Metabolomics

Biography

Biography: Anna Vanyushkina

Abstract

Bacteroides represent a major constituent of the human gut microbiota. However pathogenic strain Bacteroides fragilis BOB25 stimulates human intestinal epithelial degeneration and cause necrosis associated with inflammation and colorectal neoplasia of colon epithelial cells. An important feature of B.fragilis is an intense release of outer membrane vesicles. These vesicles could be easily absorbed by the colon epithelium. We shown the vesicles include numerous biochemical compounds, including proteins and metabolites. Considering vesicles as delivery system the question is: what components of the vesicles can affect the intestinal epithelium.

To identify qualitative and quantitative biochemistry composition of the B.fragilis vesicles we performed its proteomic and metabolomic analysis. Proteomic analysis of the vesicles we performed by mass spectrometer TripleTOF 5600 (ABSciex), combined with the HPLC system using IDA method. Metabolomic analysis of the BF vesicles we performed using HPLC-QQQ 8030 (Shimadzu) in the MRM analysis mode.

We detected more than 200 different vesicles proteins representing membranic, periplasmic, and cytosolic fractions of B.fragilis cells. It is important that among the identified proteins we identified a number of proteases, hydrolases and oxidoreductases and patatin, that potentially cause significant changes in physiology of colon epithelium cells. Metabolome analysis allowed to reveal differences in vesicles composition of nontoxigenic and toxigenic B.fragilis components of amino acids, nucleosides, nitrogen bases and cofactors metabolic pathways. This findings open a huge perspectives for studying of cell-host interactions mechanism and therapy development for inflammatory bowel diseases with high colon cancer risk caused by B.fragilis. The study was funded by RSCF grant 142400159.