Project description:<p>Small extracellular vesicles (sEVs) have been extensively investigated as a source of clinically relevant cargo. Alongside the secretion of EVs, such as exosomes, cells house a multitude of internal vesicles involved in various physiologic processes and metabolic activities. Herein, we developed protocols to isolate small intracellular vesicles (sIVs) from different cell types and investigated their biological characteristics and potential therapeutic applications. SIVs are smaller in size, have higher yield, and display a markedly greater uptake in vitro and in vivo than sEVs. The composition of proteins, RNAs, and lipids of sIVs differed from that of sEVs. Additionally, sIVs derived from mesenchymal stem cells outperformed sEVs in ameliorating retinal degeneration by mitigating endoplasmic reticulum stress and providing neuroprotective factors in an in vivo model. This study identified a distinct functional intracellular nanoparticle as a suitable alternative to sEVs in clinical translation</p>
Project description:miRNA-sequencing of grapefruit-derived extracellular vesicles and fusion nanovesicles derived from grapefruit-derived extracellular vesicles and gingival mesenchymal stem cell-derived vesicles. We then performed gene expression profiling analysis to explore the miRNAs derived from grapefruit-derived extracellular vesicles, and the retention rate of miRNAs after membrane fusion
Project description:Similar to bacterial proteins that are targeted to distinct macrophages organelles via extracellular vesicles, we propose that these vesicles also traffic small RNAs to modulate specific host factors. To test this, we aim to sequence extracellular vesicle derived sRNA, and whole bacterial small RNAs to determine selectivity, and to identify their bacterial and mammalian targets (Experimental Plan in Table-1). For this we will collect highly purified vesicles from N. gonorrhoeae (strain MS11A). We will also treat mouse derived primary macrophages with extracellular vesicles and compare their RNA response to untreated macrophages (Table-2). This will provide novel insights into how macrophages respond to N. gonorrhoeae infections. This data is part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing/
Project description:A growing body of evidence in mammalian cells indicates that secreted vesicles can be used to mediate intercellular communication processes by transferring various bioactive molecules, including mRNAs and microRNAs. Based on these findings, we decided to analyze whether T. cruzi-derived extracellular vesicles contain RNA molecules and performed a deep sequencing and genome-wide analysis of a size-fractioned cDNA library (16M-bM-^@M-^S40 nt) from extracellular vesicles secreted by noninfective epimastigote and infective metacyclic trypomastigote forms. Our data show that the small RNAs contained in these extracellular vesicles originate from multiple sources, including tRNAs. In addition, our results reveal that the variety and expression of small RNAs are different between parasite stages, suggesting diverse functions. Taken together, these observations call attention to the potential regulatory functions that these RNAs might play once transferred between parasites and/or to mammalian host cells. Small RNAs profiles (16-40 nt) of epimastigote-derived extracellular vesicles, metacyclic trypomastigote-derived extracellular vesicles and metacyclic trypomastigote parental cells.
Project description:Epithelial-mesenchymal transition (EMT) is a process by which epithelial cells lose cell-cell contact and gain cancer malignancy such as invasion, stemness, chemoresistance and metastasis. Reverse precess, mesenchymal-epithelial transition (MET) is also important for colonization. Extracellular vesicles (EVs) secreted from cancer cells are also important for cancer malignancy. To analyze RNAs from cells and EVs during EMT and MET, RNA sequencing was performed using E-cadherin-RFP/Py2T reporter system.
Project description:Primary epithelial cells isolated from fetal lungs of rat fetuses with or without lung hypoplasia induced by the administration of nitrofen to pregnant rats. Control group included epithelial cells from normal fetal lungs. Treatment with amniotic fluid stem cell derived extracellular vesicles or with mesenchymal stromal cell derived exosomes, RNA-seq of both cargos included.
Project description:Human osteogenically enhanced mesenchymal stem cells were cultured for 5 days on standard gelfoam or gelfoam coated with MSC-derived extracellular matrix To address whether mesenchymal stem cell derived extracellular matrix might stimulate the secretion of osteogenic factors by OEhMSCs, we seeded them onto standard gelfoam or ECM-coated gelfoam in vitro and cultured them under standard conditions. After 5 days, we performed transcriptome micro-array analysis to identify genes that were up-regulated >2-fold.
Project description:The discovery of cell-free micro-RNAs in body fluids has made them a promising biomarker target in the field of neurodegenerative diseases. Although they have been reported to be differentially expressed in biofluids and tissue from sporadic Parkinson’s disease patients, it remains unclear whether similar observations can be made in patients with genetic forms of the disease. Since induced pluripotent stem cell derived neurons represent a widely used research model for both sporadic and familial Parkinson’s disease, we sought to assess the usability of this model for the identification of differentially expressed cell-free micro-RNAs in the context of the Parkinson’s disease related LRRK2 G2019S mutation in a proof-of-concept study. We isolated extracellular vesicles carrying cell-free RNA from patient-derived induced pluripotent stem cell lines carrying the LRRK2 G2019S mutation and their gene corrected isogenic controls. After generation of small-RNA libraries and differential expression analysis, we validated fourteen micro-RNAs in an independent batch of cell-free and cellular RNA via RT-qPCR. Finally, we selected eleven differentially expressed micro-RNAs from our cell culture experiments and quantified their expression levels in cerebrospinal fluid derived from two LRRK2 G2019S patients and two healthy controls.
2023-10-31 | GSE243852 | GEO
Project description:miRNA sequencing of human mesenchymal stem cell-derived extracellular vesicles