INFLUENCE OF EARLY NUTRITION ON METABOLIC ...munich2014.project-earlynutrition.eu/download...Influence of Early Nutrition on Metabolic processes – Christian Hellmuth 25 1 Goichon,

INFLUENCE OF EARLY NUTRITION ON METABOLIC PROCESSES

Investigated by targeted LC/MS based metabolomics

Christian Hellmuth

02/04/2014

Note: for non-commercial purposes only

DIVISION OF METABOLIC AND NUTRITIONAL MEDICINE

DR. VON HAUNER CHILDREN’S HOSPITAL

KLINIKUM DER UNIVERSITÄT MÜNCHEN®

TABLE OF CONTENT

02/04/2014

Influence of Early Nutrition on Metabolic processes – Christian Hellmuth

2

2. EUROPEAN CHILDHOOD OBESITY PROJECT

1. METABOLOMICS




METABOLOMICS FUNCTIONAL INSIGHTS IN MOLECULAR CHANGES

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Genomics Transcriptomics

Proteomics

Metabolomics

Genotype Phenotype




METABOLOMICS ?

Determination of Metabolites

Metabolites

Small molecules (<1000 Dalton)

Substrates, intermediates and products of biological processes

Metabolome

Complete set of metabolites found within a biological sample

Understanding of biochemical changes & differences

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BENEFITS OF THE NEXT „OMICS“ FIELD

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Dynamic response of living systems to stimuli and genetics1

Information on what is actual happening2

1 Nicholson, Lindon Nature. 2008 Oct 23;455(7216):1054-6 2 Xie, Schirra et al. J Biomed Biotechnol. 2012;2012:805683




BENEFITS OF THE NEXT „OMICS“ THING

Insights into underlying molecular pathology1

Definition of biomarkers in biofluids1

Is the interplay between functional subunits changed by nutritional

changes?

Can we gain new functional insights?

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1 Heazell, Dune et al. Placenta. 2011 Mar;32 Suppl 2:S119-24




HOW TO ANALYSE THE METABOLOME?

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First: THINK




BEFORE I DETERMINE THE METABOLOME!

What do I want to know?

Background of my clinical trial

Hypothesis for metabolic differences/response

What do I have?

Sample number, material, volume

Sampling conditions

Which of the possible analytical strategies are available?

Targeted vs. untargeted metabolomics

Analytical equipment – GC, LC, MS, NMR

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CHOP - EUROPEAN CHILDHOOD OBESITY PROJECT

EARLY PROGRAMMING EFFECTS REFLECTED IN THE METABOLOME CHRISTIAN HELLMUTH, FRANCA KIRCHBERG, PETER RZEHAK, MARTINA WEBER, ANNICK XHONNEUX, NATALIA FERRE, ELVIRA VERDUCI, PIOTR SOCHA AND BERTHOLD KOLETZKO FOR THE EUROPEAN CHILDHOOD OBESITY TRIAL STUDY GROUP

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EARLY PROGRAMMING

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Conception Birth 6 Month

Maternal nutrition Maternal stress Placental development Placental transfer

Infant nutrition

Later Life

Obesity Diabetes CVD Allergy Cognition Behaviour ….

Environment and Nutrition

Genotype Phenotype

Metabolomics

PROGRAMMING




EARLY PROTEIN HYPOTHESIS

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Protein Intake

Insulin, IGF1 levels

Weight gain 0-24 month

Adipogenic activity

Later obesity risk

Metabolomics Insulinogenic amino acids

Metabolomics Changes in fat oxidation - acylcarnitines

Changes in lipid profile - phospholipids

Metabolomics Changes in energetic efficiency

Modified from Koletzko, Grote et al. Am J Clin Nutr. 2009 May;89(5):1502S-1508S




EUROPEAN CHILDHOOD OBESITY PROJECT

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Intervention groups Observational group

Lower protein formula (LP)

Higher protein formula (HP) Breastfed (BF)

263 265 163


Koletzko et al. Am J Clin Nutr. 2009 Jun;89(6):1836-45

BLOOD SAMPLING AT 6 MONTH OF AGE

RANDOMIZATION




CHOP – WEIGHT GAIN

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Koletzko, Grote et al. Am J Clin Nutr. 2009 Jun;89(6):1836-45

Data published in Socha, Koletzko et al. Am J Clin Nutr. 2011 Dec;94(6 Suppl):1776S-1784S





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Higher Protein Intake

Elevated Insulin, IGF1 levels

Increased weight gain 0-24 month

Adipogenic activity

✔

✔

✔

?

?





EUROPEAN CHILDHOOD OBESITY PROJECT

Amino acids determined by LC-MS/MS1

Polar lipids and glucose analysis by FIA-MS/MS2

21 Amino acids (AA)

41 Acylcarnitines (Carn.a)

14 Sphingomyelins (SM)

76 Phosphatidylcholins (PC)

15 Lysosphosphatidylcholins (LPC)

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1 Harder, Peissner et al. J Chromatogr B. 2011 Mar 1;879(7-8):495-504. 2 Absolute IDQ p 150 kit




CHOP – HP VS. LP

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Branched-chain amino acids




BRANCHED-CHAIN AMINO ACIDS (BCAA)

Leucine, Isoleucine, Valine

Elevated in HP group

Dietary BCAA escape first-pass liver metabolism

Oxidized in skeletal muscle

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FIRST PASS METABOLISM

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Dietary protein

GIT Portal vein Liver

Amino acids 20 % BCAA

Amino acid metabolism

Systemic circulation

BCAA Oxidation

Amino acids > 50% BCAA

Skeletal muscle

Wahren J, Felig P, Hagenfeldt L. J Clin Invest. 1976 Apr;57(4):987-99.




BRANCHED-CHAIN AMINO ACIDS (BCAA)

Leucine, Isoleucine, Valine

Elevated in HP group

Dietary BCAA escape first-pass liver metabolism

Oxidized in skeletal muscle

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BCAA OXIDATION

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Non-hepatic

4.1

Valine

5.0.oxo

Val

4.0

4.0.OH

4.0.oxo

3.0

5.1

Leucine

6.0.oxo

Leu

5.0

6.1.DC

6.0.OH.DC

5.1

Isoleucine

6.0.oxo

Ile

5.0

5.0.OH

5.0.oxo

3.0 2.0

Glutamate

α-Ketoglutarate

Glu

NAD+

NADH/H+

NAD+

NADH/H+ NAD+

NADH/H+

Shimomura Y, Murakami T, Harris RA et al. J Nutr. 2004 Jun;134(6 Suppl):1583S-1587S




CHOP – LP VS. HP

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Branched-chain amino acids

Short-chain acylcarnitines




BCAA OXIDATION

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Non-hepatic

4.1

Valine

5.0.oxo

Val

4.0

4.0.OH

4.0.oxo

3.0

5.1

Leucine

6.0.oxo

Leu

5.0

6.1.DC

6.0.OH.DC

5.1

Isoleucine

6.0.oxo

Ile

5.0

5.0.OH

5.0.oxo

3.0 2.0

NAD+

NADH/H+

BCKD Rate limiting

Brosnan, Brosnan J Nutr. 2006 Jan;136(1 Suppl):207S-11S




LEUCINE OXIDATION

Physiological level - linear stimulation

of degradation pathway

Regulation of BCAA level

Excess BCAA BCAA exceed

degradation pathway

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BCAA EFFECTS

Hydrophobic substrates for proteins1

Stimulation of insulin secretion2

Inhibition of glucagon secretion2

Reducing protein breakdown2

Enhancing protein synthesis (mTOR)3

Brain-uptake of glutamate1

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Signal for dietary protein intake

1 Brosnan, Brosnan J Nutr. 2006 Jan;136(1 Suppl):207S-11S 2 Nair, Short J Nutr. 2005 Jun;135(6 Suppl):1547S-52S. 3 Anthony, Kimball et al. J Nutr. 2000 Oct;130(10):2413-9.




IMPACT OF BCAA ON FAT METABOLISM

Leucine slows-down beta-oxidation1,2

Leucine decreases lipolysis2

Leucine increases lipogenesis2

Leucine, Isoleucin and Valin deprivation reduce fat mass3

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1 Goichon, Coeffier et al. J Proteomics. 2013 Jan 14;78:535-44. 2 Cheng, Guo et al. Diabetes. 2010 Jan;59(1):17-25. 3 Du, Guo et al. Amino Acids. 2012 Aug;43(2):725-34.




BETA-OXIDATION

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Even-chain acylcarnitines





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Protein Intake

Insulin, IGF1 levels

Weight gain 0-24 month

Adipogenic activity

✔

✔

✔


Elevation of insulinogenic amino acids (BCAA)

Reduction of beta-oxidation substrates(long-chain acylcarnitines)




TAKE HOME MESSAGE

Metabolomics provides insights into pathways and metabolic changes

Higher Protein intake results especially in elevated BCAA levels

Excess BCAA levels overcome BCAA degradation

BCAA may effect beta-oxidation and fat storage

Reduced long-chain acylcarnitines reflect reduced beta-oxidation

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THANK YOU FOR YOUR ATTENTION

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INFLUENCE OF EARLY NUTRITION ON METABOLIC ...munich2014.project-earlynutrition.eu/download...Influence of Early Nutrition on Metabolic processes – Christian Hellmuth 25 1 Goichon,