miR-133a Levels Research Articles

Expression of Plasma miRNA-133a is Significantly Lower in Acute Coronary Syndrome (ACS) than in Healthy/Non-ACS Subjects

BACKGROUND: The current biomarker diagnostic modality for acute coronary syndrome (ACS), cardiac troponin, has several limitations. Emerging studies showed that micro-RNA (miR)-133a was released from infarcted heart to circulation, yet the diagnostic value of miR-133a in ACS demonstrated a conflicting result. Therefore, this study was conducted to investigate the potency of plasma miR-133a as a biomarker candidate of ACS.METHODS: This was a case-controlled study, involving ACS and control subjects. The sociodemographic and clinical characteristics were assessed through medical records. A final of 39 ACS and 31 control subjects (consist of healthy and non-ACS subjects) passed the selection procedure by demonstrating a high purity of RNA. miR-133a from ACS and control subjects were detected by quantitative polymerase chain reaction (qPCR). Expression of miR-133a was evaluated for sensitivity and specificity as an ACS biomarker diagnostic using the receiver operating characteristic (ROC) curve.RESULTS: Plasma miR-133a expression was stably found in ACS subjects. The plasma miR-133a level was lower in ACS than in control subjects. miR-133a effectively distinguished ACS subjects from healthy subjects (AUC=0.911) and exhibited high diagnostic performance, with a sensitivity of 87.1% and specificity of 100% at a cut-off value of 44.035. In an extended model including both control subjects (healthy and non-ACS with comorbid conditions), miR-133a maintained diagnostic significance (AUC=0.874), showing sensitivity of 76.9% and specificity of 100% at a cut-off value of 11.69.CONCLUSION: Plasma miR-133a is significantly lower and effectively distinguishes ACS patients from both healthy individuals and non-ACS individuals with comorbid, with a cut-off value of 11.69. Therefore, plasma miR-133a is suggested to be a good candidate for diagnostic biomarkers of ACS.KEYWORDS: circulating miRNA, miRNA-133a, acute coronary syndrome, diagnostic biomarker

Abstract P360: MicroRNAs: Potential Biomarkers and Therapeutic Targets for Hypertensive Nephropathy

Introduction: Hypertensive nephropathy (HN) is the second most common cause of end-stage renal disease after diabetic nephropathy. miRNAs are small epigenetic modulators whose levels change in many diseases. This review summarizes the diagnostic and therapeutic potential of miRNAs in HN. Methodology: We conducted a thorough literature review on PubMed, including both retrospective and prospective studies on animal and human subjects. Additional references were sourced through cross-referencing the bibliographies of the initial articles. Results: Our literature review revealed that intra-renal levels of miR-200a, miR-141, miR-200b, miR-205, miR-429, and miR-192 were elevated in patients with HN. Conversely, intra-renal miR-204-5p levels were decreased, and further reductions in animal models using anti-miR-204-5p or miR-204 gene knockout exacerbated renal injury. Exposure of renal tissue to exogenous albumin increased intra-renal miR-184 levels, prompting further investigation into whether albumin excretion from kidney injury alters miRNA expression or if miRNA dysregulation initiates kidney injury and subsequent albumin excretion. Urinary miR-146a, miR-184, and miR-34a levels were lower in patients with albuminuria than those without, whereas urinary miR-200a levels were higher. Interestingly, urinary miR-21 levels increased before the onset of albuminuria in hypertensive mice. A positive correlation was also observed between miR-208b and miR-133a levels in blood cells and urinary albumin excretion (UAE). Notably, plasma levels of miR-let-7g-5p and miR-191-5p correlated positively with eGFR but not with UAE. Since miRNA levels varied based on the sampling source, a pivotal study identified a consistent miRNA profile in HN patients, showing that miR-208a and miR-301a were upregulated in urine yet downregulated in plasma. The therapeutic potential of miRNAs was highlighted by an animal study showing that the knockdown of miR-103a-3p can nullify albuminuria in hypertensive mice. Conclusion: The role of miRNAs in HN is increasingly evident. Studies have linked varying miRNA expression profiles in patients with HN, and specific miRNAs were detectable earlier than albuminuria. Furthermore, certain miRNAs correlate with eGFR independently of UAE. Targeting specific miRNAs, using miRNA mimics and anti-miRNAs, could underpin preventive and therapeutic strategies. These findings advocate for further research into miRNAs as potential biomarkers and therapeutic targets for HN.

Establishment of a myostatin gene-knockout C2C12 cell line and evaluation of related microRNA expression

The strategy of blocking myostatin (MSTN) signal transduction has long been regarded as a promising approach in the treatment of patients with muscle loss. However, individuals taking blocking agents often encounter issues such as lack of strength, fatigue, and poor muscle proliferation due to muscle hypertrophy and the involvement of multiple receptors. To address these challenges, a series of experiments were conducted on a C2C12 cell line in this study. First, the pX601-SaCas9-sgRNA/puro vector carrying a Cas9-encoded gene was constructed and subsequently used to produce Mstn-knockout (Mstn-KO) C2C12 cell lines. The expression level of the MSTN protein and the growth characteristics of the cell lines were verified. Moreover, the expression of muscle growth-related microRNAs in the cell lines was analyzed through real-time polymerase chain reaction (PCR). The results indicate that we have successfully established a method for constructing Mstn-KO cell lines with stable passage. No expression of the MSTN protein and strong cell proliferation were observed in the cell lines. Moreover, real-time PCR experiments showed that the expression levels of miR-1, miR-431, miR-206, and miR-133a were significantly increased (P < 0.01), the expression level of miR-23a was significantly increased (P < 0.05), and the expression level of miR-486 was significantly decreased (P < 0.05). These findings indicate that multiple miRNAs are closely associated with MSTN regulation. This study lays the foundation for further investigation into the effects of the Mstn gene on the physiological function of myoblasts and the development of drugs that block the MSTN signaling pathway.

Clinical value of the expression levels of tumor protein D52 and miR-133a on prognosis assessment of pancreatic cancer surgery.

To investigate the clinical value of the expression levels of tumor protein D52 (TPD52) and miR-133a on the prognosis assessment of pancreatic cancer surgery. This was a retrospective study. Ninety-seven patients who underwent radical surgery for pancreatic cancer in Cangzhou Central Hospital from January 2018 to January 2022 were selected and divided into four groups: TPD52 high expression group, TPD52 low expression group, miR-133a high expression group and miR-133a low expression group. The relationship between the expression levels of TPD52 and miR-133a and the clinicopathological features of patients with pancreatic cancer was analyzed. The COX regression model was used to analyze the risk factors affecting the prognosis of patients with pancreatic cancer. The high expression rate of TPD52 and the low expression rate of miR-133a in pancreatic cancer tissues were higher than those in normal paracancerous tissues(P<0.05). Based on the comparison of prognosis and survival, the median survival time of patients with high expression of TPD52 and low expression of miR-133a was lower than that of patients with low expression of TPD52 and high expression of miR-133a, with a statistically significant difference(P<0.05). Moreover, multivariate Cox regression analysis showed that low differentiation of pancreatic cancer, III-IV stage of TNM, high expression of TPD52, as well as low expression of miR-133a were independent risk factors for postoperative survival of patients with pancreatic cancer(P<0.05). TPD52 is expressed at a high level whereas miR-133a at a low level in pancreatic cancer tissues, both of which together with low differentiation of pancreatic cancer and III-IV stage of TNM constitute independent risk factors affecting the surgical prognosis of patients with pancreatic cancer.

The role of miR-133a in silibinin-mediated inhibition of the PI3K/AKT/mTOR pathway in MCF-7 breast carcinoma cells.

Breast cancer is particularly severe in women. Research highlights the crucial role of miRNAs in key cellular processes, showcasing their intricate interactions with the oncogenic PI3K/AKT/mTOR (PAM) signaling pathway and underscoring their significant role as tumor suppressors. The effect of silibinin on cell growth and survival was evaluated using an MTT assay. Bioinformatics analysis identified putative miR-133a targets inside the PAM pathway. After incubating MCF-7 cells with silibinin, we measured miR-133a, EGFR, PI3K, AKT, PTEN, and mTOR expression levels using qRT-PCR. Furthermore, protein expression levels of mTOR were assessed using Western blotting. The MTT experiment displayed that silibinin effectively inhibits MCF-7 cell proliferation in a time- and dose-dependent manner. Silibinin's IC50 value, determined at 370 μM after 48 hours, was established. qRT-PCR analysis at this IC50 concentration highlighted reduced expression of EGFR, PI3K, AKT, PTEN, and mTOR mRNAs, alongside increased miR-133a expression. Notably, miR-133a exhibited a negative correlation with both EGFR and PIK3C2A expression. Furthermore, western blotting confirmed silibinin's capacity to diminish p-mTOR protein levels, the ultimate element of the PAM signaling pathway. The findings enhance comprehension of silibinin's impact on PAM signaling and miR-133a expression, offering promise for targeted therapies in disrupting oncogenic pathways in MCF-7 breast cancer cells. This insight could advance breast cancer treatment strategies.

Citronellal can alleviate vascular endothelial dysfunction by reducing ectopic miR-133a expression

Endothelial dysfunction (ED) is the initial cause of atherosclerosis (AS) and an early marker of many cardiovascular diseases (CVD). Citronellal (CT), a monoterpenoid natural product extracted from grass plant Citronella, has been shown to have anti-thrombotic, anti-hypertensive and anti-diabetic cardiomyopathy activities. The aim of this study is to investigate the effects of citronellal on vascular endothelial dysfunction and the underlying mechanisms. The left common carotid artery was subjected to one-time balloon injury to cause vascular endothelial injury, and the AS model was established by feeding with high-fat diet. Use of HUVECs H2O2 treatment induced HUVECs oxidative stress damage model. The blood lipid level, histopathology, Western blot, immunohistochemistry, RT-PCR, ELISA and in situ fluorescence hybridization of common carotid artery tissues and HUVECs were studied. CT significantly reduced vascular plate area and endothelial lipid and cholesterol deposition in the common carotid artery of mice in a dose-dependent manner. CT increased the expression of activated protein 2α (AP-2α/TFAP2A) and circRNA_102979, and inhibited the ectopic expression level of miR-133a. However, the constructed lentivirus with AP-2α silencing and circRNA_102979 silencing reversed this phenomenon. The current study verifies CT can increase the expression levels of AP-2α and circRNA_102979 in vascular endothelium, increase the adsorption effect of circRNA_102979 on miR-133a and relieve the inhibitory effect of miR-133a on target genes, thereby alleviating AS-induced ED.

The role of miR1 and miR133a in new-onset atrial fibrillation after acute myocardial infarction

BackgroundThe development of new-onset atrial fibrillation (NOAF) after acute myocardial infarction (AMI) is a clinical complication that requires a better understanding of the causative risk factors. This study aimed to explore the risk factors and the expression and function of miR-1 and miR-133a in new atrial fibrillation after AMI.MethodsWe collected clinical data from 172 patients with AMI treated with emergency percutaneous coronary intervention (PCI) between October 2021 and October 2022. Independent predictors of NOAF were determined using binary logistic univariate and multivariate regression analyses. The predictive value of NOAF was assessed using the area under the receiver operating characteristic (ROC) curve for related risk factors. In total, 172 venous blood samples were collected preoperatively and on the first day postoperatively; the expression levels of miR-1 and miR-133a were determined using the polymerase chain reaction. The clinical significance of miR-1 and miR-133a expression levels was determined by Spearman correlation analysis.ResultsThe Glasgow prognostic score, left atrial diameter, and infarct area were significant independent risk factors for NOAF after AMI. We observed that the expression levels of miR-1 and miR-133a were significantly higher in the NOAF group than in the non-NOAF group. On postoperative day 1, strong associations were found between miR-133a expression levels and the neutrophil ratio and between miR-1 expression levels and an increased left atrial diameter.ConclusionsOur findings indicate that the mechanism of NOAF after AMI may include an inflammatory response associated with an increased miR-1-related mechanism. Conversely, miR-133a could play a protective role in this clinical condition.

Selected miRNA and Psoriasis-Cardiovascular Disease (CVD)-Overweight/Obesity Network-A Pilot Study.

Psoriasis is nowadays recognized as a multifactorial systemic disease with complex and not fully understood pathogenesis. In psoriatic patients, the increased cardiovascular disease (CVD) risk and frequent comorbidities like obesity are observed. The aim of this study was to investigate differences in miRNA (miR-22-3p, miR-133a-3p, miR-146a-5p, miR-369-3p, and Let-7b-5p) involved in CVD risk among psoriatic patients with overweight/obesity and with normal weight. The study comprised 28 male psoriatic patients and 16 male healthy controls. miRNA isolated from peripheral blood mononuclear cells was reverse-transcribed and RT-qPCR was performed. We have found decreased levels of miR-22, miR-133a, miR-146a, and miR-369 among the psoriatic patients. There was a statistically significant difference in miR-22 and miR-146a levels between psoriatic patients with overweight/obesity and with normal weight. There were positive correlations between miR-22 and miR-146a levels and psoriatic arthritis (PsA) in psoriatic patients with normal weight and between the miR-133a level and PsA in the overweight/obese patients. The decreased levels of selected miRNA are consistent with the levels observed in CVD indicating their impact on the CVD risk in psoriatic patients. miR-22 and miR-146 may be recognized as one of the contributing factors in the obesity-CVD-psoriasis network.

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&gt;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.

Investigation of miR-133a, miR-637 and miR-944 genes expression and their relationship with PI3K/AKT signaling in women with breast cancer.

MicroRNAs (miRNAs) are regulatory molecules capable of positively or negatively regulating signaling pathways, and are involved in tumorigenesis as well as various aspects of cancer. The purpose of this study was to investigate the expression levels of miR-133a, miR-637, and miR-944 in serum and tumor tissues as well as their relationship with the expression level of phosphatidylinositol-3-kinase (PI3K) and protein kinase-B (AKT) genes and proteins along with their clinical significance in breast cancer. The expressions of miR-133a, miR-637, miR-944, PI3K, and AKT genes were examined in the tumor and tumor margin tissues of 40 patients with breast cancer, as well as the serum levels of miR-133a, miR-637, and miR-944 in these patients and 40 healthy groups by quantitative real-time PCR (qRT-PCR). PI3K and AKT proteins expression in tumor and tumor margin tissues were detected using immunohistochemistry (IHC). The expression levels of miR-133a and miR-637 in the tumor tissue and serum of patients were lower than those in the tumor margin tissue and serum of the healthy group, respectively. In addition, the expression level of miR-944 in the tumor tissue was lower than that in the tumor margin tissue, but its expression increased in the serum of cancer patients compared to that in the healthy group. The expression of miR-637 was correlated with tumor location and Her2 receptors, and the expression of miR-944 was correlated with tumor location and family history. PI3K and AKT mRNA and protein levels were higher in the tumor tissues than in the tumor margin tissues (p < 0.05). The results of our study revealed that miR-637 has a better diagnostic value in breast cancer than miR-133a and miR-944.

Decrease in the expression of muscle-specific miRNAs, miR-133a and miR-1, in myoblasts with replicative senescence.

Muscles that are injured or atrophied by aging undergo myogenic regeneration. Although myoblasts play a pivotal role in myogenic regeneration, their function is impaired with aging. MicroRNAs (miRNAs) are also involved in myogenic regeneration. MiRNA (miR)-1 and miR-133a are muscle-specific miRNAs that control the proliferation and differentiation of myoblasts. In this study, we determined whether miR-1 and miR-133a expression in myoblasts is altered with cellular senescence and involved in senescence-impaired myogenic differentiation. C2C12 murine skeletal myoblasts were converted to a replicative senescent state by culturing to a high passage number. Although miR-1 and miR-133a expression was largely induced during myogenic differentiation, expression was suppressed in cells at high passage numbers (passage 10 and/or passage 20). Although the senescent myoblasts exhibited a deterioration of myogenic differentiation, transfection of miR-1 or miR-133a into myoblasts ameliorated cell fusion. Treatment with the glutaminase 1 inhibitor, BPTES, removed senescent cells from C2C12 myoblasts with a high passage number, whereas myotube formation and miR-133a expression was increased. In addition, primary cultured myoblasts prepared from aged C57BL/6J male mice (20 months old) exhibited a decrease in miR-1 and miR-133a levels compared with younger mice (3 months old). The results suggest that replicative senescence suppresses muscle-specific miRNA expression in myoblasts, which contributes to the senescence-related dysfunction of myogenic regeneration.

DNA methylation-induced ablation of miR-133a accelerates cancer aggressiveness in glioma through upregulating peroxisome proliferator-activated receptor γ.

Emerging evidences suggest that miRNAs can be used as theranostic biomarkers for multiple cancers, including glioma. Thus, identification of novel miRNAs for glioma treatment and prognosis becomes necessary and urgent. Here, by analyzing miRNA expression profiles in the glioma and para-cancer tissues by miRNA microarray and verified by RT-PCR, we found that miR-133a was significantly downregulated in the cancerous tissues, and patients with low-expressed miR-133a levels predicted an unfavorable prognosis. The following functional experiments confirmed that overexpression of miR-133a restrained cell proliferation and colony formation abilities, and induced cell cycle arrest to restrain cancer progression in glioma cells. Then, the underlying mechanisms were uncovered, and the peroxisome proliferator-activated receptor γ (PPARγ, PPARG) was verified as the downstream target of miR-133a. Mechanistically, miR-133a negatively regulated PPARG expressions by binding to its 3' untranslated regions (3'UTR). The following rescuing experiments evidenced that miR-133a overexpression-induced anti-cancer effects in glioma cells were abrogated by upregulating PPARγ. Interestingly, we noticed that the promoter region of miR-133a was hypermethylated, and removal of DNA methylation by 5-Azacytidine (AZA) significantly increased the expression levels of miR-133a in glioma cells. Taken together, we concluded that DNA-methylation-induced miR-133a silence contributed to cancer progression in glioma through upregulating PPARγ, and firstly identified the DNA-methylation-regulated miR-133a/PPARG axis as the novel indicators for glioma treatment and prognosis.

Potential diagnostic role of circulating MiRNAs in colorectal cancer.

Colorectal cancer (CRC) is the third most common and fourth most deadly cancer worldwide despite its various screening method. Thus, the search for novel and better markers is continuous. This study aimed to assess the combined expression levels of miR-133a, miR-574-3p, and miR-27a in early diagnosis of colorectal cancer in comparison to traditional tumor markers (CEA and CA19.9). miR-133a, miR-574-3p, and miR-27a were assessed in sera of 120 participants categorized into healthy control group (n = 20), benign group (n = 30) and malignant group (n = 70) using real-time PCR. miR-133a, miR-574-3p, and miR-27a expressions showed significant difference among different staging, grading and tumor size of CRC. The sensitivities of the three miRNAs whether combined or individually used were better than routinely used tumor markers (CEA and CA19.9) leading to more accurate and faster diagnosis of CRC. Synergetic detection of miRNA-133a, miRNA-574-3p, and miRNA-27a may serve as better noninvasive biomarkers with higher combined sensitivity for early diagnosis of CRC than individual detection of miRNAs.

Differences in MicroRNA Expression in Pericoronary Adipose Tissue in Coronary Artery Disease Compared to Severe Valve Dysfunction.

Pericoronary adipose tissue (PCAT) is a source of microRNAs (miRs) that act as messengers for intercellular communication. We investigated whether the PCAT surrounding significant coronary atherosclerotic lesions shows specific miR expression patterns compared with PCAT surrounding plaque-free segments. We included 49 patients with 3-vessel coronary artery disease (CAD) and 19 patients with severe valvular disease but no CAD, who underwent elective cardiac surgery. The PCAT was harvested from two sites: adjacent to a significant atherosclerotic coronary lesion and from plaque-free segments. miR-133a, miR-21, miR-26b, miR-9, and miR-143 levels in PCAT cells were quantified by real-time reverse transcription polymerase chain reaction (data expressed as arbitrary units). Expression of miR-133, miR-21, and miR-26b in adipose tissue at a site without atherosclerotic lesion was much lower in patients with CAD than in those without CAD (0.82 ± 1.37 vs 1.86 ± 0.52, P < .001, 0.45 ± 1.3 vs 1.51 ± 1.11, P < .001, 0.3 ± 1.25 vs 1.2 ± 0.73, P = .02, respectively). In addition, miR-133, miR-21, and miR-143 in CAD patients showed significantly greater expression in PCAT from atherosclerotic lesion compared with plaque-free segments (1.32 ± 0.96 vs 0.82 ± 0.37 (P = .011), 0.91 ± 1.7 vs 0.3 ± 1.25 (P = .012), 1.2 ± 1.59 vs 0.43 ± 0.54 (P < .001), respectively). Our findings open new perspectives for the role of PCAT in the pathophysiology of atherosclerosis and should be further investigated.

Differences in microRNA expression in pericoronary tissue adjacent to atherosclerotic plaque and from lesion-free coronary artery

Abstract Background Pericoronary adipose tissue (PCAT) plays a key role in the pathogenesis of atherosclerosis. PCAT is a source of microRNAs (miRs) that acts as messengers for intercellular communication. In this study, we investigated whether the PCAT surrounding coronary occlusive atherosclerotic lesions shows specific miR expression patterns compared to PCAT surrounding plaque-free segments. Purpose To evaluate the expression of miRs that are known to be implicated in the pathophysiology of atherosclerosis. Methods We included 49 patients (38 men, age 65±9 years) with 3-vessel coronary artery disease, who underwent elective coronary bypass surgery. The PCAT was harvested from two sites: adjacent to an occlusive atherosclerotic coronary lesion, and a plaque-free segment. miR-133a, miR-21, miR-26b, miR-9 and miR-143 levels in PCAT cells were quantified by real-time reverse transcription polymerase chain reaction. Results miR-26b, miR-21 and miR-143 showed significantly greater expression in PCAT samples taken from around atheromatous plaque (154±295 versus 37±96, p=0.014, for miR-26b; 122±265 versus 29±79, p=0.014, for miR-21; and 181±356 versus 31±50, p=0.03, for miR-143) (Figure 1). In addition, the expression of miR-143, miR-26b and miR-21 in PCAT from atherosclerotic plaque showed robust positive correlations with body.mass index (r=0.451, p=0.001, r=0.41, p=0.03 and r=0.416, p=0.003 respectively), Conclusions Analysis of PCAT located in proximity to atherosclerotic plaque showed a different pattern of miR expression compared to PCAT in a plaque-free segment. Our findings open new perspectives for the role of PCAT in the pathophysiological mechanisms of atherosclerotic disease and should be further investigated. Funding Acknowledgement Type of funding sources: None.

Circ-NCX1 inhibits LPS-induced chondrocyte apoptosis by regulating the miR-133a/SIRT1 axis.

Osteoarthritis (OA) is a degenerative joint disease, which is characterized by the degeneration of articular cartilage, thickening of subchondral bone, and inflammation of the synovial membrane. In this study, we aimed to investigate the effects and underlying mechanisms of circ-NCX1 in lipopolysaccharide (LPS)-induced injury in SW1353 chondrocytes, an in vitro model of OA. The levels of circ-NCX1, miR-133a, and related apoptotic proteins were determined by RT-qPCR. MTT assay was used to evaluate the cell viability. The apoptosis was determined by flow cytometry, whereas the expression of apoptosis proteins was detected by Western blot. Immunofluorescence was used to detect cleaved caspase-3 expression in cells. Luciferase reporter assay was used to verify the interaction between circ-NCX1 and miR-133a, and between miR-133a and Silent information regulator 2 homolog 1 (Sirt1). The results showed that the overexpression of circ-NCX1 significantly upregulated the chondrocyte viability and proliferation, and alleviated apoptosis in LPS-induced SW1353 cells. Immunofluorescence results showed that the overexpression of circ-NCX1 significantly reduced expression of LPS-stimulated cleaved Caspase-3. The RT-qPCR results showed that the overexpression of circ-NCX1 inhibited mRNA levels of cleaved Caspase-3 and Bax, and promoted mRNA levels of Bcl-2. Luciferase reporter assay showed that circ-NCX1 targeted miR-133a, and miR-133a directly targeted the Sirt1. In addition, overexpression of circ-NCX1 inhibited chondrocyte apoptosis and promoted Akt phosphorylation via the miR-133a/Sirt1 axis in LPS-induced chondrocytes. In conclusion, circ-NCX1 may serve as an important regulator of LPS-induced chondrocyte apoptosis through the miR-133a/Sirt1 axis, and may be involved in the development of OA.

Novel miRNA Biomarkers for Patients With Duchenne Muscular Dystrophy.

Creatine kinase (CK) as a biomarker has long been expected to be replaced by other fluid biomarkers for Duchenne muscular dystrophy (DMD) because it is independent of disease severity. Growing evidence has demonstrated that muscle-specific microRNAs, known as myomiRs, can act as biomarkers for monitoring muscle pathology and disease severity of DMD patients. To gain insights into the relationship between serum myomiRs and clinical assessment, we measured serum levels of miR-1, miR-133a, miR-133b, miR-206, miR-208a, miR-208b, and miR-499 in 48 DMD patients by using real-time quantitative reverse transcription polymerase chain reaction. These were then compared with age, muscle strength, muscle functions, CK levels, cardiac manifestations, and mutation types (deletions, duplications, and small mutations). When compared to 53 controls, the expression levels of myomiRs were all significantly elevated (p < 0.05). The receiver operating characteristic curves of all seven myomiRs reflected marked differences between DMD patients and healthy controls (p < 0.05). We also showed that serum levels of myomiRs were positively correlated with lower limb distal muscle strength in patients of all age groups. The levels of miR-499, miR-208b, miR-133a, and miR-133b had significant negative correlations with the time to be upright from the supine position (Gowers' time) and the time taken to climb four stairs in DMD patients older than 7 years. Serum levels of miR-1, miR-133a, miR-133b, and miR-499 in patients with cardiac involvement were remarkably higher than those in non-cardiac-involved patients. There was no significant difference in levels of myomiRs between the different mutation groups. Our results indicated that serum myomiRs could be considered as novel biomarkers for monitoring pathology/pathophysiology of DMD patients. In particular, miR-499, miR-208b, miR-133a, and miR-133b might have the ability to reflect the extent of muscle impairment.

MICRORNA PROFILE ANALYSIS IN PERICORONARY ADIPOSE TISSUE OF DIABETIC PATIENTS WITH SIGNIFICANT CORONARY ARTERY DISEASE

Objective: Pericoronary adipose tissue (PCAT) regulates arterial homeostasis, is considered to act in paracrine manner, and plays a role in the pathogenesis of atherosclerosis. PCAT may be a source of microRNAs (miRs) that target other tissues and act as messengers for intercellular communication. In this study, we investigated whether the PCAT surrounding coronary occlusive atherosclerotic lesions shows specific miRs expression patterns in patients with diabetes type 2 compared to non-diabetic patients. Design and method: We enrolled 46 patients (30 men, aged 65 ± 12 years old) with 3-vessel coronary artery disease who underwent elective coronary bypass surgery with and without diabetes type 2. The PCAT samples were received from all participants. miR-133a, miR-21, miR-26b, miR-9 and miR-143 expression levels in PCAT cells were quantified by real-time reverse transcription polymerase chain reaction. Results: Twenty-four patients with diabetic type 2 (15 men, 64 ± 10 years old) and twenty-two non-diabetic patients (15 men, 62 ± 15 years old) were included in the study. PCAT analysis showed a significant upregulation of miR-21 levels in diabetic compared to non-diabetic patients (184 ± 77 versus 21 ± 15, p = 0.04). Diabetic patients also revealed a significant increase of miR-26b and miR-143 expression in PCAT samples compared to on-diabetics (33 ± 22 versus 16 ± 13, p = 0.02, 93 ± 42 versus 16 ± 23, p = 0.01). No significant differences between the two sites were observed in PCAT expression of miR-133a and miR-9 (73 ± 12 versus 130 ± 143, 56 ± 44 versus 34 ± 33, respectively, p = NS for both). Conclusions: miRs expression in PCAT from diabetic patients with significant coronary disease show a distinct expression profile. Our study opens new perspectives in the pathophysiologic role of PCAT in atherosclerotic complications of diabetes and should be further investigated.

Circulating MicroRNAs in Canine Congestive Heart Failure – Why Exosomes?

By influencing protein translation, microRNAs (miRNAs) have emerged as powerful regulators of a wide range of biological processes. miRNAs can be found freely circulating in blood as well as in circulating exosomes. It has been speculated that evaluating the quantity of miRNAs contained in exosomes may serve as a better diagnostic and prognostic tool than those found freely circulating.A good test case for this relationship is to look at miRNA dysregulation effecting Canine Congestive Heart Failure, resulting from Myxomatous Mitral Valve Disease (MMVD), in these two contexts. In order to do this, we are assessing levels of miR‐1, miR‐126, and miR‐133a, in nine small breed, older dogs with this disorder, compared to healthy dogs, using Realtime qPCR analysis.We hypothesize that our results will confirm the assertion that exosomes reveal a more reliable indication of MMVD than by quantifying freely circulating miRNAs.

Elevated serum miR-133a predicts patients at risk of periprocedural myocardial injury after elective percutaneous coronary intervention.

BackgroundPeriprocedural myocardial injury (PMI) is a frequent complication of percutaneous coronary intervention (PCI) associated with poor prognosis. However, no effective method has been found to identify patients at risk of PMI before the procedure. MicroRNA-133a (miR-133a) has been reported as a novel biomarker in various cardiovascular diseases. Herein, it was sought to determine whether circulating miR-133a could predict PMI before the procedure.MethodsEighty patients with negative preoperative values of cardiac troponin T (cTnT) receiving elective PCI for stable coronary artery disease (CAD) were recruited. Venous serum samples were collected on admission and within 16–24 hours post-PCI for miRNA measurements. PMI was defined as a cTnT value above the 99% upper reference limit after the procedure. The association between miR-133a and PMI was further assessed.ResultsPeriprocedural myocardial injury occurred in 48 patients. The circulating level of miR-133a was significantly higher in patients with PMI before and after the procedure (both p < 0.001). Receiver operating characteristic curve analysis of the preoperative miR-133a level revealed an area under the curve of 0.891, with a sensitivity of 93.8% and a specificity of 71.9% to predict PMI. Additionally, a decrease was found in fibroblast growth factor receptor 1 (FGFR1) in parallel with an increase in miR-133a levels in patients with PMI.ConclusionsThis study demonstrates for the first time that serum miR-133a can be used as a novel biomarker for early identification of stable CAD patients at risk of PMI undergoing elective PCI. The miR-133a-FGFR1 axis may be involved in the pathogenesis of PMI.