Dataset Information

The proteome and lipidome of extracellular vesicles from Haemonchus contortus to underpin explorations of host-para-site cross-talk

ABSTRACT: Many parasitic worms have a major adverse impact in human and animal populations worldwide due to the chronicity of their infections. There is a growing body of evidence indicating that extracellular vesicles (EVs) are intimately involved in modulating (suppressing) inflamma-tory/immune host responses and parasitism. As one of the most pathogenic nematodes of livestock animals, Haemonchus contortus is an ideal model system for EV exploration. Here, employing a multi-step enrichment process (in vitro culture, followed by ultracentrifugation, size exclusion and filtration), we enriched EVs from H. contortus and undertook the first comprehensive (qualitative and quantitative) multi-omic investigation of EV proteins and lipids using advanced liquid chro-matography–mass spectrometry and informatic methods. We identified and quantified 561 proteins and 446 lipids in EVs, and compared these molecules with those of adult worms. We identified unique molecules in EVs, such as proteins linked to lipid transportation and lipid species (i.e. glyc-erophospholipids and sphingolipids) associated with signalling, indicating an involvement of these molecules in parasite-host cross-talk. This work provides a solid starting point to explore the func-tional roles of EV-specific proteins and lipids in modulating parasite-host cross talk, and the pro-spect of finding ways of disrupting or interrupting this relationship to suppress or eliminate para-site infection.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Caenorhabditis Elegans

TISSUE(S): Stem Cell, Whole Body

DISEASE(S): Parasitic Helminthiasis Infectious Disease

SUBMITTER: Tao Wang

LAB HEAD: Robin Beat Gasser

PROVIDER: PXD043320 | Pride | 2023-08-07

REPOSITORIES: Pride

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The Proteome and Lipidome of Extracellular Vesicles from <i>Haemonchus contortus</i> to Underpin Explorations of Host-Parasite Cross-Talk.

Wang Tao T Koukoulis Tiana F TF Vella Laura J LJ Su Huaqi H Purnianto Adityas A Nie Shuai S Ang Ching-Seng CS Ma Guangxu G Korhonen Pasi K PK Taki Aya C AC Williamson Nicholas A NA Reid Gavin E GE Gasser Robin B RB

International journal of molecular sciences 20230630 13

Many parasitic worms have a major adverse impact on human and animal populations worldwide due to the chronicity of their infections. There is a growing body of evidence indicating that extracellular vesicles (EVs) are intimately involved in modulating (suppressing) inflammatory/immune host responses and parasitism. As one of the most pathogenic nematodes of livestock animals, <i>Haemonchus contortus</i> is an ideal model system for EV exploration. Here, employing a multi-step enrichment process ...[more]

PMID: 37446130

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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.

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