Project description:We carried out Massive Parallel Sequencing (MPS) to reveal miRNA expression profiles of rats with LiCl-pilocarpine induced epileptic status (SE) in adulthood (Postnatal day 60 -P60) and infancy (P12). Hippocampal tissues were collected at 3 stages of epileptogenesis: acute (24 hours), latent (7 days) and chronic (3 months after SE) from 10 animals after SE and 10 controls in each age group (120 in total). The sequencing platform identified 666 miRNA species, out of which up to 419 miRNAs were present with the coverage of more than 500 reads altogether in individual groups (onset-age epilepsy stage combinations). We discovered that age of SE induction has impact on miRNA profile throughout all stages of epileptogenesis and infantile-onset of epilepsy leads to changes in smaller number of miRNAs.
Project description:We carried out Next Generation Sequencing (NGS) to reveal miRNA expression profiles of 16 patients with mTLE+HS and 8 post mortem controls (with median coverage of 10.8 million of unprocessed reads per sample). The sequencing platform identified 1962 miRNA species, out of which 422 miRNAs were present with the coverage of more than 500 reads altogether in all samples. We discovered 9 miRNAs potentially related to mTLE+HS that have not yet been associated with epilepsy.
Project description:Using NGS approach we performed the search of multiple sclerosis-related miRNAs. We used PBMC as an informative and easily accessible biological material. To exclude bias in miRNA expression levels caused by disease modifying therapies, miRNA profiling was performed in treatment-naïve RRMS patients. Taking into account hypothetic gender specificity in disease pathogenesis we compared miRNA expression in RRMS patients and HCs separately for men and women. MiRNA profiling in men identified 32 differentially expressed miRNAs, which passed threshold for multiple corrections and may be attributed to MS-related. At the same time we did not find well-defined MS-specific miRNA expression signatures in women using NGS
Project description:Purpose: Next-generation sequencing (NGS) technology was used to map expression profile of hippocampal tissue in mouse model of Systemic Lupus Erythematosus (SLE). Methods: Total RNA was extracted from total hippocampal tissue using NucleoSpinRNA and mRNA libraries were generated using the Illumina TruSeq Sample Preparation kit. Single-end 100bp mRNA sequencing was performed on Illumina NextSeq500 platform. Quality of sequencing was assessed using FastQC software. Raw reads in fastq format were collected and aligned to the mouse genome (mm10 version) using STAR 2.6 algorithm. Gene quantification was performed using HTSeq and differential expression analysis was performed using edgeR package. Results: We defined hippocampal-specific molecular signatures of the murine lupus transcriptome. Conclusions: By the use of the mouse hippocampal-specific transcriptome and through characterization of hippocampal neurogenesis, we showed that inflammatory mediators induce neuropsychiatric changes in SLE as an early event via the disruption of hippocampal neurogenesis. These data underscore the role of brain inflammation in the pathogenesis of early disease and support the use of immunosuppressants for the management of diffuse NPSLE.
Project description:Purpose: The goals of this study are to compare next-generation sequencing-derived hippocampal transcriptome profiling from mice lacking hippocampal Acetycholine release to evaluate the role of the neurotransmitter in hippocampal gene expression.
Project description:Purpose: Next-generation sequencing technologies allow miRNA detection at an unprecedented sensitivity.Early- to mid-gestational fetal mammalian skin wounds heal rapidly and without scarring. The goals of this study are to obtain the differently expressed miRNAs between late- and mid-gestational fetal Keratinocytes for further studies. Methods: Keratinocytes were obtained from six fetal skin samples dividied into two groups: a mid-gestation group and a late-gestation group. RNA extracted from Keratinocytes was used to prepare a small RNA library for next-generation sequencing (NGS) using an Illumina Genome Analyzer IIx. To uncover potentially novel microRNA (miRNAs), the mirTools 2.0 web server was used to identify candidate novel human miRNAs from the NGS data.UCSC Genome Browser and the Vienna RNAfold web server were used to further validate the novel miRNA candidates. The repeatability of the expression levels detected by NGS was confirmed by real-time quantitative RT-PCR. Results: Using an optimized data analysis workflow, we detected a total of 99,354,224 raw reads from the six samples, 85,252,341 (85.81%) were high-quality reads (≥ 18nt). After alignment to the human genome (GRCH38), the number of genome-aligned reads was 74,678,115 (87.60% of the high-quality reads). The number of sequence reads that correspond to known miRNAs was 61,587,749 (82.47% of the genome-matching reads), as was determined by perfect sequence matching to the database of known miRNAs (miRBase release 18). After removing the matched non-coding RNAs (Release 10), 8,755,258 reads remained for identification of novel miRNA candidates by mirTools 2.0. The results revealed the existence of 202 novel miRNA candidates and 29 known miRNAs that were not listed in miRBase release 18. Of the 202 potential novel miRNAs, 106 candidates were detected by at least 10 counts, by NGS, indicating that they have a higher probability of being novel human miRNAs. Using Student's t-test, 173 known miRNAs and 23 novel miRNA candidates were found to be statistically significant (P values < 0.05). The expression of 22 novel miRNA candidates and 88 known miRNAs was changed by more than 2.0-fold (known: 15 up-regulated and 73 down-regulated; novel: two up-regulated and 20 down-regulated). Conclusions: Our study represents the first detailed analysis of human miRNAs in fetal Keratinocytes. Using mirTools 2.0 web server, we have revealed the existence of 202 novel miRNA candidates. Of the 202 potential novel miRNAs, 106 candidates were detected by at least 10 counts, by NGS, indicating that they have a higher probability of being novel human miRNAs. Taken together, our results provide compelling evidence that dynamic expression of miRNAs in fetal Keratinocytes at different gestational ages. MiRNAs presenting altered expression at different gestational ages in fetal Keratinocytes may contribute to scarless wound healing in early- to mid-gestational fetal Keratinocytes, and thus may be new targets for potential scar prevention and reduction therapies.
Project description:Aim: Hepatic fibrosis is a major worldwide medical problem and can develop into liver cirrhosis and hepatocellular carcinoma(HCC). Until now, there are no effective drugs for liver ?brosis because the molecular mechanism of progression of liver fibrosis is not fully understood. MicroRNAs (miRNAs) are an important class of small non-coding functional RNAs that play a key role in many biological processes. The purpose of this study was to clarify how the aberrant expression of miRNAs participates in development of the liver fibrosis in rat liver fibrosis model. Methods: Fibrotic and paired normal liver tissues were collected and assesssed by deep sequencing technology. MiRNA pro?ling results were validated by quantitative real-time polymerase chain reaction (qRT-PCR) and bioinformatics was used to predict miRNA targets. Results: Nine deregulated miRNAs were induced in porcine serum (PS)-induced hepatic fibrosis versus normal liver. Further analysis revealed several signaling pathways (e.g., gap junction and neuroactive ligand-receptor interaction) may be associated with hepatic fibrogenesis. Conclusion: Several miRNAs are dysregulated in PS-induced hepatic fibrosis and seem to be closely associated with hepatic fibrogenesis. These results provide an experimental basis for understanding the mechanism of hepatic fibrosis. We sequenced two samples, including case and control. Each sample has two replicates.
Project description:MicroRNAs (miRNAs) are short noncoding RNAs that shape the gene expression landscape, including during the pathogenesis of temporal lobe epilepsy (TLE). In order to provide a full catalog of the miRNA changes that happen during experimental TLE, we sequenced Argonaute 2-loaded miRNAs in CA1, CA3 and Dentate Gyrus hippocampal subfields from the rat perforant pathway stimulation (PPS) model at regular intervals between the time of the initial precipitating insult to the establishment of spontaneous recurrent seizures.
Project description:miRNAs has an important role in the diagnosis and treatment of amyotrophic lateral sclerosis. we aimed to profile dysregulation of miRNAs in ALS blood and neuromuscular junction as well as healthy blood control by Next Generation Sequencing (NGS). The expression of three up-regulated miRNAs, as miR-338-3p, miR-223-3p and miR-326, in the ALS samples compared to healthy controls, has been validated by qRT-PCR in a cohort of 45 samples collected previously. Bioinformatics tools were used to perform ALS microRNAs target analysis and to predict novel miRNAs secondary structure. The analysis of the NGS data identified 696 and 44 novel miRNAs which were differentially expressed in ALS tissues.