The essential scientific question in this project is how metabolism is linked the to the known cellular functions in cell metabolism. There are several parallel mechanisms involved in regulating cellular metabolism, and this is regulated via signalling pathways that change the metabolic landscape of cells.
Metabolic regulation is responsive to metabolite levels, but may also be regulated by signalling pathways causing the expression or suppression of metabolically active enzymes. Furthermore, recent evidence indicates that co-factors, signalling molecules and metabolites directly participate in the regulation of gene expression through modulating enzyme activities that act on chromatin. The understanding of these regulatory mechanisms is vital but currently lacking. To fully understand metabolic regulation, the mechanisms involved need to be studied at different levels. Firstly, it is essential to identify the regulation of metabolic enzyme expression. Secondly, the impact of perturbation of metabolic enzymes on cellular state must be assessed qualitatively and quantitatively. Thirdly, metabolic (de)regulation needs to be analysed at the metabolite level using different types of metabolic analyses. Finally, the link between regulatory small molecules (co-factors, signalling molecules and metabolites), chromatin and gene expression need to be more accurately defined.
The project will use a selected panel of haematological cancer cell lines to identify mechanisms of metabolic regulation. These lines were chosen since they cover a wide range of the most recurrent mutations found in AML and CML patients. Besides these cells lines, the metabolome of primary leukaemia patient samples will also be investigated using cells that are already available in biobanks of partners within HaemMetabolome. Finally, various lentiviral model systems are available which will allow a careful evaluation of metabolic changes in normal and oncogene expressing stem/progenitor cells, both during initiation as well as during maintenance of disease.
We will employ a panel of the most commonly used drugs to treat haematological cancers and expose these cells to treatment. In each case the metabolic response will be monitored and correlated to changes on the transcriptome level.
This project combines complementary expertise in haematological cancer cell biology, metabolomics and systems modelling to characterise metabolism across a range of systems, and will for the first time elucidate underlying mechanisms how cellular metabolism regulates cell maintenance, proliferation and differentiation. For this we include key technologies to prepare and purify cells, and to study cellular metabolism using mass spectrometry (MS) and Nuclear Magnetic Resonance (NMR) approaches. This highly interdisciplinary team of scientist will provide training for 10 ESRs in a variety of disciplines linking diverse skills including cancer cell biology, key ‘omics’ technologies with accompanying essential bioinformatics, mathematical modelling, and drug discovery. This breadth of technologies will offer students an excellent training programme with a variety of inter-disciplinary skills.