Metabolomics Platform

The platform is established at the Institute for Clinical Chemistry and Laboratory Medicine of the University hospital Dresden and contributes to the mission of the NCT/UCC Dresden.

Alterations in cellular metabolism are a hallmark of many cancer types. Research in this context aims at understanding the role of cellular metabolism in tumorigenesis as well as developing new tumor metabolism-related therapies.

Metabolomics represent the comprehensive analysis of small molecules (i.e., metabolites) in body fluids, cells, tissues and tissue biopsies. Therefore, to facilitate research in cancer metabolism, the Metabolomics-Platform offers metabolite analysis based on complementary methods, specifically, Nuclear Magnetic Resonance (NMR) Spectroscopy and Mass Spectrometry (MS).

NMR-Spectroscopy

NMR Spectroscopy can quantify metabolites in a sample; the 1H NMR spectrum represents the metabolic fingerprint. It allows the identification of metabolic changes in a wide range of media. NMR Spectroscopy is an automated, simple (minimal sample preparation) and highly reproducible technique that can provide a quantitative fingerprint/profile with a 20-minute measurement.
Quantitative metabolomics via NMR Spectroscopy (Figure 1):

  • Measurement in liquid samples (quantitative analysis of up to 150 metabolites in urine; 50 metabolites in serum/plasma)
  • HR-MAS (High-Resolution Magic Angle Spinning) for measurement of metabolites in semi-solid samples (e.g. intact tissues, tumor biopsies)

Figure 1: NMR-Spectroscopy pipeline

Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS)

LC-MS/MS is a powerful tool for sensitive and specific, quantitative and qualitative analysis of small molecule metabolites. Depending on the type of mass spectrometer, different processes (such as filtration of specific mass-to-charge ratios, collision-induced dissociation or fragmentation, full mass range scans) are available, allowing for accurate quantification in targeted approaches or screening and identification of unknown substances in untargeted setups. Within the Metabolomics-Platform, LC-MS/MS-based targeted approaches include analysis of e.g. tricarboxylic acid (TCA) cycle metabolites, biogenic (poly)-amines, steroids, catecholamine metabolites, or bile acids, as well as untargeted approaches for metabolomic screenings.

Metabolic flux analysis

LC-MS/MS methodology is complemented by stable isotope metabolic flux analysis for a more dynamic understanding of the metabolic pathophysiology. Stable isotope metabolic flux analysis allows the identification of the source of oncometabolites, the rate at which they are produced as well as the regulation of such reactions in cell culture models. Depending on the initial tracer used, we can study glycolysis, TCA cycle (forward and reverse reactions), glucose and fatty acid oxidation and glutamine metabolism – all pathways implicated in cancer pathophysiology (Figure 2).

Figure 2: Quantifiable metabolic fluxes using 13C-glucose (blue and purple) or 13C-glutamine tracers (red and purple).

Service

  • NMR-Spectroscopy analysis in liquid samples and tissues
  • LC-MS/MS: Targeted and untargeted profiling of small molecule metabolites in different samples
  • Consulting service for setup of experimental designs in translational research projects
  • Planning, analysis and interpretation of metabolic flux data
  • Methodology-/assay-setup for translational research projects on request

Methods:

NMR-Spectroscopy

  • Metabolite measurement in liquid samples
  • HR-MAS for measurement of metabolites in semi-solid samples


Targeted LC-MS/MS

  • TCA cycle metabolite profiling (cell culture)
  • Steroids (plasma, tissue, cell culture)
  • Catecholamine metabolites (plasma, urine, tissue, cell culture)
  • Biogenic Amines within the Arginine metabolism (cell culture, serum, urine, feces, tissue)
  • Bile acids (plasma, tissue)
  • Further methods on request


Untargeted LC-MS/MS-based Metabolomics (plasma, cell culture, tissue)

13C-Based Metabolic flux analysis (cell culture)

 

 

 

DIRECTOR METABOLOMICS-PLATFORM

Prof. Dr. Triantafyllos Chavakis, Institute for Clinical Chemistry and Laboratory Medicine of the University hospital Dresden

HEAD, EXPERIMENTAL LC-MS/MS FACILITY

Dr. Mirko Peitzsch, Institute for Clinical Chemistry and Laboratory Medicine, University Hospital Carl Gustav Carus, TU Dresden
Email: Mirko.Peitzsch(at)uniklinikum-dresden.de
Tel: +49 (0) 351 458 14578

HEAD, NMR-SPECTROSCOPY

Dr. Alexander Funk, Institute for Clinical Chemistry and Laboratory Medicine, University Hospital Carl Gustav Carus, TU Dresden
Email: alexander.funk(at)uniklinikum-dresden.de
Tel:  +49 (0) 351 458 11319

HEAD, METABOLIC FLUX

Dr. Tiago Alves, Institute for Clinical Chemistry and Laboratory Medicine, University Hospital Carl Gustav Carus, TU Dresden
Email: tiago.alves(at)uniklinikum-dresden.de
Tel: +49 (0) 351 458 11707