Hisanori Kato. Endowed Chair of Food for Life Organization for Interdisciplinary Research Projects The University of Tokyo

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1 Workshop Argentina-Japan Bioscience and Biotechnology for the Promotion of Agriculture and Food Production -August 3rd to 7th Hisanori Kato Endowed Chair of Food for Life Organization for Interdisciplinary Research Projects The University of Tokyo 1

2 Japan 日本 Life Expectancy at Birth (2008, by the Japanese Ministry of Health, Labor, and Welfare) Female: years (the longest in the world) Male: years (the 4 th in the world) 2

3 Japanese Blessed with a Variety of Healthy Traditional Foods Abundant Experience on Functional Food Factors Accumulated research data and experience will help health promotion of other countries!! 3

4 Functional Food in Japan Food for Specified Health Uses (FOSHU) Nature, 364, 180 (1993)

5 Omics Analyses in Food and Nutrition Science Transcriptomics Metabolomics Spec #1[BP = 829.7, 24053] 2.4E % Intensity Mass (m/z) Proteomics

6 Effects of Dietary Protein Deprivation and Amino Acid Deprivation on mrna Levels of IGFBP-1 Gene Rat Liver GROWTH RETARDATION 5 Hepatoma Cells Relative mrna Levels Relative mrna Levels C PF (12 % Casein Diet)(Protein-free Diet) 0 AA+ Complete Medium AA- Amino Acid- Deprived (12 h) 6

7 Control Cells Animals Transcriptome Analysis of the Effect of Food Intake of Nutrients Functional Food Factors Excessive Intake Food Comprehensive Evaluation of the Function and Safety RNA Extraction Exhaustive Analysis of Gene Expression Treated x10 x5 DNA Microarray Analysis x2 x1/2 x1/5 x1/10 Control

8 The World of OMICS Target Genes Transcripts Proteins Metabolites Omics Discipline Genomics Transcriptomics Proteomics Metabolomics Nutritional Omics Nutrigenomics (narrow sense) Nutritranscriptomics Nutriproteomics Nutrimetabolomics Nutrigenomics Nutriomics 8

9 Other OMICS Epigenomics (Chromatin Modification) Interactomics (Protein-Protein Interaction) Toponomics (Cellular Localization) (Localizomics) Fluxomics (Cellular Flux of Metabolites) Lipidomics (Lipids) Glycomics (Sugars) CAUTION: Economics and Comics Do NOT Belong to the Omics Family

10 Use of DNA Microarray 1.Nutritional Intervention 2.Known Function of a Food Factor 3. A Food Material or Component of Interest DNA Microarray and Other Omics Analyses Understanding Whole Response Understanding Mechanisms Unknown Functionality 10

11 Casein Gluten Cornstarch Mineral Mix Vitamin Mix Cellurose Choline chloride 12% Casein (12C) 140 (+Met 2) % Gluten (12G) Protein Free (PF) Essential Amino Acid Pattern of Gluten Ile Leu Lys Met+Cys Phe+Tyr Thr Trp Val Rat Body Weight 12C 12G PF c % 11

12 Object To obtain a comprehensive view of the changes in gene expression in response to the alteration of protein nutrition Method Male Wistar rats ( 5 wk old) 12% Casein 12% Casein Protein Free 12% Gluten RNA extraction (Liver, Muscle,,,) (Day) Feeding (10:00~18:00) Rat U34A Genome Array (Affymetrix) 12

13 Ingestion of Gluten Up-regulates Genes for Both Synthesis and Catabolism of Cholesterol HMG-CoA synthase (5.4) HMG-CoA reductase (4.4) Squalene synthetase (4.4) Squalene eposidase (5.8) 2 Acetyl-CoA Acetoacetyl-CoA HMG-CoA Mevalonate Squalene Squalene epoxide Lanosterol Lanosterol 14-demethylase (3.2) 14-demethyl lanosterol Dehydrocholesterol Cholesterol Dehydrocholesterol reductase (3.2) Cholesterol 7- α hydroxylase (2.5) 7α-Hydroxycholesterol

14 Protein Nutrition and Skin Gene Expression <day> 12% Casein Diet 12C Pellet Diet 12% Casein Diet 12% Casein Diet 12% Gluten + Lys +Thr (12GLT) 12GLT 12% Gluten (12G) 12G 24 h Protein- Free Diet (PF) 10:00~18:00 PF 1.5 h Body Weight Excision of Skin Day DNA Microarray (n=4)

15 Professional Program for Agricultural Bioinformatics (Tokyo Univ.) 15

16 Choice of Normalization Method ody Weight DFW MAS RMA qfarms

17 Other Ongoing Projects (transcriptomics) Deficiency and Excess of Amino Acids Specific Proteins and Pepetides Protein Intake Levels at Various Life Stage Effect of Dietary Factor on Longevity, Diabetes, Hypertension Effect of Biolgical Rhythm and Exercise on Gene Expression Insulin-mimetic Activity of Soy Isoflavone Hypocholesteroleamic Effect of Sulfur-Containing Amino Acid Anti-obesity Effect of Dietary Amino Acid Supplementation

18 Stimulation of Glucose Uptake by a Soy Isoflavone, Genistein, in L6 Myotubes Cells L6 myotubes (from rattus) Differentiation for Methods nine to eleven days Cultivation in DME medium Precultivation for 2 hours Cultivation for 4 hours Stimulation by 50 μm genistein DNA Microarray and Pathyway Analysis

19 Genistein Seems to Activate Two Pathways Leading to GLUT4 Translocation Insulin Exercise Insulin receptor The rise of the ratio of AMP to ATP Activation of some protein kinases including IRS-1 The activation of AMPK GLUT4 Translocation VAMP2 GLUT4 VAMP7 Translocation Glucose Glucose

20 A Typical Flow of DNA Microarray Data Analysis for Food and Nutrition Science Raw Data Normalization Filtering Cluster Analysis, Principal Component Analysis, etc. (Tendency, Extraction of Marker Genes) Each Responder Genes (Rank Analysis, Statistical, Fold change ) Annotation Functional Category Pathway Analysis Interpretation Discussion Comparison with Other Data, Database 20

21 21

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