Project description:RNA-Seq of cellular and extracellular samples from DiFi cells. The distribution of extracellular RNA among cells, sEV pellet, exomeres and supermeres is distinct.
Project description:Extracellular vesicles, including exosomes, and exomere nanoparticles, are under intense investigation for cargo that may serve as clinical biomarkers or therapeutic targets. Here, we report discovery of a new extracellular nanoparticle, termed supermeres. We performed LC/MS-MS proteomics analyses on gradient-purified sEVs, NVs, exomeres and supermeres. The proteomic profile of supermeres is clearly distinct from that of sEVs, NVs and exomeres. This study identifies a new functional nanoparticle replete with potential circulating biomarkers and therapeutic targets that can be exploited for clinical benefit in a host of diseases.
Project description:Glioblastoma is a grade IV glioma of heterogeneous nature which complicates disease pathophysiology and biomarker research. Thus, the aim of our meta-analysis is to identify long noncoding RNAs (lncRNAs) and protein coding genes (PCGs) that are differentially expressed over different glioblastoma tissue datasets. Small RNA-seq of glioblastoma tissues was also performed to identify differentially expressed microRNAs (miRNAs) relative to paired controls.
Project description:Extracellular vesicles, including exosomes, and exomere nanoparticles, are under intense investigation for cargo that may serve as clinical biomarkers or therapeutic targets. Here, we report discovery of a new extracellular nanoparticle, termed supermeres. We performed LC/MS-MS proteomics analyses on gradient-purified sEVs, NVs, exomeres and supermeres. The proteomic profile of supermeres is clearly distinct from that of sEVs, NVs and exomeres This study identifies a new functional nanoparticle replete with potential circulating biomarkers and therapeutic targets that can be exploited for clinical benefit in a host of diseases.
Project description:The cell seretome is diverse and contains extracellular secreted vesicles (EVs) as well as non-vesicular extracellular nanoparticles (NVEPs) called exomeres and supermeres, all of which carry different kinds of cell signaling cargos. One supermere enriched cargo is transforming growth factor beta induced (TGFBi) protein. We have created TGFBi-neon green-His tagged expressing cell lines in the microsatellite stable (MSS) colorectal cancer cell line DiFi and another one in CC-CR cells with microsatellite instability (MSI). We have developed a fast protein liquid chromatography (FPLC) method for isolating EVs, exomeres and supermeres from biofluids. We used either FPLC or FPLC and then nickel columns that bind His tags to isolate supermeres from DiFi or CC-CR cells. We utilized a proteomics approach acquiring LC-MS/MS data to compare FPLC to FPLC and His tagged TGFBi-containing purified supermeres from DiFi and CC-CR cells. This was done using hollow fiber 3D bioreactor production of conditioned media from these cell lines.
