Purpose To analyze the ability of platelet-derived miRNAs in prediction of mortality and response to antiplatelet treatment among T2DM-patients. Methods 252 diabetic subjects were enrolled and were receiving either acetylsalicylic acid (ASA) 75mg (65%) or 150 mg (15%) or clopidogrel (19%). Plasma miR-126, miR-223, miR-125a-3p and Let-7e expressions were assessed by qRT-PCR and compared between the patients who survived and those who died. Median observation time was 5.9 years. Adjusted Cox-regression analysis was used for prediction of mortality. Differential miRNAs expression due to different antiplatelet treatment was analyzed. Results ROC curve analysis revealed increasing concentrations of miR-126, Let-7e and miR-125a-3p levels had a diagnostic ability for prediction of long-term all-cause mortality (c-index=0.75, p&amp;lt;0.001; 0.72, p&amp;lt;0.001; 0.72, p=0.001, respectively). Multivariate Cox regression model revealed high miR-126 and Let-7e expressions which were strong and independent predictors of all-cause long-term mortality (HR=5.08, 95% CI: 1.92–13.43; p=0.001; HR=5.94, 95% CI: 1.98–17.79; p=0.001,respectively). After including all miRNAs into one multivariate Cox regression model, only Let-7e was predictive of future occurrence of long-term all-cause death (HR=7.83, 95% CI: 1.2–51.1; p=0.032). 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). Conclusions Let-7e expression is a strong and independent predictor of long-term all-cause mortality among patients with T2DM. MiR-223, miR-126 and Let-7e present significant interactions with antiplatelet treatment and clinical outcomes. Funding Acknowledgement Type of funding sources: None. Figure 1Table 1

Clinical significance of microRNA-126 in Coronary artery disease patients

Abstract Background Coronary artery disease (CAD) is major public health problem and the leading cause of premature death all over the world. Early diagnosis with adequate management of CAD patients can significantly reduce morbidity and mortality rate. Purpose The purpose of this study was to explore the clinical significance of plasma miR-126 as a biomarker of stable angina pectoris patients and investigated the protective effects of hypoxic exposed HUVEC cell injury. Methods Angiographically confirmed 127 stable coronary artery (CAD) patients, 55 healthy subjects and 18hours hypoxic (1% O2) HUVEC cells were included in this study. Results The expression of miR-126 levels were remarkably down-regulated in stable CAD patients and hypoxic exposed HUVEC cells as compared with controls (p&amp;lt;0.001). Circulating plasma miR-126 was able to accurately differentiated stable CAD patients from healthy subjects with high sensitivity and specificity (AUC 0.928). The caspase-3 activity, apoptosis rate, LDH levels and intracellular ROS generation were significantly increased in 24hours hypoxic exposed HUVEC cells than normal control cells (p&amp;lt;0.001). On the contrary, over expressions of miR-126 were evidently reduced caspase-3 activity, apoptosis rate, LDH levels, intracellular ROS production and significantly improved HUVEC cellular viability (p&amp;lt;0.001). Moreover, TNF receptor-associated factor 6 (TRAF6) expressions were markedly increased in Hypoxic induced HUVEC cells, whereas mimic expression of miR-126 significantly decreased TRAF6 expression. Conclusion Down-regulated miR-126 may consider as a potential risk factor for stable CAD patients and it may used as a possible biomarker for early evaluation of CAD patients. Mimic expression of miR-126 remarkably decreased caspase-3 activity, LDH secretions, ROS levels and markedly improved HUVEC cellular viability by down-regulating TRAF6 expression, suggesting a newer therapeutic target for CAD patients. Funding Acknowledgement Type of funding sources: None.

Utility of miR-155, miR-21 and miR-16 in plasma as biomarkers of atrial fibrillation

Abstract Background Atrial fibrillation (AF) is associated with increased morbidity and mortality. MicroRNAs (miRs) are post-transcriptional regulators that could be useful in the study of AF. Purpose To investigate whether miR-16, miR-21 and miR-155 may differentiate between AF and sinus rhythm. Methods 29 controls and 42 AF (June 2019-May 2020). Lipid profile, hsCRP were measured. Serum RNA extraction was performed. Hsa-miR-21, hsa-miR-16, hsa-miR-155, and cel-miR-39 were amplified RT-PCR. Quantitative results for miR levels were generated according to the 2-ΔCt method using cel-miR-39 for normalization. Correlations were assessed by Spearman rank test. Logistic regression analysis was adjusted by age, sex, and BMI vs miR's. A p&amp;lt;0.05 was set for significance. Results (controls vs AF): age (56 [52–63] vs 60 [64–67] years, p=0.122), BMI (30.52 [27.69–32.67] vs 29.65 [26.3–32.9] kg/m2 p=0.779), female 17 (58.6%) vs 19 (45.2%), p=0.268; total cholesterol (198.4 [17.2–215] vs 171.3 [150.3–216.8] mg/dL, p=0.066), LDL (126.6 [114.7–141.6] vs 99.2 [80.7–124] mg/dL, p=0.005) HDL (40.4 [36.8–45.8] vs 43.8 [36.9–48.5] p=0.326) and CRP (2.69 [1.22–4.72] vs 2.92 [1.4–4.76] mg/L p=0.835). The CHA2DS2VASC score in AF was &amp;lt;2=11 (45.2%) and ≥2= 26 (54.8%). MicroRNA levels (controls vs AF): miR-21 0.0195 (0.0095–0.0275) vs 0.042 (0.028–0.098) p&amp;lt;0.001; miR-155 0.0178 (0.0108–0.0309) vs 0.01 (0.0056–0.0169) p&amp;lt;0.001, and miR-16 0.5238 (0.2618–1.992) vs 0.1652 (0.0765–0.3816) p&amp;lt;0.001. In the logistic regression miR-21 had a positive correlation with AF, c=0.851 p=0.002, Wald=9.89. miR-16 and miR-155 a negative relation (c=0.76,0.72; p=0.036, 0.043, Wald=4.39, 4.085; respectively) (Fig. 1A, B, C). For multivariable analysis we used miR-16, miR-21, and miR-155 as previous analysis demonstrated significance with an overall model c=0.90 p=0.028 (Wald=14.2) and significance was seen in the miR-21 Wald= 9.33 (p=0.002); and miR-155 Wald=4.7 (p=0.03); miR-16 was not significant (Wald=2.7, p=0.1). A ternary plot was performed using the predicted outputs of the model. (Fig. 2) Conclusion miR-16, miR-21 and miR-155 are useful for the discrimination of AF vs sinus rhythm and may be part of important physiopathological pathways in AF. Funding Acknowledgement Type of funding sources: Public hospital(s). Main funding source(s): Instituto Nacional de Cardiología Ignacio Chávez Figure 1. Logistic regression curvesFigure 2. Multivariate ternary plot of miRs