Dataset Information

Fusarium oxysporum f. sp. vasinfectum (UP000030701) extracellular vesicle, secretome, and cell lysate proteomes

ABSTRACT: We performed a comparative study to determine the proteome of extracellular vesicles (EVs) from the cotton pathogen Fusarium oxysporum f. sp. vasinfectum (Fov), recovered from two growth conditions in vitro. Label-free quantitative protemics was used to find significant enrichment of proteins between EV samples, the secretome (secreted-soluble proteins) and the cell lysate. Our results show that some proteins were exclusive to EVs and were upregulated compared to the secretome or cell lysate.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Fusarium Oxysporum F. Sp. Vasinfectum 25433

TISSUE(S): Cell Culture, Fungal Cell

DISEASE(S): Wounds And Injuries

SUBMITTER: Donovan Garcia Ceron

LAB HEAD: Marilyn Anderson

PROVIDER: PXD023907 | Pride | 2021-03-08

REPOSITORIES: Pride

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Size-exclusion chromatography allows the isolation of EVs from the filamentous fungal plant pathogen Fusarium oxysporum f. sp. vasinfectum (Fov).

Garcia-Ceron Donovan D Dawson Charlotte S CS Faou Pierre P Bleackley Mark R MR Anderson Marilyn A MA

Proteomics 20210308 13-14

Extracellular vesicles (EVs) are nano-sized compartments involved in cell communication and macromolecule transport that are well characterized in mammalian organisms. Fungal EVs transport virulence-related cargo and modulate the host immune response, but most work has been focused on human yeast pathogens. Additionally, the study of EVs from filamentous fungi has been hindered by the lack of protein markers and efficient isolation methods. In this study we performed the isolation and proteomic ...[more]

PMID: 33609009

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Project description:Background: Acute coronary syndromes (ACS) are associated with aberrant gene expression and epigenetic mechanisms. In particular, de novo DNA methylation is typically linked to gene silencing, but its role in heart disease remains not fully understood. Extracellular vesicles (EVs) are active components in cellular communication for their ability to carry a plethora of signalling biomolecules, thus representing a promising new diagnostic/therapeutic approach in cardiovascular diseases (CVDs). Indeed, there is the need of novel biomarkers for ACS prediction and timely detection. Purpose: We hypothesized that specific epigenetic signals can be carried by EVs. In this regard, we isolated and characterized circulating EVs from ACS patients and evaluated their potential role to influence DNA methylation in target cells. Methods: Circulating EVs were recovered, by ultracentrifugation, from plasma samples of 19 ACS patients and 50 healthy subjects (HS). Nanoparticle tracking analysis (NTA) and western blot (WB) were used to confirm the EVs integrity and purity. Peripheral blood mononuclear cells (PBMCs) of volunteer donors were treated with both ACS and HS derived EVs and genomic DNA was extracted to perform epigenome wide analysis through Reduced Representation Bisulfite Sequencing. ShinyGO, PANTHER, and STRING tools were interrogated to perform GO and PPI network analyses. Flow Cytometry, qRT-PCR, and WB analysis were also performed to evaluate and validate both intra-vesicular and intra-cellular signals. Results: Plasma ACS-derived EVs showed a significant up-regulation of DNA methyltransferases (DNMTs) gene expression levels as compared to HS (P<0.001). Specifically, de novo methylation transcripts, as DNMT3A and DNMT3B were significantly increased in plasma ACS-EVs. DNA methylation analysis of PBMCs from volunteer donors treated with HS- and ACS-derived EVs showed that RNF166 and CCND3 genes resulted the most hyper- and hypo-methylated, respectively, after by ACS-EV treatment. In addition, PPI network analysis specifically evidenced the subnetwork with SRC, as a hub gene, connecting it to NOTCH1, FOXO3, CDC42, IKBKG, RXRA, DGKG, known as important genes already involved in the onset of CVDs. Surprising, other novel genes, BAIAP2, SYP, CHL1, and SHB, which were hypomethylated, were found significantly overexpressed in PBMCs (P<0.005), underlying the fundamental modulating properties of EV cargo in atherosclerosis. Conclusions: These findings support the significant role of ACS plasma-derived EVs, inducing de novo DNA methylation signals, and modulating specific signaling pathways in target cells.

			Action	DRS
	2019-07-11_uniprot-proteome_UP000030701.tab	Other
	CD_EV1_a.raw	Raw
	CD_EV1_b.raw	Raw
	CD_EV2_a.raw	Raw
	CD_EV2_b.raw	Raw