Project description:Purpose: To detect serum exosomal ncRNA profiles of proliferative diabetic retinopathy (PDR) by High-throughput sequencing Methods: serum exosomal non-coding RNA (ncRNA) profiles profiles of PDR and MH were generated by deep sequencing, only in once, using IlluminaHiSeq 3000. After analyzing the base composition and quality of the data, according to the analysis results of the original data, the data were filtered to remove the joint sequence and the contaminated part, and to remove low-quality base sequences.If it is paired-ended sequencing data, the filtered data should be further screened to retain the paired sequences and obtain clean data. Conclusions: Our study represents the first detailed analysis of serum exosomal transcriptomes from PDR patients by RNA-seq technology. The optimized data analysis workflows reported here should provide a framework for comparative investigations of expression profiles.
2021-06-24 | GSE178721 | GEO
Project description:serum exosomal small RNA sequencing
Project description:In our study, RNA-seq on serum-derived exosomes from NSCLC patients with clinically significant differences in gefitinib sensitivity was performed to explore the differences in serum exosomal RNA expression profiles between the two groups. The exosomes were isolated and the saucer shaped round vesicles of 40–200 nm in diameter were observed with TEM and nanoparticle tracing system. Then, RNA-seq was performed on serum-derived exosomal RNAs and bioinformatics analysis was performed to analyze the differentially expressed RNAs between GS and GR group. The results demonstrated that there were 289 mRNAs with significant differences between the two groups, 252 of which were significantly up-regulated and 37 significantly down regulated (GR vs GS) .
Project description:The aim of this study was to identify and evaluate exosomal miRNAs in serum as early diagnostic markers for gastric cancer (GC). Methods: Using next-generation sequencing (NGS) and bioinformatics, we identified candidate serum exosomal miRNA markers for early detection of GC in patients. The candidates were further validated by qRT-PCR in 50 newly recruited early-stage GC patients and matched healthy individuals. Results: NGS revealed that the average mappable reads in the RNA libraries were about 6.5 million per patient. A total of 66 up and 13 down-regulated exosomal miRNAs were found in the screened cohort after removal of log2 transformed read counts <5 and p >0.05. In the validation cohort, by comparing candidate exosomal miRNAs levels in early-stage GC patients and healthy individuals, higher levels of miR-92b-3p, let-7g-5p, miR-146b-5p and miR-9-5p were found to be significantly associated with GC. Diagnostic power of the combined panels of the exosomal miRNAs or the combination of exosomal miRNAs and CEA outperformed that of single exosomal miRNA marker for establishing a diagnosis of early-stage GC. In addition, serum levels of exosomal miR-92b-3p were significantly associated with low adhesion, let-7g-5p and miR-146b-5p were significantly correlated with nerve infiltration, and miR146b-5p was statistically correlated with tumor invasion depth in early-stage GC. Conclusions: Serum exosomal miR-92b-3p, -146b-5p, -9-5p, and let-7g-5p may serve as potential noninvasive biomarkers for early diagnosis of GC. Further validation of these candidate exosomal miRNAs in larger experimental cohorts are required in order to confirm the diagnostic values.
Project description:Exosomes are small membrane vesicles of endocytic origin secreted by most cells, and contain a wealthy cargo of protein and RNA species that can modulate recipient cells’ behaviors and may be used as biomarkers for diagnosis of human diseases. They have been found in blood and are valuable sources for biomarkers due to selective cargo loading and resemblance to their parental cells. The goal of this study is to identify circRNA, lncRNA and mRNA profiles in human blood by high-throughput RNA sequencing (RNA-seq). 1-4 ml plasma or serum were used to extract exosomal RNAs by exoRNeasy Serum/Plasma Maxi kit (Qiagen). The exosomal RNAs were further treated with DNAse I and subjected to ribosome minus low-input RNAseq library preparation. The libraries were sequenced by Illumina Hiseq platform.
Project description:Occupational dermatitis medicamentosa-like of TCE (ODMLT) is a complex immune process which can cause serious damage of liver function, and becoming a serious occupational health problem in China. However, the pathogenesis of ODMLT remained undefined. The aim of the present study was to identify the expression profiles of circulating exosomal miRNAs in ODMLT patients and its role in the regulation of liver injury. The exosomes were isolated from the serum of patients and healthy individuals, and characterized by electron microscopy, nanoparticle tracking analysis, and Western blot and total RNA was extracted. Expression of exosomal miRNAs was evaluated with Agilent Human miRNA microarrays and the expression of selective serum exosomal miRNA were verified by qRT-PCR.
Project description:We conducted this study to determine whether exosome regulation underlies the antimigraine mechanisms of acupuncture. By comparing serum samples from patients with migraine and healthy controls using high-throughput small RNA sequencing technology , we identified 705 exosomal microRNAs that are differentially expressed in patients with migraine, and this set of 705 microRNAs included five that are particularly well characterised (hsa-miR-369-5p, hsa-miR-1268b, hsa-miR-145-5p, hsa-miR-222-5p, and hsa-miR-4488). By comparing serum samples collected from patients with migraine before and after acupuncture treatment, we showed that acupuncture normalised the expression levels of those five well-characterised exosomal microRNAs.
Project description:Bovine mastitis causes changes in the serum exosomal miRNAs expression. Serum samples from healthy dairy cows (n = 7) were compared to those of cows with subclinical (n = 7 ) using small RAN sequencing. Three hundred fifty-five miRNAs (341 known and 14 novel ones) were identified. There were 42 miRNAs up-regulated in serum-derived EVs from cows with subclinical mastitis, including bta-miR-1246, bta-miR-2431-3p, bta-miR-126-3p, bta-miR-29a, etc. The MAPK signaling pathway was the most affected pathway by clinical mastitis. Thus, miRNA alterations in mastitis serum-derived EVs support the potential regulator role of specific miRNAs as exosomal cargo in clinical mastitis physiology.
Project description:Preoperative prediction of lymph node (LN) metastasis is accepted as an important independent risk factor for treatment decision-making for esophageal squamous cell carcinoma (ESCC) patients. This study aimed to develop a non-invasive biomarker to identify LN metastasis preoperatively in ESCC patients with serum exosomal RNA-seq.