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Bacterial proteins encapsulated in extracellular membrane vesicles of Escherichia coli-infected macrophages

ABSTRACT: Macrophages-derived extracellular membrane vesicles (EVs), which contain proteins, lipids, metabolisms, mRNA/microRNA, and DNA, are used for signaling in cell-to-cell communications. EVs are known to increase the levels of cytokine and chemokine and promote inflammation. Our object is to identify the inflammatory factors by EVs derived from Escherichia coli-infected macrophages. Therefore, we used MS analysis to examine proteins contained in mouse macrophage (RAW264.7)-derived EVs. The associated research are published in mBio 14, e0305122, 2023 (Imamiya R et al.)

ORGANISM(S): Cellular Organisms

SUBMITTER: Mayuko Osada-Oka

PROVIDER: PXD059569 | JPOST Repository | Mon Jan 20 00:00:00 GMT 2025

REPOSITORIES: jPOST

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Project description:Extracellular vesicles derived from induced pluripotent stem cells (iPSC EVs) have immunoregulatory potential with the ability to alter monocyte-derived macrophages. Macrophages function in the propagation and resolution of inflammation which is mediated by their phenotype. Macrophages are an ideal therapeutic target as modulating their phenotype towards an anti-inflammatory pro-resolving state may be beneficial in chronic inflammatory diseases such as atherosclerosis. Macrophages are naturally phagocytotic cells and readily take up iPSC EVs however the contents of iPSC EVs and their effects on macrophages are poorly understood. Here iPSC EVs were characterized and analysed by mass-spectrometry based proteomics and a targeted microRNA (miR) panel. Their immunomodulatory effects on macrophages were assessed and a monocyte transmigration assay was used to assess the chemotactic potency of the secretome from iPSC EV treated macrophages. Proteomic analysis on iPSC EVs identified Podocalyxin-like protein 1 (PODXL1), Insulin (INS) and Solute Carrier Family 2 (Facilitated Glucose Transporter), Member 3 (SLC2A3) as the most abundant proteins unique to the iPSC EVs when compared to control NT-2 EVs. Notably, thioredoxin and peroxiredoxin related proteins were detected. miR-302d-3p was the most abundant miR in these iPSC EVs. miR-25-3p, previously reported to alter the macrophage phenotype, was significantly increased in comparison to the control NT-2 EVs. iPSC EVs increased expression of the anti-inflammatory associated MRC1 and miR-21 in human primary macrophages and decreased monocyte chemoattractant protein1 (MCP-1). Mass spectrometry based proteomics revealed that treated macrophages had decreased levels of secretory proteins, some of which have chemotactic properties, these included Azurocidin 1 (AZU1), Growth Differentiation Factor 15 (GDF15), and Ribosomal Protein S19 (RPS19). There was a decrease in monocyte transmigration towards conditioned media from macrophages treated with iPSC EVs. Collectively this study provides insights into the protein content and miR cargo of iPSC EVs and highlights their capacity to inhibit chemotactic proteins in macrophages and upregulate MRC1.

			Action	DRS
	EC1.dat	Other
	EC1.mgf	Mgf
	EC1.mzid	Mzid
	EC1.raw	Raw
	EC2.dat	Other

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Bacterial proteins encapsulated in extracellular membrane vesicles of Escherichia coli-infected macrophages

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