Day 1 :
Keynote Forum
Petras Dzeja
Metabolomics NMRS Core, Mayo Clinic, USA
Keynote: 18O-assisted 31P NMR and Mass Spectrometry: from Phosphometabolomics to Fluxomics
Time : 09:01-09:30
Biography:
Petras Dzeja has completed his PhD from Kaunas Medical University, Lithuania and postdoctoral studies from Department of Biochemistry, University of Minnesota, Minneapolis MN, USA. He is the Co-director of Metabolomics NMRS Core, at Mayo Clinic, Rochester MN, a world renown Medical Center. Metabolomics NMRS Core is part of Mayo Clinic Metabolomics Resource Center which is one of six metabolomics cores in USA funded by NIH. Dr. Dzeja has developed 18O-assisted 31P NMR and mass spectrometric phosphometabolomics technology and pioneered phosphotransfer network, phosphometabolomics and system bioenergetics concepts. He has published more than 80 papers in reputed journals and has been serving on editorial board of PLOS ONE as a metabolomics expert.
Abstract:
Evaluation of metabolomics phenotypes requires knowledge not only metabolite levels but also their turnover rates from which metabolic fluxes and status of the whole metabolic system can be deducted. In this regard, stable isotope 18O-based metabolite tagging technology provides quantitative measurements of metabolite levels and turnover rates of many metabolites which metabolism include water as a reactant, most notably phosphometabolites, amino and organic acids. Using this technology dynamics of over 10 major metabolic and signaling pathways can be tracked simultaneously, including ATP turnover, oxidative phosphorylation, glycolysis/glycogenolysis, Krebs and urea cycles pentose phosphate pathway and phosphotransfer reactions. The 18O labeling methodology is based on the incorporation of the 18O nuclei (from H218O), into metabolite group with each act of enzymatic reaction, and subsequent distribution of 18O-labeled groups among different molecules. Using this approach major metabolites and their turnover rates can be quantified in cells and tissue samples and whole blood by 18O-assisted 31P NMR and 1H NMR spectroscopy and mass spectrometry. In this way obtained dynamic metabolomics profile appears to be sensitive indicator of energy and metabolic imbalances like the ones created by genetic deficiencies, myocardial ischemia, heart failure, aging and neurodegenerative disorders. Thus, 18O-assisted 31P NMR/mass spectrometry is a valuable tool for phosphometabolomic and fluxomic profiling of transgenic models of human diseases revealing system-wide adaptations in metabolic networks, as well as for biomarker identification in human diseases and metabolic monitoring of treatment efficacy and drug toxicity.
Keynote Forum
Lu Qi
Tulane University, USA
Keynote: Diet, amino acids profile, and diabetes risk
Time : 09:31-10:00
Biography:
Dr. Lu Qi has completed his PhD from Tufts University and postdoctoral studies from Harvard University School of Public Health. He is Regents Distinguished Chair and Professor; and Director of Tulane University Obesity Research Center. He has published more than 190 papers in reputed journals and has been serving as Editor-in-Chief and an editorial board member for several journals.
Abstract:
Plasma branched-chain amino acids (BCAAs, including leucine, isoleucine, and valine) were recently related to risk of type 2 diabetes (T2D). We assessed associations between cumulative consumption of BCAAs and risk of T2D among participants from three prospective cohorts: the Nurses’ Health Study (NHS; followed from 1980 to 2012), NHS II (followed from 1991 to 2011), and the Health Professionals Follow-up Study (HPFS; followed from 1986 to 2010). rnWe found higher total BCAA intake was associated with an increased risk of T2D in men and women. Recently, we examined the effects of weight-loss diets on long-term changes in plasma amino acids and the associations of these changes with weight loss and the improvement of insulin resistance in 2 randomized clinical trials, POUNDS LOST and DIRECT. In both trials, weight loss was directly related to the concurrent reduction of the BCAAs leucine and isoleucine and aromatic amino acid (AAA) tyrosine. In addition, we showed that reduction in AAA tyrosine was significantly related to improved insulin resistance, independent of weight loss, in both trials.rn
Keynote Forum
Ramon Cacabelos
EuroEspes Biomedical Research Center (CIBE), Institute of Medical Science and Genomic Medicine, Spain
Keynote: Nutritional Metabolomics in Aging and Neurodegeneration
Time : 10:30-11:00
Biography:
to be updated...
Abstract:
to be updated...
Keynote Forum
Tsutomu Masujima
The Quantitative Biology Center (QBiC), RIKEN, Japan
Keynote: Single-cell metabolomics— Past, current, and future
Time : 10:00-10:30
Biography:
Masujimas research is primarily focused on analytical development in the fields of analytical chemistry and pharmacology. Constantly performing research from new perspectives, the professor recently established a new method (1) in direct cellular analysis which uses video-imaging and mass spectrometry, simultaneously. (2)Through this new method of analysis, the professor made it possible to view the movements of a single cell, not even 1 pl (3) in volume, and to directly analyze the thousands of types of molecules that lurk inside the cell at the exact moment that a change occurs, enabling a faster and more precise method of pinpointing exactly what those moving molecules are. This new method has been praised as something which will contribute greatly to the acceleration of life sciences and medicine, and in September of this year, was awarded the 2008 Japan Society for Analytical Chemistry Award.
Abstract:
Since cells behave individually, there needs to be a method to investigate cell metabolism at the single-cell or single-organelle level individually. Metabolites generate extensive peaks in mass spectrometry (MS), thus, these are good targets for analysis. However, even with this high sensitivity, single-cell metabolomics was hard to perform due to the tiny size of cells. The size of a typical mammalian cell is 10 µm; its volume is only 1 pico liter (pL). That may be the reason why only big cells, eggs, giant axons, and big plant cells, were the targets for the past single-cell metabolomics. Now, after live single-cell mass spectrometry has been proposed, in which 1 pL or less of single-cell content, or even an organelle, is directly sucked by a nanospray tip and fed into a MS after adding the ionization solvent. Hundreds to thousands of metabolite peaks are detected. These detected molecular peaks are aligned and analyzed using a t-test or principal component analysis to discern the specific identity of the cell or the precise internal location of the molecules. The peaks are finally annotated by MS/MS and/or data bases such as KEGG and MassBank. However, we should not be satisfied with this level of single-cell metabolomics. Metabolomics is only metabolomics. Metabolism may be the final stage of cellular activity along the journey of gene expression, but it is comprised of many necessary molecular mechanisms. Thus, we should improve single-cell metabolomics to be more comprehensive, i.e. detect not only the hydrophilic molecules but also the hydrophobic and neutral molecules. Furthermore, the method should be able to detect the activities of other molecules, such as proteins, mRNA, and DNA, to find essential pieces of molecular mechanisms. When we are able to understand what metabolomics is truly saying, metabolomics will become real metabolomics.
- Track : 1 Metabolomic Profiling
Track : 8 Clinical Metabolomics & Lipidomics
Track : 9 Cancer Therapeutic Approaches
Chair
Petras Dzeja
Metabolomics NMRS Core, Mayo Clinic, USA
Co-Chair
Björn Riefke
Bayer Pharma AG, Germany
Session Introduction
Eugenia Trushina
Mayo Clinic College of Medicine, USA
Title: Metabolic and Epigenetic Alterations in Patients with Alzheimer’s Disease
Biography:
Dr. Trushina has completed her PhD at the age of 29 years from Saratov State University, Russia and postdoctoral studies from Mayo Clinic College of Medicine, USA. She is an Associate Professor in the Departments of Neurology and Pharmacology at the Mayo Clinic Rochester, USA. Dr. Trushina scientific interests include the investigation of early molecular mechanisms of neurodegeneration, the role of mitochondria in particular. Translational aspect of her work includes the development of blood-based metabolic biomarkers and mitochondria-targeted small molecule therapeutics for Alzheimer’s Disease. Dr. Trushina is a recipient of the NIH, BrightFocus, GHR, ADDF, and Mayo Clinic Research Awards.
Abstract:
The number of people living with dementia is estimated at 44 million worldwide, and is expected to rise to 76 million by 2030 and to 136 million by 2050. Alzheimer’s Disease (AD) currently affects more than 5 million Americans, with numbers expected to grow. The pathophysiological changes in AD patients begin decades before the onset of dementia, highlighting the urgent need for the development of early diagnostic methods, novel therapeutic approaches, and methods to monitor efficacy of experimental therapeutics. Targeted metabolomics with the overall goal to determine changes in mitochondria- and energy-related metabolites was conducted in CSF and plasma from males and females with mild cognitive impairment (MCI), AD and control patients involved in the Mayo Clinic Study of Aging. Panels of metabolites that include amino acids, fatty acids, lipids and components of the TCA cycle revealed that metabolic changes in females were more pronounced and correlated strongly with the disease compared to males. Analysis of metabolic fluxes in TCA cycle in human fibroblasts from late onset AD male and female patients using Glucose and Glutamine stable isotope tracers confirmed stronger inhibition of fluxes in female patients. Epigenetic changes were also more pronounced in female patients suggesting a link to altered energy metabolism. Overall, our data demonstrate that metabolic changes associated with the development and progression of AD differ between males and females where females are affected to the greater extent.
Hsien-Yeh Hsu
National Yang-Ming University, Taiwan
Title: Fucoidan inhibitory function in cancer in vivo and in vitro: Role in the development of human anti-cancer therapeutic intervention
Time : 10:41-11:00
Biography:
Hsien-Yeh Hsu, a molecular/cellular biologist, obtained Doctoral degree from Cornell University, USA. Later, he worked in various biotechnology companies in Boston, USA. In 1990, he returned to university, and later became Assistant Professor in Weill Cornell Medical College, NYC, USA. In 1997, he joined National Yang-Ming University, Taiwan. Currently, as Professor, he works at Department of Biotechnology and Laboratory Science in Medicine, and also as Research Fellow in Genomics Research Center, Academia Sinica. He is working on polysaccharides obtained from brown algae and medicinal mushroom Ganoderma lucidum, and focusing on anti-cancer functions in human clinical experiments. Moreover, he is engaged on examining anti-cancer therapeutic intervention and on developing anti-cancer drugs.
Abstract:
Fucoidan, a polysaccharide extracted from brown seaweeds, reduces proliferation in a battery of tumor cells. In recent, we demonstrated that fucoidan reduces tumor size both in LLC1 lung cancer-xenograft male C57BL/6 mice and in 4T1 breast cancer-xenograft female Balb/c mice. As it is known, transforming growth factor β receptors (TGFRs) play important roles in the regulation of epithelial-mesenchymal transition (EMT) as well as of proliferation/progression and metastasis in cancer cells. Using these cancer cells, we found that fucoidan effectively up-regulates epithelial markers, down-regulates mesenchymal markers, and also decreases expression of transcriptional repressors Snail, Slug, and Twist. In addition, we found fucoidan inhibits TGFR-mediated migration and invasion of cancer cells. Moreover, we found fucoidan decreases TGFRI and TGFRII proteins in vivo and in vitro. To elucidate the mechanism by which fucoidan decreases TGFRI/II proteins in cancer cells, we found that fucoidan enhances ubiquitination proteasome pathway (UPP)-mediated degradation/ubiquitination of TGFRs. We further demonstrated fucoidan promotes Smurf2 and Smad7 to conjugate TGFRs, resulting in TGFR degradation. Together, we are the first to identify a novel mechanism for fucoidan anti-tumor activity, namely decreasing tumor growth by modulating TGFR proteins degradation, leading to reduction of TGFR-mediated signaling, and further inhibition of cancer cells in vivo and in vitro. Our current findings indicate that fucoidan is a potential therapeutic agent or dietary supplementation for cancers, acting via Smurf2-dependent ubiquitin degradation of TGFRs affecting the TGFR/Smad/Snail, Slug, Twist, and EMT axes.
Ashkan Emadi
University of Maryland School of Medicine, USA
Title: Interference with glutamine metabolism: a novel approach for treatment of acute myeloid leukemia
Time : 11:01-11:20
Biography:
Dr. Emadi received his M.D. at Tehran University of Medical Sciences and his Ph.D. in Organic Chemistry at the Illinois Institute of Technology. He developed novel methodologies for the regiospecific synthesis of multiple naphthoquinone derivatives related to the natural product conocurvone, and was granted “Highest Standards of Academic Achievement Award”. Following completion of his Ph.D., he completed his internship and residency in Internal Medicine at the University of Kentucky and the University of Cincinnati, respectively. Subsequently, Dr. Emadi was trained in Hematology and Medical Oncology Fellowship Program at Johns Hopkins University Sidney Kimmel Comprehensive Cancer Center. Dr. Emadi joined the University of Maryland Marlene and Stewart Greenebaum Cancer Center as an Associate Professor of Medicine, Pharmacology and Experimental Therapeutics at the University of Maryland School Of Medicine and serves as Director of the ACGME-accredited Hematology and Oncology Fellowship Program. He previously served as medical officer at the Division of Hematology Products (DHP), United States Food and Drug Administration (FDA), and as visiting scientist at Division of Adult Hematology, Department of Internal Medicine, School of Medicine, Johns Hopkins University. Dr. Emadi has experience and in-depth understanding of the multiple aspects of cancer drug development including basic organic chemistry and molecular synthesis, in vitro and in vivo studies, and all phases of clinical trials as well as regulatory science.
Abstract:
In contrast to normal cells, rapidly dividing leukemia cells reprogram their nutritional requirements to match an increased metabolic demand. The two main nutrient sources for growth and survival of cancer cells are glucose and glutamine. Depletion of glutamine or interruption of its cellular processes can be detrimental to leukemia cells but not normal cells. Asparaginases deplete amino acids asparagine and glutamine and are FDA-approved drugs for the treatment of acute lymphoblastic leukemia (ALL). The anti-leukemia effect of asparaginases in acute myeloid leukemia (AML) is not established. Here we describe the preclinical effect of different asparaginase products on the survival of AML cells with or without isocitrate dehydrogenase (IDH) mutations, which culminates in glutamine depletion resulting in disruption of protein synthesis downstream of mammalian target of rapamycin (mTOR) causing strong apoptotic and autophagic responses. The clinical experience with asparaginase in AML patients will also be discussed. Moreover, the cytotoxic effect of pharmacologic inhition of glutaminase, the enzyme that converts glutamine to ammonia and glutamate, will be reported in AML cells with different mutational status.
I-Chen Peng
National Cheng Kung University, Taiwan
Title: Myc induces expression of glutamine synthetase through promoter demethylation
Time : 11:21-11:40
Biography:
I-Chen Peng is an assistant professor of Department of Life Sciences at National Cheng Kung University, Taiwan. She received her Ph.D. in the Biochemistry and Molecular Biology Graduate Program from University of California at Riverside, USA, and the post-doctoral training in the Department of Molecular Genetics & Microbiology at Stony Brook University in New York, USA. She has published several papers in reputed journals. Dr. Peng joined the faculty in the Department of Life Sciences at National Cheng Kung University in 2014. Dr. Peng’s research focuses on targeting lipid and glutamine metabolism to treat obesity and obesity-related diseases.
Abstract:
The proto-oncoprotein Myc is known to promote glutamine usage by up-regulating glutaminase (GLS), which converts glutamine to glutamate that is catabolized in the tricarboxylic acid (TCA) cycle. Here we report that in a number of human and murine cells and cancers, Myc overexpression leads to elevated expression of glutamate-ammonia ligase (GLUL), also termed glutamine synthetase (GS), which catalyzes the de novo synthesis of glutamine from glutamate and ammonia. Elevated expression of GS promotes cell survival under glutamine limitation, while silencing of GS leads to decreased cell proliferation and xenograft tumor growth. Stable isotope based metabolite tracing shows that GS overexpression increases glutamine synthesis, cataplerotic flux at the ï¡-ketoglutarate (ï¡KG) step of the TCA cycle, and contributes to nucleotide synthesis and amino acid transport. Mechanistically, Myc binds to the promoter of thymine DNA glycosylase (TDG) and upregulates its expression, which leads to active demethylation of the GS promoter and its increased expression. These results demonstrate an unexpected role of Myc in promoting glutamine synthesis, and suggest a previously uncovered molecular connection between DNA demethylation and glutamine metabolism in Myc-driven cancers.
Jian Zhi Hu
Pacific Northwest National Laboratory, USA
Title: Slow-MAS NMR Metabolomics
Time : 11:41-12:00
Biography:
Jian Zhi Hu received his Ph.D in 1994 and is currently a senior staff scientist and principal investigator of Pacific Norwest National Laboratory. He has published more than 170 papers in peer reviewed journals, delivered a large number of presentations, received two US R&D 100 awards and 10 US patents.
Abstract:
Metabolomics studies on tissues are of significance since a disease is often associated with a specific tissue or organ malfunction. It is, therefore, expected that the changes in metabolic profile is more dramatic in the diseased tissue than body fluids. It is likely that tissue specific metabolic profiling provides a unique window of investigating the biochemistry associated with a particular disease in great detail than possible using global body fluids. In this work, we will report a non-destructive magic angle spinning NMR metabolomics technique that is capable of high resolution and high sensitivity metabolic profiling on biological samples, in particular tissue samples, with sample volume from as small as 200 nanoliters (nL) to as large as a milliliter or more using a single probe and using only a few minutes. This has been achieved by combining the techniques of high resolution slow-MAS 1H NMR technique and a switchable inductively coupled static micro-RF coil-LC resonator and by rotating the specimen at a sample spinning rate of 40 to 200 Hz about the magic angle axis. The nanoliter capability has the potential to follow the metabolic changes through a continued investigation on a single small laboratory animal over a long period of time using minimally invasive blood and tissue biopsy samples. While the milliliter capability would allow minimally destructive studies of intact biological object with size as large as >1 cm3. Examples of applications will be reported.
Mats Gustav Borén
Denator, Sweden
Title: Prevention of metabolite change due to enzymatic post-sample activity using heat based enzyme inactivation
Time : 12:01-12:20
Biography:
Mats Borén completed his PhD at the Swedish University of Agricultural Sciences. He is currently the Head of Development at Denator, the heat stabilization and sample preservation company, based in Uppsala, Sweden . He has published more than 25 papers in reputed journals primarily on the subjects of sample quality and changes post-sampling.
Abstract:
The removal of a biological sample induce a cascade of reactive changes causing alterations to the molecular profile of the sample. This is particularly true for metabolomic studies where sample preservation is key for the relevance of the analytical results. In order to avoid this and enable analysis of a molecular state closer to in vivo state, heat induced protein denaturation has been introduced as a means to preserve sample conmposition and quality1 A study has been done to evaluate the inclusion of heat stabilization in the sample preparation prior to lipid analysis. The study was performed on mouse livers from 30 healthy mice, randomized in 5 groups of 6 animals designed to compare snap freezing at -80°C to 3 heat stabilization conditions (freezing just after heat stabilization, after 1 hour and after 3 hours) and to a negative control samples maintained at room temperature during 10 minutes before freezing. For each mouse livers polar metaboltes were extracted using MeOH and PBS buffers and analyzed using NMR to generate a global metabolite profile. Representative lipids profiles were generated and compared between the different treatment groups. Analysis demonstrate that for frozen samples, MeOH extraction results are intermediate between PBS extraction and heat stabilization, suggesting that whatever the method of extraction, there is metabolic changes for frozen samples, but also that results with heat stabilization are indeed “better” than freezing. A new biological sample preservation procedure based on heat stabilization for improved metabolomics profiling has been developed.
Ching-wan Lam
The University of Hong Kong, Hong Kong
Title: Clinical metabonomics for biomarker discovery of malignant pleural effusions (MPE)
Time : 12:21-12:40
Biography:
Professor Lam obtained his MBChB from The Chinese University of Hong Kong in 1991 and FRCPA in 1997 from The Royal College of Pathologists of Australasia with a double scope of practice in Chemical Pathology and Genetics. He is a Fellow of The Australasian Association of Clinical Bicohemists. He obtained his PhD in 2000 from The Chinese University of Hong Kong. He obtained FRCP(Glas) from The Royal College of Physicians and Surgeons of Glasgow in 2012. Professor Ching-Wan Lam is TheEditor, Clinical Chimica Acta, an official journal of The International Federation of Clinical Chemistry and Laboratory Medicine.
Abstract:
Aim: Clinical metabonomics has been widely used in advanced laboratory medicine for the study of disease mechanism and biomarker discovery. Here, we will extend the use of clinical metabonomics to develop new screening test for malignant pleural effusions (MPE). Methods: 32 malignant and 18 non-malignant PE samples will be analyzed using reversed-phase liquid chromatography-tandem mass spectrometry (LC-MS/MS). Biomarkers for MPE will be determined by metabolome-wide association studies (MWAS) using Receiver Operating Characteristic Curve Explorer and Tester (ROCCET). All markers will be filtered using a metabolome-wide significance level (MWSL) at p-value ≤ 2×10−5. Results: 2731 and 3137 markers were detected in positive and negative ESI spectra respectively. Free fatty acids (FFAs) 16:0, 18:1 and 18:2 were significantly increased in MPE. FFA 18:1 (oleic acid) showed the largest area-under-ROC of 0.96 (95% CI: 0.87–1.00) with sensitivity of 84% and specificity of 100.0% (p-value: 8.23×10−8). Discussion: Pleural fluid oleic acid is a novel biomarker for screening MPE.
Yonghai Lu
National University of Singapore, Singapore
Title: Mass spectrometry-based metabolomics reveals that serum lysophospholipids are associated with incident type 2 diabetes
Time : 12:41-13:00
Biography:
Yonghai Lu is a Research Fellow of National University of Singapore. He got his PhD degree from the Chinese University of Hong Kong in 2013. His researches focus on metabolomics studies of diabetes and liver cancer, and has published more than 20 papers in international journals, such as Cancer Research, Environmental Science & Technology, and Metabolomics.
Abstract:
Type 2 diabetes (T2D) usually presents years before becoming clinically apparent, and it is thus crucial to identify more sensitive and specific biomarkers for early diagnosis of this disease. In this study, we applied an integrated mass spectrometry-based untargeted metabolomics approach to characterize serum metabolic profiles in 197 incident T2D cases and 197 healthy controls matched by age and gender, in order to identify metabolic signatures associated with incident T2D. A series of 51 metabolites were found to be altered significantly in cases compared to controls. In addition to widely reported branched-chain amino acids (leucine, isoleucine, and valine) and free fatty acids (palmitic acid, stearic acid, oleic acid, and linoleic acid), we found seven lysophospholipids were significantly associated with the risk of T2D, including lysophosphatidylglycerol (LPG) (12:0) and lysophosphatidylinositol (LPI) species (16:1, 18:1, 18:2, 20:3, 20:4, and 22:6). Odds ratios for the seven lysophospholipids per standard deviation increment ranged from 1.43 to 2.10. Of these, LPI (16:1) showed a good capability for prediction of T2D in both at-risk individuals with high glycated hemoglobin (HbA1c) levels (≥6.5%) and those with normal HbA1c values (<6.5%), with area under the curve (AUC) values of 0.806 and 0.754, respectively. Our study indicates that LPI (16:1) emerges as a potential predictor for diabetes development, although the finding was observed in a single and small cohort, raising the need for further independent validation in well-designed and large-scale studies
Biography:
Yael Porat, PhD, is founder and CEO of BioGenCell Ltd (www.BioGenCell.net), a biotechnology company focusing on stem cell therapy and regenerative medicine.Dr. Porat earned her PhD in immunology from the Sackler School of Medicine, Tel-Aviv University.Prior to founding BioGenCell, she served as head of the Global Biological Development Department at Teva Pharmaceuticals and as CTO at TheraVitae, where she led the translation of innovative research into the production of stem cell-based therapies used to treat patients with severe cardiovascular diseases.BioGenCell is developing groundbreaking technology for automated production of patient-specific, blood-derived stem-cell-based therapies. The company’s proprietary technology utilizes immune-directed stem-cell specific activity to create cell-based products for a wide range of diseases. BioGenCell’s first line of products targets incurable vascular diseases, such as heart failure, stroke and Peripheral Artery Diseases.
Abstract:
Background: Vascular diseases are a major cause of morbidity and mortality, particularly in the elderly and diabetic patient population. Stem/progenitor cell (SPCs) treatments with bone-marrow-derived cells show promising outcomes, albeit not without adverse events related to cell collection and mobilization. We describe a novel technology for generating a therapeutic population (BGC101) of enriched endothelial progenitor cells (EPCs) from peripheral blood, using dendritic cells (DCs) to specifically direct SPC activity in-vitro. This one day culture process utilizes non-mobilized blood as a source for sufficient numbers of potentially therapeutic SPCs. Methods & Results: Plasmacytoid and myeloid DCs from healthy and diabetic donors were activated with anti-inflammatory and pro-angiogenic molecules to induce specific activation signals. Co-culturing of activated DCs with SPCs, from the same patient sample, within one day (12-18hours) generated 83.7±7.4×106 BGC101 cells with 97% viability from 250ml of blood. BGC101, comprising 52.4±2.5% EPCs (expressing Ulex-lectin, AcLDL uptake, Tie2, vascular endothelial growth factor receptor 1 and 2 and CD31) and 16.1±1.9% SPCs (expressing CD34 and CD184). The final product, BGC101 has demonstrated angiogenic and stemness potential and secretion of IL-8, IL-10, VEGF and osteopontin. When administered intramuscularly to nude mice with limb ischemia, BGC101 yielded a high safety profile and improved blood perfusion (x2 p<0.0002), capillary density and limb function within 21 days. Conclusions: These observations indicate that alternatively-activated DCs promote the generation of EPC-enriched SPCs within culture for one day. The resulting unique product, BGC101, has the potential for treatment of vascular conditions, including arteriosclerotic heart disease, stroke and peripheral ischemia.
- Special Session
Session Introduction
Andrea Armirotti
Istituto Italiano di Tecnologia, Italy
Title: Metabolomics in neuroscience: Old tools for new models and new tools for old models
Time : 13:46-14:45
Biography:
Andrea Armirotti joined the Istituto Italiano di Tecnologia (Genova, Italy) in 2010, where he leads the Analytical Chemistry laboratory of the Drug Discovery and Development Department (D3). He is involved in several research projects requiring MS-based metabolomics and proteomics characterization of biological and pharmacological models. He also works in the biomarker discovery field, with the aim to translate the findings from the animal models to clinical research. He has published more than 50 papers in the analytical chemistry field, spanning over a broad range of topics.
Abstract:
Over the last years, mass spectrometry-based untargeted metabolomics, in combination with advanced data analysis techniques, has reached outstanding levels of analytical performance and it now represents a valuable tool for the efficient exploration of the biochemical pathways of the brain. Using this technology, key advancements have been made in almost any field of neuroscience, with several important breakthrough discoveries. On the other hand, new technologies for data acquisition and analysis are always coming up. We recently successfully applied some of the most well-established techniques to a new model: long-term exposure of primary neurons to graphene-related materials (GRM). We deeply investigated the molecular changes occurring in the cell as natural response to GRM. Our findings clearly outline that important alterations occur in the neuron metabolome and are associated to phenotypic and functional changes. On the other hand, these well-established tools for untargeted analysis sometimes fail in detecting significant shifts in the metabolome, particularly when they occur to low abundance species. We then applied a totally new data analysis technique to the very early stages of a well-known mouse model of Parkinson’s disease (6-hydroxy-DOPA in the brain), when no phenotypic of functional changes are observed in animals. We discovered that the regulation of a particular class of large phospholipids, naturally present at ppb levels in the brain, is significantly increased immediately after the insult to dopaminergic neurons.
- Track : 10 Plant & Environmental Metabolomics
Track : 13 Food & Nutritional Metabolomics
Session Introduction
Min Guan
Chinese Academy of Sciences, China
Title: ERRα induces mitochondrial glutaminase expression guiding anaplerosis upon osteogenic differentiation of mesenchymal stem cells
Time : 14:46-15:05
Biography:
Min Guan received her Ph.D. degree from Guangzhou Institute of Biomedicine and Health, Chinese Academy of Sciences in 2009. She performed postdoctoral studies at University of California and Johns Hopkins Medical School, USA. She was appointed as Associate Professor at Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences in 2013. She focuses on studying mechanisms of metabolic diseases, and has published in reputed journals including Nature medicine, Hepatology and Stem cells.
Abstract:
Aging deteriorates osteogenic capacity of mesenchymal stem/stromal cells (MSCs), contributing to imbalanced bone remodeling and osteoporosis. Glutaminase (Gls) catabolizes glutamine into glutamate at the first step of mitochondrial glutamine-dependent anaplerosis which is essential for MSCs upon osteogenic differentiation. Estrogen-related receptor α (ERRα) regulates genes required for mitochondrial function. Here, we found that ERRα and Gls are up-regulated by osteogenic induction in human MSCs (hMSCs). In contrast, bone mass, osteogenic differentiation capacity of MSCs, ERRα, Gls and osteogenic marker genes are significantly reduced with age. We demonstrated that ERRα binds to response elements on Gls promoter and affects glutamine anaplerosis through transcriptional induction of Gls. Conversely, ERRα inverse agonist compound 29 or mTOR inhibitor rapamycin significantly decreased expression levels of ERRα and Gls, leading to deteriorated osteogenic differentiation of hMSCs. Importantly, overexpression of ERRα in hMSCs promoted osteogenic differentiation and partially restored impairment by rapamycin. Finally, we proved that compensated ERRα expression indeed potentiated osteogenesis capability of elderly mice MSCs in vitro. Together, we establish that Gls is a novel ERRα target gene and ERRα/Gls signaling pathway plays an important role in osteogenic differentiation of MSCs. Our findings suggest that restoring age-related mitochondrial glutamine-dependent anaplerosis may be beneficial for osteoporosis.
Yongyu Zhang
Shanghai University of Traditional Chinese Medicine, China
Title: Urine metabolomics study on unilateral ureteral obstruction induced renal fibrosis in rats and intervention effects of total aglycone extracts of Scutellaria baicalensis
Time : 15:06-15:25
Biography:
Yongyu Zhang got the degree of Ph.D. of Pharmacy in Meiji Pharmaceutical University, Tokyo, Japan in 2000, and now works as a doctoral supervisor in Shanghai University of Traditional Chinese Medicine,Shanghai, China. In addition, he works as the adjunct research fellow of research center for drug metabolism, Shanghai institutes of pharmacology, Chinese Academy of Sciences and the jury of the National Natural Science Funds of China. What' s more, he have been authorized two US patents and have publicated more than 40 SCI papers.
Abstract:
Renal fibrosis (RF) is the final common pathological process to chronic renal failure caused by various kidney diseases. In the present study, we investigated effects of Total Aglycone Extracts of Scutellaria Baicalensis (TAES) on some RF closely related parameters in unilateral ureteral obstruction (UUO) rats, and a urine metabolomics method, based on gas chromatography-mass spectrometry (GC/MS), was developed to monitor metabolic alterations. In the metabolomics study, the metabolic perturbations induced by UUO were reversed based on pattern recognition analysis and different metabolites associated with RF were identified. Ontology-based enrichment analysis by BiNChE aids in the interpretation of difference metabolites and relevant disturbed pathways were extracted by MetPA analysis. Our findings indicate that TAES have positive effects on UUO-induced RF in rats, meanwhile, metabolomics method coupled with metabolites enrichment analysis is useful to reveal the pathogenesis of diseases and action mechanism of medicine on the whole body.
Ewelina P Dutkiewicz
National Chiao Tung University, Taiwan
Title: Skin metabolomics approach for the development of biomarkers in psoriatic disease
Time : 16:41-17:00
Biography:
Ewelina P Dutkiewcz has obtained her MSc degree at the Department of Chemistry at Warsaw University, Poland. Currently, she is a PhD candidate at the Department of Applid Chemistry at National Chiao Tung University (NCTU), Taiwan. Her research interests include analysis of unconventional biological specimens for clinical applications by means of mass spectrometry. She works in the group of Prof. Pawel L. Urban (NCTU) and collaborates with Dr. Hsin-Yi Chiu (NTUH). She is an author of articles (Anal. Chem., 2014, 86, 2337; J. Mass Spectrom., 2015, 50, 1321) related to a novel method for profilng of skin metabolites.
Abstract:
Human skin is covered with numerous low-molecular-weight metabolites excreted by specialized glands of skin or produced during the breakdown of proteins. Those metabolites have a huge potential as disease biomarkers. However, due to the lack of convienient analytical methodology, the analysis of skin excretions has not become popular. Recently, we have introduced a facile method – hydrogel micropatch probe – for collection and mass spectrometric profiling of skin metabolites. We have applied this method to investigate the skin metabolome of patients suffering from one of the skin diseases – psoriasis. Psoriasis is an inflammatory immune-mediated disease that affects 2-3% of the world population. We have examined over 200 subjects. With aid of customized informatic tools based on C language programming and chemometric methods, the study unraveled the differences in skin metabolomes of psoriatic patients and healthy individuals. We proposed several biomarkers of disease progression, which correlate positively or negatively with plaque severity scores of examined psoriatic lesions. Currently, we work on evaluation of the relationship between skin metabolic profiles and treatment outcomes in psoriasis. In future work, we would also like to find a relation between skin and plasma metabolomes. We target patients treated with the newest type of therapy – biologic agents – which are very effective in the case of some patients. Moreover, the cost of biologic therapy is very expensive. We believe this metabolomic study will bring new insights on the psoriasis disease pathophysiology and will help to predict the treatment response.
Tao Wu
Shanghai University of Traditional Chinese Medicine, China
Title: Serum Lipid Alterations Identified in Chronic Hepatitis B, Hepatitis B Virus-Related Cirrhosis and Carcinoma Patients
Biography:
Dr.Wu has completed her PhD at the age of 35 years from Shanghai University of Traditional Chinese Medicine. She is mainly engaged in the field of the prevention and treatment of metabolic disease and application of metabonomics in clinical metabolic disease. She has published more than 20 papers in reputed journals.
Abstract:
Backgrounds&Aims: Hepatitis B virus (HBV) is a major pathogenic factor of liver diseases. The incidences of chronic hepatitis B (CHB), HBV-induced cirrhosis and carcinoma are high and increasing. This study aimed to evaluate lipid metabolite changes in the serum that are associated with disease progression from CHB to HBV-induced cirrhosis and to HBV-induced carcinoma. Methods: A targeted metabolomic assay was performed in fasting sera from 136 patients with CHB, 104 patients with HBV-Cirrhosis, and 95 patients with HBV-carcinoma using ultra-performance liquid chromatography triple quadrupole mass spectrometry (UPLC-TQMS). Results: Totally 140 metabolites were identified. A clear separation between HBV-cirrhosis and HBV-carcinoma was obtained using the PLS-DA (partial least squares discriminant analysis) scores of 9 lipid metabolites. Among the 9 metabolites, progressively lower levels of long-chain lysophosphatidylcholines (lysoPC a C18:2, lysoPC a C20:3, lysoPC a C20:4) were observed from CHB to cirrhosis to carcinoma; lower levels of lysoPC a C20:4 were also found in patients with higher Model For End-Stage Liver Disease (MELD Score) in the same disease group; and lysoPC a C20:3 levels were lower in Child-pugh Class C than in Class A and Class B in HBV-Cirrhosis and HBV-Carcinoma patients. Octadecadienylcarnitine(AC C18:2) level was higher in HBV-Cirrhosis patients than in the other two groups. Conclusions: Serum levels of selected long-chain lysoPCs are promising markers for the progression of HBV-induced liver diseases.
Pier Paolo Piccaluga
Bologna University School of Medicine, Italy
Title: Metabolism alterations in aggressive lymphomas
Biography:
Pier Paolo Piccaluga has done MD with honors from Bologna University (1997). He did Specialty in Hematology with honors (2001), PhD in Clinical and Experimental Hematology at Bologna University (2005), Specialty in Pathology with honors at Siena University (2014) and Post-doctoral fellowship at Columbia University, New York (USA). He is a Visiting fellow at Columbia University (USA, 2008) and Kiel University (Germany, 2009), Lecturer in Pathology at Bologna University: 2006-2012. Currently he is appointed as Associate Professor at the Department of Experimental, Diagnostic, and Specialty Medicine, Bologna University Medical School, Italy.
Abstract:
Lipid metabolism alteration have been recently described in human cancer with relevant pathogenetic and possibly therapeutic implications. Our group described specific lipid metabolism patterns in Burkitt lymphomas. We have now extended our analysis to the spectrum of aggressive non Hodgkin lymphomas (NHL) including BL, diffuse large B-cell lymphoma (DLBCL), post-transplant lymphoproliferative diseases (PTLD), and nodal peripheral T-cell lymphomas (PTCLs). Particularly, we performed a gene expression profile-based analysis of 250 NHL cases and 25 samples representative of normal B-cell subsets (naïve, memory, germinal center, and plasmacells). Immunohistochemistry and quantitative real time PCR (RTQ-PCR) were used for validation. We found that BL, DLBCL and PTLD presented with consistent abnormalities if compared to normal B-cell subsets. The specific patterns were associated with clinico-pathological features including proliferation rate and genetic imbalances. Concerning DLBCL, specific analyses were carried out in ABC vs GCB cases as well as based on the genetic profile (double hit lymphomas vs. others). Further, specific attention was payed to EBV-associated tumors since virus can alter the metabolic profile. In conclusion, we identified specific metabolic patterns associated with B-cell lymphomagenesis and particularly those associated with peculiar features such as genetics and EBV infection.
Song Xue
Dalian Institute of Chemical Physics-CAS, China
Title: The correlation of polar lipids changes with TAG accumulation under nitrogen deprivation in Nannochloropsis oceanica based on lipidomics
Biography:
Song Xue PhD, is Professor, Marine Bioengineering Group Leader, Dalian Institute of Chemical Physics, Chinese Academy of Sciences. She completed PhD in Biochemical Engineering in 2003 and was funded by “100 talents plan” of Chinese Academy of Sciences. From 2007 to 2009, she worked at Florida State University as research faculty. She has published papers in Science, Nature Structural & Molecular Biology, Molecular Cell, Bioresource technology and so on.
Abstract:
Nannochloropsis as one of the ocean microalgal species, has been considered as a promising resource of biodiesel feedstocks because of its capacity of TAG accumulation. TAG biosynthesis has been classified in different pathways. Polar lipids have big contribution for TAG accumulation by providing acyl group or DAG. The quantification and characterization of the contribution in microalgae are still veiled. In this study, there are total 117 polar lipid species covering eight classes (i.e. MGDG, DGDG, SQDG, DGTS, PC, PE, PG, PI) identified by UPLC/Orbitrap and quantified by UPLC/Q-TOF to study the correlation of polar lipids changes with TAG synthesis under nitrogen deprivation condition of Nannochloropsis oceanica IMET1. Through comparision of the lipids profiling of polar lipids, we proposed that C18 acyl groups are desaturated while attached to PC. PE and DGTS acts as the carrier of EPA synthesis and the donors of imported DAG to the chloroplast for major MGDG which is eukaryotic-like molecule species. Under nitrogen limitation, the 16:0 acyl chain-containing lipid species in PC and DGTS were increased while 16:1 acyl chain-containing lipid species decreased. Similarly, in PE and DGTS, 18:0, 18:1 acyl chain-containing lipid species in PC and DGTS increased while 18:2 acyl chain-containing lipid species decreased. The changes of lipid species in DGTS, PC and PE further affected photosynthetic membrane lipids profiling. Thus we suggest under nitrogen limitation, the changes in polar lipid are mainly caused by the reduction of palmitic ∆7 desaturase and oleic ∆12 desaturase activity.
Brijesh Pandey
Amity University Lucknow, India
Title: Does Zn2+ regulate structure and function of cinnamyl alcohol dehydrogenase isolated from Leucaena leucocephala?
Biography:
Brijesh Pandey has completed his PhD in Biochemistry from Univesity of Lucknow in 2010. He is the Programme Leader of BTech Biotechnology in Amity University Uttar Pradesh, Lucknow Campus from 2008 and Assistant Professor dealing with Recombinant DNA Technology. He has contributed significantly to understanding and designing strategy for Genetic improvement of Leucaena leucocephala.
Abstract:
Cinnamyl alcohol dehydrogenase (CAD) is a key enzyme of lignin biosynthetic pathway. The effect of Zn2+ on structure and function of native CAD enzyme, isolated from stem xylem of Leucaena leucocephala, was studied. The enzyme exhibited in vitro regulation by Zn2+ concentration. Inactive oligomerization was exhibited by this homodimeric enzyme of 76 kDa, at more than 2 mM Zn2+. Effect of Zn2+ was also studied on recombinant protein. At 2 mM Zn2+, the active enzyme was maximally expressed in soluble fraction and partitioned to inclusion bodies at higher concentrations. The in silico predicted structure revealed presence of two Zn2+ binding domains, one structural and other catalytic. The structural Zn2+ binding (C100,C103,C106,C114) domain was found projected out (as lobe) from main body of enzyme and speculated to cause inactive oligomerization of enzyme at higher Zn2+ concentration. Inactive oligomerization was validated by native PAGE. The structural Zn2+ may thus, be crucial, but the actual role assignment needs further experimental validation by mutagenesis of the structural Zn2+ binding domain. The role validation of structural Zn2+ would enable us to understand the mechanism by which the Zn2+ homoeostasis regulates the enzyme.
Sayyid Nooreddin Faraji
Shiraz University of Medical Science, Iran
Title: Correlation between tempers and metabolomics: A study protocol
Biography:
Sayyid Nooreddin Faraji has completed his MSc degree from Shiraz University of Medical Science and he is currently pursuing his PhD in Medical Biotechnology, Shiraz University of Medical Science, School of Medicine, Immunology departments, Recombinant Antibodies Lab.
Abstract:
According to traditional medicine, several single or combined tempers have been narrated. There are several approaches regarding to temper classifications in traditional medicine. While previous studies showed the effects of genetic and environmental factors on human metabolomics, however there is no study to evaluate how different tempers affect the human metabolomics. The first step of the study was done based on reviewing the literature for different temper approaches and their classification weather in Iran or China. Finally the details of the hypothesis into which how to make a proper statistical society for checking the molecular tests on them is offered. The genome of all humans is almost similar but there are differences in their healthy conditions. Investigating the metabolomics profiles of peoples who have different tempers and different healthy conditions and then compare them with other peoples of the statistical groups could be helpful to establish novel and molecular based classification of tempers. In these such investigations, we can make four groups to the study including: Classification of individuals based on Ibn Sina temper classification, based on other classification, a group that obey a proper diet according to their temper at least 40 days and other individuals and patients. This study will help to make molecular basis of different types of temper more clear. Also it could help people to know their body better and make proper decision for a choosing healthier diet and life style.