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detects metabolite differences between culture media of hepatitis C virus negative and positive cells after harvest (Unpublished).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.

Item Type:

Conference or Workshop Item (Poster)

Division/Institute:

08 Faculty of Science > Departement of Chemistry and Biochemistry
04 Faculty of Medicine > Department of Radiology, Neuroradiology and Nuclear Medicine (DRNN) > Institute of Diagnostic, Interventional and Paediatric Radiology > DCR Magnetic Resonance Spectroscopy and Methodology (AMSM)
04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR) > Forschungsbereich Pavillon 52 > Abt. Magnetresonanz-Spektroskopie und Methodologie, AMSM

UniBE Contributor:

Diserens, Gaëlle; Vermathen, Martina and Vermathen, Peter

Subjects:

500 Science > 570 Life sciences; biology
500 Science > 540 Chemistry

Funders:

[4] Swiss National Science Foundation

Language:

English

Submitter:

Martina Vermathen

Date Deposited:

26 Jan 2017 09:08

Last Modified:

26 Jan 2017 09:08

URI:

https://boris.unibe.ch/id/eprint/92519

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