COMPREHENSIVE ANALYSIS OF METABOLIC FLUXES IN VIVO

体内代谢通量的综合分析

• 批准号: 8171633
• 负责人: SHAWN M BURGESS
• 金额: $ 20.91万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8171633
• 关键词: Address; Animal Model; Biochemical Pathway; Chemicals; Clinical; Complex; Computer Retrieval of Information on Scientific Projects Database; Coupling; Data; Data Analyses; Detection; Disease; Enzymes; Experimental Designs; Funding; Genetic Transcription; Grant; Hepatic; High temperature of physical object; Human; Institution; Isotopes; Kinetics; Knowledge; Liver; Measurement; Measures; Metabolic; Metabolic Pathway; Metabolism; Methodology; Methods; Modeling; Molecular Biology; Population; Research; Research Personnel; Resources; Sampling; Scientist; Source; System; Techniques; Technology; Testing; Tissue Model; Tracer; United States National Institutes of Health; Vertebral column; Work; base; brain metabolism; improved; in vivo; magnetic field; mathematical model; metabolic abnormality assessment; multiplet; research study; small molecule; technology development

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Despite recent advances in molecular biology that have provided a fuller understanding of the transcriptional basis of disease, technology that allows the ultimate end product of transcription, metabolic flux, to be measured in vivo is very limited. The importance of measuring flux is highlighted by a number of examples in animal models where changes in enzyme expression do not match changes in flux through the enzyme. Thus, if the metabolic basis of disease is to be explicitly understood, techniques which measure in vivo fluxes must be further developed so that they can be easily utilized by basic and clinical scientists. NMR isotopomer analysis of small molecule metabolites is well suited to make these measurements because chemical shift and spin-spin coupling allow the isotopomer populations to be very well defined. Knowledge of isotopic distributions in metabolites is tantamount to knowing the flux through the biochemical pathways that generated the isotopomers. The continued focus of Core I is to address technology development for NMR based metabolic analysis with an emphasis on improved sensitivity of NMR isotopomer analysis of in vivo metabolism. The specific aims are: 1. Multiplexing metabolic flux. This aim seeks to expand the number of metabolic pathways measured in a single in vivo experiment by incorporating additional13C tracers of hepatic fluxes into the techniques developed in the previous funding cycle. This aim will improve the experimental efficiency of the NMR isotopomer method (compared to other methods of isotope detection) by making simultaneous measurements of multiple hepatic fluxes. 2. Mathematical analysis of tracer data from complex systems. Our aim is to more fully refine the mathematical models that describe hepatic fluxes with respect to parameter sensitivity, experimental design and extend an existing kinetic model from a simple onecompartment tissue model to a more realistic two-compartment model. This will enhance our capabilities to use 13C NMR multiplet data for analysis of liver and brain metabolism as we move toward more in vivo work on our new high field human scanners. 3. Improve NMR detection of 2H and 13C for metabolic studies. To increase the applicability of our metabolic flux measurements, it is essential that we continue to increase the sensitivity of both the 2H and 13C NMR experiments, the backbone of this methodology. We will extend 2H NMR and JHSQC experiments to 18.8T to take advantage of the increased sensitivity and dispersion of the higher magnetic field and develop the necessary methods to take maximum advantage of new micro-solenoidal probes that offer higher sensitivity for sample limited cases. Finally, we will test the suitability of new high temperature superconducting NMR probes for metabolic measurements.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 尽管分子生物学的最新进展已经提供了对疾病的转录基础的更全面的理解，但是允许在体内测量转录的最终终产物代谢通量的技术非常有限。在动物模型中，酶表达的变化与通过酶的流量变化不匹配，这突出了测量流量的重要性。因此，如果要明确了解疾病的代谢基础，必须进一步开发测量体内通量的技术，以便基础和临床科学家可以轻松使用。小分子代谢物的NMR同位素异构体分析非常适合进行这些测量，因为化学位移和自旋-自旋耦合允许同位素异构体群体非常明确。了解代谢产物中的同位素分布就等于了解产生同位素异构体的生化途径的通量。核心I的持续重点是解决基于NMR的代谢分析的技术开发，重点是提高体内代谢的NMR同位素异构体分析的灵敏度。具体目标是：1.多重代谢流。这一目标旨在通过将额外的13 C示踪剂的肝通量纳入到以前的资助周期开发的技术中，来扩大在单个体内实验中测量的代谢途径的数量。这一目标将提高实验效率的NMR同位素方法（与其他方法的同位素检测），同时测量多个肝通量。2.复杂系统示踪剂数据的数学分析。我们的目的是更充分地完善数学模型，描述肝通量的参数敏感性，实验设计和扩展现有的动力学模型从一个简单的一个ecommerchant组织模型，一个更现实的两室模型。这将增强我们使用13 C NMR多重峰数据分析肝脏和大脑代谢的能力，因为我们将在我们的新高场人体扫描仪上进行更多的体内工作。3.改进代谢研究中2 H和13 C的NMR检测。为了提高我们的代谢通量测量的适用性，我们必须继续提高2 H和13 C NMR实验的灵敏度，这是该方法的支柱。我们将把2 H NMR和JHSQC实验扩展到18.8 T，以利用更高磁场的灵敏度和分散性，并开发必要的方法来最大限度地利用新型微型螺线管探针，这些探针为样本有限的情况提供更高的灵敏度。最后，我们将测试新的高温超导核磁共振探头的代谢测量的适用性。