Stable Isotope & Metabolomics Core

稳定同位素和代谢组学核心(SIMC)的使命是提供指导性代谢物和底物 在复杂的体内代谢啮齿动物模型和用于人类临床研究的通量测定 爱因斯坦-西奈山糖尿病研究中心(ES-DRC)成员。SIMC提供了一组输入 增强体内研究的体外代谢方法。这些服务为调查人员提供了 专门测定底物通量动态和细胞器、细胞、 组织和全身水平，从而阐明了葡萄糖、蛋白质和 脂类代谢。通过与ES-DRC其他核心的这些协作努力，确定了 对药理、饮食、环境和基因改变的影响进行了彻底的表征 糖脂动态平衡、胰岛素作用和新陈代谢。候选分子在相关基因中的作用 与葡萄糖有关的组织(即神经元、肝细胞、骨骼肌、脂肪细胞和β细胞) 通过在体内和体外进行彻底和明确的分析，可以特别地描绘出体内的动态平衡 在啮齿动物和人类以及细胞系中进行循序渐进的实验。至 为了实现这些目标，SIMC将：1)进行体内稳定的同位素底物通量分析 蛋白质合成，脂肪生成，外周葡萄糖处置，肝脏葡萄糖循环， 葡萄糖-甘油循环和葡萄糖-乳酸循环；2)测定糖酵解(细胞外酸化率) 和线粒体耗氧量(线粒体呼吸)在分离的细胞、组织外植体或组织中 培养，使用海马生物科学通量分析仪，以及更全面的稳定同位素通量 评估；3)对血浆和组织代谢物情况进行有针对性的假设驱动评估 糖酵解/糖异生、磷酸戊糖和三羧酸(TCA)循环途径中的关键代谢物 和脂类代谢，包括脂肪酸、脂酰辅酶A和脂酰肉碱；4)提供指导 以及协助制定方案，该方案使用基于质谱仪的通量和代谢物分析 与控制葡萄糖和脂肪酸有关的分子生化指标的评价方法 动态平衡；以及5)与爱因斯坦的其他ES-DRC和机构核心设施协调这些努力 西奈山。所有这些服务都适用于糖尿病研究的新手，以及 致力于糖尿病相关项目的调查人员可以通过使用 这一核心的专业知识和设施。

The mission of the Stable Isotope & Metabolomics Core (SIMC) is to provide guided metabolite and substrate flux determinations in complex in vivo metabolic rodent models and for human clinical investigation for members of the Einstein-Mount Sinai Diabetes Research Center (ES-DRC). The SIMC provides an array of in vitro metabolic methodologies that augment in vivo investigations. These services provide investigators with specialized assays to determine substrate flux dynamics and metabolite profiles at the organelle, cellular, tissue and whole body level thereby elucidating the integrative network of disorders in glucose, protein and lipid metabolism. Through these collaborative efforts with the other Cores of the ES-DRC, the effects of defined pharmacological, dietary, environmental and genetic alterations are thoroughly characterized for their effects on glucose and lipid homeostasis, insulin action, and metabolism. The role of candidate molecules in relevant tissues (i.e., neurons, hepatocytes, skeletal muscle, adipocytes and beta cells) that are related to glucose homeostasis can be specifically delineated by thorough and definitive analyses using in vivo and in vitro experimentation with a step-wise guided approach in rodents and humans as well as in cell lines. To accomplish these goals, the SIMC will: 1) perform in vivo stable isotope substrate flux assays for the determination of rates of protein synthesis, lipogenesis, peripheral glucose disposal, hepatic glucose recycling, glucose-glycerol cycling and glucose-lactate cycling; 2) determine glycolysis (extracellular acidification rates) and mitochondrial oxygen consumption (mitochondrial respiration) in isolated cells, tissue explants or tissue culture, using Seahorse Biosciences Flux Analyzers, as well as more comprehensive stable isotope flux assessments; 3) perform targeted hypothesis-driven assessments of plasma and tissue metabolite profiles for key metabolites in the glycolytic/gluconeogenic, pentose phosphate, and tricarboxylic (TCA) cycle pathways, and lipid metabolism, including fatty acid, fatty acyl CoA and fatty acyl carnitine profiles; 4) provide mentorship and assistance with protocol development , which uses mass spectrometer-based flux and metabolite profiling methods for the evaluation of molecular biochemical targets relevant to the control of glucose and fatty acid homeostasis; and 5) coordinate these efforts with other ES-DRC and Institutional Core facilities at Einstein and Mount Sinai. All of these services are available to investigators new to diabetes research, as well as to investigators working on diabetes-related projects that can be enriched and extended by the use of the expertise and facilities of this Core.