Project description:Skeletal muscle maintains remarkable regenerative ability after injury, which mainly depends on the coordination and dynamic regulation between muscle stem cells and various other cell types. Recent studies have shown that fibro/adipogenic progenitors (FAP) play an important role in skeletal muscle regeneration by coordinating the interaction between muscle stem cells and macrophages. At present, the mechanism of whether FAP cells can regulate macrophages to affect skeletal muscle regeneration remains unclear. By isolation of mouse skeletal muscle FAP cells damaged by CTX for 10 days and gathered exosomes from them called FAPD10-exos, and using the exosomes to induce macrophages in vitro, we found that they expressed high levels of M2-type factors.In summary, we believe that FAPD10-exos can promote the polarization of macrophages towards M2 phenotype by secreting exosomes, and participate in and promote the regulation and regulation of skeletal muscle regeneration to enter the stage of damage repair in the early stage of regeneration.
Project description:To explore the potential exosomal miRNAs regulating macrophage M1 polarization, we isolated and characterized inf-Exos or un-Exos, ad the miRNA profiling between inf-Exos and un-Exos was then compared by using miRNA-seq. We then performed miRNAs expression profiling analysis using data obtained from RNA-seq of inf-Exos (n = 5) and un-Exos (n = 5)
Project description:Previously, we reported that human primary (SW480) and metastatic (SW620) colorectal (CRC) cells secrete (shed) midbody remnants (MBRs), exosomes (Exos), and microparticles (MPs) are molecularly distinct at the protein level. To gain further biochemical insights into MBRs, Exos, and MPs and their emerging role in CRC, we performed, and report here for the first time, a comprehensive transcriptome and long noncoding RNA sequencing analysis and fusion gene identification of these three EV classes using the next-generation RNA sequencing technique. Differential transcript expression analysis revealed that MBRs have a distinct transcriptomic profile compared to Exos and MPs with a high enrichment of mitochondrial transcripts lncRNA/pseudogene transcripts that are predicted to bind to ribonucleoprotein complexes, spliceosome and RNA/stress granule proteins. A salient finding from this study is a high enrichment of several fusion genes in MBRs compared to Exos, MPs and cell lysates from their parental cells such as MSH2 (gene encoded DNA mismatch repair protein MSH2). This suggests potential EV-liquid biopsy targets for cancer detection. Importantly, the expression of cancer progression-related transcripts found in EV classes derived from SW480 (EGFR) and SW620 (MET and MACCA1) cell lines reflects their parental cell types. Our study is the first discovery of RNA and fusion gene compositions within MBRs (including Exos and MPs) that could have an impact on EV functionality in cancer progression and detection using EV-based RNA/ fusion gene candidates for cancer biomarkers
Project description:We have identified a number of miRNAs that are differentially expressed in LPS and IFNγ treated RAW264.7 cells, as compared to untreated cells. Microarray and quantitative real-time PCR (qRT-PCR) results showed that miR-103 decreased while the STAT1 expression increased substantially in RAW264.7 derived M1 macrophage, and there was a significant negative correlation between miR-103 and STAT1.Overexpression of miR-103 suppressed M1 polarization by inhibiting STAT1/IRF1 signaling pathway and vice versa in vitro. STAT1 is a direct target of of miR-103.
Project description:RNA-seq analysis of RAW264.7 cells treated for 24 hours with CM-PEG-PLGA@DOX/R848 nanoparticles (equivalent DOX 2μg/ml) compared to PBS-treated controls. The study aims to identify transcriptomic changes induced by the nanomaterial treatment to to investigate the mechanism of immunomodulatory activity of CM-PEG-PLGA@DOX/R848 nanoparticles.
Project description:Mouse RAW264.7 macrophages were treated with LPS, IFNb, poly(rI:rC), poly(dA:dT), VSV, HSV, Sendai virus. Genes identified by Human Innate Immunity Interactome for type I Interferon (HI5) were examined for expression. qPCR gene expression profiling. RAW264.7 macrophages were used and treated separately as indicated in the summary. Equal amount total RNA from each group was used for gene expression analysis.
Project description:Transcriptional profiling of differential miRNA expression in mouse RAW264.7 preosteoclast cells comparing control untreated cells with RAW264.7 cells treated for 6 days. Treatment conditiones tested included 50ng/mL RANKL or indicated bone metastasis tumor cell conditioned media from 4T1, 4T1.2, TSU-PR1, and TSU-PR1-B2 cell lines.