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Escherichia coli msbB-deficient mutant extracellular vesicle proteome

ABSTRACT: Bacteria actively secrete extracellular vesicles (EVs), spherical nano-sized proteolipids into the extracellular milieu. Bacterial EVs have gained wide interests as non-living complex vaccines or delivery vehicles. However, no studies have used bacterial EVs in treating cancer so far. Our results showed remarkable capability of EVs derived from Escherichia coli W3110 msbB-deficient mutant to effectively induce long-term anti-tumor immune responses that can fully eradicate established tumors without notable adverse effects. This anti-tumor effect was IFN-γ-dependent, and we present a comprehensive proteome of E. coli W3110 msbB-deficient EVs to investigate which vesicular components induce the anti-tumor effects.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Escherichia Coli

TISSUE(S): Cell Culture

SUBMITTER: Yong Song Gho

LAB HEAD: Yong Song Gho

PROVIDER: PXD006912 | Pride | 2018-11-22

REPOSITORIES: Pride

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Bacterial outer membrane vesicles suppress tumor by interferon-γ-mediated antitumor response.

Kim Oh Youn OY Park Hyun Taek HT Dinh Nhung Thi Hong NTH Choi Seng Jin SJ Lee Jaewook J Kim Ji Hyun JH Lee Seung-Woo SW Gho Yong Song YS

Nature communications 20170920 1

Gram-negative bacteria actively secrete outer membrane vesicles, spherical nano-meter-sized proteolipids enriched with outer membrane proteins, to the surroundings. Outer membrane vesicles have gained wide interests as non-living complex vaccines or delivery vehicles. However, no study has used outer membrane vesicles in treating cancer thus far. Here we investigate the potential of bacterial outer membrane vesicles as therapeutic agents to treat cancer via immunotherapy. Our results show remark ...[more]

PMID: 28931823

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Staphylococcus aureus RN4220 wild-type extracellular vesicle proteome

Project description:Bacteria actively secrete extracellular vesicles (EVs), spherical nano-sized proteolipids into the extracellular milieu. Bacterial EVs have gained wide interests as non-living complex vaccines or delivery vehicles. However, no studies have used bacterial EVs in treating cancer so far. Our results showed remarkable capability of EVs derived from Staphylococcus aureus RN4220 wild-type to effectively induce long-term anti-tumor immune responses that can fully eradicate established tumors without notable adverse effects. This anti-tumor effect was IFN-γ-dependent, and we present a comprehensive proteome of S. aureus RN4220 wild-type EVs to investigate which vesicular components induce the anti-tumor effects.

2018-11-22 | PXD006913 | Pride

Mycobacterium tuberculosis extracellular vesicle proteome

Project description:Tuberculosis is a common infectious disease mostly caused by Mycobacterium tuberculosis. M. tuberculosis have been shown to release extracellular vesicles (EVs) containing immunologically active molecules. However, only limited number of proteins has been identified in M. tuberculosis EVs. We present a comprehensive proteome of M. tuberculosis EVs to elucidate the pathogenesis of M. tuberculosis EVs and suggest several potential biomarker candidates.

2015-07-30 | PXD001160 | Pride

Quantitative proteomic analysis of trypsin-treated extracellular vesicles to evaluate non-vesicular contaminants and surface vesicular proteins.

Project description:Extracellular vesicles (EVs) are lipid bilayered vesicles released into extracellular milieu by most of cells. Currently, various purification methods of EVs have been established but most of studies use ultracentrifugation-based purification of EVs. However, these methods have the problem of co-purification with non-vesicular contaminants including various different types of particles, small apoptotic vesicles, protein aggregates, and other derived from dead cells or abundant extracellular proteins. Therefore, evaluation of non-vesicular contaminants gives valuable information in EV research. Here, we purified EVs with high purity using density gradient ultracentrifugation and then purified EVs were treated by trypsin to discriminate extra-vesicular proteins from intra-vesicular proteins.

2021-08-30 | PXD001355 | Pride

A dysregulated Acetyl/SUMO switch of FXR promotes hepatic inflammation in obesity

Project description:Acetylation of transcriptional regulators is normally dynamically regulated by nutrient status but is often persistently elevated in nutrient-excessive obesity conditions. We investigated the functional consequences of such aberrantly elevated acetylation of the nuclear receptor FXR as a model. Proteomic studies identified K217 as the FXR acetylation site in diet-induced obese mice. In vivo studies utilizing acetylation-mimic and -defective K217 mutants and gene expression profiling revealed that FXR acetylation increased proinflammatory gene expression, macrophage infiltration, and liver cytokine and triglyceride levels, impaired insulin signaling, and increased glucose intolerance. Mechanistically, acetylation of FXR blocked its interaction with the SUMO ligase PIASy and inhibited SUMO2 modification at K277, resulting in activation of inflammatory genes. SUMOylation of agonist-activated FXR increased its interaction with NF-κB but blocked that with RXRα, so that SUMO2-modified FXR was selectively recruited to and trans-repressed inflammatory genes without affecting FXR/RXRα target genes. A dysregulated Acetyl/SUMO switch of FXR in obesity may serve as a general mechanism for diminished anti-inflammatory response of other transcriptional regulators and provide potential therapeutic and diagnostic targets for obesity-related metabolic disorders. FXR-WT or the FXR-K217Q mutant was expressed in lean mice and FXR-WT or the FXR-K217R mutant was expressed in obese mice by adenoviral infection. One week after infection, mice were treated with GW4064 (30 mg/kg in corn oil) overnight before sacrifice and hepatic expression was analyzed by Illumina microarray.

2014-10-17 | E-GEOD-62414 | biostudies-arrayexpress

Fluidigm Biomark 96.96 nanofluidic arrays for qRT-PCR of 93 miRNAs: salivary extracellular vesicles

Project description:Crude extracellular vesicles (EVs) from eight healthy volunteers were separated into 6 fractions based on their densities by using the iodixanol-based density gradient centrifugation method. To determine the distribution of miRNAs among these fractions, quantities of 93 miRNAs were quantified by the TaqMan real time PCR method using the BioMark HD system (Fluidigm) equipped with 96.96 dynamic array (Fluidigm). Six samples were fractionated from a crude EVs by density gradient centrifugation. Total of 48 samples were prepared from 8 healthy volunteers. Technical replicate of 4 gave 8 x 6 x 4 x 93 = 17,856 data. As control Tris-HCl EDTA buffer (TE) was used.

2016-08-01 | E-GEOD-78763 | biostudies-arrayexpress

Extracellular vesicles isolated from human induced pluripotent stem cell-derived neurons contain a transcriptional network

Project description:Healthy brain function is mediated by several complementary signalling pathways, many of which are driven by extracellular vesicles (EVs). EVs are heterogeneous in both size and cargo and are constitutively released from cells into the extracellular milieu. They are subsequently trafficked to recipient cells, whereupon their entry can modify the cellular phenotype. Here, in order to further analyse the mRNA and protein cargo of neuronal EVs, we isolated EVs by size exclusion chromatography from human induced pluripotent stem cell (iPSC)-derived neurons. Electron microscopy and dynamic light scattering revealed that the isolated EVs had a diameter of 30-100 nm. Transcriptomic and proteomics analyses of the EVs and neurons identified key molecules enriched in the EVs involved in cell surface interaction (integrins and collagens), internalisation pathways (clathrin- and caveolin-dependent), downstream signalling pathways (phospholipases, integrin-linked kinase and MAPKs), and long-term impacts on cellular development and maintenance. Overall, we show that key signalling networks and mechanisms are enriched in EVs isolated from human iPSC-derived neurons.

2020-08-20 | E-MTAB-8254 | biostudies-arrayexpress

Supermeres, a new functional extracellular nanoparticle replete with disease biomarkers and therapeutic target

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.

2021-11-02 | PXD025213 | Pride

EBV Latent membrane protein1 modifies EV content

Project description:Extracellular vesicles (EV) mediate intercellular communication events and alterations in normal vesicle content contribute to function and disease initiation or progression. The ability to package a variety of cargo and transmit molecular information between cells renders EVs important mediators of cell to cell crosstalk. Latent membrane protein 1 (LMP1) is a chief viral oncoprotein expressed in most Epstein-Barr virus (EBV)-associated cancers and is released from cells at high levels in EVs. LMP1 containing EVs have been demonstrated to promote cell growth, migration, differentiation, and regulate immune cell function. Despite these significant changes in recipient cells induced by LMP1 modified EVs, the mechanism how this viral oncogene modulates the recipient cells towards these phenotypes is not well understood. We hypothesize that LMP1 alters EV content and following uptake of the LMP1-modified EVs by the recipient cells results in the activation of cell signaling pathways and increased gene expression which modulates the biological properties of recipient cell towards a new phenotype. Our results show that LMP1 expression alters the EV protein and microRNA content packaged into EVs.

2020-11-03 | PXD021914 | Pride

Biophysical and proteomic analyses suggest functions of Pseudomonas syringae pv tomato DC3000 extracellular vesicles in bacterial growth during plant infection

Project description:Vesiculation is a process employed by Gram-negative bacteria to release extracellular vesicles (EVs) into the environment. Bacterial EVs contain molecular cargo from the donor bacterium and play important roles in bacterial survival and growth. Here, we describe EV production in plant-pathogenic Pseudomonas syringae pv. tomato DC3000 (Pto DC3000), the causal agent of bacterial speck disease. Cultured Pto DC3000 exhibited EV structures both on the cell surface and in the vicinity of bacterial cells, observed as outer membrane vesicle (OMV) release. We used in-solution trypsin digestion coupled to mass spectrometry to identify 369 proteins enriched in EVs recovered from cultured Pto DC3000. The predicted localization profile of EV proteins supports the production of EVs also in the form of outer-inner-membrane vesicles (OIMVs). EV production varied slightly between bacterial lifestyles and also occurred in planta. The potential contribution of EVs to Pto DC3000 plant infection was assessed using plant treatments and bioinformatic analysis of the EV-enriched proteins. While these results identify immunogenic activities of the EVs, they also point at roles for EVs in bacterial defences and nutrient acquisition by Pto DC3000.

2023-10-24 | PXD023971 | Pride

Melanoma-derived extracellular vesicles mediate lymphatic remodeling and impair tumor immunity in draining lymph nodes

Project description:Transcriptional comparison of B16F10 cells with B16F10 cell-derived extracellular vesicles (EVs) to identify transcripts enriched or de-enriched in EVs compared to their donor cells.

2022-03-01 | E-MTAB-11025 | biostudies-arrayexpress

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