An important goal of basic biomedical research is understanding the molecular mechanisms that cause disease in order to enable the development of new therapeutic treatments. Towards this goal, the proposal aims to analyze cellular models of diabetes by discovery metabolite profiling (DMP) to identify dysregulated networks that promote diabetes. Given that DMP interrogates a portion of biomolecular space (metabolites) that is inaccessible by other methods, this approach is likely to uncover important metabolic pathways involved in the onset and maintenance of type 2 diabetes. DMP quantifies differences in metabolite levels between two samples using an untargeted liquid chromatography-mass spectrometry (LC-MS) method. Current protocols for DMP, which are limited to lipid analysis, will be expanded to include peptides and carbohydrates, two important metabolite classes associated with diabetes. These protocols will then be applied to identify changes in metabolic pathways that occur during adipogenesis and insulin resistance in the 3T3-L1 cell line. Both known and undiscovered metabolites will be identified and the pathways regulating the levels of these compounds examined. We anticipate that this approach will provide a unique view - one that differs substantially from genomics - of the molecular pathways involved in diabetes. Additionally, DMP will afford key insights that will guide subsequent hypothesis driven research aimed at understanding the specific biochemical, cellular, and physiological functions of the newly discovered metabolites and metabolic pathways. Lastly, this research can eventually lead to the development of better therapeutic treatments for type 2 diabetes by gaining a clearer understanding of the dysregulated molecular pathways that cause the disease. Diabetes affects millions of people world-wide and leads to many serious long-term health problems including heart disease, blindness, kidney failure and nerve damage. This proposal aims to elucidate the mechanism of the onset and maintenance of this growing epidemic in order to enable the development of new therapeutic treatments. ? ? ?
