Microfluidics meets metabolomics to reveal the impact of Campylobacter jejuni infection on biochemical pathways

Abstract

Microfluidic devices that are currently being used in pharmaceutical research also have a significant potential for utilization in investigating exposure to infectious agents. We have established a microfluidic device cultured with Caco-2 cells, and utilized metabolomics to investigate the biochemical responses to the bacterial pathogen Campylobacter jejuni. In the microfluidic devices, Caco-2 cells polarize at day 5, are uniform, have defined brush borders and tight junctions, and form a mucus layer. Metabolomics analysis of cell culture media collected from both Caco-2 cell culture systems demonstrated a more metabolic homogenous biochemical profile in the media collected from microfluidic devices, compared with media collected from transwells. GeneGo pathway mapping indicated that aminoacyl-tRNA biosynthesis was perturbed by fluid flow, suggesting that fluid dynamics and shear stress impacts the cells translational quality control. Both microfluidic device and transwell culturing systems were used to investigate the impact of Campylobacter jejuni infection on biochemical processes. Caco-2 cells cultured in either system were infected at day 5 with C. jejuni 81-176 for 48 hours. Metabolomics analysis clearly differentiated C. jejuni 81-176 infected and non-infected medias collected from the microfluidic devices, and demonstrated that C. jejuni 81-176 infection in microfluidic devices impacts branched-chain amino acid metabolism, glycolysis, and gluconeogenesis. In contrast, no distinction was seen in the biochemical profiles of infected versus non-infected media collected from cells cultured in transwells. Microfluidic culturing conditions demonstrated a more metabolically homogenous cell population, and present the opportunity for studying host-pathogen interactions for extended periods of time.

Porcine circovirus type 2 (PCV2) and Campylobacter infection induce diarrhea in piglets: Microbial dysbiosis and intestinal disorder

“…Here, PCV2 infection did induce the inflammation of colon accompanied by a significant increase in colonic permeability in terms of reduction of Claudin-1, Occludin, and ZO-1, consistent with the previous studies on piglets ( Kim et al., 2004 ; Meng, 2013 ). Similarly, Campylobacter infection significantly increased intestinal permeability by affecting the tight junctions, in accordance with previous findings in mouse and humans ( Liu et al., 2018 ; Mortensen et al., 2016 ). And thus, the increase in intestinal permeability in turn would further aggravate the invasion of PCV2 and Campylobacter , and then led to the diarrhea in piglets.…”

A Metabolomics Pilot Study on Desmoid Tumors and Novel Drug Candidates

“…Data acquisition, statistics, and pathway analysis were performed as previously described 17 , 70 , 71 . Three NMR spectra were acquired for each of the individual study samples and the pooled samples.…”

Single-Cell Omics Analyses Enabled by Microchip Technologies

“…Microfluidic devices offer great potential as powerful tools for higher-throughput metabolomic studies on single cells. Although microfluidic devices have been used as a culturing platform to investigate metabolites, few studies have applied these devices to investigate the metabolome at the single-cell level (175)(176)(177). Another challenge remains in that most single-cell devices have arisen only for specific metabolites, restricting their application to niche specialties and failing to achieve highly multiplexed metabolite analysis.…”