Metabolomic profiles of childhood obesity

Wei Perng, Matthew W. Gillman, Abby F. Fleisch, Ryan D. Michalek, Steven Watkins, Elvira Isganaitis, Mary-Elizabeth Patti, and Emily Oken

1

Obesity Prevention Program Department of Population Medicine

Harvard Medical School & Harvard Pilgrim Health Care Institute Boston, MA, USA

wei.perng@gmail.com

Note: for non-commercial purposes only

Background

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The role of metabolomics

↑Insulin resistance ↑Inflammation ↑Leptin, ↓Adiponectin ↑HDL, ↓LDL, ↑TG

Classic biomarkers of risk

Obesity

Disturbed metabolite profiles

• Type 2 DM • CVD • MetS

Disease

? ?

Research aims

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Obese vs. lean Difference in metabolite profiles?

Metabolite profiles Classic biomarkers

Maternal peripartum characteristics Child metabolite profile

obesity ● excessive GWG gestational diabetes

HOMA-IR ● leptin, adiponectin triglycerides ● CRP, IL-6

Aim 1

Aim 2

Aim 3

4

2128 live singleton births

262 mother-child pairs

Viva A Study of Health for

the Next Generation

P R O J E C T

648 eligible

Mid-childhood

Criteria: • Fasting blood from child • Maternal characteristics

Target sample size: 300 • Include: all child obesity & GDM • Criteria: adequate serum volume

Study population

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Mean ± SD or % (N) Child

Child's age (years) 8.0 ± 0.9

Female 50% (131)

Obesity (BMI ≥95th percentile) 32.1% (84)

Race/ethnicity

White 56.3% (147)

African American 24.1% (63)

Hispanic 6.9 %(18)

Mother

Mother's age at enrollment (years) 32.0 ± 5.8

Pre-pregnancy obesity (BMI >30 kg/m2) 22.5% (59)

Excessive gestational weight gain 58.8% (154)

Gestational diabetes 8.4% (22)

Characteristics of 262 Project Viva mother-child pairs

Study population

6

345 metabolites

Serum (n = 262)

Step 1: Untargeted

metabolomic profiling

Step 2: Principal components

analysis (PCA)

Step 3: Compare factor scores

VS.

Obese (BMI≥95th %ile) Lean (BMI<85th %ile)

n = 84 n = 150

18 factors

Step 4: Multivariable linear regression

Metabolite factors scores

Classic biomarkers

Maternal peripartum characteristics

Metabolite factor scores

Methods

-1,5

-1,0

-0,5

0,0

0,5

1,0

1,5

2,0

Dif

fere

nce

(9

5%

CI)

in f

acto

r sc

ore

s*

F18 F1 F2 F3 F4 F5 F6 F7 F8 F9 F10 F11 F12 F13 F14 F15 F16 F17

*Estimates are adjusted for child’s sex, age, and race/ethnicity; 95% CIs reflect Bonferroni’s correction.

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Factor 4: Branched-chain amino acids (BCAA) •Valine •Leucine •Phenylalanine •Isoleucine •Proprionylcarnitine (C3) •2-methylbutyrylcarnitine (C5) •Isovalerylcarnitine •Isobutyrylcarnitine •Tryptophan •3-methyl-2-oxovalerate • Kynurenine •Tyrosine •Gamma-glutamylleucine •4-methyl-2-oxopentanoate

Factor 9: Androgen hormones •4-androsten-3beta,17beta-diol disulfate 1 •DHEA-S •Epiandrosterone sulfate •Androsterone sulfate •4-androsten-3beta,17beta-diol disulfate 2 •Pregn steroid monosulfate •Pregnen-diol disulfate •Pregnenolone sulfate •Andro steroid monosulfate 2

Aim 1: Comparison of factor scores between obese & lean children

Results

-0,20

-0,10

0,00

0,10

0,20

0,30

0,40

0,50

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HOMA-IR

Triglycerides

(mg/dL) Leptin

(ng/mL) Adiponectin

(µg/mL) CRP

(mg/L) IL-6

(pg/mL)

Dif

fere

nce

(9

5%

CI)

in b

iom

arke

r*

Aim 2: Associations of BCAA & androgen metabolite patterns with cardiometabolic risk biomarkers

*Adjusted for maternal education and child age, sex, race/ethnicity, and fast-food intake. All biomarkers are ln-transformed.

BCAA-related pattern

Androgen pattern

Results

*Adjusted for maternal education and child age, sex, race/ethnicity, and fast-food intake.

-1,30

-0,80

-0,30

0,20

0,70

1,20

Pre-pregnancy obesity

Excessive gestational weight gain

Gestational diabetes mellitus

Aim 3: Associations of maternal peripartum characteristics with offspring metabolite patterns

Dif

fere

nce

(9

5%

CI)

in f

acto

r sc

ore

*

BCAA-related pattern

Androgen pattern

+ child BMI

Results

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10

Childhood obesity ↑ BCAA

↑ Androgen hormones

↑ BCAA

↑ Androgen hormones

Worse cardiometabolic profile

Maternal obesity ↑ BCAA in offspring

Conclusions

Aim 1

Aim 2

Aim 3

Dr. Emily Oken

Funding/Support: This study was funded by the US NIH (K24 HD069408, R37 HD 034568, P30 DK092924).

Sheryl L. Rifas-Shiman for data management and preparation.

The mothers and children of Project Viva.

Past and present Project Viva staff.

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Acknowledgements

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Questions?

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-1,30

-0,80

-0,30

0,20

0,70

1,20

Pre-pregnancy obesity

Excessive gestational weight gain

Gestational diabetes mellitus

Associations of maternal peripartum characteristics with offspring metabolite patterns, accounting for child BMI

Dif

fere

nce

(9

5%

CI)

in f

acto

r sc

ore

*

*Adjusted for maternal education and child age, sex, race/ethnicity, fast-food intake, and child’s BMI z-score

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Factor 4: Plasma metabolite concentrations of obese versus lean children Metabolite concentration z-score

Pb Obese Lean

Factor BMI ≥95th %ile BMI <85th %ile

loading n = 84 n = 150

Factor 4: mean ± SD = 0.00 ± 1.52 Valine 0.83 1.17 ± 0.30 -0.54 ±0.27 <0.0001 Leucine 0.76 1.38 ± 0.32 -0.37 ±0.29 <0.0001 Phenylalanine 0.72 1.23 ± 0.33 -0.27 ± 0.29 <0.0001 Isoleucine 0.71 1.18 ± 0.31 -0.37 ±0.27 <0.0001 Proprionylcarnitine (C3) 0.66 0.95 ± 0.29 -0.44 ±0.25 <0.0001 2-methylbutyrylcarnitine (C5) 0.63 0.80 ± 0.29 -0.03 ±0.26 0.0002 Isovalerylcarnitine 0.60 1.34 ± 0.30 -0.33 ±0.26 <0.0001 Isobutyrylcarnitine 0.56 1.19 ± 0.35 0.33 ± 0.31 0.001 Tryptophan 0.54 1.07 ± 0.33 0.21 ± 0.29 0.0007 3-methyl-2-oxovalerate 0.52 0.92 ± 0.30 -0.11 ± 0.26 <0.0001 Kynurenine 0.52 0.56 ± 0.32 -0.19 ± 0.28 0.002 Tyrosine 0.51 1.54 ± 0.28 0.04 ± 0.25 <0.0001 Gamma-glutamylleucine 0.51 0.53 ± 0.32 -0.23 ± 0.28 0.002 4-methyl-2-oxopentanoate 0.51 0.73 ± 0.30 -0.23 ± 0.26 <0.0001

a Estimates represent mean ± SE and are adjusted for child's sex, age, and race/ethnicity.

b From a t-test.

* Indicates tier 2 identification in which no commercially available authentic standards could be found, however annotated based on accurate mass, spectral and chromatographic similarity to tier 1 identified compounds.

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Factor 9: Plasma metabolite concentrations of obese versus lean children

Metabolite concentration z-score

Pb Obese Lean

Factor BMI ≥95th %ile BMI <85th %ile

loading n = 84 n = 150

Factor 9: mean ± SD: 0.00 ± 1.62

4-androsten-3beta,17beta-diol disulfate 1* 0.86 0.61 ± 0.31 -0.49 ± 0.27 <0.0001

Dehydroepiandrosterone sulfate (DHEA-S) 0.84 0.45 ± 0.27 -0.41 ± 0.23 <0.0001

Epiandrosterone sulfate 0.79 0.14 ± 0.32 -0.70 ± 0.29 0.0007

Androsterone sulfate 0.79 0.30 ± 0.29 -0.38 ± 0.26 0.002

4-androsten-3beta,17beta-diol disulfate 2* 0.78 0.48 ± 0.35 -0.74 ± 0.31 <0.0001

Pregn steroid monosulfate* 0.76 0.70 ± 0.30 -0.22 ± 0.26 <0.0001

Pregnen-diol disulfate* 0.70 0.30 ± 0.30 -0.14 ± 0.26 0.05

Pregnenolone sulfate 0.65 0.71 ± 0.33 -0.09 ± 0.29 0.002

Andro steroid monosulfate 2* 0.61 0.41 ± 0.29 -0.46 ± 0.26 <0.0001

a Estimates represent mean ± SE and are adjusted for child's sex, age, and race/ethnicity.

b From the Wald chi-squared test.

* Indicates tier 2 identification in which no commercially available authentic standards could be found, however annotated based on accurate mass, spectral and chromatographic similarity to tier 1 identified compounds.