Project description:Coagulation protease factor VIIa (FVIIa) is shown to induce anti-inflammatory and barrier protective effects via endothelial cell protein C receptor (EPCR)-dependent, protease-activated receptor-1 (PAR1)-mediated cell signaling. FVII-EPCR-PAR1 signaling also induces the release of extracellular vesicles from endothelial cells. To obtain clues on whether microRNA (miR) carried out by FVIIa-released EEVs contribute to anti-inflammatory and barrier protective effects, we analyzed miR expression in control- and FVIIa-released EEVs by deep sequencing. These data revealed that several anti-inflammatory miR expression was higher (more than 2-fold) in FVIIa-released EEVs compared to control EEVs, the most predominant being miR10a-5p. The differential expression of miR10a-5p and several other abundant miRs were validated by qRT-PCR. Subsequent in vitro and in vivo experiments showed that miR10a in FVIIa-released EEVs contribute to anti-inflammatory and barrier protective effects.
Project description:Coagulation protease signaling within the tumor-microenvironment (TME) causes cancer immune evasion and impairs immune checkpoint-inhibitor therapy. Tumor-associated macrophages produce factor X and drive immune-evasive protease activated receptor-2 (PAR2) signaling. Myeloid FXa-PAR2 signaling promotes the enrichment of resident-like macrophages displaying an immunosuppressive repair phenotype within the TME of mice with genetically induced spontaneous breast cancer development. Conversely, loss of FXa-PAR2 signaling favors the expansion of monocyte-derived macrophage subsets relevant for DNA-sensing, autophagy and cGAS-STING-IFNβ mediated stimulation of anti-tumor immunity. Thereby FXa suppresses the function of CD103+ dendritic cells relevant for priming, reactivation, and expansion of CD8+ T-cells, including antigen-experienced progenitor exhausted T-cells. Our findings thus emphasize the translational potential of FXa inhibition to synergize with immunotherapy.ompacted abstract
Project description:Various culture media that can rapidly expand bone marrow stromal cells (BMSCs) are currently available. However, the effects of those culture media on the contents of extracellular vesicles released by bone marrow stromal cells have not been fully understood. Using BMSCs from 6 healthy donors were cultured in two different culture media and characterized the small RNA profiles in extracellular vesicles.
Project description:Determine if transcripts are released by protease treatment During total RNA isolation cell free lysates from stationary-phase cultures or exponential cultures were treated with one of three proteases or buffer alone. All experimental samples are over a common reference. There are two replicates for each sample.
Project description:In order to identify small RNAs that are enriched in T-cell extracellular vesicles (EVs) upon T cell activation in a nSMase-dependent manner, we used a two-step ultracentrifugation procedure to isolate EVs released by T cells followed by small RNA profiling of EVs and T cells under resting, stimulated conditions and stimulated conditions in the presence of the vehicle control, DMSO, or nSMase inhibitor, GM4869.
Project description:We have isolated and characterized the extracellular vesicles released by Naegleria fowleri (ATCC 30808) and determined their effect on MDCK cells and erythrocytes.
Project description:Osteoblast-like cells isolated from the calvariae of wildtyope and protease-activated receptor-2 null were treated with unconditioned medium (Control) or medium conditionedby the prostate cancer cell line MDA-PCa-2b (MDA). RNAseq analysis was performed to identify differntially gene expression in wildtype and protease-activated receptor-2 null in response to factors released by prostate cancer cells.
Project description:Extracellular vesicles (EVs) are membrane vesicles released by all cell types and contain proteins and non-coding RNAs, which are transported into recipient cells to regulate their signal transduction and functions. Increasing evidence has demonstrated that EV shuttling is an effective means of bio-molecule transportation among various cell types in the tumor microenvironment, and thus plays a critical role in regulating cancer cell biology. Previous studies have shown that TAMs are an important source of extracellular vesicles and the extracellular vesicles released by TAMs can promote the invasiveness of breast cancer cells. In this study, we studied the differential expression of TAM EV and the donor cells.