Hisanori Kato Endowed Chair of Food for Life Organization for - - PowerPoint PPT Presentation

Endowed Chair of “Food for Life” Organization for Interdisciplinary Research Projects The University of Tokyo

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Workshop Argentina-Japan “Bioscience and Biotechnology for the Promotion of Agriculture and Food Production”

• August 3rd to 7th 2009-

Hisanori Kato

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Life Expectancy at Birth

(2008, by the Japanese Ministry of Health, Labor, and Welfare)

Female: 86.05 years (the longest in the world) Male: 79.29 years (the 4th in the world)

Japan 日本

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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!!

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

Omics Analyses in Food and Nutrition Science

700 1260 1820 2380 2940 3500 Mass (m/z) 2.4E 10 20 30 40 50 60 70 80 90 100 % Intensity

Spec #1[BP = 829.7, 24053]

829.7285 881.7770 1378.3245 2191.8588 2222.0081 1799.7345 785.6865 1014.0126 1060.6566 833.5817 2279.1555 1927.8366 2145.8169 1088.0897 1696.5836 871.5511 2198.8981 1051.0375 2483.3095 2869.5020 1679.5564 1881.8104 709.6359 1412.3932 2175.0235

Transcriptomics Proteomics Metabolomics

12C PF

1 2 3 4 5 6

Effects of Dietary Protein Deprivation and Amino Acid Deprivation on mRNA Levels of IGFBP-1 Gene

（12 % Casein Diet）（Protein-free Diet）

1 2 3 4 5

Relative mRNA Levels

AA+ AA-

Complete Medium Amino Acid- Deprived (12 h)

Rat Liver Hepatoma Cells

Relative mRNA Levels GROWTH RETARDATION

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x1/2 x2 x5 x10 x1/5 x1/10

Transcriptome Analysis

• f the Effect of Food

DNA Microarray Analysis RNA Extraction

Exhaustive Analysis of Gene Expression

Treated Control Control

Animals Cells

・Intake of Nutrients ・Excessive Intake ・Functional Food Factors

Food

Comprehensive Evaluation

• f the Function and Safety

The World of OMICS

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Omics Discipline

Genomics Transcriptomics Proteomics Metabolomics

· · · · ·

Nutritional Omics

Nutrigenomics (narrow sense) Nutritranscriptomics Nutriproteomics Nutrimetabolomics · · · · ·

Target

Genes Transcripts Proteins Metabolites

· · · · ·

Nutrigenomics Nutriomics

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

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Use of DNA Microarray

1.Nutritional Intervention Understanding Whole Response 2.Known Function

• f a Food Factor

Understanding Mechanisms

• 3. A Food Material or

Component of Interest Unknown Functionality DNA Microarray and Other Omics Analyses

Ile Leu Lys

Met+Cys Phe+Tyr

Thr Trp Val

100 150% 50

Essential Amino Acid Pattern of Gluten

8 6 4 2

• 2
• 4

120 140 160 180 200 220 240

PF 12C 12G

c Rat Body Weight

12% Casein （12C) 12% Gluten （12G) Protein Free （PF) Casein Gluten Cornstarch Mineral Mix Vitamin Mix Cellurose

Choline chloride 140 (+Met 2) 656 50 40 10 100 2 150 648 50 40 10 100 2 798 50 40 10 100 2

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12% Casein Protein Free 12% Gluten

Object Method

Male Wistar rats ( 5 wk old)

RNA extraction (Liver, Muscle,,,)

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7 (Day) Feeding (10:00~18:00) To obtain a comprehensive view of the changes in gene expression in response to the alteration of protein nutrition 12% Casein Rat U34A Genome Array (Affymetrix)

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Ingestion of Gluten Up Ingestion of Gluten Up-

• regulates Genes for

regulates Genes for Both Synthesis and Catabolism of Cholesterol Both Synthesis and Catabolism of Cholesterol

2 Acetyl-CoA Acetoacetyl-CoA HMG-CoA Mevalonate Squalene Squalene epoxide

HMG-CoA synthase (5.4) HMG-CoA reductase (4.4) Squalene synthetase (4.4) Squalene eposidase (5.8) Lanosterol 14-demethylase (3.2) Dehydrocholesterol reductase (3.2) Cholesterol 7-α hydroxylase (2.5)

Dehydrocholesterol Lanosterol 14-demethyl lanosterol

Cholesterol

7α-Hydroxycholesterol

• 4
• 3
• 2

0 1 7 8

12% Casein Diet 12C 12% Gluten + Lys +Thr (12GLT) 12GLT 12% Gluten (12G) 12G

Pellet Diet 12% Casein Diet 12% Casein Diet

Protein- Free Diet (PF) PF

<day> Day

10:00～18:00

Excision of Skin DNA Microarray (n=4)

1.5 h 24 h

Protein Nutrition and Skin Gene Expression Protein Nutrition and Skin Gene Expression

Body Weight

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Professional Program for Agricultural Bioinformatics (Tokyo Univ.)

DFW RMA qFARMS MAS

Body Weight

Choice of Normalization Method Choice of Normalization Method

Other Ongoing Projects (transcriptomics)

Anti-obesity Effect of Dietary Amino Acid Supplementation 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

【Cells】 L6 myotubes (from rattus) 【Methods】

Differentiation for nine to eleven days Precultivation for 2 hours Cultivation for 4 hours Stimulation by 50 μM genistein Cultivation in DME medium

Stimulation of Glucose Uptake by a Soy Isoflavone, Genistein, in L6 Myotubes

DNA Microarray and Pathyway Analysis

Insulin Activation of some protein kinases including IRS-1 GLUT4 VAMP2 Translocation Glucose The activation of AMPK GLUT4 VAMP7 Translocation Glucose Insulin receptor Exercise The rise of the ratio of AMP to ATP

Genistein Seems to Activate Two Pathways Leading to GLUT4 Translocation

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Functional Category Pathway Analysis Comparison with Other Data, Database Normalization A Typical Flow of DNA Microarray Data Analysis for Food and Nutrition Science

Cluster Analysis, Principal Component Analysis, etc.

（Tendency, Extraction of Marker Genes）

Raw Data Each Responder Genes

（Rank Analysis, Statistical, Fold change…)

Filtering Annotation Interpretation Discussion

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