Project description:Background/Aim: We investigated alterations in the expression of serum exosomal miRNAs with the progression of liver fibrosis and evaluated their clinical applicability as biomarkers. Methods: This study prospectively enrolled 71 patients who underwent liver biopsy at an academic hospital in Korea. Exosomes were extracted from serum samples, followed by next-generation sequencing (NGS) of miRNAs and targeted real-time quantitative polymerase chain reaction. A model was derived to discriminate advanced fibrosis based on miRNA levels and the performance of this model was evaluated. Validation of the effect of miRNA on liver fibrosis in vitro was followed. Methods: This study prospectively enrolled 71 patients who underwent liver biopsy at an academic hospital in Korea. Exosomes were extracted from serum samples, followed by next-generation sequencing (NGS) of miRNAs and targeted real-time quantitative polymerase chain reaction. A model was derived to discriminate advanced fibrosis based on miRNA levels and the performance of this model was evaluated. Validation of the effect of miRNA on liver fibrosis in vitro was followed. Results: NGS data revealed that exosomal miR-660-5p, miR-125a-5p, and miR-122 expression were changed significantly with the progression of liver fibrosis, of which miR-122 exhibited high read counts enough to be used as a biomarker. The level of exosomal miR-122 decreased as the pathologic fibrosis grade progressed and patients with biopsy-proven advanced fibrosis had significantly lower levels of exosomal miR-122 (P < 0.001) than those without advanced fibrosis. Exosomal miR-122 exhibited a fair performance in discriminating advanced fibrosis especially in combination with fibrosis-4 score and transient elastography. In a subgroup of patients with a non-viral etiology of liver disease, the performance of exosomal miR-122 as a biomarker was greatly improved. Inhibition of miR-122 expression increased the proliferation of the human hepatic stellate cell line, LX-2, and upregulated the expression of various fibrosis related proteins. Conclusion: Exosomal miR-122 may serve as a useful non-invasive biomarker for liver fibrosis, especially in patients with non-viral etiologies of chronic liver disease.
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:Purpose: Cystic fibrosis (CF)-associated liver disease (CFLD) is a hepatobiliary complication of CF. Current diagnostic modalities rely on non-specific assessments, while liver biopsy is the gold standard to assess severity of disease. Using miRNA-sequencing we investigated the use of serum miRNA to diagnose and monitor CFLD progression in children with CF. Methods: This was a cross-sectional cohort study of the circulatory miRNA signature of 90 children grouped by clinical, biochemical and imaging assessments as follows: healthy controls (n=30), CF patients with no evidence of liver disease (CFnoLD, n=30) and CFLD (n=30). Serum miRNAs were analysed by miRNA-sequencing. Following total circulatory RNA extraction, RNA samples were pooled together to create three libraries (i.e., 10 patients/library) per study cohort. Libraries were prepared using the Ion Total RNA-Seq Kit v2 protocol (Thermo Fisher, Revision E) and sequenced on the Ion Proton System, using the Ion PI Chip v3 following the Ion PI Hi-Q Sequencing 200 Kit protocol (Thermo Fisher, Revision C). Selected differentially expressed serum miRNA candidates were further validated in 124 individual children serum samples by qRT-PCR and statistical analysis performed to evaluate utility to predict CFLD and hepatic fibrosis severity validated by liver biopsy. Results: The nine libraries sequenced by the Ion Proton System produced 64,658,545 total reads. During processing of the sequencing data, we noticed significant low counts mapped from the CFnoLD library 3. Following principal component analysis this library was confirmed to be an outlier and excluded from the analysis. For the remaining libraries, initial threshold filtering was performed by discarding miRNAs with 10 or less counts and normalized for compositional bias in sequenced libraries and library size. After threshold filtering, 659 miRNAs were identified with a minimum average count of 10.
Project description:Purpose: Exosome-derived microRNAs (miRNAs) are potential diagnostic biomarkers. However, little is known about their effectiveness as diagnostic biomarkers of fulminant myocarditis (FM). This study aimed to explore miRNA levels in serum exosomes of patients with FM as potential biomarkers for FM diagnosis. Methods: 10 samples were screened with a exosomal small RNA sequencing platform (RiboBio). A Mann-Whitney test was performed to discover differentially expressed miRNAs in the two pairwise comparisons: FM versus HC. Results: From the differentially expressed miRNAs, fourteen candidate miRNAs discovered via small RNA sequencing with P<0.05 and fold expression change >2 were selected for further testing Conclusions: These data suggested that the miRNA panel in serum-derived exosomes provided excellent diagnostic capability for FM.
Project description:Diffuse large B-cell lymphoma (DLBCL) is the most common subtype of non-Hodgkin Lymphoma with complex clinical symptoms. Currently, the diagnosis of DLBCL depends largely on pathologic analysis of biopsy tissue, which is an invasive procedure and often poses some risk to patients. Exosomal miRNAs have emerged as promising noninvasive biomarkers for detecting tumor or monitoring disease progress. We aim to investigate the potential of circulating exosomal miRNAs to assist with diagnosis of DLBCL.In the present research, we used the miRCURY LNA™ microRNA Array to investgate the profiles of differentially expressed miRNAs in the exosomes of peripheral blood serum. As a result, circulating exosomal miRNA profiling yielded a total of 332 differentially expressed miRNAs (157 with up-expression and 175 with down-expression) between the DLBCL patients and healthy controls.
Project description:Objective: The expression pattern of exosomal miRNAs derived from parathyroid tumor is still unknown. In the present work, we aimed to examine the differences on microRNA (miRNA) expression, present in serum exosomes, by comparing parathyroid carcinoma (PC) and parathyroid adenoma (PA). Methods: MiRNA expression profile of serum exosomes, derived from 4 PC patients and 4 PA patients, were analyzed by next-generation sequencing technology. The differential expressions of target miRNAs were further verified in both serum exosomes and tissues of PC/PA patients by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Lastly, receiver operating characteristic (ROC) curves were plotted to investigate the efficiency of target exosomal miRNAs in distinguishing PC patients from PA controls. Results: Multiple differentially expressed miRNAs of serum exosomes were screened out by sequencing. Based on this screening, hsa-miR-146b-5p (p=0.0846), hsa-miR-27a-5p (p=0.0412), hsa-miR-93-5p (p=0.73), hsa-miR-381-3p (p=0.1239) and hsa-miR-134-5p (p=0.0694) were up-regulated in the serum exosomes of PC patients. These results were validated by qPCR, where the trend on differential miRNA expression was consistent with the sequencing results. Specifically, the expression of exosomal hsa-miR-27a-5p was able to clearly distinguish PC patients from PA controls, and related analysis indicated that the area under the ROC curve was 0.8594 (p=0.0157). Conclusion: Here we present, for the first time, the miRNA expression profile of serum exosomes derived from PC patients. Based on this result, we presently suggest that the exosomal hsa-miR-27a-5p may serve as a putative tumor marker for preoperative identification of PC and PA subjects.
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.