Role Of miRNAs In Pathogenesis Research Articles

Role of miRNA in pathogenesis, diagnosis, and prognosis in hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is one of the most common cancers and is responsible for the second cancer-related death globally. Many treatment regimens have been developed to cure the disease; however, life expectancy is still low. Therefore, there is an urgent need to explore new selective, specific, and robust diagnosis markers for efficient early recognition of the ailment. Along with the diagnosis, the treatment's effectiveness can be determined by prognostic markers, and miRNAs are excellent tools for the diagnosis and prognosis of HCC. In addition, the altered expression profile of a few miRNAs promotes HCC cell migration and invasion, and selective up- or downregulation of these responsible genes may help mitigate the disorder. On one hand, few of the miRNAs have been found to enhance angiogenesis, a crucial step of tumor growth; on the other hand, upregulation of specific miRNAs is reported to suppress angiogenesis and resulting tumor growth of HCC cells. Exosomal miRNAs have significant implications in promoting angiogenesis, increased endothelial cell permeability, tube formation, and metastasis to hepatic and pulmonary tissues. miRNA also attributes to drug resistance toward chemotherapy and the prevention of autophagy also. Identifying novel miRNA and determining their differential expression in HCC tissue may serve as a potential tool for diagnosis, prognosis, and therapy to enhance the life expectancy and quality of life of HCC patients. In the present review, we have summarized the recent advances in HCC-related research.

Can Echinococcus granulosus-Derived MicroRNAs be Biomarkers for Diagnosis and Follow-up of Cystic Echinococcosis Patients?

Cystic echinococcosis (CE) is a neglected tropical disease caused by the larval stages of Echinococcus granulosus (E. granulosus). MicroRNAs (miRNAs) are small noncoding RNAs acting as mediators in host-parasite interaction. Recently, numerous studies have been conducted on miRNAs in infectious diseases; however, little data are available about the role of miRNAs in pathogenesis and early diagnosis of CE. The current study evaluated the expression of four E. granulosus-derived miRNAs, including egr-miR-125,5p, egr-let-7,5p, egr-miR-2, and egr-miR-71 in fibrotic and healthy liver tissues of 31 CE patients with active and inactive hydatid cysts by qRT-PCR. Of the 31 patients, 48.4% had active cysts (CE1 and CE2), while the remainder had transitional (16.1%) and inactive (35.5%) CE types cysts. The qRT-PCR analysis revealed a significant increase of 11.2, 9.91, 6.2, and 13.1-fold in the fibrotic tissue group for egr-miR-125,5p, egr-let-7,5p, egr-miR-2, and egr-miR-71, respectively. Among these miRNAs, egr-miR-125-5p exhibited the highest area under the curve (AUC) value of 0.8050 for predicting liver fibrosis. Our findings provide new data about the role of E. granulosus-derived miRNAs in pathogenesis of CE. The high AUC of egr-miR125,5p reflecting the possibility of using egr-miR125,5p as biomarker in CE diagnosis. Further studies on serum of CE patients are needed to confirm the potential role of circulating egr-miR-2a-3p and egr-miR-125-5p in the early diagnosis of CE.

Ultrasensitive detection of serum miRNA biomarkers related to papillary thyroid cancer using ligation-initiated phosphorothioated primer-based loop-mediated isothermal amplification

Accurately distinguishing of papillary thyroid cancer (PTC) from benign nodules, at the same time, avoiding overdiagnosis is of great significance. In consideration of the role of miRNA in pathogenesis and progression of tumors, miRNA might also be useful as diagnostic marker for cancer. In this study, a ligation-initiated phosphorothioated primer-based loop-mediated isothermal amplification (PS-LAMP) strategy was established to quantify miRNAs in serum sample, thereby achieving noninvasive diagnosis of PTC. Two linker probes designed from dumbbell-shaped amplicons of LAMP are ligated to obtain an active amplicon only when they are perfectly hybridized with target miRNA, exhibiting high specificity. The introduction of PS modification into inner primers (BIP/FIP) facilitated extra efficient hairpin formation and extension for LAMP, which significantly improved sensitivity. Then, the proposed method was successfully applied in the early diagnosis of PTC, in which circulating serum level of miR-146b-5p and miR-199b-5p could draw a clear distinction between patients and controls. This ligation-initiated PS-LAMP assay opens new avenues for highly sensitive quantification of miRNA biomarkers and thus exhibits a considerable potential utility in clinical diagnosis and prognosis.

MiRNA Expression Profile in Ileocecal Adenocarcinoma Cells Infected with Cryptosporidium

Cryptosporidium spp. is an opportunistic protozoan transmitted by fecal-oral route via oocysts. The agent may cause severe infection especially in individuals with suppressed immune system, due to its intracellular location and ability to cause auto-infection. MicroRNAs (miRNAs) are non-translated endogenous RNA molecules with an average of 22 nucleotides in length that regulate the expression of genes involved in important biological functions such as proliferation, differentiation, apoptosis and immune response. Recent studies have focused on the role of miRNAs in pathogenesis of infectious diseases and their potential to be used as biomarkers. The aim of this study was to determine the miRNA profile of human ileocecal adenocarcinoma (HCT-8) cells at 24 hours of infection with Cryptosporidium spp. In the study, the HCT-8 cell line was infected with Cryptosporidium spp. that were isolated from infected human stool samples and RNA was isolated from the cells 24 hours after infection. After this process, cDNA synthesis was performed and the expression of 95 human miRNA profiles were investigated by polymerase chain reaction (PCR) method. Fold changes of expression were determined by comparison with Cryptosporidium spp. uninfected cell lines. Sequence information of miRNAs and their target genes were performed via TargetScanHuman7.1 and miRDB websites, while gene ontology (GO) pathways of target genes were analyzed with the mirPath v.3 program. It was detected that the expression of 10 miRNAs were upregulated and 11 of them were downregulated compared with the control group. It was observed that, this 21 differentially expressed miRNAs were mainly associated with apoptosis, mitotic cell cycle, and immune response. Hsa-miR-612, hsa-miR-6763-5p, hsa-miR-188-5p, hsa-miR-664b-3p, hsa-miR-210-3p, hsa-let-7e-5p hsa-let-7b-3p, hsa-miR-4787-3p, hsa-miR-548ab, hsa-miR-3714 and hsamiR-4803 were found to be associated with apoptosis; and hsa-miR-612, hsa-miR-664b-3p, hsa-miR210-3p, hsa-let-7e-5p, hsa-let-7b-3p, hsa-miR-548ab, and hsa-miR4803 were found to be associated with mitotic cell cycle. The balance of proliferation and apoptosis is very significant in the development of infection and cancer. It is thought that determination of the effect of miRNAs on proliferation-apoptosis balance could provide information related to the etiopathogenesis and prognosis of infections, and on the role of microorganisms in carcinogenesis. In this study, 12 differentially expressed miRNAs were found to be associated with immune response. This may emphasize the role of miRNAs in the prevention and treatment of infections. It was concluded that, miRNAs could be used in the diagnosis, treatment and prevention of infections with the determination of miRNA's role in the infection mechanism as a result of the increasing number of studies.

Understanding Pharmaco-Epigenomic Response of Antipsychotic Drugs Using Genome-Wide MicroRNA Expression Profile in Liver Cell Line.

Interindividual variability in drug response is a major concern among patients undergoing antipsychotic drug treatment. Apart from genetic and physiological factors, this variability in drug response could also be attributed to epigenetic mechanisms. The microRNAs (miRNAs) are key epigenetic markers that play an important role in pathogenesis and drug response. Several studies have shown that miRNAs are implicated in regulating the expression of various genes involved in drug metabolism and transport. In a conventional clinical setup, it is extremely difficult to distinguish the role of miRNA in pathogenesis and drug response as it is difficult to obtain drug naïve patients. To resolve this issue, we aimed to identify the role of antipsychotic drug treatment in inducing miRNA expression under an in vitro condition using a hepatic cell line. A liver cell line was treated with a maximum tolerable drug dosage model for haloperidol, clozapine in monotherapy, and their combination in polytherapy. Genome-wide miRNA profiling was performed using 60,000 miRNA probes in the microarray format in different treatment groups. Several miRNAs were observed to be differentially expressed impacting the pharmacokinetic, pharmacodynamics, and epigenomics properties of antipsychotic drug treatment. Interestingly, some of these miRNA expression patterns were similar to reported miRNA observations on schizophrenia pathogenesis. This study unravels the potential role of miRNAs in the mechanism of action of the antipsychotic drug and could also reflect in drug-induced side effects. This study also signifies the importance of pharmacoepigenomics approach while evaluating the role of miRNAs in pathogenesis.

Altered microRNAs in C3H10T1/2 cells induced by p.E95K mutant IHH signaling

BackgroundIndian Hedgehog (IHH), an important cell signaling protein, plays a key regulatory role in development of cartilage and chondrogenesis. Earlier studies have shown that heterozygous missense mutations in IHH gene may cause brachydactyly type A1 (BDA1), an autosomal dominant inheritance disease characterized by apparent shortness or absence of the middle phalanges of all digits. MicroRNAs (miRNAs) have been found to be significant post-transcriptional regulators of gene expression and significantly influence the process of bone-development. Therefore, it is possible that miRNAs are involved in the mechanism underlying the development of BDA1. However, the relationship between miRNAs and the pathogenesis of BDA1 remains unclear.MethodsIn this study, we used microarray-based miRNA profiling to investigate the role of miRNAs in BDA1 by characterization of differentially expressed miRNAs in C3H10T1/2 cell line induced by wild type (WT) and p.E95K mutant (MT) IHH signaling.ResultsOur results identified 6 differentially expressed miRNAs between WT and control (CT) group and 5 differentially expressed miRNAs between MT and CT groups. In particular, miR-135a-1-3p was found to be a significantly differentially expressed miRNA between WT and CT group. Results of dual-luciferase reporter gene experiment successfully discovered Hoxd10 was one of the target gene of miR-135a-1-3p. Additionally, our pathway analysis revealed that the targets of these miRNAs of interest were highly involved with Runx1/2, Notch and collagen-related pathways.ConclusionsTaken together, our findings provided important clue for future study of the process of miRNA-regulation in IHH signaling and novel insights into the regulatory role of miRNA in pathogenesis of BDA1.

Application and progress of miRNA in chronic rhinosinusitis

Chronic rhinosinusitis(CRS) is one of upper respiratory inflammatory diseases, with a high incidence. It affects the patient's work efficiency and quality of life seriously. However, the pathogenesis of disease is not definite. In recent years, some reports have proved miRNA play an important role in the development and occurrence of CRS. This paper reviewed the role of miRNA in pathogenesis,diagnosis and treatment of chronic rhinosinusitis.

Identification of a Specific miRNA Profile in HIV-Exposed Seronegative Individuals.

MicroRNAs (miRNAs) are small noncoding RNAs involved in the posttranscriptional regulation of gene expression that play important roles in viral infections. Alterations of specific miRNAs are described in HIV infection, suggesting a role for miRNAs in pathogenesis of this disease. We verified whether a particular miRNA signature could be identified in natural resistance to HIV-1. Expression level of 84 miRNAs was analyzed by RT-qPCR in plasma and unstimulated peripheral blood mononuclear cell (PBMC) of 30 seronegative individuals repeatedly exposed to HIV-1 (HESN), 30 HIV seropositive subjects (HIV+), and 30 healthy controls (HC). Results were confirmed by individual RT-qPCR in in vitro HIV-1-infected PBMC and in their cell culture medium. Dicer and Drosha expression was analyzed in basal PBMC. Whereas Dicer and Drosha expression was comparable in HESN, HIV+ and HC, several miRNAs were upregulated both in HESN and HIV+ compared with HC. Furthermore, miRNA-29a and miR-223 were upregulated in both unstimulated PBMC and plasma of HESN alone; their expression was reduced upon in vitro HIV-1 infection of HESN PBMC indicating that, upon infection, they are secreted in the extracellular milieu. These results were confirmed by individual qPCR. Our studies demonstrate that HIV-1 exposure modifies miRNAs expression even in the absence of productive infection. Because those miRNAs that are specifically increased only in HESN have been known to reduce HIV-1 replication, their modulation could represent an important mechanism in resistance to HIV-1 infection.

Genome technologies in pulmonology: a role of microRNA in asthma and COPD development

MicroRNAs (miRNAs) are small noncoding RNA molecules that affect gene expression and thus take part in the epigenetic regulation of almost all physiological and pathological processes. About 1,800 human miRNAs have been discovered to date; however, biological functions and protein targets for the majority remain to be unknown. Within the respiratory system, miRNAs contribute to the lung growth and lifelong maintenance of pulmonary homeostasis. Recently, the leading role of miRNAs in pathogenesis of various pulmonary diseases has been found, including asthma, chronic obstructive pulmonary disease (COPD) and lung cancer. Due to a significant progress in studying interactions between genes and their products and environmental factors, a great role of epigenetic variability, which is gene expression change not related to DNA damage, but could be inherited consistently, became apparent. There are three levels of epigenetic regulation corresponding to three main mechanisms: genomic (DNA methylation), proteomic (histone modification) and transcriptomic (regulation through RNA, primarily miRNA). Extending our knowledge on a role of miRNAs for the respiratory system could open new therapeutic targets and diagnostic markers for respiratory diseases, particularly asthma and COPD.

Stress response factors as hub-regulators of microRNA biogenesis: implication to the diseased heart.

MicroRNAs (miRNAs) are important regulators of heart function and then an intriguing therapeutic target for plenty of diseases. The problem raised is that many data in this area are contradictory, thus limiting the use of miRNA-based therapy. The goal of this review is to describe the hub-mechanisms regulating the biogenesis and function of miRNAs, which could help in clarifying some contradictions in the miRNA world. With this scope, we analyse an array of factors, including several known agents of stress response, mediators of epigenetic changes, regulators of alternative splicing, RNA editing, protein synthesis and folding and proteolytic systems. All these factors are important in cardiovascular function and most of them regulate miRNA biogenesis, but their influence on miRNAs was shown for non-cardiac cells or some specific cardiac pathologies. Finally, we consider that studying the stress response factors, which are upstream regulators of miRNA biogenesis, in the diseased heart could help in (1) explaining some contradictions concerning miRNAs in heart pathology, (2) making the role of miRNAs in pathogenesis of cardiovascular disease more clear, and therefore, (3) getting powerful targets for its molecular therapy.

Identification of microRNAs from Atlantic salmon macrophages upon Aeromonas salmonicida infection

Computational approach was used in to identify potent macrophage specific miRNAs involved in basic biological process of Salmo salar. Analysis of 1119 ESTs from macrophages of Atlantic salmon (Salmo salar) infected with Aeromonas salmonicida revealed expression of 3 miRNAs. Phylogenetic analysis of both the pre-miRNA sequence revealed its evolutionarily conserved nature among various species. Identified targets of the predicted miRNAs revealed the role of miRNA in pathogenesis, stress response and allosteric exchange of histones. Further detailed studies of these miRNAs will help in revealing its function in different biological process necessary for the action of macrophages upon pathogen infection.

The Role of MicroRNAs in the Pathophysiology of Neuroblastoma and Th eir Possible Use in Dia gnosis, Prognosis and Therapy

Neuroblastoma (NBL) is a typical childhood tumor developing from the precursor cells of the sympathetic nervous tissue and accounting for approximately 7% of total malignancies in pediatrics and 15% of deaths associated with this malignancy. MicroRNAs (miRNAs) are small single-stranded RNA molecules that are involved in posttranscriptional regulation of gene expression, whereas the pathophysiology of neuroblastoma tumor growth involves both upregulation of the protooncogenic miRNAs as well as downregulation of the tumor-suppresor ones. Comparison of the expression profiles of miRNAs in specific subtypes of neuroblastoma seems to be a useful tool adding to the classification of the diseases, and the assessment of the levels of specific miRNAs may be useful for estimation of the individual treatment response as well as prognosis of the patient. This paper provides the basic review of the studies focused on the role of miRNAs in pathogenesis of neuroblastoma and provides a survey of current/ possible use of these miRNAs in diagnostics, therapy or prognosis estimation in the neuroblastoma patients.

Role of miRNA in pathogenesis of inflammatory bowel disease

Role of miRNA in pathogenesis of inflammatory bowel disease

MicroRNAs: Possible role in pathogenesis of Parkinson’s disease

Parkinson's disease is one of the most common human neurodegenerative disorders caused by the loss of dopaminergic neurons from the substantia nigra pars compacta of human brain. However, causes and mechanisms of the progression of the disease are not yet fully clarified. To date, investigation of the role of miRNAs in norm and pathology is one of the most intriguing and actively developing areas in molecular biology. MiRNAs regulate expression of a variety of genes and can be implicated in pathogenesis of various diseases. Possible role of miRNAs in pathogenesis of Parkinson's disease is discussed in this review.

Overexpression of microRNA-29b induces apoptosis of multiple myeloma cells through down regulating Mcl-1

MicroRNAs (miRNAs) are small, noncoding ribonucleic acids (ncRNAs), which regulate gene expression by targeting mRNAs for translational repression and degradation. Several lines of evidences have indicated that miRNAs act as tumor suppressors and oncogenes. However, the role of miRNAs in pathogenesis of multiple myeloma (MM) remains unclear. In this study, we examined the profile of miRNA expression of primary MM cells, using miRNA microarray and quantitative real-time polymerase chain reaction (qPCR) techniques. These results showed that in the bone marrow specimens analyzed, miRNA-29b was significantly downregulated. Similar results were also observed in human myeloma cell lines (HMCLs). Adenovirus-mediated overexpression of miR-29b induced apoptosis and elevated caspase-3 activation in HMCLs. Using a bioinformatics approach, we found a perfect complementarity between miRNA-29b and the 3′UTR of myeloid-cell-leukemia 1(Mcl-1). It is further confirmed that miRNA-29b downregulated the level of Mcl-1 without effect on the mRNA level using both qRT-PCR assays and Western blot analyses. Moreover, we observed that enforced miR-29b expression by using a retarget miRNA-29b expression vector (Ad5F11p-miR-29b) could induce apoptosis and elevate caspase-3 activation in HMCLs. Our results also indicated that miRNA-29b-induced apoptosis acted antagonistically with IL-6 in HMCLs. These findings suggest that miRNA-29b may play an important role in MM as a tumor suppressor.

MiRNA-Biomarker aus Blut – Auf dem Weg zur minimalinvasiven Diagnostik

Biomarker gewinnen in der modernen Medizin zunehmend an Bedeutung. Vor allem Biomarkeranalysen auf Genom- und Transkriptionsebene erlauben dank innovativer Technologien eine frühzeitige Diagnostik, Prognose sowie Risikoeinschätzung und können somit helfen, Patienten rechtzeitig und zuverlässig zu klassifizieren. MicroRNAs (miRNAs), eine Klasse kleiner, nicht proteincodierender RNAs, und ihrer deregulierten Expression wird eine Rolle bei der Entstehung und dem Verlauf von Krankheiten wie Krebs zugesprochen. Aufgrund des Einflusses, den sie offenbar auf die Pathogenese verschiedenster Erkrankungen haben, sowie der Möglichkeit, Veränderungen der Expression mit Krankheitsstadien zu assoziieren, eignen sich miRNAs als wertvolle diagnostische Biomarker. In aktuellen Studien mit Microarray-Technologie wurde gezeigt, dass miRNA-Biomarkeranalysen auf Grundlage minimalinvasiv zugänglichen Probenmaterials wie Blut oder anderer Körperflüssigkeiten möglich sind und einen vielversprechenden Ansatz für diagnostische Biomarkertests darstellen. miRNA-Expressionsanalysen mittels etablierter Biochips und einer automatisierten Geniom-RT-Analyzer-Plattform dienen hierbei der Entwicklung von Biomarkersignaturen für eine akkurate Patientenklassifikation. Die biomarkerbasierte Patientenuntersuchung und Risikoerfassung kann entscheidend zur frühzeitigen Detektion und spezifischen Diagnose von Krebs, wie beispielsweise dem Ovarialkarzinom, oder anderen Erkrankungen beitragen. Damit kann krankheitsübergreifend ein neuer Standard in der Diagnostik, bei Verlaufbeobachtungen und Prognose etabliert werden.

MiRNA-Biomarker aus Blut – Auf dem Weg zur minimalinvasiven Diagnostik

Biomarker gewinnen in der modernen Medizin zunehmend an Bedeutung. Vor allem Biomarkeranalysen auf Genom- und Transkriptionsebene erlauben dank innovativer Technologien eine frühzeitige Diagnostik, Prognose sowie Risikoeinschätzung und können somit helfen, Patienten rechtzeitig und zuverlässig zu klassifizieren. MicroRNAs (miRNAs), eine Klasse kleiner, nicht proteincodierender RNAs, und ihrer deregulierten Expression wird eine Rolle bei der Entstehung und dem Verlauf von Krankheiten wie Krebs zugesprochen. Aufgrund des Einflusses, den sie offenbar auf die Pathogenese verschiedenster Erkrankungen haben, sowie der Möglichkeit, Veränderungen der Expression mit Krankheitsstadien zu assoziieren, eignen sich miRNAs als wertvolle diagnostische Biomarker. In aktuellen Studien mit Microarray-Technologie wurde gezeigt, dass miRNA-Biomarkeranalysen auf Grundlage minimalinvasiv zugänglichen Probenmaterials wie Blut oder anderer Körperflüssigkeiten möglich sind und einen vielversprechenden Ansatz für diagnostische Biomarkertests darstellen. miRNA-Expressionsanalysen mittels etablierter Biochips und einer automatisierten Geniom-RT-Analyzer-Plattform dienen hierbei der Entwicklung von Biomarkersignaturen für eine akkurate Patientenklassifikation. Die biomarkerbasierte Patientenuntersuchung und Risikoerfassung kann entscheidend zur frühzeitigen Detektion und spezifischen Diagnose von Krebs, wie beispielsweise dem Ovarialkarzinom, oder anderen Erkrankungen beitragen. Damit kann krankheitsübergreifend ein neuer Standard in der Diagnostik, bei Verlaufbeobachtungen und Prognose etabliert werden.