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Exosome of plasma and CSF from AD patients

ABSTRACT: 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.

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Tatsuo Mano

PROVIDER: PXD029767 | JPOST Repository | Sat Jan 06 00:00:00 GMT 2024

REPOSITORIES: jPOST

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CSF exosome proteome of Alzheimer's disease

Project description:Exosome fractions were purified from spinal fluid collected from Alzheimer's disease patients and control subjects 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.

2024-01-06 | PXD029789 | JPOST Repository

RNA-Seq of fibroblasts influenced by exosomes derived from melanoma cell line G-361

Project description:In this in-vitro study, we demonstrate the different effects of exosomes produced by melanoma cells on the functional properties of normal dermal fibroblasts and fibroblasts prepared from skin metastasis of cutaneous malignant melanoma (CMM). Both normal and cancer-associated fibroblast were prepared by a)DMEM with 10% EDS (control); b) DMEM with 10% Exosome-depleted serum (EDS) + 10 microg/ml G-361 exosomes (EXO_G361); c) DMEM with 10% EDS + 10 microg/ml exosomes from FBS (EXO_FBS). The cells were cultured for 72 hours. Transcriptome analysis was performed 72 hours after exosome application. Total RNA was isolated using RNeasy Micro Kit (Qiagen).

2022-12-31 | E-MTAB-10278 | biostudies-arrayexpress

Melanoma-derived exosomes educate bone marrow progenitor cells toward a pro-metastatic phenotype through upregulation of the MET oncoprotein.

Project description:Tumor-derived exosomes are emerging as mediators of tumorigenesis with a tissue-specific address and message. We explored the function of melanoma-derived exosomes in formation of primary tumors and metastatic progression in both murine models and patients. Whereas exosomes from highly metastatic melanoma cells increased the metastatic behavior of primary tumor cells by educating bone marrow (BM) progenitor cells via the MET receptor, exosomes from low metastatic melanoma cells did not alter the incidence of metastases. Melanoma-derived exosomes induced vascular leakiness at pre-metastatic sites, and reprogrammed BM progenitor cells towards a pro-vasculogenic phenotype (c-Kit+Tie2+MET+). Reducing MET expression in tumor-derived exosomes diminished the pro-metastatic behavior of BM cells. Importantly, MET expression was upregulated in circulating BM progenitor cells (CD45-CD117low and CD45-CD117lowTIE2+) isolated from stage III and stage IV melanoma patients. Rab1a, Rab5b, Rab7, and Rab27a were highly expressed in melanoma and Rab27a RNA interference decreased exosome production and/or soluble angiogenic factors in melanoma cells, thereby preventing mobilization of BM progenitor cells, tumor growth and metastasis. Finally, we identified a melanoma signature in exosomes isolated from metastatic melanoma patients, comprised of TYRP2, VLA-4, Hsp70, an Hsp90 isoform and MET oncoprotein, which together with Rab proteins, appear to represent exosome-specific proteins with prognostic potential, and may provide new therapeutic targets. Identification of molecular finger associated to exosome effects in metastatic organs Microarray analysis of genes differentially expressed in the lungs 24 and 48 hours after B16-F10 exosome tail vein injection compared to control.

2012-05-26 | E-GEOD-36584 | biostudies-arrayexpress

Single-cell RNA-seq of fibroblasts influenced by exosomes derived from melanoma cell line G361

Project description:In this in-vitro study, we demonstrated the different effects of exosomes produced by G361 melanoma cells on the functional properties of normal dermal fibroblasts and fibroblasts prepared from skin metastasis of cutaneous malignant melanoma. Both normal and cancer-associated fibroblasts were cultivated in either DMEM with 10% EDS (exosome-depleted serum) (control) or DMEM with 10% EDS + 10 microg/ml G361 exosomes (EXO). The cells were harvested after 24 hours of cultivation.

2021-12-31 | E-MTAB-10290 | biostudies-arrayexpress

Detection of miRNAs in exosomes released by mouse immature and mature dendritic cells

Project description:Dendritic cells (DCs) are the most potent antigen (Ag)-presenting cells. Whereas immature DCs down-regulate T cell responses to induce/maintain immunological tolerance, mature DCs promote immunity. To amplify their functions, DCs communicate with neighboring DCs through soluble mediators, cell-to-cell contact and vesicle exchange. Transfer of nanovesicles (<100nm) derived from the endocytic pathway (termed exosomes) represents a novel mechanism of DC-to-DC communication. The facts that exosomes contain exosome-shuttle microRNAs (miRNAs), and DC functions can be regulated by exogenous miRNAs, suggest that DC-to-DC interactions could be mediated through exosome-shuttle miRNAs, an hypothesis that remains to be tested. Importantly, the mechanism of transfer of exosome-shuttle miRNAs from the exosome lumen to the cytosol of target cells is unknown. Here, we demonstrate that DCs release exosomes with different miRNAs depending on the maturation of the DCs. By visualizing spontaneous transfer of exosomes between DCs, we demonstrate that exosomes fused with the target DCs, the latter followed by release of the exosome content into the DC cytosol. Importantly, exosome-shuttle miRNAs are functional, as they repress target mRNAs of acceptor DCs. Our findings unveil a mechanism of transfer of exosome-shuttle miRNAs between DCs and its role as a means of communication and post-transcriptional regulation between DCs. The study has analyzed the microRNA content of 4 samples of immature exosomes, 4 samples of matures exosomes, 2 samples of immature bone-marrow-derived DCs, and 2 samples of mature bone marrow-derived DCs.

2011-10-24 | E-GEOD-33179 | biostudies-arrayexpress

Endothelial cells require miR-214 to secrete exosomes that suppress senescence and induce angiogenesis in human and mouse endothelial cells

Project description:Signalling between endothelial cells, endothelial progenitor cells and stromal cells is crucial for the establishment and maintenance of vascular integrity and involves exosomes, among other signalling pathways. Exosomes are important mediators of intercellular communication in immune signalling, tumour survival, stress responses and angiogenesis. The ability of exosomes to incorporate and transfer mRNAs encoding for ‘acquired’ proteins or miRNAs repressing ‘resident’ mRNA translation suggests that they can influence the physiological behaviour of recipient cells. We here demonstrate that miR-214, a miRNA that controls endothelial cell function and angiogenesis, plays a dominant role in exosome-mediated signalling between endothelial cells. Endothelial cell-derived exosomes stimulated migration and angiogenesis in recipient cells, whereas exosomes from miR-214 depleted endothelial cells failed to stimulate these processes. Exosomes containing miR-214 repressed the expression of Ataxia Telangiectasia Mutated in recipient cells, thereby preventing senescence and allowing blood vessel formation. Concordantly, specific reduction of miR-214 content in exosome-producing endothelial cells abolishes the angiogenesis the angiogenesis stimulatory function of the resulting exosomes. Collectively our data indicate that endothelial cells release miR-214 containing exosomes to stimulate angiogenesis through silencing of Ataxia Telangiectasia Mutated in neighbouring target cells. Gene expression analysis of HMEC endothelial cells exposed to supernatant containing either HMEC derived exosomes (miR-214 high), HMEC derived exosomes depleted of miR-214 (miR-214 low) or containing no exosomes (no exosomes). Each sample was analysed in duplo.

2013-03-22 | E-GEOD-45375 | biostudies-arrayexpress

Study of transcriptional content in exosomes derived from untreated and growth factor-treated cultured cardiomyocytes (HL-1)

Project description:Introduction: Exosomes are nano-sized extracellular vesicles, released from various cells, which can stimulate or repress responses in target cells. We have recently shown that cultured cardiomyocytes release exosomes and that they, in turn, are involved in facilitating events in target cells by alteration of gene expression. We investigated whether external stimuli of the cardiomyocyte might influence the released exosome characteristics. Material and Methods: Exosomes were isolated from media collected from cultured cardiomyocyte (HL-1) cells with or without growth factor treatment (TGF-beta2 and PDGF-BB), with a series of differential centrifugations. The exosomes were characterized with dynamic light scattering (DLS) and Western blot and analysed with Illumina whole genome microarray gene expression. Results: An average size of 50-80 nm in diameter with no difference between treatment groups was found. Analysis of the mRNA content revealed 623 transcripts in the control group, 691 in the TGF-beta2-treated group and 362 in the PDGF-BB-treated group. 235 transcripts were common for all three groups. Conclusion: We conclude that there is a difference in mRNA content between exosomes derived from cultured cardiomyocytes stimulated with growth factors. We also conclude that all exosomes contain a basic package consisting of ribosomal transcripts and mRNAs coding for proteins with functions within the energy supply system. To study if the transcriptional content in exosomes derived from untreated and growth factor-treated cultured cardiomyocytes (HL-1) differ, and if so, can this difference be explained, 4 control (untreated) exosome samples, 4 TFG-beta2-treated cardiomyocyte-derived exosome samples and 4 PDGF-BB-treated cardiomyocyte-derived exosomes were studied.

2013-06-20 | E-GEOD-40503 | biostudies-arrayexpress

Identification of a circulating miRNA that is involved in meat yield and connects exosomes and longissimus dorsi muscle in Korean cattle steers

Project description:MicroRNAs (miRNAs) regulate gene expression. The aim of this study was to identify circulating miRNAs that are involved with meat yield and connect exosomes and longissimus dorsi (LD) muscle in Korean cattle steer. Thus, we performed analyses of the carcass characteristics, exosome extraction, bovine miRNA array, mature miRNA qPCR, bioinformatics, and qPCR. Our analysis of the carcass characteristics relative to the yield grade (YG) showed that the yield index (YI) and rib eye area were the highest, whereas the backfat thickness was the lowest for YG A cattle among the three YGs. We performed a miRNA microarray and a Venn diagram analysis to sort the circulating miRNAs that connect exosomes and LD muscle. Mature miRNA qPCR showed that miR-15a (r = 0.84), miR-26b (r = 0.91), and miR-29c (r = 0.92) had positive relationships with exosomes and LD muscle. In YG A cattle, miR-26b was considered to be a circulating miRNA connecting exosomes and LD muscle because the expression pattern of miR-26b was similar in the microarray and miRNA qPCR for miR-26b-targeted genes, where the expression levels of DIAPH3 and YOD1 were lower than those in YG C cattle. Our results suggest that circulating miR-26b may participate in cell maintenance by regulating DIAPH3 and YOD1 in the LD muscle of Korean cattle. 29 or 30 month Korean cattle steers were classified by yield grade(YG) A, B, and C. miRNA expression patterns for exosome and LD muscle were compared among YGs.

2016-03-01 | E-GEOD-73389 | biostudies-arrayexpress

Characterization of serum small extracellular vesicles and their small RNA contents across human, rat, and mouse

Project description:In the present study, the isolated serum exosomes from healthy adults, rats and mice were comparatively characterized; the small RNA compositions were analyzed; the serum exosome miRNA profiles and exosome depleted serum miRNA profiles were obtained by small RNAseq, and the distributions of miRNA component inside and outside of the serum exosomes were also compared.

2020-03-13 | E-MTAB-8376 | biostudies-arrayexpress

Characterization of serum small extracellular vesicles and their small RNA contents across human, rat, and mouse (data for rat and mouse)

Project description:In the present study, the isolated serum exosomes from healthy adults, rats, and mice were comparatively characterized; the small RNA compositions were analyzed; the serum exosome miRNA profiles and exosome depleted serum miRNA profiles were obtained by small RNAseq, and the distributions of miRNA component inside and outside of the serum exosomes were also compared.

2020-03-13 | E-MTAB-8377 | biostudies-arrayexpress

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