This proposal intends to establish a Metabolomics Junior Research Group located at the Luxembourg Center of Systems Biomedicine (LCSB). A major focus of the new group will be the installation of cutting-edge metabolomics technologies in Luxembourg and their direct application to address prevailing problems in the field of cancer metabolism. Using software (MetaboliteDetector) and experimental protocols that I already developed, the initial system will provide a platform for the measurement of several hundred metabolites present in various types of samples including plasma, urine, tissue and bacterial or eukaryotic cells. Moreover, the novel Non-targeted Tracer Fate Detection (NTFD) algorithm for the quantitative and non-targeted analysis of intracellular fluxes and thus enzyme activity – developed by me at the Massachusetts Institute of Technology (MIT) – will be installed. Metabolomics stands for the comprehensive analysis of intracellular and extracellular metabolite concentrations. With up to 400 identified metabolites in one experiment an overall picture of the metabolism emerges. The measurement of cellular metabolite concentrations plays an important role in understanding the rapid response of metabolic flux changes via allosteric or feedback inhibition mechanisms to intra- or extracellular signals. Since the set of all metabolites is directly linked to the actual state of a cell and thus to the phenotype, the metabolome is optimally suited for the determination of biomarkers that are typical for certain genotypes or diseases. In combination with stable isotopic tracers and sophisticated mathematical algorithms a dynamic dimension can be added: the measurement of fluxes through the metabolic reaction network. In addition to genomics, transcriptomics, and proteomics, metabolomics techniques are essential to get a systems level understanding of living systems and disease formation. The proposed project is structured in two technological and three applied modules: 1. Metabolomics Technologies: The technological modules of the project aim to establish an up-to-date metabolomics analytical platform in Luxembourg. GC/MS based metabolomics techniques allow for the detection of metabolites of the central carbon metabolism including most of the amino acids, sugars, organic acids and nucleotides. In addition HPLC/MS complements the detection of the aforementioned metabolites with phosphorylated- and coenzyme coupled metabolites. Starting with gas chromatography coupled to mass spectrometry (GC/MS) the analytical pipeline will be extended to high performance liquid chromatography coupled to mass spectrometry (HPLC/MS). In parallel, the MetaboliteDetector software will be further developed and adopted for the analysis of HPLC/MS based data. Finally, the NTFD technology will be extended for the non-targeted reconstruction of metabolic reaction networks based solely on stable isotopic labeled metabolomics data. 2. Applications for analyzing cancer metabolism: The three applied modules of this proposal address prevailing questions in cancer metabolism. In this context, we will analyze the effect of increased sarcosine levels on the metabolism of prostate cancer cells. Increased sarcosine levels have been shown to induce an invasive phenotype in these cells. The second module intends to systematically and comparatively analyze the metabolism of several cancer cell lines. Finally, the third module aims to determine metabolic effects of different PDGF receptor a mutations. I trust, the integration of experimental and computational methodologies in combination with their direct application in the medically relevant field of cancer metabolism makes this project competitive and attractive for Luxembourg’s research environment.