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Using small RNA-Seq to identify differential small RNA content of bacterial outer membrane vesicles (OMVs) secreted by Pseudomonas aeruginosa in response to tobramycin exposure

ABSTRACT: Using small RNA-Seq to identify differential small RNA content of bacterial outer membrane vesicles (OMVs) secreted by Pseudomonas aeruginosa in response to tobramycin exposure

PROVIDER: PRJNA762296 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR15838100.fastq.gz	Fastqsanger.gz
	SRR15838101.fastq.gz	Fastqsanger.gz
	SRR15838102.fastq.gz	Fastqsanger.gz
	SRR15838103.fastq.gz	Fastqsanger.gz
	SRR15838104.fastq.gz	Fastqsanger.gz

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Using small RNA-Seq to identify differential small RNA content of bacterial outer membrane vesicles (OMVs) secreted by Pseudomonas aeruginosa in response to tobramycin exposure

Project description:Characterization of the sRNA content of OMVs harvested from Pseudomonas aeruginosa strain PA14 LB cultue with and without tobramycin (1ug/mL)

2021-09-12 | GSE183895 | GEO

short RNA-Seq of Pseudomonas aeruginosa outer membrane vesicles (OMVs) and whole cells

Project description:Characterization of the sRNA content of P. aeruginosa OMVs compared to whole cells. Result: OMVs contain differentially packaged sRNAs. Whole cell PA14 and OMVs from 3 separate preparations.

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Using small RNA-Seq to verify that 5' tRNA-fMet1 half is over-expressed in outer membrane vesicles (OMVs)

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mRNA transcript distribution bias between Borrelia burgdorferi bacteria and their outer membrane vesicles

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Porphyromonas gingivalis outer membrane vesicles promote apoptosis via msRNA-regulated DNA methylation in periodontitis

Project description:The outer membrane vesicles (OMVs) produced by Porphyromonas gingivalis (P. gingivalis) contain a variety of bioactive molecules that may be involved in the progression of periodontitis. However, the participation of P. gingivalis OMVs in the development of periodontitis has not been elucidated. Here we isolated P. gingivalis OMVs and confirmed their participation in periodontitis both in vivo and in vitro. Microcomputed Tomography (micro-CT) and histological analysis showed that under the stimulation of P. gingivalis OMVs, the alveolar bone of rats was significantly resorbed in vivo. We found that P. gingivalis OMVs were taken up by hPDLCs (human Periodontal Ligament Cells, hPDLCs) in vitro, then subsequently resulting in apoptosis and inflammatory cytokines releasing which was accomplished by the microRNA-size small RNAs (msRNAs) sRNA45033 in the P. gingivalis OMVs. Through bioinformatics analysis and screening of target genes, Chromobox 5 (CBX5) was identified as the downstream target of screened-out small RNA s45033. Using dual-luciferase reporter assay, overexpression, and knockdown methods, s45033 was confirmed to target CBX5 to regulate hPDLCs apoptosis. In addition, Cleavage Under Targets and Tagmentation (Cut&Tag) analysis confirmed the mechanism that CBX5 regulates apoptosis through the methylation of p53 DNA. Collectively, these findings indicate that the role of P. gingivalis OMVs is immunologically relevant and related to bacterial virulence in the development of periodontitis.

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Using small RNA-Seq to verify that 5' tRNA-fMet1 half is over-expressed in outer membrane vesicles (OMVs)

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Project description:Asthma is the most common chronic respiratory disease. Asthma that cannot be well controlled by steroid treatment is called steroid-resistant asthma. Steroid-resistant asthma accounts for only 5% of all asthma cases, but it accounts for 80% of asthma healthcare costs. Nontypeable Haemophilus influenzae (NTHi), as a Gram-negative bacterium, can release outer membrane vesicles (OMVs) and transfer biomolecules to host cells and the external environment by carrying lipopolysaccharides, proteins, peptidoglycans, outer membrane proteins, cell wall components, proteins, nucleic acids, ion metabolites, and signaling molecules. Thus, it plays a role in obtaining nutrition, stress, toxin delivery, adhesion, host immune surveillance evasion, and host immune response regulation. It becomes an essential way in bacterial pathogenesis. To further clarify whether NTHi OMVs could be inhaled to induce steroid-resistant asthma, we isolated and purified NTHi OMVs. In vivo experiments showed that NTHi OMVs could be inhaled and enter airway epithelial cells. Cosensitization with OVA induces steroid-resistant asthma in mice. Furthermore, through high-throughput sequencing, we found that the NTHi OMVs and OVA co-sensitized mice had significantly enriched inflammatory and immune-related signaling pathways, and the transcription and secretion of IL-1β were increased was the potential cause of SRA.

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Project description:This study is the first to show transfer of regulatory bacterial sRNAs from bacterial OMVs to host cells. Demonstration of transfer of bacterial sRNA from Pseudomonas aeruginosa OMVs to host cells in two cell types. RNA isolated from PA14 OMV exposed primary human bronchial epithelial cells or CFBE41o- bronchial epithelial cells as well as unexposed control cells was analyzed by small RNA-Seq. Primary cell exposures were performed on two donors and exposures of CFBE41o- cells were done in triplicate.

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