miRNAs In Patients Research Articles

Differential Expression of Serum Exosome microRNAs and Cytokines in Influenza A and B Patients Collected in the 2016 and 2017 Influenza Seasons.

MicroRNAs (miRNAs) have remarkable stability and are key regulators of mRNA transcripts for several essential proteins required for the survival of cells and replication of the virus. Exosomes are thought to play an essential role in intercellular communications by transporting proteins and miRNAs, making them ideal in the search for biomarkers. Evidence suggests that miRNAs are involved in the regulation of influenza virus replication in many cell types. During the 2016 and 2017 influenza season, we collected blood samples from 54 patients infected with influenza and from 30 healthy volunteers to identify the potential role of circulating serum miRNAs and cytokines in influenza infection. Data comparing the exosomal miRNAs in patients with influenza B to healthy volunteers showed 76 miRNAs that were differentially expressed (p < 0.05). In contrast, 26 miRNAs were differentially expressed between patients with influenza A (p < 0.05) and the controls. Of these miRNAs, 11 were commonly expressed in both the influenza A and B patients. Interferon (IFN)-inducing protein 10 (IP-10), which is involved in IFN synthesis during influenza infection, showed the highest level of expression in both influenza A and B patients. Influenza A patients showed increased expression of IFNα, GM-CSF, interleukin (IL)-13, IL-17A, IL-1β, IL-6 and TNFα, while influenza B induced increased levels of EGF, G-CSF, IL-1α, MIP-1α, and TNF-β. In addition, hsa-miR-326, hsa-miR-15b-5p, hsa-miR-885, hsa-miR-122-5p, hsa-miR-133a-3p, and hsa-miR-150-5p showed high correlations to IL-6, IL-15, IL-17A, IL-1β, and monocyte chemoattractant protein-1 (MCP-1) with both strains of influenza. Next-generation sequencing studies of H1N1-infected human lung small airway epithelial cells also showed similar pattern of expression of miR-375-5p, miR-143-3p, 199a-3p, and miR-199a-5p compared to influenza A patients. In summary, this study provides insights into the miRNA profiling in both influenza A and B virus in circulation and a novel approach to identify the early infections through a combination of cytokines and miRNA expression.

Serum exosomal miR-638 is a prognostic marker of HCC via downregulation of VE-cadherin and ZO-1 of endothelial cells.

Hepatocellular carcinoma (HCC) is the second leading cause of cancer‐related death. High recurrence rates after curative resection and the lack of specific biomarkers for intrahepatic metastases are major clinical problems. Recently, exosomal microRNAs (miRNAs) have been reported to have a role in the formation of the pre‐metastatic niche and as promising biomarkers in patients with malignancy. Here we aimed to clarify the molecular mechanisms of intrahepatic metastasis and to identify a novel biomarker miRNA in patients with HCC. A highly intrahepatic metastatic cell line (HuH‐7M) was established by in vivo selection. HuH‐7M showed increased proliferative ability and suppression of apoptosis and anoikis. HuH‐7M and the parental cell (HuH‐7P) showed the similar expression of epithelial‐mesenchymal transition markers and cancer stem cell markers. In vivo, mice treated with exosomes derived from HuH‐7M showed increased tumorigenesis of liver metastases. Exosomes from HuH‐7M downregulated endothelial cell expression of vascular endothelial‐cadherin (VE‐cadherin) and zonula occludens‐1 (ZO‐1) in non‐cancerous regions of liver and increased the permeability of FITC‐dextran through the monolayer of endothelial cells. The miRNAs (miR‐638, miR‐663a, miR‐3648, and miR‐4258) could attenuate endothelial junction integrity by inhibiting VE‐cadherin and ZO‐1 expression. In patients with HCC, higher serum exosomal miR‐638 expression was associated with tumor recurrence. In conclusion, the miRNAs secreted from a highly metastatic cancer cell can promote vascular permeability via downregulation of endothelial expression of VE‐cadherin and ZO‐1. Serum exosomal miR‐638 expression holds potential for serving as a significant and independent prognostic marker in HCC.

Diagnostic and therapeutic potential of exosomal miRNAs in Alzheimer's disease.

Diagnostic and therapeutic potential of exosomal miRNAs in Alzheimer's disease.

Circulating miRNA in Patients Undergoing Total Pancreatectomy and Islet Autotransplantation

Circulating microRNAs (miRNAs) can be biomarkers for diagnosis and progression of several pathophysiological conditions. In a cohort undergoing total pancreatectomy with islet autotransplantation (TPIAT) from the multicenter Prospective Observational Study of TPIAT (POST), we investigated associations between a panel of circulating miRNAs (hsa-miR-375, hsa-miR-29b-3p, hsa-miR-148a-3p, hsa-miR-216a-5p, hsa-miR-320d, hsa-miR-200c, hsa-miR-125b, hsa-miR-7-5p, hsa-miR-221-3p, hsa-miR-122-5p) and patient, disease and islet-isolation characteristics. Plasma samples (n = 139) were collected before TPIAT and miRNA levels were measured by RTPCR. Disease duration, prior surgery, and pre-surgical diabetes were not associated with circulating miRNAs. Levels of hsa-miR-29b-3p (P = 0.03), hsa-miR-148a-3p (P = 0.04) and hsa-miR-221-3p (P = 0.01) were lower in those with genetic risk factors. Levels of hsa-miR-148a-3p (P = 0.04) and hsa-miR-7-5p (P = 0.04) were elevated in toxic/metabolic disease. Participants with exocrine insufficiency had lower hsa-miR-29b-3p, hsa-miR-148a-3p, hsa-miR-320d, hsa-miR-221-3p (P < 0.01) and hsa-miR-375, hsa-miR-200c-3p, and hsa-miR-125b-5p (P < 0.05). Four miRNAs were associated with fasting C-peptide before TPIAT (hsa-miR-29b-3p, r = 0.18; hsa-miR-148a-3p, r = 0.21; hsa-miR-320d, r = 0.19; and hsa-miR-221-3p, r = 0.21; all P < 0.05), while hsa-miR-29b-3p was inversely associated with post-isolation islet equivalents/kg and islet number/kg (r = −0.20, P = 0.02). Also, hsa-miR-200c (r = 0.18, P = 0.03) and hsa-miR-221-3p (r = 0.19, P = 0.03) were associated with islet graft tissue volume. Further investigation is needed to determine the predictive potential of these miRNAs for assessing islet autotransplant outcomes.

The importance of dysregulated miRNAs on ovarian cysts and epithelial ovarian cancer

Objective: Benign ovarian cysts (BOC) are the most common tumors in women of reproductive age. Usually, these cysts are harmless, but, a small number of them occasionally progress to malignancy. Among ovarian malignancies, epithelial ovarian cancer (EOC) comprises 90% and is the most important cause of gynecologic cancer-related deaths. We aimed to identify dysregulated miRNAs in patients with benign ovarian cysts (n = 11) compared to EOC (n = 10) and to healthy individuals (HI) (n = 15). Methods: The serum samples from EOC and BOC patients were collected before operation. We studied three different sample groups (serum of EOC (n = 8), HI (n = 8), and BOC (n = 8) patients) that contained the highest-quality of RNA. Microarray data were analyzed according to expression of miRNAs and target genes by bioinformatics tools. Results: When compared to EOC samples, 75 miRNAs were dysregulated in BOC samples. Sixty-six miRNAs from BOC were dysregulated when compared to HI samples. Bioinformatics analysis of BOC vs. EOC and BOC vs. HI showed that 46 miRNAs were congruent and their expression alterations were similar (up- or down-regulated). Further analysis showed that these 46 miRNAs are associated to one of three pathways involved in cancer pathogenesis. Conclusion: Several miRNAs might play a role in BOC formation and/or malignant transformation. These dysregulated miRNAs could potentially be a biomarker to distinguish between a completely BOC and one that is malignant or has potential for malignant transformation.

Plasma circulating microRNAs in patients with stable coronary artery disease - impact of different cardiovascular risk profiles and glomerular filtration rates

Plasma circulating microRNA (miRNA)-126, -145, and -155 are associated with vascular remodeling, atherosclerotic lesion formation, and plaque vulnerability. In this study, we evaluated the levels of plasma circulating miRNAs in patients with stable coronary artery disease (CAD), different cardiovascular risk profiles, and different glomerular filtration rates (GFR). Forty patients with stable CAD admitted for elective percutaneous coronary intervention (PCI) were enrolled in a prospective study. Before PCI, fasting blood samples were obtained to evaluate clinical parameters and miRNA-126 and miRNA-155 expression. The GFR was calculated by the MDRD and CKD-EPI formulas, and the severity of CAD was calculated according to the SYNTAX score. All these parameters were correlated with miRNAs. The association between miRNA levels and clinical characteristics was evaluated. The expression of miRNA-126 positively correlated with a higher SYNTAX score (r = 0.337; p=0.034); however, no significant correlations between miR-126, GFR, and clinical characteristics were observed. Higher plasma levels of miRNA-155 correlated with increased levels of triglycerides (r = 0.317; P = 0.049), C-peptide (r = 0.452; P = 0.011), and the HOMA index (r = 0.447; P = 0.012) and a higher body mass index (BMI) (r = 0.385; P = 0.015). GFR and miRNA-155 (MDRD - Rho=0.353; P = 0.027. CKD-EPI - Rho=0.357; P = 0.026) were found to have a moderate correlation, although miRNA-155 had no correlation with the SYNTAX score. Plasma circulating miRNA-126 levels were increased in patients with severe atherosclerosis as determined by the SYNTAX score. Elevated miRNA-155 expression was observed in patients with Stage 1 GFR but was lower in patients with Stages 2 and 3 GFR. Plasma circulating miRNA-155 had positive correlations with higher levels of BMI, HOMA index, C-peptide, and triglycerides. Although further investigations are needed to confirm the role of miRNA-155 and miRNA-126, they may serve as potential biomarkers detecting severity of CAD, lowering of kidney function and metabolic syndrome.

Discovery of extracellular vesicles derived miR-181a-5p in patient's serum as an indicator for bone-metastatic prostate cancer.

Purpose: To identify extracellular vesicle (EV)-delivered microRNAs in the patient's serum as indicators for bone-metastatic prostate cancer.Methods: First, the profiling change of serum EV-delivered miRNAs in patients with either benign prostatic hyperplasia (BPH), non-bone metastatic prostate cancer or bone-metastatic prostate cancer was detected by microRNA deep sequencing assay and microRNA-chip array assay, respectively. Second, the candidates were further confirmed using TaqMan microRNA assay in two independent validation cohorts of total 176 patients with either BPH, non-bone metastatic prostate cancer or bone metastatic prostate cancer to seek the most valuable microRNA(s).Results: Through microRNA deep sequencing and microRNA-chip array, we found 4 prospective EV-delivered miRNAs including miR-181a-5p with significantly upregulated expression in bone metastatic groups than in non-bone metastatic prostate cancer groups (p < 0.05). In the validation cohorts, logistic regression analysis was performed to evaluate the diagnostic association of candidates with bone metastasis, which indicated that miR-181a-5p was significantly associated with bone metastatic prostate cancer. Furthermore, accuracy estimate of each candidate for the diagnosis of bone metastatic prostate cancer was quantified using the area under the receiver-operating characteristic curve (AUC), which identified miR-181a-5p as the best biomarker with the AUCs of 85.6% for diagnosis of prostate cancer and 73.8% for diagnosis of bone metastatic prostate cancer.Conclusion: EV-delivered miR-181a-5p from patient's serum is a promising diagnostic biomarker for bone metastatic prostate cancer.

Assessment of plasma microRNA potentials as a non-invasive biomarker in patients with axial spondyloarthropathy.

It is assumed that abnormally expressed MicroRNAs (miRNAs) may be present in the plasma of patients with radiographic axial spondyloarthropathy (rad-AxSpA). Thus, the present study was conducted with the aim of investigating the expression profile of miRNAs in patients with rad-AxSpA. A total of 15 patients diagnosed with rad-AxSpA according to the Assessment of the SpondyloArthritis International Society (ASAS) classification criteria and nine healthy controls matched for age and gender were included in the study. Demographic data were collected, and disease activity was evaluated using the Bath Ankylosing Spondylitis Disease Activity Index (BASDAI). Peripheral blood samples were collected, and miRNAs were extracted. The expression of microRNAs was analyzed using quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) by the miScript miRNA PCR Array Human Inflammatory Response and Autoimmunity. A total of 84 miRNA profiles were evaluated, and expressions in the study and control groups were compared. When compared to the control group, 6 miRNAs (miR-125b-5p, miR-144-3p, miR-19a-3p, miR-20a-5p, miR-29c-3p, miR-30b-5p) were detected to be upregulated, and 42 miRNAs were detected to be downregulated in the rad-AxSpA group. A p-value < 0.05 was accepted as statistically significant. A significant association was found between miR-145-5p and BASDAI (p = 0.04941). MiR-144-3p, miR-302b-3p, miR-381-3p, miR-497-5p, miR-511-5p, and miR-9-5p were found to be significantly upregulated in the HLA-B27+ patients (p = 0.03063). Abnormal miRNA expressions were detected in the plasma of the patients with rad-AxSpA. It was concluded that comprehensive studies should be continued to define these miRNAs as diagnostic biomarkers for rad-AxSpA in order to detect its association with Ankylosing Spondylitis disease activity.

The prognostic impact of circulating miRNAs in patients with advanced esophagogastric cancer during palliative chemotherapy

The prognosis of patients with advanced oesophageal cancer (EC) and gastric cancer (GC) is poor. Circulating microRNAs (ci-miRNAs) may have prognostic and predictive value to improve patient selection for palliative treatment. The purpose of this study is to assess the prognostic and predictive value of specific ci-miRNAs in plasma of patients with EC and GC treated with first-line palliative gemcitabine and cisplatin. Droplet digital PCR (ddPCR) was used to quantify miR-200c-3p, miR-375, miR-21-5p, miR-148a-3p, miR-146a-5p, miR-141-3p and miR-218-5p in plasma from 68 patients. ci-miRNA expression was analyzed in relation to overall survival (OS), progression-free survival (PFS), and response to chemotherapy. ci-miRNA levels were detectable in 36 baseline (71%) samples and in 14 (47%) follow-up samples. Increased circulating miR-200c-3p in GC showed a trend (p = 0.06) towards a shorter OS. High circulating miR-375 was associated with a longer OS (p = 0.02) in patients with esophageal adenocarcinoma (EAC). No significant difference was observed in ci-miRNA expression between paired pre- and on-treatment samples. ci-miRNA expression was not associated with response to chemotherapy. ci-miRNAs can be measured in plasma samples of patients treated with first-line palliative chemotherapy using ddPCR despite prolonged storage in heparin. Elevated circulating miR-375 might be a prognostic marker for patients with EAC.

Decreased bone marrow regulatory innate lymphoid cells show a distinctive miRNA profiling in aplastic anemia

ABSTRACT Objectives A new regulatory subpopulation of innate lymphoid cells (ILCs), regulatory innate lymphoid cells (ILCregs), has been identified with both innate lymphoid cells and regulatory cells characteristics. The purpose of this study is to explore ILCregs and its associated miRNAs in patients with aplastic anemia (AA) by evaluating ILCregs frequency, associated miRNA quantification, and their significance. Methods Using 4 color combinations of surface and intracellular antibody staining, the CD45+Lin−CD127+IL-10+ ILCregs from 30 healthy donors and 30 patients newly diagnosed with AA were measured by flow cytometry. Bone marrow cells were studied by next-generation sequence miRNAs quantification. Results Our results showed that the frequency of ILCregs in bone marrow cells from healthy donors (HD) and AA patients were 0.703 ± 0.941 and 0.171 ± 0.233%, respectively. The frequencies of ILCregs in AA patients were significantly lower than that in HD (p <0.05). miRNA detection results showed different expression patterns in the AA patient group comparing with HD. Comparing with HD, there were 52 miRNAs up-regulated and 130 miRNAs down-regulated from AA patients. Analysis of miRNAs from ILCregs associated genes demonstrated different miRNAs expression patterns between HD and AA patient. Conclusion The present study demonstrated the deficiency of ILCregs and differential expression pattern of ILCregs gene-related miRNA in patients with AA. Further studies need to be done to explore the clinical significance of ILCregs and related miRNAs in patients with AA with large samples size and clinical follow-up.

MiRNA microarray profiling in patients with androgenic alopecia and the effects of miR-133b on hair growth

ObjectiveAndrogenetic alopecia (AGA), a common alopecia, is often accompanied by abnormal expression of multiple miRNAs. This study aims to investigate abnormally expressed miRNAs in patients with AGA and their specific molecular mechanism. MethodsmiRNA microarray profiling and qRT-PCR validation were used to screen and verify abnormally expressed miRNAs in patients with AGA. Human hair follicles (HFs) were treated with different concentrations of dihydrotestosterone (DHT, 10−5, 10−6, 10−7 and 10−8 mol/L) for 10 days. The effects of DHT on HF growth, proliferation, and miRNA expression in cultured HFs were investigated using immunofluorescence staining and qRT-PCR. Moreover, human dermal papilla cells (HDPCs) were treated/transfected with a Wnt/β-catenin pathway activator and/or miR-133b mimic, and then the CCK-8 assay was used to evaluate HDPC proliferation. qRT-PCR and Western blotting were used to measure the expression of Versican, ALP and β-catenin ResultsmiRNA microarray profiling identified 43 miRNAs that were significantly differentially expressed in AGA patients, and qRT-PCR verified that 8 miRNAs were significantly differentially expressed. The expression of miR-133b was abnormally high in AGA patients. DHT (10−5 mol/L) inhibited human HF growth and upregulated miR-133b expression, and DHT (10−7 mol/L) induced human HF growth and downregulated miR-133b expression. HDPC proliferation was inhibited, and the expression of β-catenin was downregulated in the miR-133b mimic-transfected group compared with the control group (P < 0.05). Wnt/β-catenin pathway activator treatment significantly promoted HDPC proliferation and upregulated the expression of β-catenin (P < 0.05). In addition, the proliferation of HDPCs was not significantly different between the group cotreated with a Wnt/β-catenin pathway activator and miR-133b mimic, and the control group (P > 0.05), but the expression of Versican and ALP was suppressed in the cotreatment group (P < 0.05) ConclusionOur data indicated that patients with androgenic alopecia have specific miRNA expression profiles and that the abnormal expression of miR-133b may inactivate the Wnt/β-catenin pathway and ultimately regulate hair growth.

Abstract 13092: Regulation of SARS-CoV-2, Viral Entry Proteins ACE2 and TMPRSS2, and Interferon Genes by Host MicroRNAs

Introduction: Infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the virus responsible for coronavirus disease 2019, can cause myocardial injury, heart failure, and death. MicroRNAs (miRNAs) have emerged as important regulators of viral pathogenesis, but the impact of host miRNAs on SARS-CoV-2 infectivity remains unknown. Hypothesis: Bioinformatic analysis of host miRNAs targeting the SARS-CoV-2 genome, viral entry proteins ACE2 and TMPRSS2, and their upstream regulators the interferons (IFNs) will reveal high-probability miRNAs that may regulate myocardial injury during SARS-CoV-2 infection. Methods: Using validated bioinformatic prediction algorithms, we generated a prioritized atlas of host miRNAs predicted to bind to SARS-CoV-2 genome, ACE2, TMPRSS2, and IFNs. We validated the biologic plausibility of these binding interactions by investigating their expression in established miRNA profiling datasets from cell lines susceptible or resistant to SARS-CoV-2 infection and in human tissue samples. Results: Phylogenetic analysis of 820 SARS-CoV-2 genomes revealed high homology and, after iterative filtering, 288 predicted binding miRNAs. Overlay of these miRNAs in susceptible and resistant cell lines identified miRNA binders to ORFs (miR-23a (1ab), miR-29a, -29c (1ab, N), miR-151a, -151b (S spike), miR-4707-3p (S spike), miR-298 (5’-UTR), miR-7851-3p (5’-UTR), miR-8075 (5’-UTR)) that may be important for viral infectivity. We also highlight miRNAs predicted to regulate critical host factors during infection including the receptor ACE2 3’-UTR (miR-9-5p, miR-218-5p), the serine protease TMPRSS2 3’-UTR (let-7d-5p, -7e-5p, miR-494-3p, miR-382-3p, miR-181c-5p), and the IFNs such as IFN-α 3’-UTR (miR-361-5p, miR-410-3p). Given the known roles for several of these miRNAs in cardiovascular injury (e.g. miR-23a, -29a, -29c, -494, and let-7), we discuss a framework for further interrogation of these miRNAs in patients and disease-based models. Conclusions: Overall, this study provides insight into potential novel regulatory mechanisms of SARS-CoV-2 by host miRNAs and lays the foundation for future investigation of these miRNAs as potential therapeutic targets or biomarkers for myocardial injury.

Effect of cardiac rehabilitation on circulating microRNA expression in heart failure: a preliminary study.

There are benefits of exercise-based cardiac rehabilitation (CR) in patients with heart failure (HF), but their underlying molecular mechanisms remain elusive. The effect of CR on the expression profile of circulating microRNAs (miRNAs), which are short noncoding RNAs that regulate posttranscriptional expression of target genes, is unknown. If miRNAs respond to changes following CR for HF, then serum profiling of miRNAs may reveal cardioprotective mechanisms of CR. This study enrolled three hospitalized patients with progressed systolic HF and three normal volunteer controls. In patients, CR was initiated after improvement of HF, which included 2 weeks of bicycle ergometer and resistance exercises. Genome-wide expression profiling of circulating miRNAs was performed using microarrays for the patients (mean±SD age, 60.0±12.2 years) and controls (58.7±0.58 years). Circulating miRNA expression profiles were compared between patients with HF before and after CR and the controls. Expression levels of two miRNAs were significantly different in patients before CR compared with controls and patients after CR. The expression of hsa-miR-125b-1-3p was significantly downregulated and that of hsa-miR-1290 was significantly upregulated in patients before CR. When performing CR, expression of certain circulating miRNAs in patients with HF is restored to nonpathological levels. The benefits of CR for HF may result from regulation of miRNAs through multiple effects of gene expression.

Differentially expressed microRNAs in peripheral blood mononuclear cells of non-segmental vitiligo and their clinical significance.

BackgroundVitiligo is a frequent acquired depigmentation skin disease due to a loss of melanocytes. This study sought to characterize the expression pattern of microRNA (miRNA) in the peripheral blood mononuclear cells (PBMCs) of non‐segmental vitiligo (NSV) patients. We also screened for molecular markers that can be used to evaluate the clinical stages of NSV.MethodsThe miRNA expression profile in the PBMCs of four patients with progressive NSV and four healthy controls was determined using high‐throughput RNA sequencing. The divergently expressed miRNA was verified via qRT‐PCR in 26 progression, 26 stable NSV, and 26 healthy controls.ResultsOur findings posited that 323 miRNAs were differentially expressed in the PBMCs of NSV patients. The top 10 up‐regulated miRNAs in patients were hsa‐miR‐335‐5p, hsa‐miR‐20a‐5p, hsa‐miR‐514a‐3p, hsa‐miR‐144‐5p, hsa‐miR‐450b‐5p, hsa‐miR‐369‐3p, hsa‐miR‐101‐3p, hsa‐miR‐142‐5p, hsa‐miR‐19b‐3p, and hsa‐miR‐340‐5p. The top 10 down‐regulated miRNAs in patients were hsa‐miR‐4443, hsa‐miR‐1248, hsa‐miR‐6859‐3p, hsa‐miR‐668‐3p, hsa‐miR‐7704, hsa‐miR‐323a‐5p, hsa‐miR‐1237‐3p, hsa‐miR‐3127‐3p, hsa‐miR‐6735‐3p, and hsa‐miR‐127‐3p. The expressions of hsa‐miR‐20a‐5p in PBMCs of progressive and stable NSV were remarkably elevated relative to the healthy controls. In the characteristics curve analysis of hsa‐miR‐20a‐5p for differentiating progressive and stable NSV from normal subjects in PBMCs, the area under curve (AUC) was 0.92 and 0.81. Compared with patients in stable NSV, the hsa‐miR‐20a‐5p was markedly increased in PBMCs of progressive NSV patients, and the AUC was 0.81.ConclusionOur results showed that divergently expressed miRNAs contribute to the pathogenesis of NSV and that hsa‐miR‐20a‐5p can be applied as a biosignature for stage assessment in PBMCs of patients with NSV.

MiR-125b Loss Activated HIF1α/pAKT Loop, Leading to Transarterial Chemoembolization Resistance in Hepatocellular Carcinoma.

Transarterial chemoembolization (TACE) is a standard locoregional therapy for patients with hepatocellular carcinoma (HCC) patients with a variable overall response in efficacy. We aimed to identify key molecular signatures and related pathways leading to HCC resistance to TACE, with the hope of developing effective approaches in preselecting patients with survival benefit from TACE. Four independent HCC cohorts with 680 patients were used. MicroRNA (miRNA) transcriptome analysis in patients with HCC revealed a 41-miRNA signature related to HCC recurrence after adjuvant TACE, and miR-125b was the top reduced miRNA in patients with HCC recurrence. Consistently, patients with HCC with low miR-125b expression in tumor had significantly shorter time to recurrence following adjuvant TACE in two independent cohorts. Loss of miR-125b in HCC noticeably activated the hypoxia inducible factor 1 alpha subunit (HIF1α)/pAKT loop in vitro and in vivo. miR-125b directly attenuated HIF1α translation through binding to HIF1A internal ribosome entry site region and targeting YB-1, and blocked an autocrine HIF1α/platelet-derived growth factor β (PDGFβ)/pAKT/HIF1α loop of HIF1α translation by targeting the PDGFβ receptor. The miR-125b-loss/HIF1α axis induced the expression of CD24 and erythropoietin (EPO) and enriched a TACE-resistant CD24-positive cancer stem cell population. Consistently, patients with high CD24 or EPO in HCC had poor prognosis following adjuvant TACE therapy. Additionally, in patients with HCC having TACE as their first-line therapy, high EPO in blood before TACE was also noticeably related to poor response to TACE. MiR-125b loss activated the HIF1α/pAKT loop, contributing to HCC resistance to TACE and the key nodes in this axis hold the potential in assisting patients with HCC to choose TACE therapy.

Expression level of miRNAS in patients with gestational diabetes

Abstract Background Gestational diabetes mellitus (GDM), which has been identified for the first time during pregnancy and is defined as glucose intolerance of varying degrees, is one of the prediabetes states that are at risk for developing type 2 diabetes. MicroRNAs (miRNAs) are associated with pregnancy complications and are even necessary for the regulation of pancreatic development and glucose-induced insulin secretion. This study aims to investigate miRNAs expressed in maternal plasma to detect the availability as a biomarker in the diagnosis of GDM. Methods In this study, 30 pregnant women with GDM diagnosed according to GDM American Diabetes Association criteria and 30 healthy pregnant women were included and miRNA expression levels were investigated. Circulating 39 miRNAs were analyzed by qRT-PCR using 96.96 Dynamic Array IFCs. Results The findings showed that the expression level of miR-7-5p significantly upregulated when compared with healthy pregnant women with gestational diabetes (fold change:2.3, p&lt;0.05). Conclusions There is controversy about what is the most appropriate test for GDM screening. Therefore, supported by further studies, miR-7-5p can be used for the diagnosis of GDM.

Serum microRNA as indicators of Wolfram syndrome’s progression in neuroimaging studies

IntroductionPatients with the ultra-rare Wolfram syndrome (WFS) develop insulin-dependent diabetes and progressive neurodegeneration. The aim of the study was to quantify microRNAs (miRNAs) in sera from patients with WFS, correlate...

Analysis of circulating miRNA levels in coronary heart disease patients with varying degrees of cardiovascular complications risk. correlations with the MSCT-CA data

Rationale. Cardiovascular diseases remain the leading cause of human death in the world. Studying the role of regulatory non-coding RNAs, which include short single-stranded miRNA molecules, allows a more detailed understanding of the pathological processes underlying the progression of atherosclerosis. &#x0D; Objective to compare the levels of circulating miRNAs in patients with coronary heart disease, confirmed by multislice computed tomography-coronarography (MSCT-CA), with risks of cardiovascular complications and clinical and demographic characteristics. To compare the profiles of circulating miRNAs in groups of patients with stable and unstable atherosclerotic plaques.&#x0D; Methods. MicroRNA levels in the plasma of peripheral blood of patients with coronary heart disease were determined using the miScript miRNA PCR Array MIHS-105Z kit (Qiagen). The significance of differences in miRNA levels between the compared groups was determined using the MannWhitney U-test. The correlations of the levels of circulating miRNAs with clinical and demographic parameters were evaluated using the Spearman correlation coefficient. Risk assessment of cardiovascular complications in these patients was carried out using validated scales (ACC/AHA, Framinghm, SCORE, MESA). Atherosclerotic plaque stability was evaluated using MSCT-CA.&#x0D; Results. The study showed a significant (p 0.05) decrease in miR-16, miR-211, miR-195 miRNA levels in the plasma of patients with coronary heart disease, which correlated with an increase in cardiac vascular risk (CVR) according to ACC/AHA, Framingham and MESA. When comparing groups of patients with stable and unstable atherosclerotic plaques, the latter revealed an increase in the level of let-7b-5p circulating microRNA (p 0.05).&#x0D; Conclusion. Significant associations of the three studied microRNAs with the estimated risk of CVR were identified. It is important to find circulating let-7b-5p in a group of patients with unstable atherosclerotic plaques. Correlations were established between the levels of circulating microRNAs and clinical and demographic characteristics of patients. The study shows the involvement of some microRNAs in the regulation of atherosclerosis.

The role of microRNA in degeneration of the intervertebral disc

MicroRNAs (miRNAs) are a class of small noncoding RNA molecules that negatively regulate gene expression at posttranscriptional levels. MiRNAs regulate many normal physiological processes, and also play an important role in the development of most disorders. The expression levels of miRNAs are characterized by endogenous properties and tissue specificity. These characteristics increase the likelihood that miRNAs can serve as useful clinical biomarkers in the diagnosis of certain diseases. Chronic lower back pain is usually associated with degeneration of the intervertebral disc (IDD), which is closely associated with apoptosis, impaired extracellular matrix, cell proliferation, and an inflammatory response. This process is characterized by a cascade of molecular, cellular, biochemical, and structural changes. Currently, there is no clinical therapy that shows the pathophysiology of disk degeneration. The presence of unregulated expression of miRNA in patients with degenerative disk disease indicates a vital role of miRNAs in the pathogenesis of IDD. It becomes apparent that epigenetic processes affect the evolution of IDD as much as the genetic background. Deregulated phenotypes of pulp nucleus cells, including differentiation, migration, proliferation, and apoptosis, are involved in all stages of the progression of human IDD. In this review, we will focus on the role and therapeutic value of miRNAs in IDD.

MiR‑31 promotes tumorigenesis in ulcerative colitis‑associated neoplasia via downregulation of SATB2.

Ulcerative colitis (UC) features chronic, non-infectious inflammation of the colon. The risk of ulcerative colitis-associated neoplasia (UCAN) increases in direct association with the duration of this disease. Whether miRNAs exert a regulatory effect on the pathogenesis of UCAN has remained to be elucidated. In the present study, differentially expressed genes (DEGs) and microRNAs (miRNAs/miRs) were identified using bioinformatics analysis of Gene Expression Omnibus datasets. Enrichment analyses were performed to determine the function of the DEGs. The target genes of key miRNAs were predicted using miRWalk. Validation of DEGs and miRNAs in patients with UC, UC with low-grade dysplasia and UC with high-grade dysplasia (UC-HGD) was performed using reverse transcription-quantitative PCR analysis. A total of 38 differentially expressed miRNAs and 307 mRNAs were identified from the profiles and miR-31 was validated as being overexpressed in UCAN tissues, particularly in the UC-HGD samples. Furthermore, special AT-rich DNA-binding protein 2 (SATB2) was validated as a target gene of miR-31 and SATB2 expression was negatively correlated with miR-31 expression. Therefore, miR-31 is upregulated in UCAN and it may promote tumorigenesis through downregulation of SATB2.