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Circulating microRNAs as candidate biomarkers in patients with systemic lupus erythematosus
Abstract
Aberrant expression of microRNAs (miRNAs) has been identified in various diseases. Recent studies demonstrated that miRNAs can be detected in the circulation and serve as potential biomarkers of various diseases. Moreover, the detection of circulating miRNAs can provide important novel information concerning diseases. In this study, a miRNA profile was used to determine the aberrantly expressed circulating miRNAs in patients with systemic lupus erythematosus (SLE) compared with patients with rheumatoid arthritis (RA) and healthy controls (HCs). To further confirm the microarray data, we identified 8 miRNAs (miR-126, miR-21, miR-451, miR-223, miR-16, miR-125a-3p, miR-155, and miR-146a) by real-time quantitative PCR (qRT-PCR) in 20 healthy controls and in 55 patients, of whom 30 patients were diagnosed with SLE and 25 were diagnosed with RA. Consistent with the microarray data, miR-126 was specifically enriched only in the blood of the SLE patients, but 4 other miRNAs (miR-21, miR-451, miR-223, and miR-16) were upregulated in the patients with SLE and were also significantly increased in the patients with RA. In contrast, miR-125a-3p, miR-155, and miR-146a showed a trend toward significantly reduced levels in the patients with SLE. In addition, to further estimate the potential roles of these differentially expressed circulating miRNAs in the pathogenesis of SLE, we used a bioinformatics exploratory analysis and identified a number of significantly enriched pathways, which implied that most dysregulated circulating miRNAs might be involved in various signal transduction pathways and cell interactions, particularly the mitogen-activated protein kinase signaling pathway. Based on these findings, we postulate that aberrantly expressed plasma miRNAs could be attractive as candidates for putative biomarkers of SLE and may help elucidate the possible pathogenesis of SLE.
... In addition, epigenetic changes play a key role in the onset of the disease such as changes in microRNAs [2][3][4]. Several studies have suggested microRNAs as a new biomarker for the diagnosis of lupus [5]. ...
... Studies indicated changes in the number of microRNA expressions in the lupus of peripheral blood mononuclear cells (PBMCs) [19,20]. Several studies identified miR-146a and miR-125a as inflammatory disease markers in autoimmune diseases, such as lupus [5,21] . Earlier studies indicated a decreased expression of miR-146a in PBMCs of lupus patients [22]. ...
... Earlier studies indicated a decreased expression of miR-146a in PBMCs of lupus patients [22]. Furthermore, miR-146a as a negative regulator of inflammation targeting STAT1 and IRF-5 causes an impairment in the IFN-I pathway, which finally helps its pathogenicity [5,19,23] . Also, miR-146a can activate TLR-4, TLR-2, and TLR-5 in monocytes [24] and increase the pro-inflammatory cytokines such as IL-1β [25]. ...
The microRNAs are non-coding RNA molecules involved in physiological and pathological processes, causing autoimmune diseases such as systemic lupus erythematosus (SLE). Probiotics are living microorganisms that possess beneficial effects on the host immune system and modulate it. The effect of Lactobacillus rhamnosus and Lactobacillus delbrueckii on the expression of miR-125a and miR-146a was studied in peripheral blood mononuclear cells (PBMCs) from newly diagnosed lupus patients in this in vitro study. During this study, 20 recently diagnosed SLE patients and 20 healthy individuals participated. Ficoll method was used to isolate the PBMCs from whole blood, which were cultured for 48 h with Lactobacillus rhamnosus and Lactobacillus delbrueckii. In the next step, total RNA containing microRNA was extracted. cDNA was synthesized for miR-125a and miR-146a genes and analyzed by real-time PCR. Results were presented as fold changes. As compared to healthy controls, SLE patients expressed lower levels of miR-125a and miR-146a. PBMCs treated with Lactobacillus rhamnosus, Lactobacillus delbrueckii, or both probiotics had significantly higher levels of miR-125a and miR-146a compared to the untreated group. Treatment of PBMCs with both L. rhamnosus and L. delbrueckii upregulated the expression of miR-125a and miR-146a in treated cells compared with untreated cells in SLE patients (p = 0.02, p = 0.001). Lactobacillus rhamnosus and Lactobacillus delbrueckii modify lupus patients’ immune responses and disease effects by regulating miR-125a and miR-146a.
... MiR-21, which is significantly overexpressed in CD4+ T-cells and is associated with DNA hypomethylation, was shown to be upregulated in the plasma of SLE patients (121). According to Chen X et al, circulating miRNA are mostly derived from circulating blood cells (115), and this result proposes that the increased level of circulating miR-21 might be due to selective secretion of upregulated intracellular miR-21 in CD4+ T-cells and the exposure of CD4+ T-cells to circulating plasma in inflammatory conditions. ...
... However, the specific mechanism of selective secretion remains unclear. Moreover, circulating miR-21 was also reported to be increased in patients with RA, indicating that it might not be specific for SLE (121). Plasma miR-21 level was highly correlated with SLE disease activity score, and therefore may be used as a biomarker to distinguish the severence of SLE (122,123). ...
... According to Wang et al, the serum and plasma levels of miR-146a in SLE patients were down regulated compared to healthy controls and negatively correlated with SLEDAI (121,125). Furthermore, serum exosomal miR-146a have shown to be decreased in SLE patients, and previous studies have reported that serum exosomal miR-146a negatively regulates senescence of SLE bone marrow mesenchymal stem cells by suppressing the TRAF6/NF-KB signaling pathway (182,183). These are consistent with the report that miR-146a, which is known to contribute to the disease by regulating the type 1 IFN pathway, is significantly decreased in the PBMC of SLE patients (126). ...
Systemic lupus erythematosus is a chronic autoimmune disease of which clinical presentation is vastly heterogeneous, ranging from mild skin rashes to severe renal diseases. Treatment goal of this illness is to minimize disease activity and prevent further organ damage. In recent years, much research has been done on the epigenetic aspects of SLE pathogenesis, for among the various factors known to contribute to the pathogenic process, epigenetic factors, especially microRNAs, bear the most therapeutic potential that can be altered unlike congenital genetic factors. This article reviews and updates what has been discovered so far about the pathogenesis of lupus, while focusing on the dysregulation of microRNAs in lupus patients in comparison to healthy controls along with the potentially pathogenic roles of the microRNAs commonly reported to be either upregulated or downregulated. Furthermore, this review includes microRNAs of which results are controversial, suggesting possible explanations for such discrepancies and directions for future research. Moreover, we aimed to emphasize the point that had been overlooked so far in studies regarding microRNA expression levels; that is, which specimen was used to assess the dysregulation of microRNAs. To our surprise, a vast number of studies have not considered this factor and have analyzed the potential role of microRNAs in general. Despite extensive investigations done on microRNA levels, their significance and potential role remain a mystery, which calls for further studies on this particular subject in regard of which specimen is used for assessment.
... Indeed, miR-16 was shown to target the 3 0 untranslated region of TNF-a [22,29] and thereby, miR-16 might regulate TNF-a signalling [12], which is crucial for RA pathogenesis. Interestingly, miR-16 has been found at significantly lower levels in persons with early RA [30], but upregulated in established RA [30][31][32]. Nevertheless, miR-16 remains a reliable marker of disease activity in people with RA [12,22]. ...
... Nevertheless, miR-16 remains a reliable marker of disease activity in people with RA [12,22]. miR-21 levels were also elevated in plasma of people with RA vs healthy adults [32]. It has been observed that miR-21 is expressed at higher levels in Treg vs Th17 cells. ...
... However, in RA there is an absence of TRAF6 and IRAK1 regulation by miR-146a, which could contribute to the sustained production of TNF-a and thus amplify inflammation [22]. miR-146 expression is low in people with RA [30,32] and is inversely correlated with CRP, ESR and TNF-a levels [12]. ...
MicroRNAs (miRNAs) are endogenously generated single-stranded RNAs that play crucial roles in numerous biological processes, such as cell development, proliferation, differentiation, metabolism and apoptosis. They negatively regulate target gene expression by repressing translation of messenger RNA into a functional protein. Several miRNAs have been implicated in the development and progression of RA. They are involved in inflammatory and immune processes and are associated with susceptibility to RA and disease activity. They are also considered to be potential markers of disease activity or even therapeutic targets. Likewise, several miRNAs are affected acutely by exercise and regulate exercise-related adaptations in the skeletal muscle and cardiovascular system and aerobic fitness. Interestingly, some miRNAs affected by exercise are also important in the context of RA. Investigating these might increase our understanding of the effects of exercise in RA and improve exercise prescription and, potentially, disease management. In this review, we focus on the miRNAs that are associated with both RA and exercise and discuss their roles in (and potential interactions between) RA and exercise-induced adaptations.
... MiRNA is a class of small (approximately 22-25 nucleotides) noncoding RNAs involved in the post-transcriptional regulation of expression of many genes [35,36] and the regulation of transcription and neuroinflammation [35][36][37]. In MS and SLE, specific miRNAs are found in blood and cerebrospinal fluid, which are characterized by the increased expression [37][38][39][40]. The extracellular functions of miRNAs include signaling between cells and the regulation of angiogenesis, neurogenesis, and cell proliferation [41]. ...
... As mentioned above, miRNAs have important functions in living organisms. Therein, the same miRNAs were found to be important in schizophrenia [48][49][50][51][52][53][54][55], MS [56][57][58], and systemic lupus erythematosus [40][41][42]. miR-137 is responsible for the proliferation and differentiation of embryonic and neurons stem cells, as well as the maturation of the synapses [59]; it inhibits AMPA-receptor-mediated synaptic transmission by reduction of the expression of the GluA1 protein of this receptor [60] and disrupts synaptic plasticity [61]. In the regulation of dopamine, D2 receptor expression miR-9-5p is involved, whereas miR-206 is associated in the site-specific regulation of NT5C2 [62]. ...
... The blood of patients with MS and SLE contains abzymes that recognize and hydrolyze several of their own antigens: MBP, DNA, histones, and oligosaccharides [8][9][10][11][13][14][15][16][17][18][19][20]. In schizophrenia [37][38][39][48][49][50][51][52][53][54][55], SLE [40][41][42], and MS [56][57][58] patients, some specific miRNAs are characterized by increased expression and participate in signaling between cells, the regulation of angiogenesis, neurogenesis, and cell proliferation. The change in miRNA expression in the extracellular compartment is associated with specific pathological processes. ...
Autoantibodies-abzymes hydrolyzing DNA, myelin basic protein, and oligosaccharides have been revealed in the sera of patients with multiple sclerosis (MS). In MS, specific microRNAs are found in blood and cerebrospinal fluid, which are characterized by increased expression. Autoantibodies, specifically hydrolyzing four different miRNAs, were first detected in the blood of schizophrenia patients. Here, we present the first evidence that 23 IgG antibodies of MS patients effectively recognize and hydrolyze four neuroregulatory miRNAs (miR-137, miR-9-5p, miR-219-2-3p, and miR-219-5p) and four immunoregulatory miRNAs (miR-21-3p, miR-146a-3p, miR-155-5p, and miR-326). Several known criteria were checked to show that the recognition and hydrolysis of miRNAs is an intrinsic property of MS IgGs. The hydrolysis of all miRNAs is mostly site-specific. The major and moderate sites of the hydrolysis of each miRNA for most of the IgG preparations coincided; however, some of them showed other specific sites of splitting. Several individual IgGs hydrolyzed some miRNAs almost nonspecifically at nearly all internucleoside bonds or demonstrated a combination of site-specific and nonspecific splitting. Maximum average relative activity (RA) was observed in the hydrolysis of miR-155-5p for IgGs of patients of two types of MS—clinically isolated syndrome and relapsing-remitting MS—but was also high for patients with primary progressive and secondary progressive MS. Differences between RAs of IgGs of four groups of MS patients and healthy donors were statistically significant (p < 0.015). There was a tendency of decreasing efficiency of hydrolysis of all eight miRNAs during remission compared with the exacerbation of the disease.
... Published evidence indicates that miRNAs participate in different immune responses (5,6), which makes them crucial candidates for use as diagnostic markers and treatment targets (7,8). Increasing evidence suggests that the expression characteristics of serum miRNAs makes them possible biomarkers of autoimmune diseases, including SLE, and therefore they should be exploited for the betterment of humankind (9,10). ...
... Recent studies have indicated that miR-451a was involved in the regulation of various human physiological and pathological processes (26). Furthermore, previous studies revealed that the levels of circulating miR-451a, which is important for erythropoiesis, were abnormal in the plasma of patients with SLE relative to normal controls, and miR-451a might be linked to abnormal erythropoiesis in SLE (9). Therefore, the present study evaluated the association between the levels of exosomal miR-451a in serum and the patient SLEDAI scores ( Figure 4A) or 24-h urine protein quantity ( Figure 4B). ...
... In this study, it was found that the difference in expression of exosomal miR-451a in CD4 + T or B cells and in serum of normal individuals and patients with SLE patients was markedly large, which might be associated with the selection mechanism of exosomes encapsulating miRNA (27). Emerging evidence indicated that miR-451a was involved in the regulation of various human physiological and pathological processes (26) and the level of circulating miR-451a was closely correlated with erythropoiesis formation, it was possible that miR-451a could be linked to the abnormal erythropoiesis of SLE (9). According to our current results, we used multiple international authoritative databases such as Mirbase/Targetscans/PicTar to predict the potential target gene of miRNA-451a, which hinted that MIF may act as a target gene. ...
Systemic lupus erythematosus (SLE) is a multi-system autoimmune disease characterized by continuous inflammation and the production of autoantibodies. Exosomes, acting as a critical tool for communication between cells, are involved in the pathogenesis of SLE, particularly in inflammation and immune imbalance. In this study, we aimed to extract and confirm the pro-inflammatory effect of serum exosomes in SLE. Then, we attempted to find differentially expressed exosomal microRNAs in the serum of healthy subjects and SLE patients via miRNA microarray analysis and validated the target exosomal microRNA, exosomal miR-451a, which expression level decreased in serum of SLE patients by RT-qPCR. Furtherly, we analyzed the correlation between exosomal miR-451a and disease activity, kidney damage and typing, and traditional medicine therapy. Finally, we investigated the intercellular communication role of exosomal miR-451a in SLE by co-culture assay in vitro. Taken together, our study demonstrated that downregulated serum exosomal miR-451a expression correlated with SLE disease activity and renal damage as well as its intercellular communication role in SLE which provided potential therapeutic strategies.
... Interestingly, many evidences show that miRNAs are playing main role in the induction of pro-inflammatory cytokines, which mediate the secretion of IL-1β-induced extracellular matrix degradation of cartilage [16][17][18]. In addition to that, the other evidences prove that miRNAs promoting inflammation and maintenance of RA synovium through the upregulation of the monocyte chemokine production and pro-inflammatory chemokine receptors [19]. ...
... A study by Hu SL, et al. 2017, demonstrated that downregulation of miR-21-5p and induction of IL-1β production via the ALK receptor, JNK, ERK, and AP-1 signaling pathways mediated by myostatin suggesting that mir-21-5p plays an important role in the treatment of RA [15]. Interestingly, a study shows that miR-21 is the most commonly upregulated miRNA in various cancers and it is upregulated in the blood and T cells of patients with RA [19,20]. Recent study also reported that miR-21 can induce the RA progression through the NF-κB nuclear translocation and NF-κB signaling pathway activation, which in turn accelerating fibroblast-like synoviocytes (FLS) proliferation [21]. ...
... It has been reported that miR-21 may be involved in the activation of the JAK/STAT signaling pathway in patients with systemic juvenile idiopathic arthritis [37]. As we know that systemic lupus erythematosus (SLE) is a kind of autoimmune disorder characterized by B cell homeostasis imbalance, which leads to tissue inflammation and it has been reported that miR-21 expressed higher levels in SLE [19,38]. So miR-21 plays an important role in SLE, but it requires more validated results and experiments. ...
Arthritis is a kind of autoimmune disease, which includes many circumstances that affect joints, the tissues surrounding the joints, and other connective tissues. Osteoarthritis (OA) and rheumatoid arthritis (RA) are the common arthritis seen in many populations. Researchers have made extensive studies on all types of arthritis, novel drugs are being developed by many laboratories, but yet no treatment option is available for these diseases and need new insight into the molecular pathways and pathophysiology of all types of arthritis. MicroRNAs (miRNAs), a class of non-coding RNAs, have shown to be played a plenty of roles in both a suppressive and a promoting role in disease pathogenesis and progression. Among the classes of microRNAs, miR-21 is a widespread miRNA commonly upregulated in many diseases and suggesting that it plays an important role in cell proliferation, apoptosis, and invasion. It is highly expressed in osteoclast precursors and the pro-osteoclastogenic nature of miR-21 makes it a promising candidate as a therapeutic target to treat bone-related disorders. Up to now, there are few papers that demonstrate the role of miR-21 in arthritis and related bone disorders and the number of studies related to different types of arthritis is sparse. Therefore, the main thrust of this paper is to provide an overview of the current clinical evidence and significance of miR-21 in arthritis and bone-related inflammation disorders. We summarize the important research findings surrounding the role of miR-21 and its involvement in the treatment of different types of arthritis.
... Antecedently a study conducted on profile of mRNAs in SLE patients identified 8 miRs to be deregulated in SLE patients, among them miR-16, miR-21, mir-451 and miR-223 were overexpressed in the subjects, whereas miR-125a-3p, miR-155 and miR-146a were downregulated. MiR-125a-3p was potentially involved in the progression and development of SLE [28]. ...
... Previously some of the reports had focused on deregulation of adaptive immunity in LN, however miRs have been identified to be the potential regulators in development and progression of disease. Furthermore, a report recently has presented the potential of miRs such as mir-16, miR-21, mir-451 and miR-223 were up-regulated, whereas miR-155, miR-125a-3p and miR-146a were down-regulated in LN [28]. However, more studies are needed in search of new therapies. ...
... We discovered that, the levels of miR-125a-3p were downregulated in kidneys of MRL/MPJ-Fas lpr /J mice compared to BALB/C mice ( Figure 1C). In a study reported earlier, 8 miRs were found to be deregulated in SLE subjects [28], among which miR-125a-3p was involved in cascades related to SLE. Thus, we supposed that the regulation of miR-125a-3p is essential for the development and progression of SLE in MRL/MPJ-Fas lpr /J mice. ...
Lupus nephritis (LN) is an autoimmune disorder mediated by systemic lupus erythematosus (SLE). Micro RNAs also called as miRs act potentially in the development and progression of SLE. MiR-125a-3p is reported to be down-regulated inpatients of SLE. Aim of the study was to evaluate the function of miR-125a-3p and its effect on regulation of fibrosis and interleukin (IL)-17 in LN. The Renal physiology in MRL/MPJ-Fas lpr/J mice was done by Hematoxylin and eosin staining; expression of miR-125a-3p in renal tissues by RT-PCR, Immunoblotting analysis was done expression of proteins. For in vitro studies, rat mesangial SV40MES13 cells were used and were transfected with vectors. Luciferase activity was done for studying the potential binding of miR-125a-3p with IL-17. It was found that, the expression of miR-125a-3p in kidney tissues of experimental mice were towards lower side versus the control; on the contrary the levels of IL-17 were up-regulated in experimental mice. Luciferase activity suggested that miR-125a-3p binds potentially on the 3'UTR region of IL-17. The assay also suggested up-regulation of miR-125a-3p and suppressed levels of IL-17 in SV40MES13 cells. The up-regulation of miR-125a-3p suppressed the levels of collagen I/II and transforming growth factor-β1 (TGF-β1) in SV40MES13 cells. MiR-125a-3p could be a important factor in the pathogenesis of LN which causes decrease in expression of IL-17 by potentially binding to the 3'UTR region causing suppression of fibrosis via down-regulating TGF-β1 in the SV40MES13 rat mesangial cells.
... Previous microarray studies indicated that miR-574-5p is among the miRNAs that was significantly upregulated in the plasma from SLE patients and rheumatoid arthritis patients [29]. To ascertain the aberrant upregulation of miR-574-5p in SLE, we assessed the levels of serum and tissues miR-574-5p in lupus patients and lupus-prone mice. ...
... Consistent with previous studies of SLE and rheumatoid arthritis human patients [29,64,65], we showed that miR-574-5p was not only significantly elevated in the serum of SLE patients but also in serum and a number of tissues in the lupus-prone B6.Fas lpr mice (Fig. 1). Furthermore, the up-regulation in miR-574-5p expression appears to be positively correlated with the development of lupus or lupus nephritis in both humans and mice. ...
Endosomal single-stranded RNA-sensing Toll-like receptor-7/8 (TLR7/8) plays a pivotal role in inflammation and immune responses and autoimmune diseases. However, the mechanisms underlying the initiation of the TLR7/8-mediated autoimmune signaling remain to be fully elucidated. Here, we demonstrate that miR-574-5p is aberrantly upregulated in tissues of lupus prone mice and in the plasma of lupus patients, with its expression levels correlating with the disease activity. miR-574-5p binds to and activates human hTLR8 or its murine ortholog mTlr7 to elicit a series of MyD88-dependent immune and inflammatory responses. These responses include the overproduction of cytokines and interferons, the activation of STAT1 signaling and B lymphocytes, and the production of autoantigens. In a transgenic mouse model, the induction of miR-574-5p overexpression is associated with increased secretion of antinuclear and anti-dsDNA antibodies, increased IgG and C3 deposit in the kidney, elevated expression of inflammatory genes in the spleen. In lupus-prone mice, lentivirus-mediated silencing of miR-574-5p significantly ameliorates major symptoms associated with lupus and lupus nephritis. Collectively, these results suggest that the miR-574-5p-hTLR8/mTlr7 signaling is an important axis of immune and inflammatory responses, contributing significantly to the development of lupus and lupus nephritis.
... Results miR-574-5p is upregulated in SLE patients and lupus-prone mice and correlates with disease severity Previous microarray studies indicated that miR-574-5p is among the miRNAs that was signi cantly upregulated in the plasma from SLE patients and rheumatoid arthritis patients (29). To ascertain the aberrant upregulation of miR-574-5p in SLE, we assessed the levels of serum and tissues miR-574-5p in lupus patients and lupus-prone mice. ...
... Consistent with previous studies of SLE and rheumatoid arthritis human patients (29,62, 63), we showed that miR-574-5p was not only signi cantly elevated in the serum of SLE patients but also in serum and a number of tissues in the lupus-prone B6.Fas lpr mice (Fig. 1). Furthermore, the up-regulation in miR-574-5p expression appears to be positively correlated with the development of lupus or lupus nephritis in both humans and mice. ...
Endosomal single-stranded RNA-sensing Toll-like receptor-7/8 (TLR7/8) plays a pivotal role in inflammation and immune responses and autoimmune diseases. However, the mechanisms underlying the initiation of the TLR7/8-mediated autoimmune signaling remain to be fully elucidated. Here, we demonstrate that miR-574-5p is aberrantly upregulated in tissues of lupus prone mice and in the plasma of lupus patients, with its expression levels correlating with the disease activity. miR-574-5p binds to and activates human hTLR8 or its murine ortholog mTlr7 to elicit a series of MyD88-dependent immune and inflammatory responses. These responses include the overproduction of cytokines and interferons, the activation of STAT1 signaling and B lymphocytes, and the production of autoantigens. In a transgenic mouse model, the induction of miR-574-5p overexpression is associated with increased secretion of antinuclear and anti-dsDNA antibodies, increased IgG and C3 deposit in the kidney, elevated expression of inflammatory genes in the spleen. In lupus-prone mice, lentivirus-mediated silencing of miR-574-5p significantly ameliorates major symptoms associated with lupus and lupus nephritis. Collectively, these results suggest that the miR-574-5p-hTLR8/mTlr7 signaling is an important axis of immune and inflammatory responses, contributing significantly to the development of lupus and lupus nephritis.
... According to most of the literature reports, miR-574-5p are highly expressed in cancer such as: thyroid cancer, esophageal cancer, colorectal cancer, non-small cell lung cancer and other types of tumors [27,28]. Other articles have reported that miR-574-5p is associated with eclampsia [29], coronary syndromes [30], sepsis [31], and autoimmune diseases [32][33][34]. ...
... Circulating miR-574-5p expression was up-regulated in patients both in SLE and RA. [32,37]. ...
Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized with aggressive arthritis, accompanied by extra articular and systemic manifestations. The pathogenesis of RA is still unclear. MicroRNA (miRNA) is single stranded, noncoding, small molecule RNA that is actively involved in inflammatory diseases and may play a role in RA. A prospective cohort study involving 66 patients with RA and 16 normal controls (NC) was conducted. All patients （49 females and 17 males）were divided into three groups （low disease activity 21/66, moderate disease activity 18/66, high disease activity 27/66）based on DAS28-ESR. The expression of miR-574-5p and the concentration of proinflammatory cytokines TNF-α, IL-1β and IL-6 in serum of the patients and normal controls were detected. Compared with the NC, the serum levels of miR-574-5p, TNF-α, IL-1β and IL-6 were significantly increased (P < 0.05). In addition, the fold of increase was correspondent to the level of disease activity. Moreover, the expression of miR-574-5p appeared to correlate to the concentration of individual cytokine. The expression of miR-574-5p in the serum of patients with RA was correlated with disease activity and proinflammatory cytokines, suggesting that it may be involved in the pathogenesis of RA.
... Heat shock proteins DOX-induced oxidative stress and ATP depletion induce high expression of heat shock proteins (HSPs), which regulate the activity of multiple signaling intermediates involved in the execution of apoptotic signaling pathways [75]. For example, HSP10 and HSP60 overexpression led to an increase in the post-translational modification of Bcl-2 proteins induced by DOX [129], HSP20 reduced DOX-associated oxidative stress, and cardiotoxicity via interacting with AKT phosphorylation [76]. A significant 3,9-fold upregulation of the HSP90 gene (HSPCB) was detected in the rat hearts at 48 hours post-DOX administration [85], a finding that correlated with the 16-fold HSP90 induction by cardiac ischemia [77], due to ROS accumulation [78] and reduction of ATP concentration [79]. ...
... Elevated miR-23b suppressed IL-17-, TNF-αand IL-1β-induced NF-κB activation, which correlated with its downregulation in inflammatory disorders, such as SLE and rheumatoid arthritis [126]. Several of DemiRs have been associated with inflammatory and autoimmune diseases such as elevated miR-16, miR-486, miR 92, miR-532, miR-140 [127][128][129]. None of these miRNAs have been reported as potential candidate biomarkers for the prediction of DOXinduced cardiotoxicity. ...
Doxorubicin (DOX) is a highly effective chemotherapy agent that often causes cardiotoxicity. Despite a number of extensive studies, the risk for DOX cardiotoxicity remains unpredictable. The majority of the studies on DOX-induced cardiotoxicity have been focused on the effects on cardiomyocytes that lead to contractile dysfunction. The roles of systemic inflammation, endothelial injury and neutrophil recruitment, all induced by the DOX, are increasingly recognized as the mechanisms that trigger the development and progression of DOX-induced cardiomyopathy. This review explores recent data regarding the possible mechanisms and biomarkers of early subclinical DOX-associated cardiotoxicity.
... Plasma levels of miR-16, miR-21, miR-223, miR-451 and miR-126 were higher in systemic lupus patinets than in HC. The first 4 miRNAs were higher also in RA patients compared to HC. 79 Another study showed that upregulated plasma levels of miR-24, miR-26a, and miR-125a-5p distinguished patients with RA from HC with high specificity. The combination of miR-24 and miR-125a-5p was described as a potential diagnostic marker of RA. ...
... Next, bioinformatics analysis confirmed that these miRNAs regulate transduction pathways and cellular interaction in SLE. 79 A previous study on CD4 þ T cells from patients with SLE showed that miR-126 downregulated DNA methylation and contributed to higher T cell autoreactivity by targeting DNA methyltransferase 1 (Dnmt1), which causes DNA methylation. 108 MiR-21 as a direct target of inhibitor PDCD4 (programmed cell death protein 4) contributes to aberrant T cell phenotype in human SLE patients. ...
MicroRNAs (miRNAs) are small non-coding single-stranded RNAs of about 22 nucleotides in length that act as post-transcriptional regulators of gene expression. Depending on the complementarity between miRNA and target mRNA, cleavage, destabilization, or translational suppression of mRNA occurs within the RISC (RNA-induced silencing complex). As gene expression regulators, miRNAs are involved in a variety of biological functions. Dysregulation of miRNAs and their target genes contribute to the pathophysiology of many diseases, including autoimmune and inflammatory disorders. MiRNAs are also present extracellularly in their stable form in body fluids. Their incorporation into membrane vesicles or protein complexes with Ago2, HDL, or nucleophosmin 1 protects them against RNases. Cell-free miRNAs can be delivered to another cell in vitro and maintain their functional potential. Therefore, miRNAs can be considered mediators of intercellular communication. The remarkable stability of cell-free miRNAs and their accessibility in body fluid makes them potential diagnostic or prognostic biomarkers and potential therapeutic targets. Here we provide an overview of the potential role of circulating miRNAs as biomarkers of disease activity, therapeutic response, or diagnosis in rheumatic diseases. Many circulating miRNAs reflect their involvement in the pathogenesis, while for plenty, their pathogenetic mechanisms remain to be explored. Several miRNAs described as biomarkers were also shown to be of therapeutic potential, and some miRNAs are already tested in clinical trials.
... Moreover, persons with serious co-morbidities or mental health problems have also been excluded. Written informed consent from each participant was obtained after the study approval by the institutional review board of the Medical University of Graz (EC Nr. 24-224 ex [11][12]. The BioPersMed study is conducted in compliance with Good Clinical Practice Guidelines Procedures (GCP) and carried out according to the principles of the Declaration of Helsinki. ...
... The study was conducted in accordance with the Declaration of Helsinki, and approved by the Ethics Committee of the Medical University of Graz (EC Nr. 24-224 ex [11][12]. ...
Hashimoto’s thyroiditis (HT) is the most prevalent autoimmune disorder of the thyroid (AITD) and characterized by the presence of circulating autoantibodies evoked by a, to date, not fully understood dysregulation of the immune system. Autoreactive lymphocytes and inflammatory processes in the thyroid gland can impair or enhance thyroid hormone secretion. MicroRNAs (miRNAs) are small noncoding RNAs, which can play a pivotal role in immune functions and the development of autoimmunity. The aim of the present study was to evaluate whether the expression of 9 selected miRNAs related to immunological functions differ in patients with HT compared to healthy controls. MiRNA profiles were analysed using quantitative reverse transcription polymerase chain reaction (qRT-PCR) in 24 patients with HT and 17 healthy controls. Systemic expressions of miR-21-5p, miR-22-3p, miR-22-5p, miR-142-3p, miR-146a-5p, miR-301-3p and miR-451 were significantly upregulated in patients with HT (p ≤ 0.01) and were suitable to discriminate between HT and healthy controls in AUC analysis. Altered expressions of miR-22-5p and miR-142-3p were associated with higher levels of thyroid antibodies, suggesting their contribution to the pathogenesis of HT.
... Moreover, modulating the abundance of miRNAs is considered as promising therapeutic strategies 16,17 . A study has shown that miR-125a-3p was upregulated in serum of patients with RA, while few studies have illustrated the regulation mechanism of miR-125a-3p on RA 18 . In this study, we explored the role and mechanism of the miR-125a-3p on RA. ...
... For instance, miR-338-3p was significantly upregulated in RA tissues, and miR-338-3p could enhance the inflammatory response via targeting SPRY1 22 . One study has found that downregulated miR-125a-3p has been previously identified as a landmark event in RA tissues 18 . In this study, we have illustrated the regulation mechanism involved mediating the antiinflammatory effects of miR-125a-3p in RA-FLS. ...
Objectives:
To explore the role and mechanism of miR-125a-3p in rheumatoid arthritis (RA) progression.
Methods:
The RA-tissues and fibroblast-like synovial cells in rheumatoid arthritis (RA-FLS) were used in this study. qRT-PCR, western blot and ELISA assay were performed to detect the expression levels of IL-6, IL-β and ΤΝF-α. Dual-luciferase reporter gene assay was used to observe the binding effect of miR-125a-3p and MAST3, and CCK-8 was used to observe the effect of miR-125a-3p on the proliferation of RA-FLS.
Results:
miR-125a-3p was significantly downregulated in the RA-tissues and RA-FLS, and miR-125a-3p could inhibit the proliferation and reduce the inflammation response of RA-FLS. Besides, MAST3 was found as a target of miR-125a-3p, and increased MAST3 could reverse the effects of miR-125a-3p on RA-FLS including decreased proliferation, reduced inflammation level and the inactivation of Wnt/β-catenin and NF-κB pathways.
Conclusions:
This study suggests that miR-125a-3p could inactivate the Wnt/β-catenin and NF-κB pathways to reduce the proliferation and inflammation response of RA-FLS via targeting MAST3.
... Because of the diversity of the proposed pathogenic mechanisms, SLE is considered the prototype of autoimmune diseases 28,29 . Recently, variations in miRNA expression have been found in certain autoimmune diseases, such as SLE, rheumatoid arthritis (RA), and primary Sjogren's syndrome (pSS) 30,31 . Therefore, these molecules may be considered as new biomarkers that are specific for diverse autoimmune disorders. ...
... To understand this finding, critical attention should be paid towards the role of mic-RNA 155 in the development of anti-dsDNA Ab, it's noteworthy that miR-155 is encoded within a region known as the B cell integration cluster gene 60 . miR-155 was shown to regulate antibody production in SLE patients 30 . Furthermore, Bic/miR-155-deficient mice showed a decrease in B cell immunity 61 . ...
Objective:
Systemic Lupus Erythematosus (SLE) is an autoimmune inflammatory disease. miR-155 and miR-146a were expressed in many autoimmune diseases such as rheumatoid arthritis. The aim of this study was to examine miR-155 rs767649 and miR-146a rs57095329 polymorphisms in SLE susceptibility in an Egyptian cohort and to investigate the correlation between them and clinical data and disease activity.
Patients and methods:
The two SNPs were analyzed in 120 patients with SLE and 100 healthy controls using RT-PCR.
Results:
The TT genotype and T allele of miR-155 rs767649 were associated with a significant increase in the risk of SLE, particularly in females. On the other hand, miR-146a (rs57095329) polymorphism was not associated with SLE risk. The AT/TT genotypes of miR-155 rs767649 showed higher distributions among patients with higher SLEDAI and nephritis.
Conclusions:
This study had demonstrated for the first time the association between miR-155 rs767649 and the risk of development of SLE in an Egyptian cohort, mostly in females.
... The current study showed that there was a statistically significant difference in fold change of microRNA-125a gene expression inSLE patients compared to controls, and it had a significant reduction in activity by 2.7 times compared to controls.This finding was consistent with other studies which also reported a down-regulation of this microRNA in SLE patients in comparison to controls (6,19) . MiR-125a was down regulated in the serum of SLE patients, and this down regulation of miR-125a leads to the upregulation of its predicted target gene Kruppel-like factor 13 (KLF13), which is a critical transcription factor in the regulation of the expression of the chemokine RANTES (CCL5) (20) , this, in turn,leads to the elevation of the expression of theproinflammatory chemokine RANTES (21) . ...
... In addition, the current study revealed that microRNA-146a gene expression was significantly different in fold change in the serum of SLE patientsin comparison to controls, it had a reduction in activity by 2.4 times compared to controls, andthese results are consistent with other studies (19,26) . The present work showed that the downregulation of miR-146a in patients with SLE is associated negatively with clinical disease activity, the presence of renal involvement, and proteinuria, these results are consistent with several other studies (27,28), the strong association between the level of miR-146a and clinical disease activity indicate that this microRNA play a role in lupus pathogenesis and it may serve as disease biomarker. ...
Systemic lupus erythematosus is a heterogeneous autoimmune disease associated with a high rate of morbidity and mortality. Identifying new SLE biomarkers might be a useful tool to facilitate early and precise diagnosis of the disease, identify the risk of organ involvement and decide the most appropriate treatment. To identify the role of microRNAs (miR-125a, and miR-146a) and their target cytokines (RANTES, and IFN-γ) in the immune-pathogenesis of SLE. This case-control study was conducted during the period from December 2018 to November 2019 which included 35 SLE patients who were among patients attending the Rheumatology Clinic, and Rheumatology Ward in Baghdad Teaching Hospital, and Private Nursing Home Hospital in Baghdad Medical City. In addition, we were included 35 healthy individuals as controls. The serum levels of miR-125a and miR-146a were determined by the two-step reverse transcription-polymerase chain reaction (RT-PCR), while the serum levels of the cytokines RANTES and IFN-γ were detected by the ELISA technique in 35 SLE patients and 35 healthy controls. The levels of miR-125a and miR-146a were significantly down-regulated in SLE patients in comparison to controls (p<0.001, p=0.005 respectively). Serum miR-146a was inversely associated with proteinuria (p=0.018) and the SLE Disease Activity Index (p=0.008). This study also revealed an increased serum level of RANTES (p=0.03), and IFN-γ (p=0.049) in SLE patients compared to controls. The results of the current study suggested that serum miR-125a and miR-146a and the cytokines (RANTES, and IFN-γ) may participate in the immune-pathogenesis of SLE. How to cite this article: Al-hasso IKQ, Al-Derzi AR, et al (2020): The role of microRNAs (MiR-125a and MrR-146a), RANTEs, and IFN-yin systemic lupus erythematosus, Ann Trop Med & Public Health; 23(S13B): SP231382.
... 22 Several specific miRNAs are characterized by increased expression in the blood of SLE patients. 24,25 The extracellular miRNAs possess several functions: cell proliferation, signaling between cells, regulation of neurogenesis, and angiogenesis. 26 Pathological processes resulting in inflammatory processes in SLE patients are associated with the change in miRNAs expression. ...
... 45 However, other studies showed a decrease in miR-146a expression in PBMCs and plasma of SLE patients. 24,46 At the same time, the expression of pro-inflammatory miR-155-5p and miR-326 was significantly increased in PBMCs of SLE patients. 45 Besides, the expression level of miR-326 was higher in regulatory T (Treg) cells isolated from SLE patients compared to healthy individuals. ...
Background and objectives:
Systemic lupus erythematosus (SLE) is an inflammatory disease. The sera of SLE patients contain antibodies-abzymes hydrolyzing myelin basic protein (MBP), DNA, nucleotides, and oligosaccharides. The blood of SLE patients contains an increased amount of some specific miRNAs. This study aimed to analyze a possible hydrolysis of eight microRNAs found in the blood of SLE patients with high frequency by blood antibodies-abzymes.
Patients and methods:
Using affinity chromatography of the serum proteins of SLE patients and healthy donors on protein G-Sepharose and following FPLC gel filtration, electrophoretically homogeneous IgG preparations containing no impurities of canonical RNases were obtained. These preparations were used to analyze their activity in the hydrolysis of eight miRNAs.
Results:
It was shown that SLE IgGs hydrolyze very efficiently four neuroregulatory miRNAs (miR-219-2-3p, miR-137, miR-219a-5p, and miR-9-5p) and four immunoregulatory miRNAs (miR-326, miR-21-3p, miR-155-5p, and miR-146a-3p). To demonstrate that the miRNAs hydrolysis is an intrinsic property of SLE IgGs, several rigid criteria were checked. Only some IgGs of healthy donors showed very weak, but reliably detectable activity in the hydrolysis miRNAs. The average activity of SLE patients IgGs according to median values is statistically significant 84.8-fold higher than that of healthy donors. The maximum and comparable average activity (RA) was observed in the hydrolysis of three miRAs: miR-9-5p, miR-155-5p, and miR-326. MiR-9-5p plays an important role in the development of lupus nephritis, while miR-326 activates the production of antibodies by B cells. The major and moderate specific sites of the hydrolysis of each miRNA were revealed. The hydrolysis of eight microRNAs was mostly site specific. Several SLE IgGs hydrolyzed some miRNAs demonstrating a combination of site-specific and non-specific splitting.
Conclusion:
Since inflammatory processes in SLE are associated with the change in miRNAs expression, the decrease in their concentration due to hydrolysis by autoantibodies-abzymes may be important for SLE pathogenesis.
... The current study showed that there was a statistically significant difference in fold change of microRNA-125a gene expression inSLE patients compared to controls, and it had a significant reduction in activity by 2.7 times compared to controls.This finding was consistent with other studies which also reported a down-regulation of this microRNA in SLE patients in comparison to controls (6,19) . MiR-125a was down regulated in the serum of SLE patients, and this down regulation of miR-125a leads to the upregulation of its predicted target gene Kruppel-like factor 13 (KLF13), which is a critical transcription factor in the regulation of the expression of the chemokine RANTES (CCL5) (20) , this, in turn,leads to the elevation of the expression of theproinflammatory chemokine RANTES (21) . ...
... In addition, the current study revealed that microRNA-146a gene expression was significantly different in fold change in the serum of SLE patientsin comparison to controls, it had a reduction in activity by 2.4 times compared to controls, andthese results are consistent with other studies (19,26) . The present work showed that the downregulation of miR-146a in patients with SLE is associated negatively with clinical disease activity, the presence of renal involvement, and proteinuria, these results are consistent with several other studies (27,28), the strong association between the level of miR-146a and clinical disease activity indicate that this microRNA play a role in lupus pathogenesis and it may serve as disease biomarker. ...
... The current study showed that there was a statistically significant difference in fold change of microRNA-125a gene expression inSLE patients compared to controls, and it had a significant reduction in activity by 2.7 times compared to controls.This finding was consistent with other studies which also reported a down-regulation of this microRNA in SLE patients in comparison to controls (6,19) . MiR-125a was down regulated in the serum of SLE patients, and this down regulation of miR-125a leads to the upregulation of its predicted target gene Kruppel-like factor 13 (KLF13), which is a critical transcription factor in the regulation of the expression of the chemokine RANTES (CCL5) (20) , this, in turn,leads to the elevation of the expression of theproinflammatory chemokine RANTES (21) . ...
... In addition, the current study revealed that microRNA-146a gene expression was significantly different in fold change in the serum of SLE patientsin comparison to controls, it had a reduction in activity by 2.4 times compared to controls, andthese results are consistent with other studies (19,26) . The present work showed that the downregulation of miR-146a in patients with SLE is associated negatively with clinical disease activity, the presence of renal involvement, and proteinuria, these results are consistent with several other studies (27,28), the strong association between the level of miR-146a and clinical disease activity indicate that this microRNA play a role in lupus pathogenesis and it may serve as disease biomarker. ...
Systemic lupus erythematosus is a heterogeneous autoimmune disease associated with a high rate of morbidity and mortality. Identifying new SLE biomarkers might be a useful tool to facilitate early and precise diagnosis of the disease, identify the risk of organ involvement and decide the most appropriate treatment. To identify the role of microRNAs (miR-125a, and miR-146a) and their target cytokines (RANTES, and IFN-γ) in the immune-pathogenesis of SLE. This case-control study was conducted during the period from December 2018 to November 2019 which included 35 SLE patients who were among patients attending the Rheumatology Clinic, and Rheumatology Ward in Baghdad Teaching Hospital, and Private Nursing Home Hospital in Baghdad Medical City. In addition, we were included 35 healthy individuals as controls. The serum levels of miR-125a and miR-146a were determined by the two-step reverse transcription-polymerase chain reaction (RT-PCR), while the serum levels of the cytokines RANTES and IFN-γ were detected by the ELISA technique in 35 SLE patients and 35 healthy controls. The levels of miR-125a and miR-146a were significantly down-regulated in SLE patients in comparison to controls (p<0.001, p=0.005 respectively). Serum miR-146a was inversely associated with proteinuria (p=0.018) and the SLE Disease Activity Index (p=0.008). This study also revealed an increased serum level of RANTES (p=0.03), and IFN-γ (p=0.049) in SLE patients compared to controls. The results of the current study suggested that serum miR-125a and miR-146a and the cytokines (RANTES, and IFN-γ) may participate in the immune-pathogenesis of SLE. How to cite this article: Al-hasso IKQ, Al-Derzi AR, et al (2020): The role of microRNAs (MiR-125a and MrR-146a), RANTEs, and IFN-yin systemic lupus erythematosus, Ann Trop Med & Public Health; 23(S13B): SP231382.
... This was also taking into consideration the biological relevance of these candidates to RA, as we previously reported on changes in expression of IFITM1 (hypo-methylated and over-expressed) and IRF8 (hyper-methylated and underexpressed) in CD4+ T-cells in early RA compared to HC [39] (Supplementary File), while for TNF-α, changes were observed but were more limited (hypo-methylated but nonsignificantly over-expressed). MIR21 has been reported as well in RA, with >5-fold higher expression levels, but in the serum rather than cells [41]. ...
Unlabelled:
Biomarkers for the classification of rheumatoid arthritis (RA), and particularly for anti-citrullinated peptide antibody (ACPA)-negative patients, remain an important hurdle for the early initiation of treatment. Taking advantage of DNA-methylation patterns specific to early RA, quantitative methylation-specific qPCR (qMSP) offers a robust technology for the development of biomarkers. We developed assays and established their value as RA classification biomarkers.
Methods:
DNA-methylation data were screened to select candidate CpGs to design qMSP assays. Eight assays were developed and tested on two early inflammatory arthritis cohorts. Logistic regression and bootstrapping were used to demonstrate the added value of the qMSP assays.
Result:
Differentially methylated CpG data were screened for candidate CpG, thereby meeting the qMSP assay requirements. The top CpG candidate was in the TNF gene, for which we successfully developed a qMSP assay. Significantly lower DNA-methylation levels were observed in RA (p < 4 × 10-9), with a high predictive value (OR < 0.54/AUC < 0.198) in both cohorts (n = 127/n = 157). Regression using both datasets showed improved accuracy = 87.7% and AUC = 0.944 over the model using only clinical variables (accuracy = 85.2%, AUC = 0.917). Similar data were obtained in ACPA-negative patients (n = 167, accuracy = 82.6%, AUC = 0.930) compared to the clinical variable model (accuracy = 79.5%, AUC = 0.892). Bootstrapping using 2000 datasets confirmed that the AUCs for the clinical+TNF-qMSP model had significant added value in both analyses.
Conclusion:
The qMSP technology is robust and can successfully be developed with a high specificity of the TNF qMSP assay for RA in patients with early inflammatory arthritis. It should assist classification in ACPA-negative patients, providing a means of reducing time to diagnosis and treatment.
... 11,28,36 One study, however, has shown a significant reduction in the circulating level of miR-155 in RA patients compared to controls, and another study has demonstrated significant upregulation of miR-155 levels in the plasma of RA patients. 37,38 Finally, the expression level of miR-16-5p was investigated in this study as the last miRNA. It has been stated that miR-16 is involved in RA, and it is suggested as a suitable candidate marker of RA disease activity. ...
Rheumatoid arthritis is a chronic autoimmune disease that causes inflammation and destruction of the joints. The objective of the current study was to evaluate the expression of microRNA (miR)-155-5p, miR-210-3p, and miR-16-5p in the plasma of patients with rheumatoid arthritis in comparison with a healthy control group to attain an expression profile for earlier diagnosis and treatment.
To carry out this study, 100 individuals were chosen as two equally sized groups of rheumatoid arthritis patients and healthy controls. Five milliliters of blood were drawn from each individual, and plasma RNA was extracted using Trisol solution. Complementary DNAs were synthesized using the Moloney leukemia virus (MMLV) and deoxynucleoside triphosphate (dNTP). Finally, real-time polymerase chain reaction (PCR) was implemented using the SYBR Green kit.
The mean expression of miR-155-5p, miR-210-3p, and miR-16-5p were 2.46±2.79, 1.97±1.90, and 69.62±88.44 in the rheumatoid arthritis group, and 0.34±0.33, 9.82±9.34, and 7.94±7.09 in the healthy group, respectively. Additionally, significant differences were revealed in the relative
expression of the selected microRNAs in 4 subgroups of rheumatoid arthritis patients with different disease activities based on the disease activity score 28 (DAS28). ROC curve analysis showed that miR-155-5p (area under the curve, AUC=0.90, sensitivity=80%, specificity=81%), miR-210-3p (AUC=0.75, sensitivity=66%, specificity=71%), and miR-16-5p (AUC=0.96, sensitivity=89%, specificity=82%) could be potential biomarkers for rheumatoid arthritis diagnosis.
Increased expressions of miR-16-5p and miR-155-5p and decreased expression of miR-210-3p in rheumatoid arthritis patients compared with healthy individuals demonstrate the effectiveness of these microRNAs in disease incidence and progression. Thus, the expression levels of these microRNAs can be used for diagnostic and therapeutic purposes.
... Moreover, knockout mice for miR-146a exhibited hyper-reactive and pro-inflammatory circulating neutrophils (36). Besides, there is some evidence that miR-146a participates in the pathogenesis of several autoimmune diseases, such as rheumatoid arthritis, multiple sclerosis and systemic lupus erythematosus (37,38,39). Thus, overall miR-21 and miR-146a appear to act as negative regulators of inflammation, induced by inflammation itself. ...
Objective:
Thyroid eye disease (TED) is an immune-mediated disorder of the eye. Intravenous glucocorticoid (GC) is the first-line treatment for patients with active moderate-to-severe TED. However, the response rate is between 50 and 80%. There are still no simple and reliable markers of responsiveness to GC therapy. We aimed to explore the possible role of miR-146a and miR-21 as predictors of responsiveness to GC treatment in TED.
Methods:
We carried out a prospective longitudinal study on 30 consecutive adult patients with active moderate-to-severe TED and eligible for GC therapy. All patients received the standard GC treatment with methylprednisolone iv. In cases of progressive worsening of Gorman Score for diplopia or with duction restriction <30° in at least two consecutive controls patients also underwent orbital radiotherapy. Response to GC treatment was defined as a decrease of 2 or more points in the clinical activity score (CAS) or CAS <4/10 at 24 weeks. Circulating miRNAs were extracted from patients' serum and quantified by real time PCR.
Results:
Twenty-three (77%) patients responded to GC. Thyroid surgery, higher CAS, greater proptosis and higher pre-treatment circulating levels of miR-146a emerged as predictive factors of responsiveness to GC. A ROC analysis revealed that miR-146a could predict responsiveness to GC with a positive predictive value of 100%.
Conclusion:
This is the first study investigating the role of pre-treatment circulating miR-21 and miR-146a to predict responsiveness to GC in TED. miR-146a emerged as a simple, objective, new marker of GC sensitivity that could be used to avoid ineffective administration of GC therapy to TED patients.
... Other molecules that are being studied as potential biomarkers for several diseases are miRNAs (miR) [19,20]. Recent studies performed in RA patients have discovered that an increased expression of some miRNAs, such as let-7d, miR-24, miR-126, miR-130a, miR-221, or miR-431, could be predictive as a biomarker for this disease [21]. ...
Rheumatoid factor (RF) and anti-citrullinated protein antibodies (ACPAs) are the most frequently used rheumatoid arthritis (RA) diagnostic markers, but they are unable to anticipate the patient’s evolution or response to treatment. The aim of this study was to identify possible severity biomarkers to predict an upcoming flare-up or remission period. To address this objective, sera and anticoagulated blood samples were collected from healthy controls (HCs; n = 39) and from early RA (n = 10), flare-up (n = 5), and remission (n = 16) patients. We analyzed leukocyte phenotype markers, regulatory T cells, cell proliferation, and cytokine profiles. Flare-up patients showed increased percentages of cluster of differentiation (CD)3+CD4− lymphocytes (p < 0.01) and granulocytes (p < 0.05) but a decreased natural killer (NK)/T lymphocyte ratio (p < 0.05). Analysis of leukocyte markers by principal component analysis (PCA) and receiver operating characteristic (ROC) curves showed that CD45RO+ (p < 0.0001) and CD45RA+ (p < 0.0001) B lymphocyte expression can discriminate between HCs and early RA patients, while CD3+CD4− lymphocyte percentage (p < 0.0424) and CD45RA+ (p < 0.0424), CD62L+ (p < 0.0284), and CD11a+ (p < 0.0185) B lymphocyte expression can differentiate between flare-up and RA remission subjects. Thus, the combined study of these leukocyte surface markers could have potential as disease severity biomarkers for RA, whose fluctuations could be related to the development of the characteristic pro-inflammatory environment.
... miRNAs in patient biological samples are also currently under study for their potential as novel biomarkers [77][78][79][80][81][82]. In particular, recent reports have indicated the diagnostic/prognostic value of dysregulated miRNAs in patients with different types of rheumatic diseases [83][84][85][86]. However, miRNA investigation and application in clinical practice are hindered by the lack of consensus regarding the technique to be used for their isolation [87][88][89] and quantification [90,[90][91][92][93]. ...
Juvenile Idiopathic Arthritis (JIA) represents the most common chronic pediatric arthritis in Western countries and a leading cause of disability in children. Despite recent clinical achievements, patient management is still hindered by a lack of diagnostic/prognostic biomarkers and targeted treatment protocols. MicroRNAs (miRNAs) are short non-coding RNAs playing a key role in gene regulation, and their involvement in many pathologies has been widely reported in the literature. In recent decades, miRNA’s contribution to the regulation of the immune system and the pathogenesis of autoimmune diseases has been demonstrated. Furthermore, miRNAs isolated from patients’ biological samples are currently under investigation for their potential as novel biomarkers. This review aims to provide an overview of the state of the art on miRNA investigation in JIA. The literature addressing the expression of miRNAs in different types of biological samples isolated from JIA patients was reviewed, focusing in particular on their potential application as diagnostic/prognostic biomarkers. The role of miRNAs in the regulation of immune responses in affected joints will also be discussed along with their potential utility as markers of patients’ responses to therapeutic approaches. This information will be of value to investigators in the field of pediatric rheumatology, encouraging further research to increase our knowledge of miRNAs’ potential for future clinical applications in JIA.
... Jacob et al. [190] reported that IL-17 plays a priming role in murine LN. Wang et al. [191] further identified that miR-125a-3p is involved in the progression and development of SLE. Zhang et al. [192], in a recent study, found that this miR was downregulated in patients with SLE and could decrease IL17 levels and suppress renal fibrosis via down-regulating TGF-β1 in LN mice. ...
Lupus nephritis (LN) is one of the most severe complications in patients with systemic lupus erythematosus (SLE). Traditionally, LN is regarded as an immune complex (IC) deposition disease led by dsDNA–anti-dsDNA-complement interactions in the subendothelial and/or subepithelial basement membrane of glomeruli to cause inflammation. The activated complements in the IC act as chemoattractants to chemically attract both innate and adaptive immune cells to the kidney tissues, causing inflammatory reactions. However, recent investigations have unveiled that not only the infiltrating immune-related cells, but resident kidney cells, including glomerular mesangial cells, podocytes, macrophage-like cells, tubular epithelial cells and endothelial cells, may also actively participate in the inflammatory and immunological reactions in the kidney. Furthermore, the adaptive immune cells that are infiltrated are genetically restricted to autoimmune predilection. The autoantibodies commonly found in SLE, including anti-dsDNA, are cross-reacting with not only a broad spectrum of chromatin substances, but also extracellular matrix components, including α-actinin, annexin II, laminin, collagen III and IV, and heparan sulfate proteoglycan. Besides, the glycosylation on the Fab portion of IgG anti-dsDNA antibodies can also affect the pathogenic properties of the autoantibodies in that α-2,6-sialylation alleviates, whereas fucosylation aggravates their nephritogenic activity. Some of the coexisting autoantibodies, including anti-cardiolipin, anti-C1q, anti-ribosomal P autoantibodies, may also enhance the pathogenic role of anti-dsDNA antibodies. In clinical practice, the identification of useful biomarkers for diagnosing, monitoring, and following up on LN is quite important for its treatments. The development of a more specific therapeutic strategy to target the pathogenic factors of LN is also critical. We will discuss these issues in detail in the present article.
... miR-146a-3p is expressed by leukocytes and its function is clearly associated with inflammation and innate immunity [23,24]. Studies have shown a decrease in miR-146a expression in peripheral blood mononuclear cells (PBMCs) and the plasma of systemic lupus erythematosus (SLE) patients, suggesting that miR-146a has antiinflammatory properties [25,26]. It has been shown that miR-146a is upregulated in the skin and keratinocytes of atopic dermatitis (AD) patients, which alleviates chronic inflammation in a mouse model of AD [27][28][29]. ...
Th17 (T-helper 17) cells subtype of non-T2 (non-type 2) asthma is related to neutrophilic infiltration and resistance to inhaled corticosteroids (ICS), so is also known as severe asthma. Methyl-CpG binding domain protein 2 (MBD2) regulates the differentiation of the Th17 cells, tending to show a therapeutic target in severe asthma. miR-146a-3p is associated with anti-inflammatory characteristics and immunity. Moreover, bioinformatic analysis showed that MBD2 may be a target gene of miR-146a-3p. However, the role of miR-146a-3p in the differentiation of Th17 cells via MBD2 in severe asthma remains unknown. Here, we aimed to explore how miR-146a-3p interacts with MBD2 and affects the differentiation of Th17 cells in severe asthma. First, we recruited 30 eligible healthy people and 30 patients with severe asthma to detect the expression of miR-146a-3p in peripheral blood mononuclear cells (PBMCs) by qRT-PCR. Then, we established a HDM/LPS (house dust mite/lipopolysaccharide) exposure model of bronchial epithelial cells (BECs) to evaluate the expression of miR-146a-3p, the interaction between miR-146a-3p and MBD2 using western blot and luciferase reporter analysis and the effect of miR-146a-3p regulated Th17 cells differentiation by flow cytometry in BECs in vitro. Finally, we constructed a mouse model of Th17 predominant neutrophilic severe asthma to assess the therapeutic potential of miR-146a-3p in severe asthma and the effect of miR-146a-3p regulated Th17 cells differentiation via MBD2 in vivo. Decreased miR-146a-3p expression was noted in severe asthma patients, in the BECs and in the animal severe asthma models. Moreover, we demonstrated that miR-146a-3p suppressed Th17 cells differentiation by targeting the MBD2. miR-146a-3p overexpression significantly reduced airway hyperresponsiveness, airway inflammation and airway mucus secretion, while also inhibiting Th17 cells response in vivo, which relieved severe asthma. By targeting MBD2 to suppress Th17 cells differentiation, miR-146a-3p provides a potential novel therapeutic for Th17 predominant neutrophilic severe asthma.
... In MS and SLE, specific miRNAs characterized by increased expression are revealed in the cerebrospinal fluid and blood [28][29][30][31]. The extracellular miRNAs participate in signaling between cells and regulating neurogenesis, angiogenesis, and cell proliferation [32]. The change in miRNA expression in many cases is associated with pathological processes. ...
It was shown that the spontaneous development of experimental encephalomyelitis (EAE) in C57BL/6 mice occurs due to changes in the profile of bone marrow stem cells differentiation. This leads to the appearance of lymphocytes producing antibodies-abzymes that hydrolyze DNA, myelin basic protein (MBP), and histones. The activity of abzymes in the hydrolysis of these auto-antigens slowly but constantly increases during the spontaneous development of EAE. Treatment of mice with myelin oligodendrocyte glycoprotein (MOG) leads to a sharp increase in the activity of these abzymes with their maximum at 20 days (acute phase) after immunization. In this work, we analyzed changes in the activity of IgG-abzymes hydrolyzing (pA)23, (pC)23, (pU)23, and six miRNAs (miR-9-5p, miR-219a-5p, miR-326, miR-155-5p, miR-21-3p, and miR-146a-3p) before and after mice immunization with MOG. Unlike abzymes hydrolyzing DNA, MBP, and histones, the spontaneous development of EAE leads not to an increase but to a permanent decrease of IgGs activity of hydrolysis of RNA-substrates. Treatment of mice with MOG resulted in a sharp but transient increase in the activity of antibodies by day 7 (onset of the disease), followed by a sharp decrease in activity 20–40 days after immunization. A significant difference in the production of abzymes against DNA, MBP, and histones before and after mice immunization with MOG with those against RNAs may be since the expression of many miRNAs decreased with age. This can lead to a decrease in the production of antibodies and abzymes that hydrolyze miRNAs with age mice.
... Similar decrease was observed in serum-derived EVs from SLE patients [62], and in serum of patients with CLE [63]. However, another study involving plasma samples showed that miR-150 expression was slightly upregulated [64]. These discrepancies may result from the differences in stages and manifestations of the disease between enrolled cohorts. ...
At present, special efforts are being made to develop the strategies allowing for activation of long-lasting antigen-specific immune tolerance in therapy of allergic and autoimmune diseases. Some of these therapeutic approaches are aimed at modulating cell functions at genetic level by using miRNA-based and miRNA-targeting treatments. Simultaneously, the crucial role of extracellular vesicles as natural miRNA conveyors is highlighted for induction of antigen-specific immune tolerance, especially that they appear to be easily manipulatable for therapeutic applications. Among other immune-related miRNAs, miR-150 is getting special attention as it is differently expressed by immune cells at various stages of their maturation and differentiation. In addition, miR-150 is involved in different signaling cascades orchestrating humoral and cell-mediated mechanisms of both innate and adaptive immune responses. Therefore, miR-150 is considered a master regulator of immunity in mammals. Currently, physiological miR-150-dependent regulatory circuits and causes of their malfunctioning that underlie the pathogenesis of allergic and autoimmune disorders are being unraveled. Thus, present review summarizes the current knowledge of the role of miR-150 in the pathogenesis and complications of these diseases. Furthermore, the involvement of miR-150 in regulation of immune responses to allergens and self-antigens and in induction of antigen-specific immune tolerance is discussed with the special emphasis on the therapeutic potential of this miRNA.
... It is necessary to mention about extracellular concentration may not be correlated with intracellular. The previous studies regarding to another molecule -miR-21 -showed that the plasma concentration may be elevated (25), although concentration in peripheral blood mononuclear cells (PBMC) or CD4 + cells may be reduced (26). In turn, the gene expression is not regulated only by one miRNA and biological effect of plasma miRNA concentration may not be adequate to transcription silencing in cells. ...
Objective Micro-RNAs (miRNAs) are an endogenous small, single-stranded, non-coding RNAs with a 18-25 nucleotide long and have been reported as potential extracellular biomarkers of various diseases. They mainly decrease the gene expression by inhibiting the translation or cause mRNA destabilisation. The aim of our study was to identify miRNAs whose concentration may be associated with severity of rheumatoid arthritis (RA). Methods A total of 74 unrelated individuals, 50 with RA and 24 in a control group were enrolled to the study. Real-time PCR was used to evaluate the plasma concentration levels of 8 miRNAs: miR-26a, miR-125b, miR-20b, miR-22, miR-221, miR-17, miR-93, miR-106b. Results The logistic regression results showed that miR-22 (p=0.0003) and miR-26a (p=0.049) may be the most important molecules distinguishing RA patients and healthy controls. Moreover, the quantity of miR-22 was different between rheumatoid factor (RF)-positive and RF-negative patients (p=0.04). Conclusion In this study we demonstrated for the first time that plasma concentration of miR-22 may be considered as a potential molecular marker associated with disease activity.
... Compared to RA patients, the levels of miR-24, -125a-5p, and ePRAM in systemic lupus erythematosus (SLE) and osteoarthritis (OA) patients were lower [152]. [155]. Significantly increased expression of miR-24, along with six other miRs were observed in IL-1β-stimulation OA synovial explants [156]. ...
MiRNAs (miRs) have been proven to be well-validated therapeutic targets. Emerging evidence has demonstrated that intricate, intrinsic and paradoxical functions of miRs are context-dependent because of their multiple upstream regulators, broad spectrum of downstream molecular targets and distinct expression in various tissues, organs and disease states. Targeted therapy has become an emerging field of research. One key for the development of successful miR-based/targeted therapy is to acquire integrated knowledge of its regulatory network and its association with disease phenotypes to identify critical nodes of the underlying pathogenesis. Herein, we systematically summarized the comprehensive role of miR-24-3p (miR-24), along with its passenger strands miR-24-1-5p* (miR-24-1) and miR-24-2-5p* (miR-24-2), emphasizing their microenvironment, intracellular targets, and associated gene networks and regulatory phenotypes in 18 different cancer types and 13 types of other disorders. MiR-24 targets and regulates numerous genes in various cancer types and enhances the expression of several oncogenes (e.g., cMyc, BCL2 and HIF1), which are challenging in terms of druggability. In contrast, several tumor suppressor proteins (p21 and p53) have been reported to be downregulated by miR-24. MiR-24 also regulates the cell cycle and is associated with numerous cancer hallmarks such as apoptosis, proliferation, metastasis, invasion, angiogenesis, autophagy, drug resistance and other diseases pathogenesis. Overall, miR-24 plays an emerging role in the diagnosis, prognosis and pathobiology of various diseases. MiR-24 is a potential target for targeted therapy in the era of precision medicine, which expands the landscape of targetable macromolecules, including undruggable proteins.
... miRNAs can modulate 90% of protein-coding genes in several biological processes, such as proliferation, apoptosis, differentiation, immune cell lineage commitment, and maintenance of the immune system homeostasis (6). Some abnormalities in the immune system and the development of autoimmune diseases are highly related to the alteration of miRNAs' transcription (7,8). Studies indicated that these small molecules could be used as a biomarker to diagnose and monitor various autoimmune diseases. ...
MicroRNAs (miRNAs) are small noncoding conserved RNAs containing 19 to 24 nucleotides that are regulators of post-translational modifications and are involved in the majority of biological processes such as immune homeostasis, T helper cell differentiation, central and peripheral tolerance, and immune cell development. Autoimmune diseases are characterized by immune system dysregulation, which ultimately leads to destructive responses to self-antigens. A large body of literature suggests that autoimmune diseases and immune dysregulation are associated with different miRNA expression changes in the target cells and tissues of adaptive or innate immunity. miR-155 is identified as a critical modulator of immune responses. Recently conducted studies on the expression profile of miR-155 suggest that the altered expression and function of miR-155 can mediate vulnerability to autoimmune diseases and cause significant dysfunction of the immune system.
... Recent studies have shown that various autoimmune diseases have distinct miRNAs expression profiles. A microarray analysis plus real-time quantitative PCR identified that miR-126 was specifically enriched only in the blood of the SLE patients instead of healthy donors and RA patients (130). In another study, TaqMan Low Density Arrays were used to detect 365 miRNAs in PBMC of SLE patients and healthy controls. ...
T helper 17 (Th17) cells are characterized by the secretion of the IL-17 cytokine and are essential for the immune response against bacterial and fungal infections. Despite the beneficial roles of Th17 cells, unrestrained IL-17 production can contribute to immunopathology and inflammatory autoimmune diseases, including multiple sclerosis, rheumatoid arthritis, and inflammatory bowel disease. Although these diverse outcomes are directed by the activation of Th17 cells, the regulation of Th17 cells is incompletely understood. The discovery that microRNAs (miRNAs) are involved in the regulation of Th17 cell differentiation and function has greatly improved our understanding of Th17 cells in immune response and disease. Here, we provide an overview of the biogenesis and function of miRNA and summarize the role of miRNAs in Th17 cell differentiation and function. Finally, we focus on recent advances in miRNA-mediated dysregulation of Th17 cell fate in autoimmune diseases.
... The upregulation of miR-451 in PBMC might be either constitutive and represent a biomarker of the preclinical phase or might be increased in response to mild systemic inflammation, as evidenced here by higher CRP levels and supported by in vitro stimulation with CXCL16. However, we did not verify any difference in miR-451 expression in plasma samples from our validation cohort, although circulating miR-451 expression was previously shown to be increased in patients with established RA 15 . This discrepancy might be explained by the established clinical arthritis in patients with an advanced phase of RA. ...
Individuals carrying anti-citrullinated protein antibodies (ACPA) are considered at high risk of developing rheumatoid arthritis (RA). The altered expression of miRNAs contributes to the pathogenesis of RA. We aimed to identify differentially expressed miRNAs in the peripheral blood of ACPA-positive individuals with arthralgia at risk of RA compared to healthy controls (HC) and to determine their implications in the preclinical phase of RA. A comprehensive analysis of miRNAs revealed the dysregulation of miR-451 in peripheral blood mononuclear cells (PBMC) and plasma from RA-risk individuals. Higher miR-451 expression in PBMC from RA-risk individuals was further validated. Notably, miR-451 was previously shown to regulate CXCL16, a protein involved in RA pathogenesis. The expression of miR-451 in PBMC positively correlated with the CXCL16 mRNA, which could be secondary to the inflammation-induced expression of miR-451. Transfection of monocytes with pre-miR-451 in vitro resulted in the downregulation of CXCL16. Moreover, flow cytometry revealed a lower count of CXCL16-positive monocytes in RA-risk individuals. We propose that the constitutive or inflammation-induced upregulation of miR-451 in PBMC downregulates the expression of CXCL16, reduces the inflammatory milieu and thereby strives to delay the shift from the preclinical phase to the clinical manifestation of RA. This hypothesis warrants further investigation.
... In SLE patients, Mir223 has been only explored as immunological biomarkers for disease pathophysiology. Although upregulation of Mir223 in peripheral plasma was reported (10,11), the expression of Mir223 in peripheral plasma was significantly decreased in SLE patients with active nephritis (12). S1pr1 has been known to be expressed in several cell types of the immune system. ...
Objective
The micro RNAs (miRNAs) and their target mRNAs are differentially expressed in various immune-mediated cells. Here, we investigated the role of Mir223 and sphingosine-1-phosphate receptor 1 ( S1pr1 ) in the pathogenesis of systemic lupus erythematosus.
Methods
We analyzed miRNA and mRNA profiling data of CD4 ⁺ splenic T cells derived from MRL/MpJ- Fas lpr /J mice. We performed 3′ untranslated region (UTR) luciferase reporter gene assay using human umbilical vein endothelial cells (HUVECs). We generated the B6- Mir223 −/− Fas lpr/lpr mice and the lupus phenotypes were analyzed.
Results
In CD4 ⁺ splenic T cells, we identified upregulation of miR-223-3p and downregulation of the possible target, S1pr1 by RNA sequencing of MRL/MpJ- Fas lpr /J mice. The transfection with miR-223-3p mimic significantly suppressed a luciferase activity in HUVEC treated with a Lentivirus vector containing 3′ UTR of S1pr1 . The mRNA levels of S1pr1 were significantly decreased after miR-223-3p overexpression. In B6- Mir223 −/− Fas lpr/lpr mice, the proportion of CD3 ⁺ T cells, CD3 ⁺ CD4 ⁻ CD8 ⁻ cells, B cells, plasma cells, and S1PR1 ⁺ CD4 ⁺ T cells in the spleen was significantly increased compared with that in B6- Mir223 +/+ Fas lpr/lpr mice by flow cytometry. B6- Mir223 −/− Fas lpr/lpr mice demonstrated the elevation of glomerular and renal vascular scores associated with enhanced intraglomerular infiltration of S1PR1 ⁺ CD4 ⁺ T cells.
Conclusion
Unexpectedly, the deletion of Mir223 exacerbated the lupus phenotypes associated with increased population of S1PR1 ⁺ CD4 ⁺ T in spleen and the enhanced infiltration of S1PR1 ⁺ CD4 ⁺ T cells in inflamed kidney tissues, suggesting compensatory role of Mir223 in the pathogenesis of lupus nephritis.
... hsa-miR-1825, hsa-miR-4769-3p, hsa-miR-1228-3p, hsa-miR-1972 are all down-regulated after infusion with Ig in CVID patients. In accordance with our results, hsa-miR-1825 has been associated with immune disorders as systemic lupus erythematosus and rheumatoid arthritis where it is down-regulated 21 . ...
Common variable immunodeficiency (CVID) is a complex primary immunodeficiency disorder characterized by a high clinical and genetic heterogeneity. The molecular underlying causes of CVID are not still now clear and the delays in diagnosis and treatment worsen the prognosis of the patients. MicroRNAs are non-coding, endogenous small RNAs often deregulated in human diseases, such as autoimmune and other immune-based disorders. In the present study, we aimed to evaluate miRNAs associated with the CVID and, in particular, with the response to the first Ig replacement therapy. To this aim, we compared miRNA profile obtained by serum samples of treatment-naïve CVID patients before and 24 h after the first Ig replacement therapy. For the first time, using a microarray assay followed by an integrated bioinformatics/biostatistics analysis, we identified five microRNAs (hsa-miR-6742, hsa-miR-1825, hsa-miR-4769-3p, hsa-miR-1228-3p, hsa-miR-1972) differently modulated in CVID patients by Ig infusion. All of them were down-regulated, excepted miR-6742 which was up-regulated. The latter may be of particular interest, since its functions are related to pathways involving Class I MHC mediated antigen processing and adaptive as well as innate Immune System. In conclusion, this study shows for the first time the modulation of miRNAs involved in CVID patients after the first Ig replacement therapy. Further studies are needed to assess whether such miRNAs could represent novel potential biomarkers in management and therapy of CVID patients.
... Studies have shown that miR-223 deficiency results in spontaneous inflammation, which is characterized by massive neutrophil proliferation and hyperactivation, hence providing direct evidence for the contribution of this miRNA in granulopoiesis (21)(22)(23). Moreover, aberrant miR-223 expression was shown to be associated with pathophysiology of a number of human diseases (17,(24)(25)(26)(27)(28)(29). Regarding inflammatory diseases, elevated levels of miR-223 expression were found in the lungs of patients with chronic obstructive pulmonary disease (30). ...
Group B streptococcus (GBS) is a human pathogenic bacteria inducing a strong inflammatory response that may be detrimental for host tissues if not finely regulated. The inflammatory response can be modulated by different molecular mechanisms, among which growing evidences point toward the crucial role of microRNAs. Regarding innate inflammatory response, studies have reported that miR-223 is essential for the control of granulocyte proliferation and activation. Moreover, number of investigations on miRNA expression profiling performed in various inflammatory settings reveal that miR-223 is among the top differentially expressed miRNA. Yet, the dynamic pattern of expression of miR-223 in vivo with respect to the evolution of the inflammatory process, especially in microbial infection, remains elusive. In this study, we analyzed the kinetic expression of miR-223 in an inflammatory model of GBS-induced murine pneumoniae and looked for correlates with inflammatory markers including innate cell infiltrates. We found that miR-223 expression is rapidly induced at very early time points (3-6 h post infection) mainly by lung infiltrating neutrophils. Interestingly, the level of miR-223 accumulating in the lungs remains higher at later stages of infection (24h and 48h pi) and this correlates with reduced expression of primary inflammatory cytokines and chemokines and with a shift in infiltrating monocyte and macrophage subtypes toward regulatory phenotype. Transient inhibition of miR223 by an antagomir resulted in significant increase of CXCL2 expression and partial enhancing of infiltrating neutrophils in GBS infected lung tissues. This suggests the potential contribution of miR-223 to the resolution phase of GBS-induced acute inflammation.
... However, there is evidence supporting that miR-21 expression is upregulated in PBMC of SLE patients leading to PDCD4 expression inhibition and aberrant T-cell responses [23]. Considering miR-21 as candidate circulatory biomarker in the plasma of SLE patients [24], some studies have investigated miR-21 expression in the plasma samples instead of blood cells; for example, Tang et. al. studied 44 untreated SLE patients' plasma samples and found that miR-21 expression in SLE is higher than control and positively correlated with SLEDAI score [25]. ...
Background: Systemic lupus erythematosus (SLE) is an autoimmune disease with variable clinical manifestations mostly affecting the skin, joints, hematopoietic system, kidney, and the central nervous system. The micro-RNAs (miR) are small, non-coding, endogenous RNAs, with key roles in many biological processes. The aim of this study was to investigate the cellular expression of miR-21 and miR-155 in SLE patients and healthy controls.
... According to their in vitro investigations, miR-146a negatively regulates the IFN-I pathway, through targeting interferon regulatory factor 5 (IRF5) and STAT1 [81] (Fig. 2). Reduced serum and plasma levels of miR-146a have been observed in patients with SLE and they are negatively associated with disease activity [88][89][90][91] (Table 1). Nevertheless, in another research, higher expression of miR-146a plasma level has been reported in patients with LN along with its positive and negative association with anti-double strand DNA (anti-dsDNA) autoantibodies and proteinuria, respectively [92] (Table 1). ...
MicroRNAs (miRNAs) are single-strand endogenous and non-coding RNA molecules with a length of about 22 nucleotides, which regulate genes expression, through modulating the translation and stability of their target mRNAs. miR-146a is one of the most studied miRNAs, due to its central role in immune system homeostasis and control of the innate and acquired immune responses. Accordingly, abnormal expression or function of miR-146a results in the incidence and progression of immune and non-immune inflammatory diseases. Its deregulated expression pattern and inefficient function have been reported in a wide spectrum of these illnesses. Based on the existing evidence, this miRNA qualifies as an ideal biomarker for diagnosis, prognosis, and activity evaluation of immune and non-immune inflammatory disorders. Moreover, much attention has recently been paid to therapeutic potential of miR-146a and several researchers have assessed the effects of different drugs on expression and function of this miRNA at diverse experimental, animal, besides human levels, reporting motivating results in the treatment of the diseases. Here, in this comprehensive review, we provide an overview of miR-146a role in the pathogenesis and progression of several immune and non-immune inflammatory diseases such as Rheumatoid arthritis, Systemic lupus erythematosus, Inflammatory bowel disease, Multiple sclerosis, Psoriasis, Graves' disease, Atherosclerosis, Hepatitis, Chronic obstructive pulmonary disease, etc., discuss about its eligibility for being a desirable biomarker for these disorders, and also highlight its therapeutic potential. Understanding these mechanisms underlies the selecting and designing the proper therapeutic targets and medications, which eventually facilitate the treatment process.
Systemic lupus erythematosus (SLE), as an immunological illness, frequently impacts young females. Both vulnerabilities to SLE and the course of the illness's clinical symptoms have been demonstrated to be affected by individual differences in non-coding RNA expression. Many non-coding RNAs (ncRNAs) are out of whack in patients with SLE. Because of the dysregulation of several ncRNAs in peripheral blood of patients suffering from SLE, these ncRNAs to be showed valuable as biomarkers for medication response, diagnosis, and activity. NcRNAs have also been demonstrated to influence immune cell activity and apoptosis. Altogether, these facts highlight the need of investigating the roles of both families of ncRNAs in the progress of SLE. Being aware of the significance of these transcripts perhaps elucidates the molecular pathogenesis of SLE and could open up promising avenues to create tailored treatments during this condition. In this review we summarized various non-coding RNAs and Exosomal non-coding RNAs in SLE.
Liquid biopsy is a rapidly evolving diagnostic technique used to analyze tissue-derived information found in the blood or other bodily fluids. It represents a new way to guide therapeutic decisions, mainly in cancer, but its application in other fields of medicine is still growing. Here, we discuss how liquid biopsy has been used in autoimmune rheumatic diseases such as rheumatoid arthritis, systemic lupus erythematosus, or primary Sjögren's syndrome. Additionally, in aspect of liquid biopsy, we analyze the molecular biomarkers utilized in the field of rheumatology, including circulating cell-free DNA, microRNA, and proteomic content.
Background
Systemic lupus erythematosus (SLE) is a common autoimmune disease, and its pathogenesis remains unclear. The alteration of genetic materials is believed to play a role in SLE development. This study evaluated the association between the genetic variants of microRNA‐21 (miR‐21) and microRNA‐155 (miR‐155) and SLE.
Methods
The SNaPshot genotyping method was used to detect the genotypes of selected SNPs in patients and controls. The expression of miR‐21 and miR‐155 was analyzed using reverse transcription‐quantitative polymerase chain reaction (RT‐qPCR). The functional annotation and the biological effects of SNPs were assessed by HaploReg V4.1 and Regulome DB V2.0 software. The Hardy–Weinberg equilibrium test was used to gather statistics, and odds ratios (ORs) and 95% confidence intervals (CIs) were evaluated by logistic regression.
Results
The distribution difference of TA genotype in rs767649 was observed (TA vs. T/T: OR = 0.68, 95%CI, 0.48–0.95, p = 0.026). There was a significant difference in the T/A + A/A (T/A + A/A vs. T/T: OR = 0.68, 95%CI, 0.49–0.94, p = 0.020). A significant difference in T allele distribution was found in the depressed complement of SLE (T vs. A: OR = 0.67, 95%CI, 0.47–0.95, p = 0.026). There were significant differences in genetic variants of rs13137 between the positive and the negative SSB antibodies (Anti‐SSB) (T vs. A: OR = 0.67, 95%CI, 0.47–0.95, p = 0.026; T/A + T/T vs. AA: OR = 2.23, 1.18–4.49, p = 0.013). The expression levels of miR‐21 and miR‐155 were significantly higher in patients than in controls (p < 0.001).
Conclusions
This study provides novel insight that genetic variants of rs767649 and rs13137 are associated with susceptibility to SLE.
Autoimmune diseases are a group of different inflammatory disorders characterized by systemic or localized inflammation, affecting approximately 0.1–1% of the general population. Several studies suggest that genetic risk loci are shared between different autoimmune diseases and pathogenic mechanisms may also be shared. The strategy of performing differential gene expression profiles in autoimmune disorders has unveiled new transcripts that may be shared among these disorders. Microarray technology and bioinformatics offer the most comprehensive molecular evaluations and it is widely used to understand the changes in gene expression in specific organs or in peripheral blood cells. The major goal of transcriptome studies is the identification of specific biomarkers for different diseases. It is believed that such knowledge will contribute to the development of new drugs, new strategies for early diagnosis, avoiding tissue autoimmune destruction, or even preventing the development of autoimmune disease. In this review, we primarily focused on the transcription profiles of three typical autoimmune disorders, including type 1 diabetes mellitus (destruction of pancreatic islet beta cells), systemic lupus erythematosus (immune complex systemic disorder affecting several organs and tissues), and multiple sclerosis (inflammatory and demyelinating disease of the central nervous system).KeywordsAutoimmune diseasesDifferential gene expressionMicroarrayBioinformaticsBiomarkersType 1 diabetes mellitusSystemic lupus erythematosusMultiple sclerosisImmune systemImmunodeficiencySelf-antigensTolerance lossT cellsCell proliferationGenetic lociImmunological primingSex hormonesImmune responsesArbovirusCoronavirusZikaChikungunyaDengueRheumatological complicationsMultiple sclerosisNeuromyelitis opticaGuillain-Barré syndromeSARS-CoV2HLA haplotypesChromosomal locationLong noncoding RNA (lncRNA)B-cell insulin secretionmiRNAGlucose metabolismGender-related factorsBacterial infectionsUltra-violet radiationGenome-wide associationLinkageCytokine signaturesSusceptibility signatureNephritisInterferonPeripheral blood mononuclear cells (PBMC)Autoreactive T cellsMyelin-oligodendrocyte complexCNSMacrophage infiltrationDemyelinationGliosisAxonal lossGWAS studies
Background
Inhibiting Tfh cell overexpansion prevents autoimmune responses and disease flares in systemic lupus erythematosus (SLE). miR-21 is highly expressed in SLE CD4⁺ T cells, but whether inhibiting miR-21 can reduce Tfh cell expansion and alleviate the disease progression of lupus is unclear.
Aim of the study
To address the role and molecular mechanism of miR-21 in regulating Tfh cell expansion and its therapeutic effect on SLE.
Methods
We treated 12-week-old MRL/lpr mice with Antagomir-21, which specifically inhibited miR-21 in vivo. After 12 weeks of treatment, we examined the proportions of Tfh cells and germinal center (GC) B cells and serum levels of autoantibodies and evaluated disease severity by histological scoring and albuminuria. We determined the level of intracellular free iron in CD4⁺ T cells by PGSK probe and examined the expression of the Fth and Tfrc genes by qPCR. Immunohistochemistry (IHC) was used to assess the 5-hmC level in the draining lymph nodes (dLNs) and spleen.
Results and Conclusions
Inhibiting miR-21 significantly reduced the expansion of Tfh cells and GC B cells. Furthermore, Antagomir-21 highly improved skin lesions and nephritis in MRL/lpr mice. Inhibiting miR-21 reduced intracellular iron accumulation and DNA hydroxymethylation in T cells. In conclusion, inhibiting miR-21 in vivo improves intracellular iron homeostasis and inhibits Tfh cell overexpansion, contributing to reduced autoimmune responses and the remission of disease symptoms in murine lupus.
Rheumatoid arthritis (RA) and ankylosing spondylitis (AS) belong to the most common inflammatory rheumatic diseases. MicroRNAs (miRNAs) are small 18–22 RNA molecules that function as posttranscriptional regulators. They are abundantly present within extracellular vesicles (EVs), small intercellular communication vesicles that can be found in bodily fluids and that have key functions in pathological and physiological pathways. Recently, EVs have gained much interest because of their diagnostic and therapeutic potential. Using NanoString profiling technology, the miRNA repertoire of serum EVs was determined and compared in RA and AS patients before and after anti-TNF therapy to assess its potential use as a diagnostic and prognostic biomarker. Furthermore, possible functional effects of those miRNAs that were characterized by the most significant expression changes were evaluated using in silico prediction algorithms. The analysis revealed a unique profile of differentially expressed miRNAs in RA and AS patient serum EVs. We identified 12 miRNAs whose expression profiles enabled differentiation between RA and AS patients before induction of anti-TNF treatment, as well as 4 and 14 miRNAs whose repertoires were significantly changed during the treatment in RA and AS patients, respectively. In conclusion, our findings suggest that extracellular vesicle miRNAs could be used as potential biomarkers associated with RA and AS response to biological treatment.
Skin disorders are a large family of various complex diseases and dysfunctions. With the development of epigenetics in human diseases, more and more skin disorders were proved to be associated with epigenetic modifications. Defined as heritable changes in gene expression that do not involve a change in the genomic DNA sequence, epigenetics may play a significant role and open a new window in skin disorders.
In this chapter, we summarize the epigenetic modifications of a number of skin disorders, including immunologic skin diseases, infectious skin diseases, skin tumors, and skin aging. We also review a number of candidate epigenetic markers and potential therapeutic strategies. Understanding the concepts of epigenetics can provide insights into the pathogenesis of skin disorders, and may lead to the development of biomarkers, improve prognosis, and support the development of novel molecular targets in therapies, especially refractory skin diseases. Epigenetics will serve as the next large area of medicine with the potential to treat and possibly prevent skin diseases in the future.
Epigenetic modifications have been well documented in autoimmune diseases. MicroRNAs (miRNAs), in particular, have long intrigued scientists in the field of autoimmunity. Owing to its central role in the development of the immune system, microRNA-155 (miR-155) is deeply involved in systemic lupus erythematosus (SLE). Despite the advancements made in treating SLE, the disease still remains incurable. Therefore, recent attention has been drawn to the manipulation of epigenetics in the development of curative treatments. In fact, it is a widely held view that miRNA-targeted therapy is a new glimmer of hope in the treatment of autoimmune diseases. However, the duplicity of miRNAs should not be overlooked. A single miRNA can target several mRNAs, and some mRNAs may possess opposing functions. In this review, we highlight the role of miR-155 as a biomarker and review its functions in SLE patients and animal models while discussing possible reasons behind inconsistencies across studies.
miRNA-21 is among the most abundant and highly conserved microRNAs (miRNAs) recognized. It is expressed in essentially all cells where it performs vital regulatory roles in health and disease. It is also frequently claimed to be a biomarker of diseases such as cancer and heart disease in bodily-fluid based miRNA studies. Here we dissociate its contributions to cellular physiology and pathology from its potential as a biomarker. We show how it has been claimed as a specific predictive or prognostic biomarker by at least 29 diseases. Thus, it has no specificity to any one disease. As a result, it should not be considered a viable candidate to be a biomarker, despite its continued evaluation as such. This theme of multiple assignments of a miRNA as a biomarker is shared with other common, ubiquitous miRNAs and should be concerning for them as well.
Background
MicroRNAs (miRNAs) contribute to the development and progression of systemic lupus erythematosus (SLE) via affecting a wide range of targeted genes and facilitating the development of lupus nephritis (LN).
Aim
To analyze the serum expression of miR‐181a and miR‐223 in SLE patients and to assess whether they could serve as novel biomarkers for SLE diagnosis and to distinguish LN.
Subject & methods
This study included 70 control subjects and 116 patients with SLE (67 non‐LN and 49 LN groups). Circulating miR‐181a and miR‐223 expression levels were analyzed among Egyptian population using real time polymerase chain reaction.
Results
Up‐regulation of miR‐181a was detected among SLE patients compared to healthy controls and higher values were reported among LN group compared to non‐LN group. Down‐regulation of miR‐223 was reported among SLE patients compared to controls and lower values were reported among LN group compared to non‐LN group. The higher miR‐181a expression and the lower miR‐223 expression were associated with higher stages of LN. SLE disease activity index (SLEDAI), proteinuria and serum creatinine were independently correlated with miR‐181a and miR‐223 among SLE patients by linear regression analysis. ROC curve analysis revealed that combined miR‐181a and miR‐223 expressions increased the sensitivity and specificity for the diagnosis of SLE and further to distinguish LN from non‐LN patients.
Conclusion
MiR‐181a and miR‐223 could play a role in evaluating SLE disease progression and prognosis. Combined miR‐181a and miR‐223 expression analysis could serve as novel serum based biomarkers in the diagnosis of SLE and predicting LN among Egyptians.
Toll-like receptors 7 and 8 (TLR7/8) are transmembrane receptors that recognize single-stranded RNA. Activation of these receptors results in immune cell stimulation and inflammatory cytokine production which is normally a protective host response. However, aberrant activation of TLR7/8 is potentially pathogenic and linked to progression of certain autoimmune diseases such as lupus. Thus, we hypothesize that an inhibitor that blocks TLR7/8 would be an effective therapeutic treatment. Prior efforts to develop inhibitors of TLR7/8 have been largely unsuccessful due to the challenge of producing a small molecule inhibitor for these difficult targets. Here, we report the characterization of M5049 and Compound 2, molecules which were discovered in a medicinal chemistry campaign to produce dual TLR7/8 inhibitors with drug-like properties. Both compounds showed potent and selective activity in a range of cellular assays for inhibition of TLR7/8 and block molecule synthetic ligands and natural endogenous RNA ligands such as microRNA and Alu RNA. M5049 was found to be potent in vivo and TLR7/8 inhibition efficaciously treated disease in several murine lupus models and, interestingly, was efficacious in a disease context where TLR7/8 activity has not previously been considered a primary disease driver. Furthermore, M5049 had greater potency in disease models than expected based on its in vitro potency and pharmacokinetic/pharmacodynamic properties. With preferential accumulation in tissues, and the ability to block multiple TLR7/8 RNA ligands, M5049 may be efficacious in treating autoimmunity and has the potential to provide benefit to a variety of patients with varying disease pathogenesis. Significance Statement We report discovery of a novel toll-like receptor 7 and 8 (TLR7/8) inhibitor (M5049), characterize its binding mode, potency/selectivity, pharmacokinetic and pharmacodynamic properties, and demonstrate its potential for treating autoimmune diseases in two mouse lupus models. TLR7/8 inhibition is unique in that it may block both innate and adaptive autoimmunity and thus we believe that M5049 has the potential to benefit patients with autoimmune diseases.
Objective:
To determine the circulatory miRNA expression levels in patients with Hashimoto thyroiditis (HT) at the time of diagnosis and follow-up period compared with healthy controls.
Methods:
We collected blood samples from 34 patients with HT (4 males and 30 females) at the time of first diagnosis (Group P) and euthyroid period (Group E). Thirty-three healthy controls (Group H) blood samples were also included in the study. Expression levels of five different circulating miRNAs (miR-22, miR-141, miR-155, miR-375, miR-451) were evaluated using real-time polymerase chain reaction.
Results:
There was a significant difference in miR-375 levels between the P group and the H. Also, for miR-451, there was a significant difference between the P and E groups. Finally, there was a moderate positive correlation between thyroidstimulating hormone values and miR-22 expression levels for the P group.
Conclusion:
miRNAs have important roles at all stages of the diseases. More studies must be performed in all thyroid diseases and autoimmune diseases, including HT.
Many patients with systemic autoimmune diseases have signs of a continuous production of type I interferon (IFN) and display an increased expression of IFN-α-regulated genes. The reason for the on-going IFN-α synthesis in these patients seems to be an activation of plasmacytoid dendritic cells (pDCs) by immune complexes (ICs), consisting of autoantibodies in combination with DNA or RNA-containing autoantigens. Such interferogenic ICs are internalized via the FcγRIIa expressed on pDCs, reach the endosome, and stimulate Toll-like receptor (TLR)-7 or -9, which subsequently leads to IFN-α gene transcription. Variants of genes involved in both the IFN-α synthesis and response have been linked to an increased risk to develop systemic lupus erythematosus (SLE) and other autoimmune diseases. Among these autoimmunity risk genes are IFN regulatory factor 5 (IRF5), which is involved in TLR signaling, and the signal transducer and activator of transcription 4 (STAT4) that interacts with the type I IFN receptor. Several other gene variants in the IFN signaling pathway also confer an increased risk to develop an autoimmune disease. The observations that IFN-α therapy can induce autoimmunity and that many autoimmune conditions have an on-going type I IFN production suggest that the type I IFN system has a pivotal role in the etiopathogenesis of these diseases. Possible mechanisms behind the dysregulated type IFNsystem in autoimmune diseases and how the IFN-α produced can contribute to the development of an autoimmune process will be reviewed.
Systemic lupus erythematosus (SLE) is a complex autoimmune disease with a strong genetic predisposition, characterized by an upregulated type I interferon pathway. MicroRNAs are important regulators of immune homeostasis, and aberrant microRNA expression has been demonstrated in patients with autoimmune diseases. We recently identified miR-146a as a negative regulator of the interferon pathway and linked the abnormal activation of this pathway to the underexpression of miR-146a in SLE patients. To explore why the expression of miR-146a is reduced in SLE patients, we conducted short parallel sequencing of potentially regulatory regions of miR-146a and identified a novel genetic variant (rs57095329) in the promoter region exhibiting evidence for association with SLE that was replicated independently in 7,182 Asians (P(meta) = 2.74×10(-8), odds ratio = 1.29 [1.18-1.40]). The risk-associated G allele was linked to reduced expression of miR-146a in the peripheral blood leukocytes of the controls. Combined functional assays showed that the risk-associated G allele reduced the protein-binding affinity and activity of the promoter compared with those of the promoter containing the protective A allele. Transcription factor Ets-1, encoded by the lupus-susceptibility gene ETS1, identified in recent genome-wide association studies, binds near this variant. The manipulation of Ets-1 levels strongly affected miR-146a promoter activity in vitro; and the knockdown of Ets-1, mimicking its reduced expression in SLE, directly impaired the induction of miR-146a. We also observed additive effects of the risk alleles of miR-146a and ETS1. Our data identified and confirmed an association between a functional promoter variant of miR-146a and SLE. This risk allele had decreased binding to transcription factor Ets-1, contributing to reduced levels of miR-146a in SLE patients.
Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by the production of autoantibodies against a host of nuclear antigens. The pathogenesis of lupus is incompletely understood. Environmental factors may play a role via altering DNA methylation, a mechanism regulating gene expression. In lupus, genes including CD11a and CD70 are overexpressed in T cells as a result of promoter hypomethylation. T-cell DNA methyltransferase expression is regulated in part by the extracellular signal-regulated kinase (ERK) signaling pathway. In this study, we investigate the effects of decreased ERK pathway signaling in T cells using transgenic animals. We generated a transgenic mouse that inducibly expresses a dominant-negative MEK in T cells in the presence of doxycycline. We show that decreased ERK pathway signaling in T cells results in decreased expression of DNA methyltransferase 1 and overexpression of the methylation-sensitive genes CD11a and CD70, similar to T cells in human lupus. Our transgenic animal model also develops anti-dsDNA antibodies. Interestingly, microarray expression assays revealed overexpression of several interferon-regulated genes in the spleen similar to peripheral blood cells of lupus patients. This model supports the contention that ERK pathway signaling defects in T cells contribute to the development of autoimmunity.
MicroRNAs (miRNAs) circulate in the bloodstream in a highly stable, extracellular form and are being developed as blood-based biomarkers for cancer and other diseases. However, the mechanism underlying their remarkable stability in the RNase-rich environment of blood is not well understood. The current model in the literature posits that circulating miRNAs are protected by encapsulation in membrane-bound vesicles such as exosomes, but this has not been systematically studied. We used differential centrifugation and size-exclusion chromatography as orthogonal approaches to characterize circulating miRNA complexes in human plasma and serum. We found, surprisingly, that the majority of circulating miRNAs cofractionated with protein complexes rather than with vesicles. miRNAs were also sensitive to protease treatment of plasma, indicating that protein complexes protect circulating miRNAs from plasma RNases. Further characterization revealed that Argonaute2 (Ago2), the key effector protein of miRNA-mediated silencing, was present in human plasma and eluted with plasma miRNAs in size-exclusion chromatography. Furthermore, immunoprecipitation of Ago2 from plasma readily recovered non-vesicle-associated plasma miRNAs. The majority of miRNAs studied copurified with the Ago2 ribonucleoprotein complex, but a minority of specific miRNAs associated predominantly with vesicles. Our results reveal two populations of circulating miRNAs and suggest that circulating Ago2 complexes are a mechanism responsible for the stability of plasma miRNAs. Our study has important implications for the development of biomarker approaches based on capture and analysis of circulating miRNAs. In addition, identification of extracellular Ago2-miRNA complexes in plasma raises the possibility that cells release a functional miRNA-induced silencing complex into the circulation.
Systemic lupus erythematosus is a chronic autoimmune disease characterized by loss of tolerance to self-Ags and activation of autoreactive T cells. Regulatory T (Treg) cells play a critical role in controlling the activation of autoreactive T cells. In this study, we investigated mechanisms of potential Treg cell defects in systemic lupus erythematosus using MRL-Fas(lpr/lpr) (MRL/lpr) and MRL-Fas(+/+) mouse models. We found a significant increase in CD4(+)CD25(+)Foxp3(+) Treg cells, albeit with an altered phenotype (CD62L(-)CD69(+)) and with a reduced suppressive capacity, in the lymphoid organs of MRL strains compared with non-autoimmune C3H/HeOuj mice. A search for mechanisms underlying the altered Treg cell phenotype in MRL/lpr mice led us to find a profound reduction in Dicer expression and an altered microRNA (miRNA, miR) profile in MRL/lpr Treg cells. Despite having a reduced level of Dicer, MRL/lpr Treg cells exhibited a significant overexpression of several miRNAs, including let-7a, let-7f, miR-16, miR-23a, miR-23b, miR-27a, and miR-155. Using computational approaches, we identified one of the upregulated miRNAs, miR-155, that can target CD62L and may thus confer the altered Treg cell phenotype in MRL/lpr mice. In fact, the induced overexpression of miR-155 in otherwise normal (C3H/HeOuj) Treg cells reduced their CD62L expression, which mimics the altered Treg cell phenotype in MRL/lpr mice. These data suggest a role of Dicer and miR-155 in regulating Treg cell phenotype. Furthermore, simultaneous appearance of Dicer insufficiency and miR-155 overexpression in diseased mice suggests a Dicer-independent alternative mechanism of miRNA regulation under inflammatory conditions.
To date, there are no highly sensitive and specific minimally invasive biomarkers for detection of breast cancer at an early stage. The occurrence of circulating microRNAs (miRNAs) in blood components (including serum and plasma) has been repeatedly observed in cancer patients as well as healthy controls. Because of the significance of miRNA in carcinogenesis, circulating miRNAs in blood may be unique biomarkers for early and minimally invasive diagnosis of human cancers. The objective of this pilot study was to discover a panel of circulating miRNAs as potential novel breast cancer biomarkers.Using microarray-based expression profiling followed by Real-Time quantitative Polymerase Cycle Reaction (RT-qPCR) validation, we compared the levels of circulating miRNAs in plasma samples from 20 women with early stage breast cancer (10 Caucasian American (CA) and 10 African American (AA)) and 20 matched healthy controls (10 CAs and 10 AAs). Using the significance level of p
MicroRNAs (miRNA) have emerged as an important new class of modulators of gene expression. In this study we investigated miRNA that are differentially expressed in lupus nephritis. Microarray technology was used to investigate differentially expressed miRNA in peripheral blood mononuclear cells (PBMCs) and Epstein-Barr Virus (EBV)-transformed cell lines obtained from lupus nephritis affected patients and unaffected controls. TaqMan-based stem-loop real-time polymerase chain reaction was used for validation. Microarray analysis of miRNA expressed in both African American (AA) and European American (EA) derived lupus nephritis samples revealed 29 and 50 differentially expressed miRNA, respectively, of 850 tested. There were 18 miRNA that were differentially expressed in both racial groups. When samples from both racial groups and different specimen types were considered, there were 5 primary miRNA that were differentially expressed. We have identified 5 miRNA; hsa-miR-371-5P, hsa-miR-423-5P, hsa-miR-638, hsa-miR-1224-3P and hsa-miR-663 that were differentially expressed in lupus nephritis across different racial groups and all specimen types tested. Hsa-miR-371-5P, hsa-miR-1224-3P and hsa-miR-423-5P, are reported here for the first time to be associated with lupus nephritis. Our work establishes EBV-transformed B cell lines as a useful model for the discovery of miRNA as biomarkers for SLE. Based on these findings, we postulate that these differentially expressed miRNA may be potential novel biomarkers for SLE as well as help elucidate pathogenic mechanisms of lupus nephritis. The investigation of miRNA profiles in SLE may lead to the discovery and development of novel methods to diagnosis, treat and prevent SLE.
Systemic lupus erythematosus is a complex autoimmune disease caused by genetic and epigenetic alterations. DNA methylation abnormalities play an important role in systemic lupus erythematosus disease processes. MicroRNAs (miRNAs) have been implicated as fine-tuning regulators controlling diverse biological processes at the level of posttranscriptional repression. Dysregulation of miRNAs has been described in various disease states, including human lupus. Whereas previous studies have shown miRNAs can regulate DNA methylation by targeting the DNA methylation machinery, the role of miRNAs in aberrant CD4+ T cell DNA hypomethylation of lupus is unclear. In this study, by using high-throughput microRNA profiling, we identified that two miRNAs (miR-21 and miR-148a) overexpressed in CD4+ T cells from both patients with lupus and lupus-prone MRL/lpr mice, which promote cell hypomethylation by repressing DNA methyltransferase 1 (DNMT1) expression. This in turn leads to the overexpression of autoimmune-associated methylation-sensitive genes, such as CD70 and LFA-1, via promoter demethylation. Further experiments revealed that miR-21 indirectly downregulated DNMT1 expression by targeting an important autoimmune gene, RASGRP1, which mediated the Ras-MAPK pathway upstream of DNMT1; miR-148a directly downregulated DNMT1 expression by targeting the protein coding region of its transcript. Additionally, inhibition of miR-21 and miR-148a expression in CD4+ T cells from patients with lupus could increase DNMT1 expression and attenuate DNA hypomethylation. Together, our data demonstrated a critical functional link between miRNAs and the aberrant DNA hypomethylation in lupus CD4+ T cells and could help to develop new therapeutic approaches.
MicroRNAs (miRNAs), endogenous small noncoding RNAs regulating the activities of target mRNAs and cellular processes, are present in human plasma in a stable form. In this study, we investigated whether miRNAs are also stably present in synovial fluids and whether plasma and synovial fluid miRNAs could be biomarkers of rheumatoid arthritis (RA) and osteoarthritis (OA).
We measured concentrations of miR-16, miR-132, miR-146a, miR-155 and miR-223 in synovial fluid from patients with RA and OA, and those in plasma from RA, OA and healthy controls (HCs) by quantitative reverse transcription-polymerase chain reaction. Furthermore, miRNAs in the conditioned medium of synovial tissues, monolayer fibroblast-like synoviocytes, and mononuclear cells were examined. Correlations between miRNAs and biomarkers or disease activities of RA were statistically examined.
Synovial fluid miRNAs were present and as stable as plasma miRNAs for storage at -20 degrees C and freeze-thawing from -20 degrees C to 4 degrees C. In RA and OA, synovial fluid concentrations of miR-16, miR-132, miR-146a, and miR-223 were significantly lower than their plasma concentrations, and there were no correlation between plasma and synovial fluid miRNAs. Interestingly, synovial tissues, fibroblast-like synoviocytes, and mononuclear cells secreted miRNAs in distinct patterns. The expression patterns of miRNAs in synovial fluid of OA were similar to miRNAs secreted by synovial tissues. Synovial fluid miRNAs of RA were likely to originate from synovial tissues and infiltrating cells. Plasma miR-132 of HC was significantly higher than that of RA or OA with high diagnosability. Synovial fluid concentrations of miR-16, miR-146a miR-155 and miR-223 of RA were significantly higher than those of OA. Plasma miRNAs or ratio of synovial fluid miRNAs to plasma miRNAs, including miR-16 and miR-146a, significantly correlated with tender joint counts and 28-joint Disease Activity Score.
Plasma miRNAs had distinct patterns from synovial fluid miRNAs, which appeared to originate from synovial tissue. Plasma miR-132 well differentiated HCs from patients with RA or OA, while synovial fluid miRNAs differentiated RA and OA. Furthermore, plasma miRNAs correlated with the disease activities of RA. Thus, synovial fluid and plasma miRNAs have potential as diagnostic biomarkers for RA and OA and as a tool for the analysis of their pathogenesis.
Molecular diagnostics in cancer should provide the highest specificity and sensitivity in classification, prognostic stratification and early detection. miRNAs could contribute to hitting the mark, or at least to come nearer, by virtue of their cancer-specific expression and stability. Indeed, different to other RNA classes, miRNAs can be detected and quantified not only in frozen tissues, but also in formalin-fixed paraffin-embedded tissues, as well as serum/plasma samples. Thus, miRNA studies have quickly moved from research on the molecular basis of cancer to areas of clinical application. This review summarizes the potential role of miRNAs as molecular markers for cancer classification, prognostic stratification and drug-response prediction. It also summarizes their potential as circulating markers and cancer-predisposing genes. If we consider that studies on miRNAs in cancer therapy have already given important contributions, miRNAs have an impact in all cancer areas. Whether this will translate into clinical applications is still too early to say. However, in the diagnostic field, miRNAs may already represent an improvement over presently available approaches; for example, their expression profile is effective in the identification of tissue of origin of metastasis. In addition, circulating miRNAs are expected to provide improved specificity and/or sensitivity over presently available markers.
Laser capture microdissection (LCM) has successfully isolated pure cell populations from tissue sections and the combination of LCM with standard genomic and proteomic methods has revolutionized molecular analysis of complex tissue. However, the quantity and quality of material recovered after LCM is often still limited for analysis by using whole genomic and proteomic approaches. To procure high quality and quantity of RNA after LCM, we optimized the procedures on tissue preparations and applied the approach for cell type-specific miRNA expression profiling in colorectal tumors.
We found that the ethanol fixation of tissue sections for 2 hours had the maximum improvement of RNA quality (1.8 fold, p = 0.0014) and quantity (1.5 fold, p = 0.066). Overall, the quality (RNA integrity number, RIN) for the microdissected colorectal tissues was 5.2 +/- 1.5 (average +/- SD) for normal (n = 43), 5.7 +/- 1.1 for adenomas (n = 14) and 7.2 +/- 1.2 for carcinomas (n = 44). We then compared miRNA expression profiles of 18 colorectal tissues (6 normal, 6 adenomas and 6 carcinomas) between LCM selected epithelial cells versus stromal cells using Agilent miRNA microarrays. We identified 51 differentially expressed miRNAs (p <= 0.001) between these two cell types. We found that the miRNAs in the epithelial cells could differentiate adenomas from normal and carcinomas. However, the miRNAs in the stromal and mixed cells could not separate adenomas from normal tissues. Finally, we applied quantitative RT-PCR to cross-verify the expression patterns of 7 different miRNAs using 8 LCM-selected epithelial cells and found the excellent correlation of the fold changes between the two platforms (R = 0.996).
Our study demonstrates the feasibility and potential power of discovering cell type-specific miRNA biomarkers in complex tissue using combination of LCM with genome-wide miRNA analysis.
Systemic lupus erythematosus is a poorly understood autoimmune disease, characterized by autoantibodies to nuclear antigens and immune complex deposition in organs like the kidney. Current evidence indicates that a pathologic CD4+T cell subset, characterized by impaired extracellular signal-regulated kinase (ERK) pathway signaling, DNA hypomethylation, and consequent aberrant gene expression contributes to disease pathogenesis. Hydralazine is a lupus-inducing drug that also decreases T cell DNA methylation by inhibiting the ERK signaling pathway, replicating the defect found in lupus T cells. These observations suggest that defective ERK pathway signaling alters gene expression in T cells by inhibiting DNA methylation, contributing to lupus pathogenesis. The signaling defect in hydralazine-treated and lupus T cells has now been mapped to protein kinase C delta. Understanding the mechanism causing decreased ERK pathway signaling in lupus may shed light on mechanisms contributing to disease development in genetically predisposed people.
MicroRNAs (miRNAs) are a class of small noncoding RNAs that regulate gene expression at the posttranscriptional level by degrading or blocking translation of messenger RNA (mRNA) targets. MiRNAs play important regulatory roles in a variety of cellular functions as well as in several diseases, including cancer. MiRNA-specific expression profiles have been reported for several pathological conditions. Recently, the discovery of miRNAs in serum opens up the possibility of using miRNAs as biomarkers of disease. In this review, we discuss the potential use of miRNAs as clinically diagnostic biomarkers of various cancers and other diseases as well as the approaches used to detect these molecules in serum and plasma.
MicroRNA have recently been identified as regulators that modulate target gene expression and are involved in shaping the immune response. This study was undertaken to investigate the contribution of microRNA-146a (miR-146a), which was identified in the pilot expression profiling step, to the pathogenesis of systemic lupus erythematosus (SLE).
TaqMan microRNA assays of peripheral blood leukocytes were used for comparison of expression levels of microRNA between SLE patients and controls. Transfection and stimulation of cultured cells were conducted to determine the biologic function of miR-146a. Bioinformatics prediction and validation by reporter gene assay and Western blotting were performed to identify miR-146a targets.
Profiling of 156 miRNA in SLE patients revealed the differential expression of multiple microRNA, including miR-146a, a negative regulator of innate immunity. Further analysis showed that underexpression of miR-146a negatively correlated with clinical disease activity and with interferon (IFN) scores in patients with SLE. Of note, overexpression of miR-146a reduced, while inhibition of endogenous miR-146a increased, the induction of type I IFNs in peripheral blood mononuclear cells (PBMCs). Furthermore, miR-146a directly repressed the transactivation downstream of type I IFN. At the molecular level, miR-146a could target IFN regulatory factor 5 and STAT-1. More importantly, introduction of miR-146a into the patients' PBMCs alleviated the coordinate activation of the type I IFN pathway.
The microRNA miR-146a is a negative regulator of the IFN pathway. Underexpression of miR-146a contributes to alterations in the type I IFN pathway in lupus patients by targeting the key signaling proteins. The findings provide potential novel strategies for therapeutic intervention.
Dysregulated expression of microRNAs (miRNAs) in various tissues has been associated with a variety of diseases, including cancers. Here we demonstrate that miRNAs are present in the serum and plasma of humans and other animals such as mice, rats, bovine fetuses, calves, and horses. The levels of miRNAs in serum are stable, reproducible, and consistent among individuals of the same species. Employing Solexa, we sequenced all serum miRNAs of healthy Chinese subjects and found over 100 and 91 serum miRNAs in male and female subjects, respectively. We also identified specific expression patterns of serum miRNAs for lung cancer, colorectal cancer, and diabetes, providing evidence that serum miRNAs contain fingerprints for various diseases. Two non-small cell lung cancer-specific serum miRNAs obtained by Solexa were further validated in an independent trial of 75 healthy donors and 152 cancer patients, using quantitative reverse transcription polymerase chain reaction assays. Through these analyses, we conclude that serum miRNAs can serve as potential biomarkers for the detection of various cancers and other diseases.
The two most commonly used methods to analyze data from real-time, quantitative PCR experiments are absolute quantification and relative quantification. Absolute quantification determines the input copy number, usually by relating the PCR signal to a standard curve. Relative quantification relates the PCR signal of the target transcript in a treatment group to that of another sample such as an untreated control. The 2(-DeltaDeltaCr) method is a convenient way to analyze the relative changes in gene expression from real-time quantitative PCR experiments. The purpose of this report is to present the derivation, assumptions, and applications of the 2(-DeltaDeltaCr) method. In addition, we present the derivation and applications of two variations of the 2(-DeltaDeltaCr) method that may be useful in the analysis of real-time, quantitative PCR data. (C) 2001 Elsevier science.
Statistical Learning Theory focuses on the machine learning theory for small samples. Support vector machine (SVM) are new methods based on statistical learning theory. There are many kinds of function can be used for kernel of SVM. Wavelet function is a set of bases that can approximate arbitrary functions in arbitrary precision. So Marr wavelet was used to construct wavelet kernel. On the other hand, the parameter selection should to be done before training WSVM. Modified chaotic particle swarm optimization (CPOS) was adpoted to select parameters of SVM. It is shown by simulation that the CPOS algorithm can derive a set of optimal parameters of WSVM, and WSVM model possess some advantages such as simple structure, fast convergence speed with high generalization ability.
The regulation of gene expression at the posttranscriptional level has revealed important control levels for genes important to the immune system. MicroRNAs (miRNAs) are small RNAs that regulate gene expression by inhibiting protein translation or by degrading the mRNA transcript. A single miRNA can potentially regulate the expression of multiple genes and the proteins encoded. MiRNA can influence molecular signaling pathways and regulate many biological processes including immune function. Although the role of miRNAs in development and oncogenesis has been well characterized, their role in the immune system has only begun to emerge. During the past few years, many miRNAs have been found to be important in the development, differentiation, survival, and function of B and T lymphocytes, dendritic cells, macrophages, and other immune cell types. We discuss here recent findings revealing important roles for miRNA in immunity and how miRNAs can regulate innate and adaptive immune responses.
To identify microRNA genes with abnormal expression in the CD4+ T cells of patients with systemic lupus erythematosus (SLE) and to determine the role of microRNA-126 (miR-126) in the etiology of SLE.
MicroRNA expression patterns in CD4+ T cells from patients with SLE and healthy control subjects were analyzed by microRNA microarray and stem loop quantitative polymerase chain reaction (qPCR). Luciferase reporter gene assays were performed to identify miR-126 targets. Dnmt1, CD11a, and CD70 messenger RNA and protein levels were determined by real-time qPCR, Western blotting, and flow cytometry. CD11a, CD70, and EGFL7 promoter methylation levels were detected by bisulfite sequencing. IgG levels in T cell-B cell cocultures were determined by enzyme-linked immunosorbent assay.
The expression of 11 microRNA was significantly increased or decreased in CD4+ T cells from patients with SLE relative to that in CD4+ T cells from control subjects. Among these, miR-126 was up-regulated, and its degree of overexpression was inversely correlated with Dnmt1 protein levels. We demonstrated that miR-126 directly inhibits Dnmt1 translation via interaction with its 3'-untranslated region, and that overexpression of miR-126 in CD4+ T cells can significantly reduce Dnmt1 protein levels. The overexpression of miR-126 in CD4+ T cells from healthy donors caused the demethylation and up-regulation of genes encoding CD11a and CD70, thereby causing T cell and B cell hyperactivity. The inhibition of miR-126 in CD4+ T cells from patients with SLE had the opposite effects. Expression of the miR-126 host gene EGFL7 was also up-regulated in CD4+ T cells from patients with SLE, possibly in a hypomethylation-dependent manner.
Our data suggest that miR-126 regulates DNA methylation in CD4+ T cells and contributes to T cell autoreactivity in SLE by directly targeting Dnmt1.
MicroRNAs (miRNAs) are newly discovered, small, noncoding ribonucleic acids (RNAs) that play critical roles in the regulation of host genome expression at the posttranscriptional level. During last 20 years, miRNAs have emerged as key regulators of various biological processes including immune cell lineage commitment, differentiation, maturation, and maintenance of immune homeostasis and normal function. Thus, it is not surprising that dysregulated miRNA expression patterns now have been documented in a broad range of diseases including cancer as well as inflammatory and autoimmune diseases. This rapidly emerging field has revolutionized our understanding of normal immunoregulation and breakdown of self-tolerance. This review focuses on the current understanding of miRNA biogenesis, the role of miRNAs in the regulation of innate and adaptive immunity, and the association of miRNAs with autoimmune diseases. We have discussed miRNA dysregulation and the potential role of miRNAs in systemic lupus erythematosus (SLE), rheumatoid arthritis, and multiple sclerosis. Given that most autoimmune diseases are female-predominant, we also have discussed sex hormone regulation of miRNAs in inflammatory responses, with an emphasis on estrogen, which now has been shown to regulate miRNAs in the immune system. The field of miRNA regulation of mammalian genes has tremendous potential. The identification of specific miRNA expression patterns in autoimmune diseases as well as a comprehensive understanding of the role of miRNA in disease pathogenesis offers promise of not only novel molecular diagnostic markers but also new gene therapy strategies for treating SLE and other inflammatory autoimmune diseases.
Recent studies showed that micro-RNA play important roles in the pathogenesis of autoimmune diseases. We studied the levels of miR-146a and miR-155 in the serum and urinary supernatant of patients with systemic lupus erythematosus (SLE).
The serum and urinary supernatant levels of miR-146a and miR-155 were determined by real-time quantitative polymerase chain reaction in 40 patients with SLE and 30 healthy controls.
Compared to controls, serum miR-146a and miR-155 levels were lower, and the urinary level of miR-146a was higher, in SLE. Estimated glomerular filtration rate (eGFR) correlated with both serum miR-146a (r = 0.519, p = 0.001) and miR-155 (r = 0.384, p = 0.014). Serum miR-146a inversely correlated with proteinuria (r = -0.341, p = 0.031) and the SLE Disease Activity Index (r = -0.465, p = 0.003). Serum miR-146a and miR-155 levels also correlated with red blood cell count, platelet count, and lymphocyte count. After treatment with calcitriol for 6 months, serum miR-146a level of SLE patients increased significantly (p < 0.001), and its change inversely correlated with the level of calcium-phosphate product (r = -0.466, p = 0.003).
The results suggested that serum miR-146a and miR-155 participate in the pathophysiology of SLE and might be used as biomarkers of SLE.
In the past several years, the importance of microRNA (miRNA) in cancer cells has been recognized. Proper control of miRNA expression is essential for maintaining a steady state of the cellular machinery. Recently, it was discovered that extracellular miRNAs circulate in the blood of both healthy and diseased patients, although ribonuclease is present in both plasma and serum. Most of the circulating miRNAs are included in lipid or lipoprotein complexes, such as apoptotic bodies, microvesicles, or exosomes, and are, therefore, highly stable. The existence of circulating miRNAs in the blood of cancer patients has raised the possibility that miRNAs may serve as a novel diagnostic marker. However, the secretory mechanism and biological function, as well as the meaning of the existence of extracellular miRNAs, remain largely unclear. In this review, we summarize the usefulness of circulating miRNA for cancer diagnosis, prognosis, and therapeutics. Furthermore, we propose a mechanism for the secretion and incorporation of miRNA into the cells.
MicroRNAs are small RNAs that control gene expression. Besides their cell intrinsic function, recent studies reported that microRNAs are released by cultured cells and can be detected in the blood.
To address the regulation of circulating microRNAs in patients with stable coronary artery disease.
To determine the regulation of microRNAs, we performed a microRNA profile using RNA isolated from n=8 healthy volunteers and n=8 patients with stable coronary artery disease that received state-of-the-art pharmacological treatment. Interestingly, most of the highly expressed microRNAs that were lower in the blood of patients with coronary artery disease are known to be expressed in endothelial cells (eg, miR-126 and members of the miR-17 approximately 92 cluster). To prospectively confirm these data, we detected selected microRNAs in plasma of 36 patients with coronary artery disease and 17 healthy volunteers by quantitative PCR. Consistent with the data obtained by the profile, circulating levels of miR-126, miR-17, miR-92a, and the inflammation-associated miR-155 were significantly reduced in patients with coronary artery disease compared with healthy controls. Likewise, the smooth muscle-enriched miR-145 was significantly reduced. In contrast, cardiac muscle-enriched microRNAs (miR-133a, miR-208a) tend to be higher in patients with coronary artery disease. These results were validated in a second cohort of 31 patients with documented coronary artery disease and 14 controls.
Circulating levels of vascular and inflammation-associated microRNAs are significantly downregulated in patients with coronary artery disease.
MicroRNA (miRNA) have received increasing attention as posttranscriptional regulators that fine-tune the homeostasis of the inflammatory response. This study aimed to clarify whether miR-125a, which was identified in a pilot expression profiling step, is involved in the inflammatory chemokine pathway in systemic lupus erythematosus (SLE).
Independent verification of miR-125a expression in amplified samples from SLE patients and normal controls was performed by TaqMan quantitative polymerase chain reaction (PCR) analysis. A combination of 3 bioinformatic prediction techniques and reporter gene assays was used to identify miR-125a targets. In vitro systems of overexpression by transfection and inducible expression by stimulation were performed to investigate the function of miR-125a, which was followed by real-time quantitative PCR and enzyme-linked immunosorbent assay.
In SLE patients, the expression of miR-125a was reduced and the expression of its predicted target gene, KLF13, was increased. Bioinformatics predicted that miR-125a base-paired with sequences in the 3'-untranslated region of KLF13. Overexpression of miR-125a led to a significant reduction in the expression of RANTES and KLF13. MicroRNA-125a inhibited endogenous KLF13 expression in a dose-dependent manner, as determined using gain- and loss-of-function methods. A luciferase reporter system confirmed the miR-125a binding sites. Notably, miR-125a expression was induced in T cells in a dose- and time-dependent manner. Finally, the introduction of miR-125a into T cells from SLE patients alleviated the elevated RANTES expression.
MicroRNA-125a negatively regulates RANTES expression by targeting KLF13 in activated T cells. The underexpression of miR-125a contributes to the elevated expression of RANTES in SLE. Our findings extend the role of miRNA in the pathogenesis of lupus and provide potential strategies for therapeutic intervention.
MicroRNAs (miRNAs) are negative regulators of gene expression that play an important role in hematopoiesis. Thalassemia, a defective globin synthesis leading to precipitate of excess unbound globins in red blood cell precursors, results in defective erythroid precursors and ineffective erythropoiesis. Expression pattern of miR-451, an erythroid-specific miRNA, was analyzed during differentiation of erythroid progenitors derived from normal and thalassemic peripheral blood CD34-positive cells, after 14 days of culture. A biphasic expression with transient up-regulation of miRNA-451 on day 3 of cultures was observed during thalassemic erythroid differentiation. In contrast, the expression pattern of the miR-451 in erythroid cells obtained from the other extravascular hemolytic anemia, i.e., hereditary spherocytosis patients showed no transient up-regulation of miR-451 on day 3 of cultures. Our results suggest that early erythroid progenitors in beta-thalassemia have a dysregulated miRNA-451 expression program, and analysis of microRNA is a relevant approach to determine abnormalities of erythropoiesis.
Aberrant expression profiles of circulating microRNAs (miRNAs) have been described in various diseases and provide high sensitivity and specificity. We explored circulating miRNAs as potential biomarkers in patients with heart failure (HF).
The goal of this study was to determine whether miRNAs allow to distinguish clinical HF not only from healthy controls but also from non-HF forms of dyspnea.
A miRNA array was performed on plasma of 12 healthy controls and 12 HF patients. From this array, we selected 16 miRNAs for a second clinical study in 39 healthy controls and in 50 cases with reports of dyspnea, of whom 30 were diagnosed with HF and 20 were diagnosed with dyspnea attributable to non-HF-related causes. This revealed that miR423-5p was specifically enriched in blood of HF cases and receiver-operator-characteristics (ROC) curve analysis showed miR423-5p to be a diagnostic predictor of HF, with an area under the curve of 0.91 (P<0.001). Five other miRNAs were elevated in HF cases but also slightly increased in non-HF dyspnea cases.
We identify 6 miRNAs that are elevated in patients with HF, among which miR423-5p is most strongly related to the clinical diagnosis of HF. These 6 circulating miRNAs provide attractive candidates as putative biomarkers for HF.
Inflammatory bowel disease (IBD) consists of Crohn's disease (CD) and ulcerative colitis (UC). It is thought to be caused by genetic, abnormal immune response of the intestinal immune system and dysfunction of intestinal mucosal barrier against enteric bacteria. Mutational genes can affect the development of IBDs via certain signaling pathways. The abnormal signaling pathways play an important role in the inflammatory process and can lead to dysregulation of the inflammatory response and are crucial in the pathogenesis of IBDs. The signaling pathways mainly include P38 MAPK, JNK MAPK, PI3K/Akt, NF-kappaB signaling pathways. Intestinal microorganisms play a key role in the initiation and maintenance of disease. Disorders of signaling pathways including TLR, NF-kappaB can act on the intestinal barrier, and cause uninhibitedly release of effector T cells which are the central cells mediating inflammation in CD. This review highlights relevant patents and a new insight of signaling pathways associated with IBDs will help to develop better therapeutic approaches.
miRNAs have recently emerged as key regulators of the immune system, being involved in lymphocyte selection and proliferation, in T(reg) cells differentiation, and in hematopoiesis in general. Rheumatoid arthritis (RA) is an autoimmune pathology the etiology of which is still obscure. Although a multifactorial pathogenesis has been hypothesized, the precise mechanisms leading to the disease are still poorly understood at the molecular level. miRNA expression profile analysis highlighted that miR-223 is the only miRNA that is strikingly deregulated in peripheral T-lymphocytes from RA patients compared with healthy donors. Further analysis by quantitative reverse transcription-polymerase chain analysis confirmed that miR-223 is overexpressed in T-lymphocytes from RA patients (n = 28) compared with healthy donors (n = 10). Moreover, purification of different T-lymphocyte populations from RA patients highlights that miR-223 is expressed at higher levels in naive CD4(+) lymphocytes, whereas its expression is barely detectable in T(h)-17 cells. In summary, our data provide a first characterization of the miRNA expression profiles of peripheral T-lymphocytes of RA patients, identifying miR-223 as overexpressed in CD4(+) naive T-lymphocytes from these individuals. A deeper analysis of the biologic functions and effects of the expression of miR-223 in T-lymphocytes is needed to clarify the exact link between our observation and the disease.
The concept of endothelial progenitor cells (EPCs) has attracted considerable interest in cardiovascular research, but despite a decade of research there are still no specific markers for EPCs and results from clinical trials remain controversial. Using liquid chromatography-tandem mass spectrometry, we analyzed the protein composition of microparticles (MPs) originating from the cell surface of EPC cultures. Our data revealed that the conventional methods for isolating mononuclear cells lead to a contamination with platelet proteins. Notably, platelets readily disintegrate into platelet MPs. These platelet MPs are taken up by the mononuclear cell population, which acquires "endothelial" characteristics (CD31, von Willebrand factor [VWF], lectin-binding), and angiogenic properties. In a large population-based study (n = 526), platelets emerged as a positive predictor for the number of colony-forming units and early outgrowth EPCs. Our study provides the first evidence that the cell type consistent with current definitions of an EPC phenotype may arise from an uptake of platelet MPs by mononuclear cells resulting in a gross misinterpretation of their cellular progeny. These findings demonstrate the advantage of using an unbiased proteomic approach to assess cellular phenotypes and advise caution in attributing the benefits in clinical trials using unselected bone marrow mononuclear cells (BMCs) to stem cell-mediated repair.
MicroRNAs (miRNAs) are small conserved non-coding RNA molecules that post-transcriptionally regulate gene expression by targeting the 3' untranslated region (UTR) of specific messenger RNAs (mRNAs) for degradation or translational repression. miRNA-mediated gene regulation is critical for normal cellular functions such as the cell cycle, differentiation, and apoptosis, and as much as one-third of human mRNAs may be miRNA targets. Emerging evidence has demonstrated that miRNAs play a vital role in the regulation of immunological functions and the prevention of autoimmunity. Here we review the many newly discovered roles of miRNA regulation in immune functions and in the development of autoimmunity and autoimmune disease. Specifically, we discuss the involvement of miRNA regulation in innate and adaptive immune responses, immune cell development, T regulatory cell stability and function, and differential miRNA expression in rheumatoid arthritis and systemic lupus erythematosus.
Purpose of review:
There has been a tremendous interest to understand the mechanisms that regulate gene expression in autoimmunity and their effect on disease phenotypes by exploring different epigenetic mechanisms. In this review, we will introduce lupus epigenetics through reviewing historical key findings, then we will focus on the most recent and relevant findings in this field reflecting our own and naturally biased opinion.
Recent findings:
In addition to uncovering more methylation-sensitive loci in critical genes and proposing a role for these genes in the pathogenesis of lupus, there has been a great interest in high-resolution unbiased genome-wide epigenetic studies to investigate aberrant methylation and histone code patterns, the two major epigenetic markers. In recent years, we have also witnessed an increasing interest in the role of microRNAs in the pathogenesis of lupus, and as candidate molecules with intriguing therapeutic potentials.
Summary:
Epigenetics is an exciting field that is serving as a link, as we currently understand, between genetic susceptibility and the environment in predisposing to lupus. Certainly, epigenetic aberrancies play a fundamental role in propagation of the lupus phenotype. Whether the available epigenetic-modifying agents would be useful treating human lupus is still an open question, but is unlikely in our opinion. Indeed, gene-specific epigenetic modifiers will be a challenging and an exciting area for future research.
MicroRNAs (miRNAs) are noncoding RNA molecules of 21-24 nt that regulate the expression of target genes in a post-transcriptional manner. Evidence indicates that miRNAs play essential roles in embryogenesis, cell differentiation, and pathogenesis of human diseases. This study describes a comparison between the miRNA profile of kidney biopsies from lupus nephritis (LN) patients and the controls, to develop further understanding of the pathogenesis of LN. Kidney biopsies were taken from five LN patients detected LN Class II and three normal controls. The miRNA microarray chip analysis identified 66 miRNAs differentially expressed in LN. The chip results were confirmed by QRT-PCR. This work indicates that miRNAs are potential diagnosis biomarkers and probable factors involved in the pathogenesis of LN.
Human systemic lupus erythematosus (SLE) is an autoimmune disease characterized by autoantibodies to nuclear components with subsequent immune complex formation and deposition in multiple organs. A combination of genetic and environmental factors is required for disease development, but how the environment interacts with the immune system in genetically predisposed hosts to cause lupus is unclear. Recent evidence suggests that environmental agents may alter T cell chromatin structure and gene expression through effects on DNA methylation, a repressive epigenetic mechanism promoting chromatin inactivation, to cause lupus in people with the appropriate genetic background. DNA methylation is regulated by ERK pathway signaling, and abnormalities in ERK pathway signaling may contribute to immune dysfunction in lupus through epigenetic effects on gene expression. This article reviews current evidence for epigenetic abnormalities, and in particular DNA demethylation, in the pathogenesis of idiopathic and some forms of drug-induced lupus, and how impaired ERK pathway signaling may contribute to the development of human lupus through effects on T cell DNA methylation.
The two most commonly used methods to analyze data from real-time, quantitative PCR experiments are absolute quantification and relative quantification. Absolute quantification determines the input copy number, usually by relating the PCR signal to a standard curve. Relative quantification relates the PCR signal of the target transcript in a treatment group to that of another sample such as an untreated control. The 2(-Delta Delta C(T)) method is a convenient way to analyze the relative changes in gene expression from real-time quantitative PCR experiments. The purpose of this report is to present the derivation, assumptions, and applications of the 2(-Delta Delta C(T)) method. In addition, we present the derivation and applications of two variations of the 2(-Delta Delta C(T)) method that may be useful in the analysis of real-time, quantitative PCR data.
MicroRNAs (miRNAs) are endogenous approximately 22 nt RNAs that can play important regulatory roles in animals and plants by targeting mRNAs for cleavage or translational repression. Although they escaped notice until relatively recently, miRNAs comprise one of the more abundant classes of gene regulatory molecules in multicellular organisms and likely influence the output of many protein-coding genes.
microRNAs (miRNAs) are an abundant class of newly identified endogenous non-protein-coding small RNAs. They exist in animals, plants, and viruses, and play an important role in gene silencing. Translational repression, mRNA cleavage, and mRNA decay initiated by miRNA-directed deadenylation of targeted mRNAs are three mechanisms of miRNA-guided gene regulation at the post-transcriptional levels. Many miRNAs are highly conserved in animals and plants, suggesting that they play an essential function in plants and animals. Lots of investigations indicate that miRNAs are involved in multiple biological processes, including stem cell differentiation, organ development, phase change, signaling, disease, cancer, and response to biotic and abiotic environmental stresses. This review provides a general background and current advance on the discovery, history, biogenesis, genomics, mechanisms, and functions of miRNAs.
MicroRNAs (miRNAs) are noncoding RNA molecules of 21-24 nt that regulate the expression of target genes in a post-transcriptional manner. Evidence indicates that miRNAs play essential roles in embryogenesis, cell differentiation and pathogenesis of human diseases. This study describes a comparison between the miRNA profile of the systemic lupus erythematosus (SLE) patients and the controls to develop further understanding of the pathogenesis of SLE. Peripheral blood mononuclear cells were isolated from blood samples of 23 SLE patients, 10 idiopathic thrombocytopenic purpura patients and 10 healthy controls. The miRNA microarray chip analysis identified 16 miRNAs differentially expressed in SLE. The chip results were confirmed by northern blot analysis. This work indicates that miRNAs are potential diagnosis biomarkers and probable factors involved in the pathogenesis of SLE.
MicroRNA identification in plasma and se-rum: a new tool to diagnose and monitor diseases
- Cortez Ma Calin
- Ga
Cortez MA, Calin GA. MicroRNA identification in plasma and se-rum: a new tool to diagnose and monitor diseases. Expert Opin Biol Ther 2009;9:703–11.