NYR-Diabetes Research Center (NYR-DRC)

Abstract The mission of the Stable Isotope & Metabolomics Core (SIMC) is to provide NYR-DRC research investigators cost effective, efficient and robust platforms for the determination of metabolites/lipids in cells, tissues and biofluids. This consists of steady-state metabolite and lipid species levels, metabolic flux determined by stable isotope labeling, which include spatial profiling at cellular resolution within tissues using a variety of biochemical, analytical and mass spectroscopy approaches. The SIMC mass spectrometric platform is designed to cover intersecting metabolism pathways, to improve our understanding of conditions which change fuel utilization and biosynthesis that occur both in the cytosolic and mitochondria, i.e., glycolysis, pentose and tricarboxylic acid (TCA) cycles, their interaction with one carbon metabolism, TCA metabolism yielding acetyl, malonyl, propionyl, succinyl CoAs (and others) and their linkage to epigenetic modifications (methylation and histone modifications by the mentioned CoA species) as well as phospholipid and triglyceride recycling pathways. Spatial mass spectroscopy imaging (MSI) at cellular resolution can be related/projected onto other spatial methodologies such as Visium spatial transcriptomics, or spatial proteomic imaging to assess multi-omic cellular atlases which include metabolic flux, metabolites and lipid profiles. Differences in cellular function within organs adds to the complexity for understanding physiological regulation or pathogenesis of metabolic diseases. To accomplish these goals, SIMC utilizes state-of-the-art liquid chromatography for targeted (Sciex 6500+ QTRAP and Waters Absolute TQ triple quadrapole) mass spectrometry and untargeted (Waters Cyclic Ion Mobility Time of Flight MS) and gas chromatography (Agilent-- -single quadrapole MS) based metabolite and lipid analyses. Additionally, specific changes in metabolic flux in carbohydrate (i.e. glucose production and disposal) and lipid (i.e. lipogenesis) metabolism are determined using a variety of stable isotope tracers for in vitro (cell culture or organoids) and in vivo (whole animal and spatial cellular localization within organs). Both tissue culture, and tissue derived (islet cells, adipocytes, muscle tissue) experiments can also be coupled with Seahorse Flux analyzers for the measurements of mitochondrial function and rates of glycolysis. Metabolomic/lipidomic assays are applied to a variety of rodent and human studies determined from various biological fluids and materials (plasma, urine, synovial, lymph, fecal). The overall strategy is to provide set of metabolomic/lipidomic data analyses, that together with Seahorse, and stable isotope assessment of metabolic rates, provide complementary information to synergize with the Animal Physiology, Human Islet & Adenovirus, and Translational Research Cores of the NYR-DRC to inform detailed understanding of experimental research questions. SIMC works in coordination with the Translational Research Core for human studies by establishing the experimental design and quantitative analyses of plasma isotope levels and metabolite profiles.