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Day 2

Thursday, December 8

8:30 Morning Coffee, Poster and Exhibit Viewing

 

Disease Applications

9:00 Comments by Session Chairperson
Dr. Rima Kaddurah Daouk

9:15 Metabolite Profiling of Intracellular Parasites
Dr. Malcolm McConville, Associate Professor, Biochemistry and Molecular Biology, University of Melbourne
We have used GC-TOF-MS and HPLC metabolite profiling approaches to identify new and unanticipated metabolic pathways in pathogenic stages of the protozoan parasite Leishmania spp, which proliferates within human macrophages. We show that the intracellular stages of these parasites accumulate a novel class of mannose-containing oligosaccharides and display an increased reliance on lipid and amino acid metabolism. The importance of these pathways for intracellular survival has been validated using genetic approaches. Metabolite profiling approaches are currently being extended to other intracellular pathogens.

9:45 Metabolite Profiling on Urine Samples from Autistic and Control Children 
Dr. Ana C. Soria, Chemistry, University of York
Results on application of a metabolite profiling approach in an unbiased strategy for finding endogenous urinary markers for autism will be presented. Having used state-of-the-art analytical techniques (Capillary Electrophoresis and NMR) to collect data, statistical data analysis was employed in a data-fusion approach to elucidate and combine difference features from autistic and control populations. MS/MS and NMR are to be used to obtain structural information and identification of compounds responsible for any difference features.

10:15 A Functional Analysis of Cardiac Disease through Metabolic Profiling
Dr. Jules Griffin, Department of Biochemistry, University of Cambridge
Metabolic profiling of cardiac tissue through high resolution NMR spectroscopy and GC-MS in conjunction with multivariate statistics provides a powerful high- throughput functional genomic tool for monitoring disease processes in animal models. We are currently investigating a range of mouse models of cardiac disease including models of Duchenne muscular dystrophy, cardiac arrhythmia, cardiac hypertrophy, mitochondrial dysfunction and metabolic syndrome using this metabolomic approach. The metabolic profiles demonstrate that strain background has a profound impact on the global metabolic phenotype of a mouse, providing insight into how a given gene deletion may result in very different responses in diverse populations. Furthermore, we will present data showing the cross correlation of metabolite profiles with perturbations in the proteome to understand Duchenne Muscular dystrophy and how a combined LC-MS, GC-MS and NMR spectroscopic study of the metabolic syndrome can be used to monitor dysfunction in lipid metabolism.

10:45 Coffee Break, Poster and Exhibit Viewing

11:15 Microbial Metabolite and Flux Analysis in Relation to Gut Health
Dr. Albert de Graaf, Department of Surgery, Wageningen Centre for Food Sciences/University Maastricht
To characterize microbial metabolic conversions 13C-labeled substrates were supplied to colonic bacterial populations and the 13C-contents of metabolites were analyzed using NMR and mass spectrometry techniques. This gave a quantitative overview of the substrate-specific metabolite spectrum. Exploiting the isotopomeric labeling patterns in the metabolites gave further insight into the operative bacterial metabolic routes. This combined metabolomic/fluxomic approach has the potential to yield plasma metabolic biomarkers for colonic health status.

11:30 HTS, Metabolomics and Discovery Pharma
Dr. George G. Harrigan, HTS, Pfizer Corporation
Metabolomics can be defined as the study of comprehensive changes in cellular, systemic or body metabolism induced by genetic, chemical or environmental modulation or understanding, defining, and interpreting any change (metabol) in any biological system in terms of changes in metabolism. As such, it has the potential to impact drug discovery and development at all stages in the pharmaceutical value chain. It is argued here that incorporating metabolomics in the early (most expensive!) stages of drug discovery allows a greater emphasis on mechanistic-based approaches and greater opportunities for impact. Case studies on transgenic animal model characterization and on a mechanism-of-action investigation of a lipid-modifying chemical are presented.

12:00 Serum Markers of Caloric Restriction
Dr. Bruce S. Kristal, Burke Medical Research Institute and Weill Medical College of Cornell University
Dietary or caloric restriction (DR) is the most potent and reproducible known means of increasing longevity and reducing morbidity in mammals. Exploratory studies previously identified 93 redox-active small molecules from sera (measured by HPLC coupled with coulometric detector arrays) with potential to distinguish dietary groups in both male and female rats. Classification and predictive power were addressed using megavariate data analysis approaches. Data processing choices of transformation, scaling, and winsorizing (outlier removal) each affected strength of the models, and, in some cases, revealed distinct metabolites to be of importance in building these models, often in gender-specific ways. We have now adapted these markers for use in humans. We will present these modeling approaches, the models and their ability to distinguish sera based on caloric intake, and the potential for moving these markers to epidemiological studies in human sera.

12:15 Lunch on Your Own 
(Sponsored Technology Workshops Available)

1:30 Comments by Session Chairperson
Dr. Rima Kaddurah Daouk, Duke University Medical Center

1:45 Phenotype Characterization of Animal Models using Metabolomics
Dr. Matej Oresic, chief Research Scientist, Biomolecules, VTT Biotechnology
Functional characterization of animal models in studies of complex diseases is an on-going challenge. The interventions lead to distributed and multifactorial responses, which are associated both with disease mechanisms as well as with compensatory physiological adaptation. Metabolomics as a phenotyping tool facilitates such studies. We will introduce our analytical, statistical, and bioinformatics approaches to phenotype characterization using metabolomics, as well as demonstrate them with applications in domains of diabetes and cancer research.

2:15 The Choline Metabolite Profile in Tumor Invasion and Metastasis
Dr. Kristine Glunde, Assistant Professor, Radiology Department - NMR Research - Oncology Section, Johns Hopkins University School of Medicine
The increase of cellular phosphocholine and total choline-containing compounds is one of the most widely established metabolic characteristics of cancer. This elevation is closely related to malignant transformation, invasion, and metastasis. Clinical magnetic resonance spectroscopy (MRS) of total-choline containing compounds is attracting a high level of interest as a method to detect the extent of invasion of brain, prostate, and breast tumors, and to differentiate between recurrence or necrosis following treatment. This presentation will highlight some of the recently discovered relationships between the aberrant choline metabolite profile in cancer cells and the underlying molecular mechanisms, whose identification can further our understanding of tumor biology and uncover unique targets to exploit for therapy. 

2:45 Metabolic Profiling for Understanding of Muscle Physiology and Insulin Action 
Dr. Deborah Muoio, Assistant Professor, Sarah W. Stedman Nutrition and Metabolism Center, and Department of Medicine, Duke University Medical Center
Feeding of a high fat diet causes circulating hyperlipidemia, leading to muscle insulin resistance. This condition is reversed by exercise. We seek to understand changes in metabolic regulation occurring in response to these maneuvers, via a combination of targeted MS-based metabolic profiling and metabolic flux analysis. Application of these and other tools has led to an understanding that lipid-induced insulin resistance is a condition in which elevated beta-oxidation of fatty acids is not supported by coordinate upregulation of TCA cycle flux. Understanding of this "disconnect" and its molecular basis suggests new therapeutic targets for insulin resistance of obesity and diabetes. 

3:15 Quantitative 1H-NMR Metabolomics to Monitor Graft Outcome After Transplantation 
Dr. Natalie J. Serkova, Assistant Professor, Anesthesiology Department, Biomedical MRI/MRS Cancer Center Core, University of Colorado Health Sciences Center
Severe pathophysiological conditions (such as ischemia/reperfusion, organ failure, cancer, drug toxicity) result in disturbances in concentrations, binding, or fluxes of the body’s endogenous metabolites. This occurs either by direct chemical reaction or by up/ down-regulation of key enzymes or nucleic acids that control metabolism. High-resolution 1H-NMR (nuclear magnetic resonance spectroscopy) on human and animal biosamples, such as tissue biopsies, blood, urine, cerebrospinal fluids, and saliva, allows for the simultaneous detection of hundreds of low molecular weight (max. 20 kDa) metabolites within a biological matrix at concentrations above 10 µM. Because nearly all major classes of endogenous metabolic intermediates (e.g. carbohydrates, ketone bodies, amino acids, high-energy phosphates, phospholipids, fatty acids, etc.) have characteristic NMR spectra, the technique is very useful for pattern identification and recognition of disease-related and toxicity-induced metabolic variations. Another important advantage of 1H-NMR based metabolomics is ability to quantify.

3:45 Afternoon Refreshment Break, Poster and Exhibit Viewing

4:15 Industry Scale Metabolite Profiling for Toxicology & Drug Development
Dr. Arno Krotzky, metanomics GmbH & Co. KGaA
Based on its leading technology platform and multi-year experience, metanomics has advanced metabolite profiling at an unparalleled scale, metabolite range and quality of results to highly successfully support of drug development, toxicology, health and nutrition, diagnostics and plant biotechnology. The power of mass spec-based metabolite profiling coupled to state-of-the-art bioinformatics for direct, reliable identification and interpretation of biomarkers, modes of action and physiological status in the context of biological pathways will be shown. We will demonstrate the scale, sensitivity and resolution at which metanomics has advanced metabolite profiling to classify toxicological mode-of-action to support better and accelerated product decisions. We will demonstrate the content, operation and value of our new metabolite profiles for a mode-of-action database which comprises the results of one of the largest and most comprehensive of such studies worldwide. In the second part of the talk we will present results from diabetes studies and other applications where metabolite profiling is applied to identify metabolic biomarkers for health diagnostics, early risk prediction and gene functions.

4:30 Metabolomics: Accelerating Drug Discovery and Development with Novel Biomarker Identification
Dr. Chris Beecher, VP Biochemistry & Technology, Biochemistry & Technology, Metabolon
Metabolon has developed a metabolomics technology that allows for the quantitative measure and identification of the repertoire of biochemicals contained in a biological sample. This discussion will cover an introduction to the metabolomics technology; results from several successful projects, including: patient stratification for Amyotrophic Lateral Sclerosis (ALS or Lou Gherig's Disease); identification of off-targeted effects for several members of a class of commercial drugs; and outcome prediction for pre-term labor which won Metabolon the 2005 March of Dimes award for Best Research.

4:45 Developing Metabolic Biomarkers by Measuring Isotopomer Ratios of Specific Metabolites 
Dr. Laszlo Boros, M.D., Associate Professor, UCLA and Chief Scientific Advisor to SiDMAP
Precise and relevant biomarkers can be developed by uniquely labeling glucose molecules with non-radioactive 13C isotopes and measuring their incorporation and distribution in metabolites throughout various metabolic pathways using GC/MS. An explanation of how the technology (SiDMAP) is used for biomarker discovery, as well as examples of actual biomarker discoveries, will be provided. 

5:00 Close of Metabolic Profiling Conference

5:00-5:30 Early Registration for Analytical Methods