miR-145 Levels Research Articles

Abstract 3757: miR-200 inhibits the activation of cancer-associated fibroblasts in lung tumor microenvironment

Abstract Cancer-associated fibroblasts (CAFs) are key components of the tumor microenvironment and promote cancer progression and metastasis. CAFs are regulated by diverse processes including regulation by miRNAs. To identify miRNAs involved in the regulation of CAF activation, we profiled miRNA expression within mouse lung normal fibroblasts (LFs) and CAFs using the NanoString nCounter method. Among numerous miRNAs differentially expressed between CAFs and LFs, miR-200 was selected for further investigation, which is more highly downregulated in CAFs. Overexpression of miR-200 in CAFs suppressed the migration and invasion of lung cancer cells in co-culture systems and inhibited metastasis in a mouse model. In addition, we verified that lung cancer patients with high miR-200 levels showed a good prognosis than those with low miR-200 levels. Next, we tried to identify novel miR-200 targets based on RNA sequencing data, lung adenocarcinoma TCGA, and TargetScan. NRP2 was confirmed to be a target of miR-200 by 3΄-UTR assay. NRP2 knockdown by siRNAs in CAFs suppressed cancer cell invasion, and NRP2 addback in CAF-200 restored cancer cell invasion inhibited by miR-200. NRP2 functions as a co-receptor with VEGFR, thus, CAF-200 was less responsive to VEGF than control CAFs. Collectively, this study suggests that downregulation of miR-200 in CAFs promotes lung cancer progression in tumor microenvironment; therefore, miR-200 will be a good therapeutic target or a biomarker in lung adenocarcinoma. Citation Format: Inyoung Cheon, Sieun Lee. miR-200 inhibits the activation of cancer-associated fibroblasts in lung tumor microenvironment. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3757.

High expression of miR-107 and miR-17 predicts poor prognosis and guides treatment selection in acute myeloid leukemia.

The prognostic significance of miR-107 and miR-17 in patients with acute myeloid leukemia (AML) remains unclear. A total of 173 patients with de novo AML from the Cancer Genome Atlas database were enrolled in this study and further divided into a chemotherapy group (98 cases) and an allogeneic hematopoietic stem cell transplantation (allo-HSCT) group (75 cases) according to their therapy regimen. In the chemotherapy cohort, high miR-107 or miR-17 expression was associated with poorer overall survival (OS) and event-free survival (EFS). On the other hand, there were no significant differences in OS and EFS between the high- and low-expression subgroups in the allo-HSCT group. Next, we stratified the total number of patients with AML into high- and low-expression groups according to the median expression levels of miR-107 or miR-17. In the high miR-107 or miR-17 expression group, patients treated with allo-HSCT had longer OS than those treated with chemotherapy. In the low miR-107 or miR-17 expression group, no significant differences in OS and EFS were observed between the two therapy subgroups. When patients were further clustered into three groups (both low miR-107 and low miR-17, either high miR-107 or high miR-17, and both high miR-107 and high miR-17), patients with both high miR-107 and high miR-17 expression had the worst OS and EFS of the entire group and of the chemotherapy group. On the other hand, there were no significant differences in OS and EFS among the three subgroups in the allo-HSCT group. Cox regression confirmed the concurrence of high expression of miR-107 and miR-17 might act as an independent prognostic factor for EFS and OS in the entire group and the chemotherapy group. Bioinformatics analysis showed differentially expressed genes (DEGs) associated with miR-107 and miR-17 expression were mainly enriched in multiple metabolic processes. The combination of miR-107 and miR-17 provides prognostic significance for patients with AML and should be considered in the clinical selection of the optimal treatment regimen when deciding between chemotherapy and allo-HSCT.

Osteocalcin, miR-143, and miR-145 Expression in Long-Standing Type 1 Diabetes Mellitus and Their Correlation with HbA1c.

Inadequate management and control of hyperglycemia predisposes diabetic patients to a wide range of complications. Thus, this opens new windows for exploring and scrutinizing novel candidate biomarkers. This study was designed to scrutinize the relationship between HbA1c, osteocalcin, calcium, phosphorus, and expression levels of miR-143 and miR-145 in individuals with T1DM and explore their correlations and diagnostic potential for T1DM. 120 unrelated participants were included (i.e., 90 participants with type 1 diabetes mellitus and 30 healthy controls) and were allocated into two groups. Participants with T1DM were allocated into three subgroups (i.e., below 1year, 1-8years, and over 8years) based on diabetic duration. Participants with T1DM experienced noticeable HbA1c elevation. However, osteocalcin, phosphorus, and calcium profiles notably declined in participants with diabetes compared with those in healthy controls. Moreover, the expression levels of miR-143 and miR-145 decreased in participants with diabetes with a significant difference between participants with diabetes and healthy controls. Additionally, significant alterations in HbA1c, osteocalcin, phosphorus, and calcium profiles and expression levels of miR-143 and miR-145 were observed with increasing diabetic duration (T1DM > 8years compared with those with a diabetes duration of less than 1year). This study suggests that miR-143 and miR-145 are prospective biomarkers of diabetes mellitus, which may help predict the progression of complications.

Tan IIA mitigates vascular smooth muscle cell proliferation and migration induced by ox-LDL through the miR-137/TRPC3 axis.

Tanshinone IIA (Tan IIA) has an important role in treatment of cardiovascular diseases, including atherosclerosis. The vascular smooth muscle cells (VSMCs) are a major part of the atherosclerotic plaque. However, the biological functions of Tan IIA in regulating VSMCs function remain mostly unclear. This research aimed at identifying the explicit molecular mechanism that Tan IIA regulates oxidized low-density lipoprotein (ox-LDL)-mediated VSMC proliferation and migration. VSMCs challenged by ox-LDL were adopted as cellular model of atherosclerosis, and suffered from Tan IIA treatment. After that, cells proliferation, apoptosis or migration were measured. The expression levels of microRNA (miR)-137, transient receptor potential cation channel subfamily C member 3 (TRPC3) and proliferating cell nuclear antigen (PCNA) were measured. The targeting relationship between miR-137 and TRPC3 was determined. It was found that Tan IIA blunted VSMC proliferation, PCNA expression and migration mediated by ox-LDL. Tan IIA promoted miR-137 level, and miR-137 knockdown reversed the influences of Tan IIA on VSMC proliferation, PCNA expression and migration in the presence of ox-LDL. TRPC3 was verified to be targeted by miR-137. Moreover, TRPC3 silencing exacerbated the influences of Tan IIA on VSMC proliferation, apoptosis and migration, and it mitigated the inhibitive effects of miR-137 knockdown on function of Tan IIA. We confirmed for the first time that Tan IIA constrained ox-LDL-stimulated VSMC proliferation and migration via regulating the miR-137/TRPC3 axis, which provided a theoretical basis for the research and promotion of Tan IIA as a therapeutic drug.

Serum Exosome-Derived MiR-7 Exacerbates Chronic Obstructive Pulmonary Disease by Regulating Macrophage Differentiation.

Polarization of macrophages and miR-7 have been reported to greatly influence the progress of chronic obstructive pulmonary disease (COPD). However, the interaction is unclear. We aimed to investigate the role of miR-7 in the serum exosome of COPD, thus further revealing the underlying mechanism of COPD. The study was conducted in 2022 in The Affiliated Changzhou No.2 People's Hospital of Nanjing Medical University, Changzhou, China. COPD mouse model was established. Macrophages were sorted by flow cytometry assay. ELISA kits were used to detect the levels of TNF-α and IL-6. Exosomes were identified by confocal microscopy and PKH67 staining. RT-qPCR and western blot assay were performed to determine the mRNA and protein expressions. H&E staining assay was used to assess the tissue injury. CCK-8 assay was applied to evaluate cell viability. Luciferase reporter assay was used to confirm the binding between PIM1 and miR-7. The exosomes derived from the COPD mice serum exerted high level of miR-7, which induced M1 differentiation of macrophages and increased the secretion of proinflammatory factors in vivo and in vitro. The effects of exosomes from COPD mice could be inhibited by miR-7 inhibitor. Bioinformatic prediction, luciferase reporter assay, and western blot assay showed an interaction between miR-7 and PIM1. Further examination showed that miR-7 regulated macrophage activation and differentiation to M1 via PIM1 in vitro. miR-7 from serum exosomes might exacerbate COPD by stimulating macrophage differentiation to M1, supplying a potential therapeutic target for COPD treatment.

PTD-FGF2 Attenuates Elastase Induced Emphysema in Mice and Alveolar Epithelial Cell Injury

Aberrant communication in alveolar epithelium is a major feature of inflammatory response for the airway remodeling leading to chronic obstructive pulmonary disease (COPD). In this study, we investigated the effect of protein transduction domains (PTD) conjugated Basic Fibroblast Growth Factor (FGF2) (PTD-FGF2) in response to cigarette smoke extract (CSE) in MLE-12 cells and porcine pancreatic elastase (PPE)-induced emphysematous mice. When PPE-induced mice were intraperitoneally treated with 0.1–0.5 mg/kg PTD-FGF2 or FGF2, the linear intercept, infiltration of inflammatory cells into alveoli and pro-inflammatory cytokines were significantly decreased. In western blot analysis, phosphorylated protein levels of c-Jun N-terminal Kinase 1/2 (JNK1/2), extracellular signal-regulated kinase (ERK1/2) and p38 mitogen-activated protein kinases (MAPK) were decreased in PPE-induced mice treated PTD-FGF2. In MLE-12 cells, PTD-FGF2 treatment decreased reactive oxygen species (ROS) production and further decreased Interleukin-6 (IL-6) and IL-1b cytokines in response to CSE. In addition, phosphorylated protein levels of ERK1/2, JNK1/2 and p38 MAPK were reduced. We next determined microRNA expression in the isolated exosomes of MLE-12 cells. In reverse transcription-polymerase chain reaction (RT-PCR) analysis, level of let-7c miRNA was significantly increased while levels of miR-9 and miR-155 were decreased in response to CSE. These data suggest that PTD-FGF2 treatment plays a protective role in regulation of let-7c, miR-9 and miR-155 miRNA expressions and MAPK signaling pathways in CSE-induced MLE-12 cells and PPE-induced emphysematous mice.

ASSOCIATION BETWEEN TWENTY SERUM MIRNAS AND CLINICOPATHOLOGICAL VARIABLES IN PATIENTS WITH BREAST CANCER

Determining that miRNAs play a role in breast cancer pathogenesis suggests that it may be useful in the diagnosis and treatment of breast cancer. In this study, 39 patients with invasive breast cancer were diagnosed with serum levels of 20 miRNAs(miR-105, miR-21, miR-141, miR-200a, miR-200b, miR-200c, miR-203, miR-210, miR-375, miR-34a, miR-133a, miR-155, miR-139-5p, miR-143, miR-145, miR-365, miR-299-5p, miR-411, miR-452, miR-17) were analyzed during diagnosis and after treatment. In the analysis results, it is detected that serum levels of miR-200c(p=0.030), miR-375(p=0.045), miR-34a(p=0.042) were significantly higher in the local advanced/metastatic group. Serum level of miR-141(p=0.062) was lower in patients with positive lymph node involvement, whereas miR-133a(p=0.037) levels were higher in the same patient group. MiR-105(p=0.015), miR-203(p=0.015), miR-375(p=0.033), miR-145(p=0.025) serum levels were significantly higher in the progesterone receptor negative group, likewise miR-105(p=0.053) levels were high in the estrogen receptor negative patient group. The high levels of miR-375 and miR-133a were noticeable in human epidermal growth factor receptor-2 positive patients(p=0.037 and p=0.014, respectively). MiR-143(p=0.009) and miR-145(p=0.017) levels were observed to be higher in the patient group with a ki-67 index>20%. It was found that 2 miRNAs(miR-133a(p=0.018) and miR-139-5p(p=0.004)) were significantly higher in patients in the luminal B group, which were separated by molecular subgroups. Nine of miRNAs that evaluated(miR-105(p=0.0001), miR-21(p=0.001), miR-141(p=0.041), miR-200a(p=0.003), miR-200b(p=0.0001), miR-200c(p=0.0001), miR-203(p=0.0001), miR-34a(p=0.0001), miR-452(p=0.018)) significantly increased after treatment and 5 of the miRNAs(miR-155(p=0.0001), miR-143(p=0.0001), miR-145(p=0.0001), miR-365(p=0.0001), miR-299-5p(p=0.0001)) were significantly reduced after treatment. We think that miRNAs may help in evaluating the follow-up and prognosis of invasive breast cancer.

Prognostic utility of microRNA-145 and CD 133 in oral squamous cell carcinoma: A pilot study from Northern India.

Multiple factors alter the microRNAs (miRNAs) at cellular level and promote oncogenesis in oral mucosa. The present study was performed with an aim to find any expression of miRNA-145 in oral squamous cell carcinoma and correlate it with CD-133 immuno-expression, clinicopathological, and demographical variables in oral squamous cell carcinoma (OSCC) with respect to controls. In a prospective observational study 50 samples from patients of OSCC and 20 from unremarkable oral mucosa were studied. After initial detailed histology, miRNA-145 profiling was performed using qRT-PCR, followed by CD-133 immunohistochemistry (IHC). The mean age of patients with oral cancer was 47.5±10.25 years. Mean miR-145 levels in OSCC were 0.4312±0.32026 and mean in healthy controls was 0.99±0.21771. There was significant downregulation of miRNA-145 in cases with respect to controls. Significant reduced levels of miRNA-145 with respect to higher clinical tumor size, pathological pT tumor, nodal status, and resultant clinical tumor stage was observed. As far as presence and absence of stem cells was concerned it was seen that tumors displaying presence of stem cells highlighted by CD-133 had lower levels of miRNA-145 as compared to tumors with absent CD-133 staining. miRNA-145 is significantly altered in OSCC in our patient population and its reduced values carry a poor prognosis. Its interaction with CSCs may not be significant but mean miRNA-145 levels are lower in tumors with CSCs. There should be further studies on the larger sample size for these two biomarkers, to know its value.

Treatment of Acute Respiratory Distress Syndrome Caused by COVID-19 with Human Umbilical Cord Mesenchymal Stem Cells

This study aimed to identify the impact of mesenchymal stem cell transplantation on the safety and clinical outcomes of patients with severe COVID-19. This research focused on how lung functional status, miRNA, and cytokine levels changed following mesenchymal stem cell transplantation in patients with severe COVID-19 pneumonia and their correlation with fibrotic changes in the lung. This study involved 15 patients following conventional anti-viral treatment (Control group) and 13 patients after three consecutive doses of combined treatment with MSC transplantation (MCS group). ELISA was used to measure cytokine levels, real-time qPCR for miRNA expression, and lung computed tomography (CT) imaging to grade fibrosis. Data were collected on the day of patient admission (day 0) and on the 7th, 14th, and 28th days of follow-up. A lung CT assay was performed on weeks 2, 8, 24, and 48 after the beginning of hospitalization. The relationship between levels of biomarkers in peripheral blood and lung function parameters was investigated using correlation analysis. We confirmed that triple MSC transplantation in individuals with severe COVID-19 was safe and did not cause severe adverse reactions. The total score of lung CT between patients from the Control and MSC groups did not differ significantly on weeks 2, 8, and 24 after the beginning of hospitalization. However, on week 48, the CT total score was 12 times lower in patients in the MSC group (p ≤ 0.05) compared to the Control group. In the MSC group, this parameter gradually decreased from week 2 to week 48 of observation, whereas in the Control group, a significant drop was observed up to week 24 and remained unchanged afterward. In our study, MSC therapy improved lymphocyte recovery. The percentage of banded neutrophils in the MSC group was significantly lower in comparison with control patients on day 14. Inflammatory markers such as ESR and CRP decreased more rapidly in the MSC group in comparison to the Control group. The plasma levels of surfactant D, a marker of alveocyte type II damage, decreased after MSC transplantation for four weeks in contrast to patients in the Control group, in whom slight elevations were observed. We first showed that MSC transplantation in severe COVID-19 patients led to the elevation of the plasma levels of IP-10, MIP-1α, G-CSF, and IL-10. However, the plasma levels of inflammatory markers such as IL-6, MCP-1, and RAGE did not differ between groups. MSC transplantation had no impact on the relative expression levels of miR-146a, miR-27a, miR-126, miR-221, miR-21, miR-133, miR-92a-3p, miR-124, and miR-424. In vitro, UC-MSC exhibited an immunomodulatory impact on PBMC, increasing neutrophil activation, phagocytosis, and leukocyte movement, activating early T cell markers, and decreasing effector and senescent effector T cell maturation.

Exosome-Encapsulated miR-31, miR-192, and miR-375 Serve as Clinical Biomarkers of Gastric Cancer.

In recent years, microRNAs (miRNAs) derived from exosomes have been attracting attention as novel clinical biomarkers in a variety of cancers. In this study, plasma samples from 60 gastric cancer (GC) patients and 63 healthy individuals were collected, and the exosomal microRNAs (ex-miRNAs) were isolated. We determined the specific ex-miRNAs through miRNA microarray and a database of differentially expressed miRNAs called dbDEMC. Then, the expression levels of exosomal miR-31, miR-192, and miR-375 were analyzed by quantitative polymerase chain reaction (qRT-PCR). Compared to the matched controls, exosomal miR-31, miR-375, and miR-192 were significantly upregulated in GC patients. Also, they were found to be associated with gender, with miR-192 being significantly upregulated in male GC patients. Kaplan-Meier analysis indicated that high expressions of exosomal miR-31, miR-375, and miR-192 were positively correlated with poor clinical outcomes of GC patients. Cox univariate and multivariate analysis found that ex-miR-375 expression and TNM stage were independent prognostic factors of overall survival (OS). Our findings revealed that exosomal miR-31, miR-192, and miR-375 might serve as noninvasive, sensitive, and specific biomarkers for the diagnosis and prognosis of GC patients.

Anethole suppresses the growth of human skin cancer cells by targeting microRNA498/STAT4 axis

Purpose: To investigate the effects of anethole on human skin cancer cells.
 Methods: Cell viability was determined using CCK-8 and colony formation assay, while AO/EB and Annexin V/PI assays were used to assess apoptosis. The mRNA expressions of genes were assayed by quantitative reverse transcription-polymerase chain reaction (qRT-PCR).
 Results: The growth of skin cancer cells was significantly reduced by anethole (p < 0.05). On the other hand, normal human skin cells were marginally affected by anethole. Furthermore, anethole significantly suppressed the colony formation potential of CRL-6475 skin cancer cells in a concentration-dependent manner (p < 0.05). Results from AO/EB and annexin V/PI staining assays revealed that anethole induced apoptosis in skin cancer cells, while qRT-PCR results indicate that anethole significantly up-regulated microRNA-498 in skin cancer cells (p < 0.05). However, anethole had no effect on the expression levels of miR-7, miR-9, miR-145, miR-31, miR-27, miR-498 miR-1298, miR-1299, 1179, miR-375, miR-508 and miR-23. CCK-8 assay results confirmed that miR-498 up-regulation significantly mimicked the anti-proliferative effects of anethole. Bioinformatics analysis and luciferase reporter assay led to identification of STAT4 as the regulatory target of miR-498. Furthermore, silencing of STAT4 significantly mimicked the effects of anethole administration (p < 0.05).
 Conclusion: These results indicate that anethole inhibits skin cancer cell growth by targeting the miR-498/STAT4 axis, and therefore, has potentials for use in the management of skin cancer.

Micro RNA based MSC EV engineering: Targeting the BMP2 cascade for bone repair.

Mesenchymal stem cell derived extracellular vesicles (MSC EVs) possess excellent immunomodulatory and therapeutic properties. While beneficial, from a translational perspective, extracellular vesicles with consistent functionality and target specificity are required to achieve the goals of precision medicine and tissue engineering. Prior research has identified that the miRNA composition of mesenchymal stem cell derived extracellular vesicles contributes significantly towards extracellular vesicles functionality. In this study, we hypothesized that mesenchymal stem cell derived extracellular vesicle functionality can be rendered pathway-specific using a miRNA-based extracellular vesicles engineering approach. To test this hypothesis, we utilized bone repair as a model system and the BMP2 signaling cascade as the targeted pathway. We engineered mesenchymal stem cell extracellular vesicles to possess increased levels of miR-424, a potentiator of the BMP2 signaling cascade. We evaluated the physical and functional characteristics of these extracellular vesicles and their enhanced ability to trigger the osteogenic differentiation of naïve mesenchymal stem cell in vitro and facilitate bone repair in vivo. Results indicated that the engineered extracellular vesicles retained their extracellular vesicles characteristics and endocytic functionality and demonstrated enhanced osteoinductive function by activating SMAD1/5/8 phosphorylation and mesenchymal stem cell differentiation in vitro and enhanced bone repair in vivo. Furthermore, the inherent immunomodulatory properties of the mesenchymal stem cell derived extracellular vesicles remained unaltered. These results serve as a proof-of-concept for miRNA-based extracellular vesicles engineering approaches for regenerative medicine applications.

Pharmacological reversal of post-transcriptional alterations implicated in alcohol-induced alveolar macrophage dysfunction.

Alcohol use disorders (AUD) cause alveolar macrophage (AM) immune dysfunction and increase risk of lung infections. Excessive alcohol use causes AM oxidative stress, which impairs AM phagocytosis and pathogen clearance from the alveolar space. Alcohol induces expression of NADPH oxidases (Noxes), primary sources of oxidative stress in AM. In contrast, alcohol decreases AM peroxisome proliferator-activated receptor gamma (PPARγ), a critical regulator of AM immune function. To explore the underlying molecular mechanisms for these effects of alcohol, we hypothesized that ethanol promotes CCAAT/enhancer-binding protein beta (C/EBPβ)-mediated suppression of Nox-related microRNAs (miRs), in turn enhancing AM Nox expression, oxidative stress, and phagocytic dysfunction. We also hypothesized that PPARγ activation with pioglitazone (PIO) would reverse alcohol-induced C/EBPβ expression and attenuate AM oxidative stress and phagocytic dysfunction. Cells from the mouse AM cell line (MH-S) were exposed to ethanol invitro or primary AM were isolated from mice fed ethanol invivo. Ethanol enhanced C/EBPβ expression, decreased Nox 1-related miR-1264 and Nox 2-related miR-107 levels, and increased Nox1, Nox2, and Nox 4 expression in MH-S cells invitro and mouse AM invivo. These alcohol-induced AM derangements were abrogated by loss of C/EBPβ, overexpression of miRs-1264 or -107, or PIO treatment. These findings identify C/EBPβ and Nox-related miRs as novel therapeutic targets for PPARγ ligands, which could provide a translatable strategy to mitigate susceptibility to lung infections in people with a history of AUD. These studies further clarify the molecular underpinnings for a previous clinical trial using short-term PIO treatment to improve AM immunity in AUD individuals.

P101 Circulating MicroRNA-16 in Inflammatory Bowel Disease Patients - Potential Biomarker to Assess Inflammation

Abstract Background miR-16 is important for the inflammatory processes and the signal pathways that participate in the immune system regulation. As there is an immune mucosal inflammation in IBD, miR-16 could also serve as a potential biomarker to assess inflammation. Methods The study aims to assess the expression of serum miR-16 in IBD patients and to correlate its expression with clinical parameters such as disease extent, activity and severity. 35 patients with UC and 35 patients with CD were included in the study. Serum miR-16 expression was assessed with reverse transcriptase quantitative real time PCR (RT-qPCR), and was then correlated with the miR-16 levels in a group of 30 healthy subjects. Results miR-16 expression is increased in patients with CD, localized in the small intestine (3.54±2.33) and in pancolitis Е3 of UC (2.01±1.54). There is a significant difference in miR-16 expression in inflammatory (B1), stenotic (B2) and penetrating (B3) form of CD (р=0.010) with higher expression levels in the more severe B2 and B3 forms. There is no significant difference in miR-16 expression according to the forms of evolution in patients with UC. In patients with CD, there is a moderate direct correlation between the expression of miR-16 and the Crohn’s disease activity index CDAI (r=0.424; p=0.015). miR-16 expression levels increase progressively in correlation with the activity of CD. Patients with moderate activity of their CD have significantly higher miR-16 expression (4.27±3.00) as compared to patients with mild activity (2.38±1.06) or in remission (2.41±1.23) of the disease (р=0.048). The increased expression levels of miR-16 are a risk factor for CDAI > 150 levels (OR=4.80 (0.506-45.495); p=0.015). In patients with UC, there is no correlation between the miR-16 expression levels and the severity (S) index, measured as per the Montreal classification or the endoscopic activity, measured as per the Partial Mayo score. In patients with severe disease, measured by both S index (1.38±0.89) and Partial Mayo score (1.80±1.12), there is a decrease in miR-16, close to normal controls. MiR-16 expression levels do not correlate with any treatment that was given in patients with either CD or UC. Conclusion The increased miR-16 serum expression corelates with the localization of CD in the small intestine and the stenotic and penetrating form of the disease. Increased miR-16 levels are also found in extensive UC. In patients with CD there is a positive correlation between the expression of miR-16 and the activity of the disease as measured by the CDAI.

Immunomodulatory effects of probiotic supplementation in patients with asthma: a randomized, double-blind, placebo-controlled trial

BackgroundAsthma is considered to be a chronic inflammatory disorder of the airways. Probiotics are living microorganisms that are found in the human gut and have protective effects against a wide range of diseases such as allergies. The aim of this study was to investigate the improvement of clinical asthma symptoms and changes in the expression pattern of selective microRNAs in patients with asthma and the changes in IL-4 and IFN-γ plasma levels after receiving probiotics.Materials and methodsThe present study was a randomized, double-blind, placebo-controlled trial that enrolled 40 asthmatic patients. They were treated with probiotics or placebo: 1 capsule/day for 8 weeks. Pulmonary function tests, IL-4 and IFN-γ levels, and expression of microRNAs were assessed at baseline and after treatment.ResultsThe results showed that the expression of miR-16, miR146-a and IL-4 levels in patients with asthma after receiving probiotic supplementation was significantly reduced and miR-133b expression was increased. In addition, pulmonary function tests showed a significant improvement in Forced Expiratory Volume in 1 s and Forced Vital Capacity after receiving probiotics.ConclusionIn our study, 8-week treatment with probiotic supplementation led to reduced Th2 cells-associated IL-4 and improved Forced Expiratory Volume and Forced Vital Capacity. It appears probiotics can be used in addition to common asthma treatments.

Evaluation of circulating miR-16-5p and miR-223-5p in association with musculoskeletal ultrasonography seven-joint score in the assessment of rheumatoid arthritis activity

Rheumatoid arthritis (RA) is characterized by ongoing joint destruction. MicroRNAs (miRs) are blood-based biomarkers linked to RA pathogenesis. The musculoskeletal ultrasonography seven-joint score (US7) is an objective tool to assess RA activity. We aimed to evaluate miR-223 and miR-16 roles in monitoring RA activity and to investigate if there is a link between their plasma levels and US7 score. This study enrolled 76 RA patients classified according to Disease Activity Score 28-joint count with erythrocyte sediment rate (DAS28-ESR) to inactive cases (n = 38) and active cases (n = 38). Each patient's joint was scored for synovial proliferation (gray-scale ultrasound 'GSUS7') and vascularization (power Doppler ultrasound 'PDUS7'). Real-time quantitative PCR was used to measure the expression levels of miR-16 and miR-223 in plasma. When compared to inactive group, the active group revealed significant upregulation of miR-16 and miR-223, (P = 0.001 and P = 0.02, respectively). miR-16 and miR-223 levels were correlated with synovitis PDUS7 (r = 0.34, p < 0.01 and r= 0.25, P = 0.03, respectively). miR-16 was also positively correlated with synovitis GSUS7 (r= 0.42, p < 0.001). miR-223 upregulation discriminated active from inactive RA patients at AUC = 0.64, with 76% sensitivity and 50% specificity at cutoff > 2.8-fold change), whereas miR-16 distinguished the two groups at AUC = 0.78 with 87% sensitivity and 53% specificity at cutoff >38.27-fold change. In conclusion, upregulated miR-16 may have more potential to serve as activity biomarkers than miR-223 in RA. The miR-16 level was linked to synovitis GSUS7 and synovitis PDUS7 changes but miR-223 only linked to synovitis PDUS.

Synaptosome microRNAs: emerging synapse players in aging and Alzheimer's disease.

Synaptosome microRNAs: emerging synapse players in aging and Alzheimer's disease.

From microRNA to protein, linking the neurotrophic hypothesis of depression to the Wistar Kyoto rat

Animal models of depression offer an alternative research-platform to clinical studies where limitations such as incomplete medical histories, patient heterogeneity, and difficulty tracking comorbidities leading to depression can be more easily overcome. In that context the Wistar Kyoto (WKY) rat strain has for many years been used as an endogenous model of depression. Here we want to further explore the depressive-like phenotype of the WKY rats to extend its validity as a preclinical model of depression.For this purpose, 49 microRNAs (miRNAs), which have been linked to the neurotrophic hypothesis of depression were selected and their levels were examined in hippocampus, prefrontal cortex, and blood from the WKY rats. Subsequently, miRNA/target mRNA/protein relationships were explored.Seventeen miRNAs across brain and blood exhibited significant regulation (>30%). Fifteen of 17 miRNAs were upregulated in the WKY rats, when compared to the Wistar Hannover Galas rats. This was coupled with general downregulation of corresponding mRNA targets in both brain regions. We observed decreased Trkb mRNA levels in both brain regions, which was coupled with increased miR-103-1 and miR-212 levels (targeting Trkb).While the relationship between mRNA and protein was not entirely linear, we did observe downregulation of TrkB protein in the prefrontal cortex and downregulation of fully mature glycosylated TrkB in both brain regions. This may indicate disruption of the BDNF-TrkB pathway in WKY rats.In conclusion, this study identifies several molecular alterations in brain and blood from the WKY rats and highlights the WKY rat strain as a viable model of depression.

Association of the circulating micro-RNAs with susceptible and newly diagnosed type 2 diabetes mellitus cases

BackgroundDiabetes mellitus (DM) is an array of metabolic diseases, which results from deregulation in insulin secretion or its action leading to abnormally high levels of blood sugar associated with long-term damage, organs failure, especially the eyes, heart, kidneys, blood vessels and nerves whose frequency has increased progressively worldwide. ObjectivesThe aims of this study were to evaluate the plasma levels of miRNA-126, miRNA-486, miRNA-223 and miRNA-375 in newly diagnosed T2DM susceptible and healthy control individuals. MethodsIn this study, we evaluated the T2DM-related miRNAs miRNA-126, miRNA-486, miRNA-223 and miRNA-375 in plasma of three study groups comprising healthy control (N = 36), newly diagnosed T2DM (N = 29) and T2DM susceptible individuals (N = 30) using Real Time polymerase chain reaction (RT-PCR) (Taqman®). All the statistical analysis were done using SPSS version 21software. ResultsSignificant associations were found between these parameters. The expression levels of miR-126, miR-486, miR-223, and miR-375 were downregulated in T2DM patients and further reduced in T2DM-susceptible individuals compared to healthy controls. Additionally, miR-223 and miR-375 showed an inverse correlation with HbA1c levels, while all four miRs displayed an inverse correlation with FBS levels in the combined group analysis. ConclusionThese observed associations further suggest that miR-126, miR-223, and miR-375 may be involved in the regulation of glucose metabolism and glycemic control. Their down regulation could potentially contribute to impaired insulin signaling, beta-cell dysfunction, inflammation, and disrupted glucose homeostasis, all of which are characteristics of diabetes.

Prolonged alcohol consumption influences microRNA expression in the nucleus accumbens of the rat brain.

The microRNA (miR) species analyzed in this study are involved in molecular mechanisms of TLR4 and TLR7 signaling, mediating the development of neuroinflammation and neurodegeneration. We have investigated the expression levels of miR-let7b, miR-96, miR-182, miR-155, and the mRNA content of HMGB1, TLR3, TLR4 in the nucleus accumbens (NAc) of the brain of rats exposed to long-term alcoholization. The long-term alcoholization caused a decrease in miR-let7b, miR-96, miR-182, and TLR7 mRNA levels; this was accompanied by an increase in miR-155, TLR4, and Hmgb1 mRNA levels in the NAc of rat brain. TLR7 is functionally linked to miR-let7b. The data of a simultaneous decrease in miR-let7b and TLR7 mRNA are of interest for further studies; they may indicate on the lack of functionally significant links between Hmgb1 and the miR-let7b-TLR7 system in NAc. The existing evidence of a functional relationship between TLR4 with miR-155 and miR-182 and our observations on their expression changes during chronic alcoholization are very interesting and require further investigation. The suggestion about the development of neuroinflammatory process in NAc under prolonged alcohol exposure are relevant for studying the level of TLR gene expression in NAc, as well as the expression of miR species, which may have a functional relationship with the TLR system.