Project description:The experiment is for demonstrating the miRNA profiles in plasma exosomes derived from mild cognitive impairment and Alzheimer's disease patients and healthy donors.
Project description:Exosome fractions were purified from plasma or spinal fluid collected from Alzheimer's disease patients using MagCapture. Elution of exosomes bound to affinity beads was performed with 1% SDS. Proteins were precipitated from the exosome fraction by acetone precipitation; the precipitates were dissolved in 50% trifluoroethanol and subjected to reduction with DTT and alkylation reaction with iodoacetamide. The proteins were degraded by trypsin/Lys-C, and the peptides produced were desalted and purified using MonoSpinC18. The peptides were measured by LC-MS/MS and quantified by non-label quantification method. The peak area values of peptide-proteins for each sample were normalized to the total peak area.
Project description:We sequenced plasma exosomal RNA obtained from 58 blood samples from 29 cervical cancer patients treated with concurrent chemoradiotherapy (CCRT) followed by intracavitary brachytherapy. The patients with 2018 International Federation of Gynecology and Obstetrics [FIGO] stage IB-IVB at diagnosis were included. The 29 cervical cancer patients had two blood samples for each patient drawn before and after CCRT. Next generation sequencing data from plasma exosome included small RNAs, long non-coding RNAs, and mRNAs, of which miRNAs and mRNAs were used for analysis. The log2 fold change values were calculated between the two samples from each patient to detect miRNAs as predictors of early progression and metastasis. We analysed their biological functions through miRNA-mRNA network analysis after selecting several potential exosomal miRNAs associated with clinical results.
Project description:Stroke is one of the major causes of death and long-term disability, of which acute ischemic stroke (AIS) is the most common type. Although circRNA expression profiles of AIS patients have been reported to be significantly altered in blood and peripheral blood mononuclear cells, the role of exosome-contained circRNAs after AIS is still unknown. Plasma exosomes from ten AIS patients and ten controls were isolated, and through microarray and bioinformatics analysis, profile and putative function of circRNAs in the plasma exosomes were studied. A total of 198 circRNAs were differentially quantified (|log2 FoldChange|≥1.00, P<0.05) between AIS patients and controls.The functions of host genes of differentially quantified circRNAs, including RNA and protein process, focal adhesion and leukocyte transendothelial migration, were associated with the development of AIS. As miRNA sponge, differentially quantified circRNAs had the potential to regulate pathways related to AIS, like PI3K-Akt, AMPK and chemokine pathways. Of 198 differentially quantified circRNAs, 96 circRNAs possessing strong translational ability could affect cellular structure and activity, like focal adhesion, tight junction and endocytosis. Most differentially quantified circRNAs were predicted to bind to EIF4A3 and AGO2- two RNA binding proteins (RBPs)- and play a role in AIS. In conclusion, plasma exosome-derived circRNAs were significantly differentially quantified between AIS patients and controls, and participated in the occurrence and progression of AIS by sponging miRNA/RBPs or translating into proteins, indicating that circRNAs from plasma exosomes could be crucial molecules in the pathogenesis of AIS and promising candidates as diagnostic biomarkers and therapeutic targets for the condition.
Project description:miRNAs are kown to be deeply involved in the progression of chronic inflammatory diseases. We comprehensively analyzed miRNAs in human plasma from AD(atopic dermatitis) and ACD(allergic conjunctival disease) patients. We performed a comprehensive human plasma miRNA analysis and identified significantly up-regulated miRNAs in both AD (atopic dermatitis) and ACD (allergic conjunctival disease).
Project description:827 human miRNA standard human panel Plasma derived extracellular vesicles (EV)/exosomes can serve as markers of cell damage/disease but can also have therapeutic utility depending on the nature of their cargo, such as miRNA. Currently there are challenges and lack of innovations regarding early diagnosis and therapeutic options within different aspects of management of trauma patients. We characterized the miRNA cargo of exosomes purified from trauma patients, and compared it to those from healthy participants. Methods Plasma was collected from a prospective, cohort study of trauma patients (n = 15; 7 males, 8 females) with hip and/or femur fractures and an injury severity score (ISS) ≥ 15; elective total hip arthroplasty (THA) patients (n = 8; 4 males, 4 females) served as operative controls. Exosomes were isolated from plasma with the Invitrogen Total Exosome Isolation Kit and RNA was isolated using a miRNeasy Mini Kit. Direct quantification of miRNA copies was performed by NanoString Technologies on a human miRNA gene panel and analyzed with nSolver Results Trauma plasma-derived exosomes had 60 miRNA that were significantly downregulated and 3 miR upregulated when compared to THA (p<0.05). Thirteen miR have a direct role in hematopoiesis regulation, including miR-223 and miR-451a. Further, male trauma plasma-derived exosomes demonstrated downregulation of 150 miR compared to male THA (p<0.05). While female trauma plasma-derived exosomes demonstrated downregulation of only four miR and upregulation of 2 miR compared to female THA (p<0.05). Conclusions Sex-specific differences in the miR from plasma-derived exosomes occurred following severe trauma. These miR changes in plasma-derived exosomes likely contribute to postinjury systemic inflammation, immune system dysregulation, and bone marrow dysfunction and may merit consideration as future potential therapeutic targets.
Project description:Introduction: microRNAs are promising candidate breast cancer biomarkers due to their cancer-specific expression profiles. However, efforts to develop circulating breast cancer biomarkers are challenged by the heterogeneity of microRNAs in the blood. To overcome this challenge, we aimed to develop a molecular profile of microRNAs specifically secreted from breast cancer cells. Our first step towards this direction relates to capturing and analyzing the contents of exosomes, which are small secretory vesicles that selectively encapsulate microRNAs indicative of their cell of origin. To our knowledge, circulating exosome microRNAs have not been well evaluated as biomarkers for breast cancer diagnosis or monitoring. Methods: Exosomes were collected from the conditioned media of human breast cancer cell lines, mouse plasma of patient-derived orthotopic xenograft models (PDX), and human plasma samples. Exosomes were verified by electron microscopy, nanoparticle tracking analysis, and western blot. Cellular and exosome microRNAs from breast cancer cell lines were profiled by next-generation small RNA sequencing. Plasma exosome microRNA expression was analyzed by qRT-PCR analysis. Results: Small RNA sequencing and qRT-PCR analysis showed that several microRNAs are selectively encapsulated or highly enriched in breast cancer exosomes. Importantly, the selectively enriched exosome microRNA, human miR-1246, was detected at significantly higher levels in exosomes isolated from PDX mouse plasma, indicating that tumor exosome microRNAs are released into the circulation and can serve as plasma biomarkers for breast cancer. This observation was extended to human plasma samples where miR-1246 and miR-21 were detected at significantly higher levels in the plasma exosomes of 16 breast cancer patients as compared to the plasma exosomes of healthy control subjects. Receiver Operating Characteristic (ROC) curve analysis indicated that the combination of plasma exosome miR-1246 and miR-21 levels is a better indicator of breast cancer than their individual levels. Conclusions: Our results demonstrate that certain microRNA species, such as miR-21 and miR-1246, are selectively enriched in human breast cancer exosomes and significantly elevated in the plasma of breast cancer patients. These findings indicate a potential new strategy to selectively analyze plasma breast cancer microRNAs indicative of the presence of breast cancer.
Project description:To determine the differential miRNA levels in methamphetamine addicts, we comparatively profiled plasma exosome miRNA expression of methamphetamine abusers and healthy controls using miRNA sequencing
Project description:We performed miRNA microarray analysis to find differentially expressed miRNAs between nasopharyngeal carcinoma patients and normal control subjects In this study, 20 nasopharyngeal carcinoma patients and 10 normal control subjects were selected to collect plasma samples and perform miRNA microarray profiling