Dataset Information

Protein compositions of extracellular vesicles released from Staphylococcus aurues JE2 and its lgt mutant

ABSTRACT: This project detected the protein composition of extracellular vesicles (EV) produced by S. aureus WT JE2 strain and its lgt mutant (lipoprotein deficient mutant). Our recent study found that mutation of lgt in S. aureus USA300 JE2 strain significantly increased EV production in S. aureus and decreased the pore-forming toxin content of vesicles, indicatiung that lipoproteins modulate the EV biogenesis of S. aureus. To gain a better understanding of this process, we detected the protein compositions associated with EVs purified from both WT and lgt mutant EVs by LC-MS/MS. Compared to WT EVs, more cytosoli proteins and less extracellular proteins were identified in lgt mutant EVs. A total of 180 and 198 proteins were identified from WT and lgt mutant EVs, respectively.In addition 27.2% and 33.8% of identified proteins were unique to WT or lgt mutant EVs respectively.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Staphylococcus Aureus

SUBMITTER: Xiaogang Wang

LAB HEAD: Jean C. Lee

PROVIDER: PXD014888 | Pride | 2021-09-08

REPOSITORIES: Pride

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Publications

Staphylococcus aureus Extracellular Vesicles: A Story of Toxicity and the Stress of 2020.

Wang Xiaogang X Koffi Paul F PF English Olivia F OF Lee Jean C JC

Toxins 20210120 2

Staphylococcus aureus generates and releases extracellular vesicles (EVs) that package cytosolic, cell-wall associated, and membrane proteins, as well as glycopolymers and exoproteins, including alpha hemolysin, leukocidins, phenol-soluble modulins, superantigens, and enzymes. S. aureus EVs, but not EVs from pore-forming toxin-deficient strains, were cytolytic for a variety of mammalian cell types, but EV internalization was not essential for cytotoxicity. Because S. aureus ...[more]

PMID: 33498438

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Project description:Introduction: Cell-derived extracellular vesicles ([EVs], i.e., exosomes and microparticles) have been reported to mediate the cardioprotective effects of stem cells. However, a head-to-head comparison of their effects with those of the cells from which they derive has not been reported. Hypothesis: The effects of the EV content of human embryonic stem cell (hESC)-derived cardiac progenitors may be equivalent to those of the parent cells. Methods: Two series of immunodeficient (nude) mice underwent permanent coronary artery ligation. Three weeks later, those with an echocardiographically-determined LV ejection fraction â¤ 50% were randomly allocated to receive transcutaneous echo-guided peri-infarct injections of alpha-MEM media (controls), hESC-derived SSEA-1+ cardiac progenitors (500000 cells in 30 Î¼L), or total EV secreted by those 500000 cells in 48 hours in an equivalent volume. The second series also included sham-operated animals for which the needle was inserted into the myocardium in three places without injection. EVs were collected from the cell-secreted medium by ultracentrifugation and characterized by flow cytometry, and nanosight NTS. Outcomes were assessed after 6 weeks by echocardiography, taking LV end-systolic volume (ESV) as the primary end point. Hearts were processed for the assessment of fibrosis and angiogenesis. Total RNA was extracted from carefully selected animals for gene expression analysis by Affymetrix chip. Only animals with similar VTS at baseline were used for this analysis. All data were collected and analyzed blindly. Results: Cardiac progenitors were successfully generated by exposure of the pluripotent ESC to bone morphogenetic protein-2, purified by anti-CD15 immunomagnetic sorting and then cultured on vitronectin for 48 hours, after which cells or EVs derived from the same cell batch were injected. After 6 weeks in the first series, LVESV [mÂ±SEM] in controls did not significantly differ from the pre-injection value (difference: -2.64 Â± 1.54 ÂµL, p=0.12, n= 12). Conversely, in the cell-treated group, LVESV significantly decreased by -4.20 Â± 0.96 ÂµL (p=0.0007, n=16). A similar decrease was seen in the EV-treated group: -5.73 Â± 1.21 ÂµL (p=0.0003, n=15). Similar patterns were seen for LV end-diastolic volumes: controls (-2.46 Â± 1.19 ÂµL, p=NS), cells (-4.48 Â± 1.47 ÂµL, p=0.009), EV (-4.29 Â± 1.31 ÂµL, p=0.005). For the second series, the VTD of cell- and EV-treated animals remained stable (cell: -1.85 Â± 3.27 ÂµL, p=NS, n= 4; EV: +0.05 Â± 1.53 ÂµL, p=NS, n = 4), whereas sham- and control-treated animals deteriorated significantly (sham: +2.20 Â± 0.35 ÂµL, p = 0.003, n = 5; control: +3.27 Â± 0.87 ÂµL, p = 0.03, n = 4). The trend held for VTS, though differences were not significant. Analysis of gene expression data revealed interesting pathways that were more highly expressed in cell- and EV-treated animals than in controls and sham-operated animals. Conclusions: These data support the hypothesis that EVs may be critical mediators of the paracrine effects of cell therapy, and for the sake of streamlining translational processes, deserve to be considered as potential alternatives to stem cell transplantation. Gene expression levels were compared between mouse hearts.

Dataset Information

Protein compositions of extracellular vesicles released from Staphylococcus aurues JE2 and its lgt mutant

Publications

<i>Staphylococcus aureus</i> Extracellular Vesicles: A Story of Toxicity and the Stress of 2020.

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