Stable Isotope Tracing for Folate-mediated One-Carbon Trafficking in Colon Cancer Cells and a 3-D Cell Model of embryogenesis
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Owner/Developer: Agilent Technologies
United States of America
21 July 2016
United States of America
Metabolomics is a powerful tool for probing complex biological mixtures to discover changes in metabolite levels arising from gene mutations, drug treatments, and environmental exposures. However, these metabolite profiling experiments only provide a snapshot of cellular metabolism, devoid of information on the dynamics and flux of metabolites within metabolic networks. Stable isotope tracing has been effective for elucidating metabolic fluxes through biochemical pathways and networks. However this approach has largely been limited to the study of predicted tracer-derived metabolites.
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In vitro methods
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This e-seminar describes an untargeted stable isotope tracing (USIT) approach that seeks to overcome this limitation. This technique enables the discovery of disparate temporal fates for one or more heavy isotope-labeled atoms in a precursor metabolite of interest.
Applying this approach, the presentation comprehensively describes the array of folate-dependent one-carbon (1C) trafficking reactions, which contribute to the serine addiction of rapidly proliferating cells (in models of cancer and early embryogenesis). The USIT approach does not require any preconceived knowledge of metabolites that undergo isotopic enrichment (or cognate metabolic pathways). In addition, < 1% isotopic incorporation is ample for confident discovery of metabolic fates. The folate-dependent 1C tracing findings reveal unexpected molecular determinants of 1C fate and flux. In addition to recognition of a specific 1C transfer reaction that is apparently most essential for the support of cancer cell growth and embryonic development.
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