NMR spectroscopy detects metabolite differences between culture media of hepatitis C virus negative and positive cells after harvest

Diserens, Gaëlle; Vermathen, Martina; Lévy, P; Dufour, J-F; Vermathen, Peter; Bartosch, B (27 June 2016). NMR spectroscopy detects metabolite differences between culture media of hepatitis C virus negative and positive cells after harvest (Unpublished). In: 12th Annual Conference of the Metabolomics Society. Dublin, Ireland. 27.-30.06.2016.

Introduction:
Chronic infection with hepatitis C virus (HCV) is one of the main causes of hepatocellular
carcinoma. Given the importance of glutamine in carcinogenesis, we investigated the role of
glutaminolysis in HCV infection. Here we report on a NMR substudy investigating metabolite
differences between cell culture supernatants of hepatitis C virus negative (HCV) and positive (HCV+) cells after harvest.
Methods:
HCV infected and uninfected cells were cultured in conditioned Dulbecco’s Modified Eagle’s
Medium complemented with (1) additional Glucose and Glutamine, (2) void of Glutamine or
(3) void of Glucose. Cells and supernatants were collected at days 3, 4, and 5 after addition
of conditioned cell media. For each medium composition and time point, three HCV positive and three HCV negative cell cultures were harvested.
Watersuppressed 1DNOESY spectra of 54 cell supernatants were acquired on a 400MHz
Bruker AvanceII spectrometer at 298K. Spectra were bucketed and buckets from control
culture media were subtracted from those of the supernatant spectra, and scaled to the
number of cells at the harvest point.
Results:
A complete separation in PLSDA was obtained between cell media in contact with either
HCV+ or HCVcells for all three media types. Moreover, the media separated further with
increasing time post incubation. Two media separated also well between those in contact
with HCV+ or HCVcells in unsupervised PCA. Loading plots demonstrated contributions
from numerous metabolites responsible for the separation including glutamine and glutamate.
Discussion:
HCV infected cells appear to have different nutrition and excretion pathways compared to
noninfected cells. Most importantly glutamine was reduced, while glutamate was increased
in HCV+. These NMR results support complementary measurements demonstrating that
HCV modulates the transcript levels of key enzymes of the glutamine metabolism in vitro and
in liver biopsies of chronic HCV patients. Consistently, HCV infection increased glutamine
utilization and dependence.

Interest & Impact

Downloads

0 since deposited on 26 Jan 2017
0 in the past 12 months

Citations

5 Citations in Web of Science ®
6 Citations in Scopus

Search

in Google Scholar™

Services

Actions (login required)

Edit item

Actions (login required)

Edit item