Metabolite levels and turnover rates are the most sensitive indicators of body energy and metabolic imbalances created by genetic deficiencies, myocardial ischemia, diabetes, aging and obesity. Next-generation screening of disease-related metabolic phenotypes requires development technologies for monitoring metabolites, their turnover rates and flux distribution within large-scale metabolic networks. Dynamic metabolomic profiling may uncover early changes in cell energetics systems, phosphotransfer networks and metabolic signaling circuits in human diseases.
Traditional metabolomic diagnostic platforms are typically based on measurement of static metabolite levels. The Metabolomics Nuclear Magnetic Resonance Spectroscopy (NMRS) core's developed metabolomic platform allows researchers to synchronously track energy metabolite levels and their respective turnover rates through different pathways that ultimately determine the phenotype, facilitating metabolic fingerprinting and disease diagnosis.
Metabolite level and turnover rate analysis using 31P NMR and 18O-assisted 31P NMR spectroscopy. Using this approach, up to 50 phosphorus-containing metabolites can be analyzed.
Analytical platform for dynamic phosphometabolomic analysis of metabolic phenotypes of human diseases. A. Analytical technologies; B. Static versus dynamic metabolomics metabolite levels and turnover rates by 18O-labeling as detected by 18O-induced shift in 31P NMR metabolite spectra; C. Deduction of metabolic network dynamics and enzymatic fluxes from 18O labeling data.
Phosphometabolomics is an emerging area in metabolomic analyses that targets more than 500 phosphometabolites critical in metabolic phenotyping and medical diagnostics. Phosphometabolites represent a major class of metabolites indispensable to life activities, such as genetic inheritance, signal transduction, metabolism and energy transduction.
Phosphometabolite levels and turnover rates are the most sensitive indicators of body energy imbalances in disease states. Large-scale 18O stable isotope-based metabolomic technology allows determination of dynamic metabolomic signatures of metabolic deficiencies critical for diagnosis and monitoring of human diseases. This methodology permits accurate measurements of phosphometabolite and oligophosphate turnover rates based on LC/ESI-MS and 2-D 31P NMR correlation spectroscopy of 18O/16O exchange and isotopologue statistics.
Potential applications include monitoring of:
- Adenosine triphosphate (ATP) synthesis and utilization
- Phosphotransfer enzyme rates (creatine kinase, adenylate kinase and nucleoside diphosphate kinase)
- Glycolytic and glycogenolytic metabolite levels and fluxes
- Mitochondrial Krebs and urea cycle activity
- Mitochondrial substrate (G3P) shuttle activity
- Adenosine monophosphate (AMP) metabolic signal dynamics
- Purine and pyrimidine nucleotide turnover rates
Studies demonstrate that this approach is valuable for metabolomic profiling of transgenic models simulating human disease and diagnosis of mitochondrial deficiency. Monitoring phosphometabolite turnover rates and targeted analysis of metabolic network dynamics by 18O labeling in human samples allow metabolic phenotyping for prediction, diagnosis and treatment of human diseases. Human whole blood 18O metabolic labeling methodology has the ability to detect a subtle alteration in network dynamics and metabolomic signatures of human diseases associated with risk of thrombosis and sudden death.
Beyond medical diagnostics, this technology provides a useful platform for system metabolomics studies in health and diseases, drug development, evaluation of treatment efficiency, and determination of drug side effects.